pa ge 1 pa ge 1 american journal of chemistry and pharmacy (ajcp) empagliflozin-induced acute pancreatitis and euglycemic diabetic ketoacidosis in type 2 diabetes mellitus yasser sadawey1*, hesham metwally2 volume 1 issue 2, year 2022 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v1i2.747 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: october 11, 2022 accepted: october 19, 2022 published: october 23, 2022 acute pancreatitis and euglycemic dka, edka, are uncommon but potentially fatal complications in diabetic patients on sodium-glucose cotransporter 2 inhibitors (sglt2). this case report presents the exceptional occurrence of a 49-year-old; male patient diagnosed with empagliflozin-induced edka coupled with t2dm as a precursor to acute pancreatitis. euglycemic dka may be produced after administering just one dose of sglt2. on discharge, empagliflozin was managed to stop. since the initial administration of sglt2, this diagnosis should be considered a potential consequence. appropriate medical treatment is made possible by diagnosing and treating this life-threatening condition promptly.keywords diabetic ketoacidosis, euglycemic dka, sodium-glucose cotransporter-2, empagliflozin, type 2 diabetes mellitus, pancreatitis. 1 department of medicine, mediclinic me, al jowhara hospital, al ain, uae 2 department of radiology, mediclinic me, al jowhara hospital, al ain, uae * corresponding author’s e-mail: dr.yassersadawey@gmail.com introduction the clinical trifecta of metabolic acidosis, hyperglycemia, and elevated ketone bodies in the urine and blood is known as diabetic ketoacidosis (dka). euglycemic dka (edka) is the precursor to a particular group of people whose blood glucose levels are between normal ranges (nyenwe & kitabchi, 2016). at the presentation time, 37 out of 211 dka patients had normal blood sugar levels (300 mg/dl) and a plasma bicarbonate level of 10 mmol/l. munro et al. (1973) were the first to document this phenomenon. normoglycemia was interpreted as less than 250 mg/dl. thus, when serum glucose levels are below 250 mg/dl, edka is described as a trifecta that includes high anion gap metabolic acidosis with positive urine and serum ketones (kitabchi et al., 2009). one of the most recent classes of oral drugs for the treatment of t2dm has been legalized, and it is called sodium-glucose cotransporter-2 (sglt-2) inhibitors. they may be administered to these patients alone or with other medications. they prevent glucose reabsorption in the proximal renal tubule, which lowers blood sugar levels (sarafidis et al., 2019). the us food and drug administration (fda) officially warned patients using these medications in 2015 that they are at an elevated risk of developing dka with unusually mild to moderate glucose spike, or edka (do, n.d.). therefore, the present case report explores the rare presentation of empagliflozin-induced euglycemic dka, (edka) and acute pancreatitis in a patient with t2dm just 6 days after empagliflozin introduction. abbreviations dka=diabetic ketoacidosis, edka=euglycemic dka, sglt-2=sodium-glucose cotransporter-2, fda= food and drug administration, vbg=venous blood gas, cva=cerebral vascular accident, hdu=high dependency unit, ct= computed tomography methodology case presentation a presentation from a 49-year-old man nonsmoker with a 5-year medical history and worsening type 2 diabetes mellitus due to poor compliance was made to the mediclinic al jowhara hospital in the united arab emirates. additionally, he had three days’ history of upper abdominal pain, nausea, and vomiting. he denies fever, loose motion, melena, cough, or headache. six days before the presentation, he suffered from cva with residual right-sided weakness. during his admittance, he was initiated on aspirin, clopidogrel, atorvastatin 40mg, lisinopril 5mg, linagliptin/metformin 5mg/1000mg once, and empagliflozin 25mg. the initial observational analysis revealed that he was tachypneic but otherwise vitally normal. other vital signs include his rbg as 11.1 mmol/l, dry mucous membranes, but normal capillary refill time. additionally, on clinical assessment, there was right lower limb’s minor motor impairment, intact sensory functions, normal reflexes, and normal cranial and cerebellar functions were notable. his venous blood gases (vbg) represented ph 7.121, hco3 9.6 mmol/l, pco2 25.4 mmhg, lactate 2.1 mmol/l, na 128 mmol/l, cl 101 mmol/l, and k 5.2 mmol/l and urine showed ketone +3 suggestive of high anion gap (17.4), severe metabolic ketoacidosis and his hba1c was 8.1%. the patient’s lipase and amylase were elevated at 440 u/l and 354 u/l, respectively, compatible with acute pancreatitis (table 1). ecg assessment was reported normal, as well as chest x-ray and abdominal https://doi.org/10.54536/ajcp.v1i2.747 https://journals.e-palli.com/home/index.php/ajcp mailto:dr.yassersadawey%40gmail.com?subject= pa ge 2 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(2) 1-4, 2022 figure 1: axial and coronal ct images in venous phase showing minimal peripancreatic fat stranding around the head and body as early features of pancreatitis. no peripancreatic fluid. ultrasound. abdomen ct showed mild enlargement of pancreatic head with subtle peri-pancreatic fat stranding, a picture of mild pancreatitis. the patient was admitted to hdu for management of edka, linagliptin/metformin, empagliflozin were held, and intravenous fluids and insulin infusion were initiated with repeated vbg every 2 hours. his ph normalized, ketonuria improved, and the anion gap closed. he made a good recovery with normalizing pancreatic enzymes and improved symptoms. once he could tolerate normal oral intake, he shifted to sitagliptin/metformin 100/1000 mg and gliclazide 120 mg. after dietary consultation and education, he was discharged. upon follow-up visits, he was clinically stable, tolerating his medications with good glycemic control, and his hba1c became 6.2%. using the naranjo algorithm of adverse drug reactions, we concluded that empagliflozin was the most likely culprit for his pancreatitis. table 1: initial and follow up laboratory values parameters initial day1 day 2 on discharge venous ph (7.36–7.44) 7.121 7.224 7.36 7.439 serum bicarbonate (21–28 mmol/l) 9.6 11.6 19.2 25.7 pco2 (35-45mmhg) 25.4 22.5 33 39.9 serum lactate (0.5–1.4 mmol/l) 2.1 1.7 1.6 1.7 serum sodium (135–145 meq/l) 128 132 134 134 chloride (98–107 mmol/l) 101 108 103 99 serum potassium (3.5–5.0 mmol/l) 5.2 5.7 4.1 3.8 anion gap (8–16 mmol/l) 17.4 12.4 11.8 9.3 urine ketone (negative) +++ ++ + ve hba1c (%) 8.1 lipase (0-160u/l) 440 u/l 117 110 amylase (23–85 u/l) 354u/l 123 78 discussion dka occurs in 1.34/million t2d patients annually (jensen et al., 2017). in contrast, it is unknown how often euglycemic dka will occur. the uncommon occurrence of euglycemic dka may cause a deferment in care and prognosis, which might have significant or fatal consequences(calçada et al., 2021). a recent family of anti-diabetic medications known as sglt2 inhibitors (empagliflozin, canagliflozin, and dapagliflozin) increases the likelihood of euglycemic dka discrete to the length of treatment (peters et al., 2015; somagutta et al., 2021; taylor et al., 2015). however, the prevalence of edka increased after the introduction of sodiumglucose transporter 2 (sglt2) inhibitors in some rare cases. although there have been few reports of dka in sglt2i investigations, the precise numbers of edka events have not been evaluated (wibawa et al., 2021). the results of another trial,“the empa-reg outcome (empagliflozin cardiovascular outcome event trial in type 2 diabetes mellitus patients-removing excess glucose)” study, revealed that dka occurs less frequently than 0.1% of the duration (rosenstock, 2015; sampani et al., 2020). rates of dka-related occurrences among type 2 diabetic patients on sglt2 inhibitors ranged from 0.16 to 0.76 events per 1000 patient years (goldenberg et al., 2016).sglt2 inhibitors raise the risk of dka in t2d https://journals.e-palli.com/home/index.php/ajcp pa ge 3 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(2) 1-4, 2022 patients by seven times (blau et al., 2017). according to other studies, sglt2i improves beta cells in the pancreas, increasing lipolysis and raising the concentration of ketone bodies (da silva et al., 2018). they have also been indicated to activate the pancreatic alpha cells, triggering glucagon production and inducing an imbalance in insulin and glucagon level (candelario & wykretowicz, 2016; pfützner et al., 2017). the patient, in this case, presented with acute pancreatitis while taking prescribed empagliflozin, and there was no known cause of pancreatitis, such as gallstones or alcohol consumption. the development of pancreatitis with regular exposure to the medication and the symptom treatment response after withdrawal make empagliflozin a plausible cause of pancreatitis. the molecular basis of pancreatitis caused by empagliflozin is unidentified. it is an unusual reaction similar to previous cases of druginduced pancreatitis because of the immunologic or cytotoxicity effects the drug or its metabolites have on the body. conclusion the fda’s warning is supported by multiple pieces of literature that detail many physiologically tenable possibilities for negative consequences of sglt2 (empagliflozin) inhibitor. the preponderance of the data supports the causative involvement of sglt2 inhibitors in the pathogenesis of acute pancreatitis and edka. therefore, sglt-2 inhibitor initiation should be avoided in acute conditions. since normoglycemia might mask the presence of acidosis, medical professionals should be aware of this diagnosis and immediately begin therapy. given the nature of the disease, edka should be retained in the differential even if it is rare. patients should be informed about negative consequences and stimuli to prevent recurrences. acknowledgement the authors are grateful to the mediclinic al jowhara hospital for their consistent support throughout the research. references blau, j. e., tella, s. h., taylor, s. i., & rother, k. i. (2017). ketoacidosis associated with sglt2 inhibitor treatment: analysis of faers data. diabetes/metabolism research and reviews, 33(8), e2924. calçada, m. b., fernandes, l., soares costa, r., montezinho, s., martins duarte, f., frutuoso, l., & freitas, a. r. (2021). euglycemic diabetic ketoacidosis after a single dose of empagliflozin in a patient with pancreatitis. clinics and practice, 11(2), 216-218. candelario, n., & wykretowicz, j. (2016). the dka that wasn’t: a case of euglycemic diabetic ketoacidosis due to empagliflozin. oxford medical case reports, 7, 144-146. da silva, p. n., da conceição, r. a., do couto maia, r., & de castro barbosa, m. l. (2018). sodium– glucose cotransporter 2 (sglt-2) inhibitors: a new antidiabetic drug class. medchemcomm, 9(8), 1273-1281. do, w. y. c. (n.d.). fda issues multiple warnings for newest class of diabetes drugs. goldenberg, r. m., berard, l. d., cheng, a. y., gilbert, j. d., verma, s., woo, v. c., & yale, j.-f. (2016). sglt2 inhibitor–associated diabetic ketoacidosis: clinical review and recommendations for prevention and diagnosis. clinical therapeutics, 38(12), 2654-2664. e2651. jensen, m. l., persson, f., andersen, g. s., ridderstråle, m., nolan, j. j., carstensen, b., & jørgensen, m. e. (2017). incidence of ketoacidosis in the danish type 2 diabetes population before and after introduction of sodium–glucose cotransporter 2 inhibitors—a nationwide, retrospective cohort study, 1995–2014. diabetes care, 40(5), e57-e58. kitabchi, a. e., umpierrez, g. e., miles, j. m., & fisher, j. n. (2009). hyperglycemic crises in adult patients with diabetes. diabetes care, 32(7), 1335-1343. munro, j., campbell, i., mccuish, a., & duncan, l. (1973). euglycaemic diabetic ketoacidosis. br med. j., 2(5866), 578-580. nyenwe, e. a., & kitabchi, a. e. (2016). the evolution of diabetic ketoacidosis: an update of its etiology, pathogenesis and management. metabolism, 65(4), 507-521. peters, a. l., buschur, e. o., buse, j. b., cohan, p., diner, j. c., & hirsch, i. b. (2015). euglycemic diabetic ketoacidosis: a potential complication of treatment with sodium–glucose cotransporter 2 inhibition. diabetes care, 38(9), 1687-1693. pfützner, a., klonoff, d., heinemann, l., ejskjaer, n., & pickup, j. (2017). euglycemic ketosis in patients with type 2 diabetes on sglt2-inhibitor therapy—an emerging problem and solutions offered by diabetes technology. endocrine, 56(1), 212-216. rosenstock, j, f. e. (2015). euglycemic diabetic ketoacidosis: a predictable, detectable, and preventable safety concern with sglt2 inhibitors. diabetes care, 38(9), 1638-1642. https://doi.org/ doi: 10.2337/dc15-1380. sampani, e., sarafidis, p., & papagianni, a. (2020). euglycaemic diabetic ketoacidosis as a complication of sglt-2 inhibitors: epidemiology, pathophysiology, and treatment. expert opinion on drug safety, 19(6), 673-682. sarafidis, p., ferro, c. j., morales, e., ortiz, a., malyszko, j., hojs, r., khazim, k., ekart, r., valdivielso, j., & fouque, d. (2019). sglt-2 inhibitors and glp1 receptor agonists for nephroprotection and cardioprotection in patients with diabetes mellitus and chronic kidney disease. a consensus statement by the eureca-m and the diabesity working groups of the era-edta. nephrology dialysis transplantation, 34(2), 208-230. somagutta, m. r., agadi, k., hange, n., jain, m. s., batti, e., emuze, b. o., amos-arowoshegbe, e. o., popescu, s., hanan, s., & kumar, v. r. (2021). https://journals.e-palli.com/home/index.php/ajcp pa ge 4 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(2) 1-4, 2022 euglycemic diabetic ketoacidosis and sodium-glucose cotransporter-2 inhibitors: a focused review of pathophysiology, risk factors, and triggers. cureus, 13(3). taylor, s. i., blau, j. e., & rother, k. i. (2015). sglt2 inhibitors may predispose to ketoacidosis. the journal of clinical endocrinology & metabolism, 100(8), 28492852. wibawa, k., kuhuwael, f. v., putra, c. r. j., widiastuti, s. u., & suciadi, l. p. (2021). euglycemic diabetic ketoacidosis associated with empagliflozin in patients hospitalized with acute pulmonary embolism. clinical diabetology, 10(2), 204-208. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 9 american journal of chemistry and pharmacy (ajcp) a rare type of epilepsy: a case report ali hassan salman nasser1* volume 1 issue 2, year 2022 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v1i2.1053 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: december 05, 2022 accepted: december 16, 2022 published: december 23, 2022 epilepsy is one of the most frequent severe brain disorders, impacting about 70 million individuals globally. its prevalence is bimodal, with neonates and the elderly suffering the highest risk. whether using medications or not, nearly two out of every three individuals suffering from new-onset epilepsy will achieve long-term seizures remission, although around half will experience mild to moderately severe side effects. patients with epilepsy, mainly the 20-30percent of the respondents whose seizures are not fully controlled by present treatments (drug-resistant epilepsy), have a significantly increased risk of death, mental and physical comorbidities, and severe pharmaceutical effects. here we present a case of 56-year-old man who was admitted to a private hospital and diagnosed with a rare type of epilepsy. he was unconscious, hence reported as comatose state. mental diseases, such as depression, have also been predominantly studied in our patients, although there was a rising acknowledgment of a bidirectional association between the epilepsy and depression. the patient has been introduced to pregabalin 75mg once to rouse from a comatose state. following this regime, he was declared conscious and alive. he was discharged and continued to pregabalin 75mg daily until the next follow-up. pregabalin was found to be a significant addition to the treatment of epilepsy because of its effective antiseizure action, favorable pharmacokinetic properties, and efficacy in frequent co-morbidities. antiepileptic drug development must be resuscitated rapidly in order to identify new effective anti-seizure medications for the treatment of drug-resistant epilepsy international health practitioners may continue the further investigation related to the onset of this condition to reach significant evidence and treatment. keywords epilepsy, obstructive sleep apnea, depression, hypersomnolence, insomnia, pregabalin, case study 1 avp, oman insuransce co dubai, united arab emirates * corresponding author’s e-mail: alihassan35s@outlook.com introduction the choice of whether or not to do a formal laboratory assessment of the sleeping patient is crucial to the evaluation of patients with sleep problems. many of the presenting symptoms may be related to different diseases that entail sleep disruption; however, much of the information in this section focuses on the patient with respiratory irregularities during sleep. though a more comprehensive system for categorizing sleep disorders has been devised and published elsewhere (coleman et al., 1982; thorpy, 2012), the method used here is more streamlined and focuses on symptoms. since there are more sleep labs available than ever before, this article will concentrate on how to recommend individuals who may be suffering from a sleep issue to one. table 1 provides an exhaustive list of symptoms experienced by individuals with sleeprelated breathing issues. in contrast, table 2 provides the most prevalent signs that indicate sleep disturbance as the underlying cause. patients with sleep disorders often have insomnia (trouble starting or keeping asleep) or excessive daytime drowsiness (hypersomnolence) (bixler et al., 1979; thorpy, 2012). because of the intertwined nature of sleep and wakefulness, it is not uncommon for both of these conditions to coexist. however, to treat either disease effectively, it is necessary first to identify whether insomnia or hypersomnolence is more prevalent. table 1: symptoms in patients with breathing disorders during sleep major or common symptoms awakening from slumber (occasionally with shortness of breath) abrupt cessations of breathing (as reported by bed partner) an excessive inability to stay awake throughout the day (hypersomnolence) screeching snores minor or less common symptoms sleep-related abnormal motor behavior insomnia table 2: symptoms suggesting underlying sleeping disorders insomnia hypersomnolence hypersomnolence simply asking patients may diagnose pathologic daytime drowsiness if they have experienced tiredness during typically awake activities like chatting or eating (smith, 1988). sleepiness during sedentary activities like reading or watching television is a common symptom of a https://doi.org/10.54536/ajcp.v1i2.1053 https://journals.e-palli.com/home/index.php/ajcp pa ge 10 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(2) 9-12, 2022 sleep disturbance in its early stages, but it may be easy to overlook. it is thus important to listen to the input of loved ones who may have noticed a shift in the subject’s degree of attention. hypersomnolence differs from exhaustion, fatigue, or lethargy since those states do not automatically lead to drowsiness. the social shame and fear of job loss sometimes lead individuals to conceal clinically obvious hypersomnolence (smith, 1988). moreover, sleep apnea and other sleep disorders may cause cognitive impairment, reducing the individual’s sensitivity to tiredness. therefore, it may be more important to assess how well a person sleeps than how well they can remain up when trying to provoke this symptom. inquire as to whether or not you tend to nod off when doing boring or repetitive tasks. many factors may lead to excessive sleepiness. night-time sleep problems are the first cause of daytime drowsiness because they alter the typical sleep pattern (smith, 1988). sleep disorders like sleep apnea or restless leg syndrome may drastically reduce restorative slow-wave sleep while boosting stage 1 sleep. the frequency of simultaneous partial arousals, which may follow respiratory or movement abnormalities, is more significant than any changes in the distribution of phases. a night of sleep may be severely fragmented and shortened by as many as 300 to 500 arousals, each lasting 15 to 20 seconds. despite the high frequency, patients are unaware of a sleep disorder since the arousals do not lead to complete restlessness (zarcone, 1973). insomnia insomnia, which is not a diagnosis in and of itself but rather a symptom complex defined by difficulty in beginning sleep, intermittent arousal from sleep, or early morning awakenings with an inability to return to rest, is another important presenting symptom of individuals with sleep disorders. it is common to practice dividing insomnia into two categories: temporary and chronic (thorpy, 2012). insomnia that lasts just a few nights and is brought on by external factors that often pass on their own is considered transient or episodic. psychophysiological stress is the leading cause of temporary sleeplessness (carskadon et al., 1976). though sleep lab examinations often show normal sleep architecture, chronic or persistent insomnia is typically attributable to a chronic worry state (carskadon et al., 1976). suppose a patient has real insomnia, as shown by decreased total sleep duration, fragmented sleep, or early morning awakenings. in that case, they will likely also have chronic weariness or lassitude but not hypersomnolence. insomnia is common in patients who suffer from sleep apnea or movement problems because of the numerous full awakenings that might occur as a result. the elderly may have a lower arousal threshold than younger patients, making it more difficult to fall asleep and stay asleep even when mild central or obstructive sleep apnea is present. similarly, to patients with hypersomnolence, endogenous and external stimuli may have tertiary effects on sleep. insomnia and drowsiness throughout the day are common side effects of serious medical conditions such as chronic pain, dyspnea, and others. however, normal sleep start with early morning awakenings and is more common among depressed people. significant problems with falling asleep may also be caused by exogenous causes, such as stimulant drugs or substance usage. epilepsy around 1% of the population has epilepsy, making it the second most frequent major neurological illness after stroke (stafstrom, 2006). about 50 million individuals worldwide have epilepsy, with 90 percent of those affected living in poorer nations (kandar et al., 2012). our knowledge of epilepsy has advanced in various ways during the last several years. it is a common chronic neurological condition characterized by repeated unprovoked seizures and an imbalance between brain excitability and inhibition (blume et al., 2001; holmes & ben-ari, 2001). keeping the thalamus and its cortical connections, the temporolimbic system, and the ascending reticular system in good working order is essential for maintaining consciousness. if the ascending reticular system were damaged structurally or functionally, the patient would have severe abnormalities in their ability to quantify their surroundings or perhaps go into a coma. eeg patterns in a coma have been used to predict coma outcomes (bauer et al., 2013) for decades and correlate to the degree of consciousness impairment and the depth of coma. however, the normal eeg patterns do not indicate a specific coma etiology (synek, 1989, 1990a, 1990b). these developments have led to a new taxonomy of epileptic seizures and epilepsies. the diagnosis is founded on a thorough clinical history and a credible eyewitness description of a seizure (thijs et al., 2019). over the last decade, novel medicines for the treatment of epilepsy have proliferated. with an everexpanding range of possibilities, physicians must select which therapy, or combined effect of interventions, is optimal for a specific individual. despite the existence of controlled scientific trials for each therapy option, the answers to these concerns may remain elusive(leemanmarkowski & schachter, 2017; thijs et al., 2019). ancillary investigations can aid in the determination of the etiology and prognosis (stafstrom & carmant, 2015). therefore, we report a case of a male who have suffered from a rare cognitive behavior that included epilepsy and depression with variables like insomnia, obstructive sleep apnea, and hypersomnolence. the combination of such conditions have turned the case as rare. case presentation a 56-year-old male, who is receiving treatment in a private hospital in the united arab emirates, suffers from a very strange condition for which the doctors did not find an explanation and considered it a very rare case in medical science. the patient’s chronic symptoms had a history of depression during sleeping and he used to https://journals.e-palli.com/home/index.php/ajcp pa ge 11 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(2) 9-12, 2022 take “clonazepam gabapentin” to treat depression. the patient was also diabetic for which he takes “metformin” 750mg per day. history of cardiovascular disease was also reported and “atrova statin” was used to treat congenital heart disease. along with these medications, “esomeprazole” was used to treat stomach acid 20mg once a day, “ergocalciferol” one capsule per week was given to the patient that enhances vit d level, “neurobion” 100mg to enhance vitamin b complex and magnesium oxide 400mg once a day. the acute illness of patient reported the onset symptoms 5 years ago, which was worst on the right leg and worsened over time in the form of mild irritability in the leg, causing difficulty in moving. he had difficulty and utility of the feet in both legs, which was worsening during sleep and at times of prolonged sitting. the abnormal leg movements started worsening, and he used to have wild flinging movements in sleep that resulted in fatigue and numbness of the outer aspect of the left thigh. the patient was started with sifrol, lyrica, and lidocaine to treat this condition, but no effect was measured. due to this, the doctor advised shifting towards another treatment, i.e., “pramipexole with gabapentin,” to treat the numbness and burning sensation and increased the dose of lyrica and lidocaine. insomnia was also reported from the patient’s history, and “zolpidem” was used to treat it, 5mg once a day. no history of difficulty of gait or any dystonic posturing, no history session of involvement of higher mental functions; cranial nerves, cerebellar system, autonomic nervous system. at the time of admittance, patient was unconscious for straight seven hours. no seizures were reported. according to the neurologist, this problem is not limited to his area of expertise; as the patient experiences this condition four or five times a year. there have been no warning symptoms before the onset of this condition; it occurred suddenly. doctors in the emergency department take immediate measures to bring a comatose patient back to life, but no improvement was observed initially. the physician also implanted a needle into the patient’s body, used cold water and bits of iron to contact the patient’s feet to rouse him. after ensuring that the patient’s vital organs are functioning correctly, he was taken to the medical imaging room for mri & ct scan. the patient was neither dead nor alive; and merely “attentive” to the movements and sounds in his immediate environment. after 7 hours, the patient spontaneously rouses from his coma, without the help of any medical professionals or mechanical aids, and expresses a desire to be discharged; hence the patient was considered “alive.” after regaining consciousness, the patient answered the basic questions correctly about his identity and present state. however, the patient remained silent when the doctor inquires about the events that transpired half an hour before his loss of consciousness. the patient was sent to a team of specialists, including a cardiologist, psychiatrist, oncologist, ent, and pulmonologist. moreover, the internal brain and nerves, and the result came from the doctors after the testing, from x-rays to medical tests, showed everything was normal. since he was in unconscious state for 7 hours, the neurologist prescribed him antiepileptic drugs; pregabalin and lyrica. according to the neurologist, the condition of unconsciousness could have been a possible reason of a rare type of epilepsy. discussion from the investigation it is evident that patient has a rare type of epilepsy. nowadays, several safe antiepileptic drugs are introduced limiting the indications for depression and anxiety prescription. despite the fact that there are safer ways to treat status epilepticus, the medicine has its own market. increased chloride inflow and neural inhibition are hypothesized to be responsible for the antiepileptic effects of the drugs through its impact on gammaaminobutyric acid (gaba) receptors. (olsen et al., 1986). depression is an under recognized yet prevalent comorbidity of epilepsy. the identification of mood disorders is the first and most crucial step in providing effective treatment for this group. improved detection and treatment quality may be achieved by increased clinical attention and the regular use of a validated screening instrument. people who have epilepsy, like the general community, are increasingly curious in complementary and alternative medicine treatments for persistent conditions like depression (fenton & udwin, 1965). epileptic disorders include sleep disorders, such as narcolepsy (nrclp), restless legs syndrome (rls), and obstructive sleep apnea syndrome (osas), are increasingly thought to be influenced by a set of inherited factors, and evidence for this is mounting (caylak, 2009). excessive daytime drowsiness, cataplexy, sleep paralysis, and hallucinations are characteristic symptoms of nrclp. a common symptom of rls is an incessant itch to move one’s legs, which may make it difficult to fall asleep. recurrent bouts of upper airway collapse and blockage during sleep define obstructive sleep apnea syndrome (osas), a serious sleep disorder (almbaidheen & bodur, 2021). however, the results of our research imply that antiepileptic medicine pregabalin might be useful in the treatment of individuals suffering from epilepsy, depression, and osa. pregabalin, the s-enantiomer of 3-aminomethyl5-methylhaxanoic acid, is an antiepileptic drug (aed) of the second generation, developed after gabapentin, with enhanced pharmacokinetic and pharmacological properties. pregabalin is a favorable therapy choice due to its rapid absorption, high bioavailability, and absence of transporter saturation at therapeutic levels. more than ninety percent of the drug is removed in the urine, and nearly none is metabolized by the liver. pregabalin is believed to exert its antiepileptic action through modifying calcium channel traffic and physiology by binding to the 2 subunit of p/q-type voltage-gated calcium channels, hence reducing presynaptic calcium influx and neurotransmitter release (hamandi & sander, https://journals.e-palli.com/home/index.php/ajcp pa ge 12 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(2) 9-12, 2022 2006; schulze-bonhage, 2013). currently, our group is conducting follow-up experiments to validate this rare case and to investigate the potential benefits of antiepileptic medications. moreover, the current scenario is also under study by international hospitals and medical professionals. conclusion it is generally agreed that epilepsy, with a lack of clarity in its definition, is best understood as a transitional condition. the health professionals diagnosed our patient with a very unusual kind of epilepsy. our findings indicate a strong link between epilepsy, osa, depression, and rls. moreover, medical researchers worldwide are also examining the present situation. in our case, pregabalin has proven to be effective against epilepsy as it prevents seizures in epilepsy by lowering aberrant electrical activity in the brain. it suppresses the nerves pain by influencing pain messages traveling through the brain and down the spine. it prevents brain from producing the chemicals that cause anxiety. more antiepileptogenic drugs are also required to prevent epilepsy before the first seizure in at-risk individuals, as well as disease-modifying medications to treat persistent severe seizures associated with progressing underlying illness. acknowledgement the author is thankful to the university of manchester, saudi arabia, for the continuous support of this research study. funding no funding sources are reported. conflict of interest the author does not have any conflict of interest. references almbaidheen, m., & bodur, m. (2021). case report: early onset narcolepsy initially misdiagnosed as obstructive sleep apnea syndrome. the turkish journal of pediatrics, 63(2), 334-338. bauer, g., trinka, e., & kaplan, p. w. (2013). eeg patterns in hypoxic encephalopathies (post–cardiac arrest syndrome): fluctuations, transitions, and reactions. journal of clinical neurophysiology, 30(5), 477-489. bixler, e. o., kales, a., soldatos, c. r., kales, j. d., & healey, s. (1979). prevalence of sleep disorders in the los angeles metropolitan area. the american journal of psychiatry. blume, w. t., lüders, h. o., mizrahi, e., tassinari, c., van emde boas, w., & engel jr, e. o., jerome. (2001). glossary of descriptive terminology for ictal semiology: report of the ilae task force on classification and terminology. epilepsia, 42(9), 1212-1218. carskadon, m. a., dement, w. c., mitler, m., guilleminault, c., zarcone, v. p., & spiegel, r. (1976). self-reports versus sleep laboratory findings in 122 drug-free subjects with complaints of chronic insomnia. am j psychiatry, 133(12), 1382-1388. caylak, e. (2009). the genetics of sleep disorders in humans: narcolepsy, restless legs syndrome, and obstructive sleep apnea syndrome. american journal of medical genetics part a, 149(11), 2612-2626. coleman, r. m., roffwarg, h. p., kennedy, s. j., guilleminault, c., cinque, j., cohn, m. a., karacan, i., kupfer, d. j., lemmi, h., & miles, l. e. (1982). sleep-wake disorders based on a polysomnographic diagnosis: a national cooperative study. jama, 247(7), 997-1003. fenton, g. w., & udwin, e. (1965). homicide, temporal lobe epilepsy and depression: a case report. the british journal of psychiatry, 111(473), 304-306. hamandi, k., & sander, j. w. (2006). pregabalin: a new antiepileptic drug for refractory epilepsy. seizure, 15(2), 73-78. holmes, g. l., & ben-ari, y. (2001). the neurobiology and consequences of epilepsy in the developing brain. pediatric research, 49(3), 320-325. kandar, h., das, s. k., ghosh, l., & gupta, b. k. (2012). epilepsy and its management: a review. journal of pharmascitech, 1(2), 20-26. leeman-markowski, b. a., & schachter, s. c. (2017). cognitive and behavioral interventions in epilepsy. current neurology and neuroscience reports, 17(5), 1-11. olsen, r. w., yang, j., king, r. g., dilber, a., stauber, g. b., & ransom, r. w. (1986). v. barbiturate and benzodiazepine modulation of gaba receptor binding and function. life sciences, 39(21), 1969-1976. schulze-bonhage, a. (2013). pharmacokinetic and pharmacodynamic profile of pregabalin and its role in the treatment of epilepsy. expert opinion on drug metabolism & toxicology, 9(1), 105-115. smith, p. l. (1988). evaluation of patients with sleep disorders. seminars in respiratory medicine, stafstrom, c. e. (2006). epilepsy: a review of selected clinical syndromes and advances in basic science. journal of cerebral blood flow & metabolism, 26(8), 983-1004. stafstrom, c. e., & carmant, l. (2015). seizures and epilepsy: an overview for neuroscientists. cold spring harbor perspectives in medicine, 5(6), a022426. synek, v. (1989). validity of a revised eeg coma scale for predicting survival in anoxic encephalopathy. clinical and experimental neurology, 26, 119-127. synek, v. (1990a). revised eeg coma scale in diffuse acute head injuries in adults. clinical and experimental neurology, 27, 99-111. synek, v. (1990b). value of a revised eeg coma scale for prognosis after cerebral anoxia and diffuse head injury. clinical electroencephalography, 21(1), 25-30. thijs, r. d., surges, r., o’brien, t. j., & sander, j. w. (2019). epilepsy in adults. the lancet, 393(10172), 689701. thorpy, m. j. (2012). classification of sleep disorders. neurotherapeutics, 9(4), 687-701. zarcone, v. (1973). narcolepsy. new england journal of medicine, 288(22), 1156-1166. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 5 american journal of chemistry and pharmacy (ajcp) oral acyclovir induced acute kidney injury yasser sadawey1*, yasameen abdulhameed1 volume 1 issue 2, year 2022 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v1i2.829 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: october 18, 2022 accepted: october 29, 2022 published: november 01, 2022 acyclovir is an antiviral medication often used to treat outbreaks of herpes in children. acyclovir has an exceptional safety profile; there is a possibility that it might induce severe nephrotoxicity in rare cases. in this report, we present the case of a 17-year-old girl with acute renal injury as a side effect of using acyclovir. the patient was admitted for three days due to drug-induced aki, which was reversed by discontinuing the afflicted medicine (acyclovir), initiating appropriate hydration with intravenous normal saline 0.9%, and refraining from any nephrotoxic medications. following up a week later, the patient was asymptomatic and had normal kidney function. therefore, it is critical to use acyclovir correctly to avoid possibly fatal consequences. keywords oral acyclovir, acute kidney injury, acyclovir nephrotoxicity, medicine 1 department of medicine, mediclinic me, al jowhara hospital, al ain, uae * corresponding author’s e-mail: yassersadawey12@outlook.com introduction acyclovir is now the most effective treatment for several herpes viruses, such as herpes encephalitis (balfour h. h., 1984; yy., 1984), varicella zoster virus (vzv), or herpes simplex virus (hsv) infections in children with compromised immune systems. in addition, high-dose intravenous acyclovir to combat viral infections may be beneficial (yy., 1984). the adverse effects of acyclovir are well-known; nonetheless, most people do not consider it. the most prevalent underlying mechanism that acyclovir may generate is crystal nephropathy, which is reported to have the adverse side effect of inducing acute renal injury (aki). it is hypothesized that the mechanism that causes the damage is the precipitation and crystallisation of the medicine inside the renal tubules, which results in obstruction and perhaps cellular necrosis (izzedine, 2005; mason, 2008). patients with acyclovir-induced acute kidney injury have a fast reduction in their renal function and an increase in their blood creatinine level. this usually occurs between 12 and 48 hours after administering the medicine (yildiz c, 2013; zhang y, 2016). appropriate diagnosis and management of aki caused by acyclovir are necessary for an effective prognosis(zhang y, 2016). in this report, we present a case of a 17-year-old girl who developed an acute kidney injury as a side effect of acyclovir in the management of herpes zoster infection. methodology case presentation seventeen years old female with known epilepsy was admitted to our hospital. she was diagnosed five years ago with epilepsy and started on treatment for a few weeks with poor compliance due to medication side effects. she has a five-day history of skin rash affecting her upper back and right shoulder (itchy, vesicular, painful, and affecting a dermatomal distribution of cervical 4,5 and 6) associated with nausea and vomiting at the time of admittance in another facility. during that period, she was started on acyclovir 800 mg orally three times daily for the diagnosis of herpes zoster infection, diclofenac sodium 50 mg twice and levetiracetam 500 mg orally every 12 hours. unfortunately, she was discharged one day before her current presentation to our hospital. she also gave a history of reduced urine output. upon arrival, she was vitally stable and afebrile, weighted 60 kg with normal capillary refill time but dry mucous membranes. systemic examination, apart from mild epigastric tenderness, was reported as unremarkable. neurological examination revealed unremarkable results, including motor, sensory, cranial nerves, and cerebellar functions, and no photophobia or neck stiffness. chest x-ray and ultrasound abdomen were unremarkable. ct head done earlier showed mucosal thickening and retention cyst of left maxillary sinuses. her laboratory results showed creatinine of 202wµ mol/l, normal electrolytes and ph 7.33, and hco3 18 meq/l, lactate 0.8 mmol/l. she was admitted as having acute kidney injury, probably drug-induced, using the naranjo algorithm of adverse drug reaction. the patient was admitted for three days as drug-induced aki, and the culprit drug (acyclovir) was stopped with the initiation of proper hydration with normal intravenous saline 0.9% and avoiding all nephrotoxic medications. she was started on metoclopramide 10 mg intravenous when needed, pantoprazole 40 mg intravenous once daily, and paracetamol 1 gm for pain control, daily renal function tests showed improvement of kidney functions, and she was tolerant of oral intake. upon follow-up a week later, a patient had normal kidney functions and was asymptomatic. https://doi.org/10.54536/ajcp.v1i2.829 https://journals.e-palli.com/home/index.php/ajcp mailto:yassersadawey12%40outlook.com?subject= pa ge 6 table 1: laboratory values of admitted case normal value feb 2021 baseline 2 days pre admission 1 day pre admission day 1 admission day 2 day 3 follow up in 1 week sodium 136-145 mmol/l 140 139 139 138 141 140 136 potassium 3.2-5.5 mmol/l 4.3 4.2 4.2 4.2 3.8 4.5 chloride 98-107 mmol/l 105 104 103 101.4 106.6 103.8 urea 2.8-8.1 mmol/l 6.3 4.5 6.5 9.1 6.9 3.9 3.7 creatinine 34-80 micrmol/l 56 118 231 206 148 82 65 co3 22-29 mmol/l 26 24 19 ph 7.35-7.45 na 7.35 7.35 crp <5mg/l na 5.3 28.8 35 26 10 alkaline phosphatase 48-95 u/l 109 alt 0-17 u/l 12.2 11.1 amylase 29-118 u/l 57 ast 0-23 u/l 15.8 13 calcium 2.31-2.64 mmol/l 2.35 tsh 0.5-4.5 microiu/ml 1.89 hct (pcv) 31-42% 32.8 29.4 34.9 haemoglobin 10.9-14.3 g/dl 10.6 9.5 11.2 basophils % 0.4-1.3% 0.3 0.4 0.5 eosinophils % 0.4-8.2 % 1.2 2.4 3.1 lymphocytes % 20.1-48.8 % 27.6 28.8 5.7 mch 24.7-32.8 pg 23 22.9 23.2 mchc 32.3-35.6g/dl 32.3 32.4 32 mcv 75.5-95.3 fl 71.3 70.7 72.4 monocytes % 4.10-12.90% 11.3 13 10.3 neutrophils % 42.8-75.1 % 59.6 55.4 80.4 platelet count 150-410 10³/ųl 249 237 264 red cell count 3.63-4.92 10⁶/ųl 4.6 4.15 4.82 white cell count 3.8-11.8 10⁶/ųl 6.7 6.2 7.4 urine rotine blood (rbc) negative negative negative ketone negative positive + negative protein negative negative 10 mg/dl amorphous urate nil nil nil amorphous phosphate nil nil nil bilirubin negative negative negative crystals nil nil nil fatty casts nil nil nil granular casts nil nil nil hyaline casts nil nil nil pa ge 7 rbc casts nil nil nil wbc casts nil nil nil glucose negative negative negative leukocytes 0-5 cell/hpf 10-jun 0-5 leukocyte esterase negative positive + negative nitrite negative negative negative ph 5-9 ph 5 6 bacteria nil few ++ nil specific gravity 1.003 1.035 1.01 1.009 discussion nephropathy caused by oral acyclovir is uncommon and only manifests itself at very high doses (more than 500 mg/m2) when the medication is used by patients whose volume status has significantly reduced (perazella., 1999). it is critical to know the possible nephrotoxicity for the patients being treated with acyclovir. our patient had an acute kidney injury while taking oral acyclovir and other nephrotoxic drugs and being volume depleted, but her creatinine improved when acyclovir stopped therefore, acyclovir crystalluria and subsequent intrarenal obstruction and nephropathy would be the most plausible explanation for the observed renal injury, even if the crystals were undetected on regular urine analysis. in individuals with underlying volume depletion and renal insufficiency, acyclovir dosage should be lowered. to avoid crystallisation and eventual tubular blockage, gradual medication infusion over 1-2 hours, appropriate fluid replenishment, and introduction of high urine flow rates (100-150 ml/h) should be recommended (brigden et al., 1982; sawyer et al., 1988). renal function should be monitored regularly in patients receiving high-dose intravenous acyclovir in patients diagnosed with renal impairment at any dosage level. common symptoms include unusual sickness, nausea, abdominal discomfort, vomiting, and muscle twitching while receiving treatment (wade jc, 1983). when serum creatinine levels rise, acyclovir must be managed to be discontinued. hydration should be maintained during therapy to ensure a high urine flow(gunness p, 2010). if no improvement is observed in patients with renal function soon after discontinuing acyclovir, other causes of renal toxicity should be sought. treatment may be maintained at a lower dose for secondary infections that respond to acyclovir while a renal function is closely monitored. since volume contraction may go undiagnosed in outpatients treated with acyclovir, they are more susceptible to acyclovirinduced kidney damage(chawla, 2017). conclusion it is concluded that to prevent morbidity, acute kidney injury must be diagnosed as soon as possible (ratan., 2003). since acyclovir is frequently prescribed for renal transplantation, patients with herpes simplex virus and herpes zoster infections, and patients with neurological viral infections, therefore, practitioners must be aware of the associated risk, side effects, and how drug-related issues might be addressed. ethical approval ethical approval for this study was obtained from mediclinic al jowhara hospital (reference number: uhn: 1238416). all procedures performed in the study involving the patient were by the ethical standards of the governmental guidelines and with the 1964 helsinki declaration and its later amendments or comparable ethical standards. conflict of interest the authors declare that there is no conflict of interest regarding the publication of this case report. funding any grants did not fund this work. informed consent written informed consent was obtained from the participant. reference balfour h. h., j. (1984). intravenous acyclovir therapy for varicella in immunocompromised children. . the journal of pediatrics, 104(1), 134-136. https://doi.org/ https://doi.org/10.1016/s0022-3476(84)80611-3 brigden, d., rosling, a. e., & woods, n. c. (1982). renal function after acyclovir intravenous injection. the american journal of medicine, 73(1), 182-185. chawla, l., bellomo, r., bihorac, a. et al. . (2017). acute kidney disease and renal recovery: consensus report of the acute disease quality initiative (adqi) 16 workgroup. nat rev nephrol 13, 241-257. https://doi. org/https://doi.org/10.1038/nrneph.2017.2. gunness p, a. k., koren g. (2010). the effect of acyclovir on the tubular secretion of creatinine in vitro. j transl med., 30(8), 139. https://doi.org/10.1186/1479pa ge 8 5876-8-139 izzedine, h., launay-vacher, v., & deray, g. (2005). antiviral drug-induced nephrotoxicity. american journal of kidney diseases : the official journal of the national kidney foundation, 45(5), 807-817. https://doi.org/ https://doi.org/10.1053/j.ajkd.2005.02.010 mason, w. j., & nickols, h. h. . (2008). images in clinical medicine. crystalluria from acyclovir use. the new england journal of medicine, 358(13). https://doi.org/ https://doi.org/10.1056/nejmicm066726 (14) perazella., m. a. (1999). crystal-induced acute renal failure. american journal of medicine, 106(4), 459-465. ratan., p. k. s. a. s. k. (2003). a case of chronic renal dysfunction following treatment with oral acyclovir. scandinavian journal of infectious diseases, 35(10), 770772. sawyer, m. h., webb, d. e., balow, j. e., & straus, s. e. (1988). acyclovir-induced renal failure: clinical course and histology. the american journal of medicine, 84(6), 1067-1071. wade jc, m. j. (1983). neurologic symptoms associated with parenteral acyclovir treatment after marrow transplantation. ann intern med, 98, 921-925. yildiz c, o. y., gucer s, cengiz ab, topaloglu r. (2013). acute kidney injury due to acyclovir. cen case rep, 1, 38-40. https://doi.org/10.1007/s13730-012-0035-0 yy., b. (1984). the use of acyclovir in children. pediatric infectious disease, 3(4), 575-578. https://doi.org/ https://doi.org/10.1097/00006454-19840700000017 zhang y, c. y., teng y. (2016). acute renal injury induced by valacyclovir hydrochloride: a case report. exp ther med, 6, 4025-4028. https://doi.org/10.3892/ etm.2016.3905. pa ge 1 pa ge 32 american journal of chemistry and pharmacy (ajcp) covid infection and variables affecting its severity among pregnant women in dubai kefah el debek1* volume 2 issue 1, year 2023 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v2i1.1397 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: march 16, 2022 accepted: april 27, 2023 published: april 30, 2023 the acute respiratory syndrome has produced an unprecedented global disaster since december 2019 known as coronavirus 2 (sars-cov-2). pregnant women are a particular group that needs special care during crises and infectious illnesses. compared to uninfected women, this cross-sectional study was based on pregnant women infected with covid-19. the data was collected from dha latifa hospital, medical records. the research committee approved the study in dha. the study was based on 326 patients aged 20-42 years, of which covid-infected pregnant women were 199 and 118 were uninfected. most patients were from the uae, with a mean age of 31.63+ 6.36 and an average bmi of 28.96±1.16 kg/m2 of covid-infected pregnant women. while the mean age of 31.33+ 6.23 and an average bmi of 27.06±11.05 kg/m2 of uninfected pregnant women. hemoglobin (hb), white blood cells (wbcs), and c-reactive protein(crp) showed significantly different when compared to covid-infected and uninfected pregnant women. significant results showed in comorbid diseases compared with covid-infected and uninfected pregnant women, but a high frequency was observed in uninfected women. the results showed insignificance with comorbid diseases when compared with covidinfected pregnant women and uninfected pregnant women. a decrease in wbc and crp was observed in covidinfected pregnant women keywords covid, comorbid diseases, hemoglobin, infection, pregnant women 1 dubai health authority, united arab emirates * corresponding author’s e-mail: khaldbk@dha.gov.uae introduction the severe acute respiratory syndrome has produced an unprecedented global disaster since december 2019 known as coronavirus 2 (sars-cov-2) (kamel boulos & geraghty, 2020; sohrabi et al., 2020). pregnant women are a particular group that needs special care during crises and infectious illnesses. according to the literature, pregnant women infected with covid-19 suffer a more severe form of the disease, which increases their chance of dying by up to 35.0 percent and causes nearly a quarter of them to develop pneumonia. furthermore, pregnant women affected by covid-19 infection had a higher likelihood of being hospitalized than non-pregnant people of the same age (31.5 percent versus 5.8 percent), as reported by the cdc (centers for disease control and prevention). ukoss research showed that many pregnant/expected women were admitted to the hospital in the third trimester or postpartum, and severe illness happened in later pregnancy (vousden et al., 2021). according to the literature, immunologic and physiological changes during pregnancy can be blamed for the complexity of the maternal disease and obstetric effects. these changes include reduced lung volume, cardiovascular and hemodynamic deviations, high oxygen demand, hypoxia causing respiration problems such as low maternal tolerance, structural changes with an enlarged transthoracic diameter, and restricted diaphragm expansion as previous health issues like preeclampsia or diastole (chen et al., 2020; yan et al., 2020). the epidemiology of severe covid-19-mediated pregnancy-related changes in the inflammatory response to viruses has been linked to an overt th1 and th2 response to sars-cov-2. natural killer (nk), plasmacytoid dendritic cells decline while increased progesterone levels in circulating cells during pregnancy (wastnedge et al., 2021). these changes in the mother’s immune system affect the clinical course of covid-19 and the therapy and prevention of covid-19 in pregnancy. pregnant women may be more prone to serious respiratory infections because of decreased lung capacity and difficulty removing secretions (thompson et al., 2020). regarding the potential effects of covid in early pregnancy, there is less research (up to 12 weeks gestation). seasonal influenza has been linked to a greater incidence of miscarriage (dorélien, 2019). extrapolating from the impact of other viruses in late pregnancy (more than 24 weeks gestation) (dorélien, 2019) (mosby et al., 2011), covid-19 infection might result in higher rates of unfavourable pregnancy results in fetal growth limitation, premature delivery, and perinatal death. once available, population-level data on these results must be evaluated to spot tendencies relevant to the covid-19 pandemic. this research examines the relationship between maternal severity due to covid-19 and the effect of comorbid disease. examining how demographic variables affected by covid on pregnant women. we also studied the correlation between the severity of illness and the number of dosages, comorbid diseases, and the duration of the vaccine (the time since it was given wither within 6 months or in more than 6 months). materials and methods this cross-sectional study was based on 326 patients https://doi.org/10.54536/ajcp.v2i1.1397 https://journals.e-palli.com/home/index.php/ajcp pa ge 33 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 32-37, 2023 aged 20-42 years, of which covid-infected pregnant women were 199 while 118 were uninfected. the data was collected from dha latifa hospital, medical records. this research was conducted after the ethical approval from the research committee in dha. informed consent was also obtained. the questionnaire contains data on demographic characteristics, symptoms, blood profile, and vaccination details. statistical package of social sciences (spss, v20.0) was used for data analysis. the data was represented as mean ± sd, while variable differences were analyzed as frequency (percentage). fisher and student t-tests were used to comparing covid-infected and uninfected pregnant women. a relative regression test was performed to determine the relations of participants’ characteristics such as age, bmi, and severity of covid-19 symptoms and blood profile. the statistically significant value p= <0.05 was considered. results and discussion results table 1: demographic characteristics of the study variables all patients (n=326) p-value age 31.63+ 6.36 <0.001* ethnicity, n (%) uae 182(0) 0.005* yemen 13(3.9) 0.05* pakistan 12(3.68) 0.05* oman 8(2.4) 0.55 comery 1(0.3) 0.55 kyrgyzstan 1(0.3) 0.55 comoros 4(1..22) 0.55 kuwait 1(0.3) 0.55 france 1(0.3) 0.55 pakistan 3(0.9) 0.55 sri lanka 2(0.6) 0.55 jordan 7(2.1) 0.55 tunisia 1(0.3) 0.55 vietnam 1(0.3) 0.55 tajikistan 1(0.3) 0.55 azerbaijan 1(0.3) 0.55 filipi0 14(4.29) 0.05* sudan 1(0.3) 0.55 bangladesh 1(0.3) 0.55 saudi arabia 2(0.6) 0.55 india 15(4.6) 0.05* leba0n 1(0.3) 0.55 syrian 8(2.4) 0.35 iran 12(3.6) 0.05* iraq 1(0.3) 0.55 egypt 11(3.3) 0.05* turkey 1(0.3) 0.55 china 3(0.9) 0.55 bahrain 2(0.6) 0.55 morocco 6(1.8) 0.55 united kingdom 1(0.3) 0.55 turkey 1(0.3) 0.55 leba0n 1(0.3) 0.55 south africa 1(0.3) 0.55 kazakhstan 2(0.6) 0.55 kenya 2(0.6) 0.55 somalia 1(0.3) 0.65 *significance at p-value<0.05. table 2: variation in the covidinfected and uninfected pregnant women related to symptoms, vaccination and blood variables. variables covidinfected pregnant women uninfected pregnant women n 199 118 age 31.63+ 6.36 31.33+ 6.23 bmi (kg/m2) 28.96±1.16 27.06±11.05 https://journals.e-palli.com/home/index.php/ajcp pa ge 34 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 32-37, 2023 gestational age (weeks) 25.06±3.5 27.06±11.05 vaccinated 1±0.4 0.737+0.4 hb 11±0.141 11.20± 1.34 wbc 7 ± 3.25 8.37 + 2.862 crp 18 ± 10.1 17.79+3.722 procalcitonin 0±0.049 0.489+3.27 fever 1±0.7 0±0.3 sore throat 0±0.0 0±0.15 shortness of breath 0±0.7 0±0.0 cough 1±0 0±0.2 dehydration (acetone in urine) 1±0.7 1±0.41 lethargy 0±0. 0±0. figure 1: differences between demographic characteristics of covid-infected and uninfected pregnant women figure 2: differences between blood variables of covid-infected and uninfected pregnant women. https://journals.e-palli.com/home/index.php/ajcp pa ge 35 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 32-37, 2023 figure 3: difference between comorbid disease in covid-infected and uninfected pregnant women demographic characteristics showed in table 1. most patients were from the uae, with a mean age of 31.63+ 6.36 and an average bmi of 28.96±1.16 kg/m2 of covid-infected pregnant women. while the mean age of 31.33+ 6.23 and an average bmi of 27.06±11.05 kg/m2 of uninfected pregnant women. variation in the covid-infected and uninfected pregnant women related to symptoms, vaccination and blood variables (table 2). this study showed that the age, bmi, and gestational age were insignificant compared to the covid-infected and uninfected pregnant women (fig 1). hemoglobin (hb), white blood cells (wbcs), and c-reactive protein (crp) showed significantly different when compared to covid-infected and uninfected pregnant women (fig 2). significant results showed in comorbid diseases compared with covid-infected and uninfected pregnant women, but a high frequency was observed in uninfected women (fig 3). table 3: the severity of illness and its correlates severity of illness type of vaccine number of doses received less than 6 month more than 6 months severity of illness pearson correlation 1 -0.679 -0.75 -0.108 -0.29 type of vaccine pearson correlation -0.679 1 .828** .778** .618** number of doses received pearson correlation -0.75 .828** 1 .823** .833** less than 6 month pearson correlation -0.108 .778** .823** 1 .511** more than 6 months pearson correlation -0.29 .618** .833** .511** 1 **. correlation is significant at the 0.01 level (2-tailed). table 3. shows the correlation between the severity of the vaccine and the number of doses received, dosage received in less than 6 months, and more than 6 months. it can be seen that a negatively moderate correlation was found between the severity of illness and the type of vaccine, and a negatively strong correlation was found between the dosage and severity of illness. in contrast, a negatively weak correlation was seen in the severity of illness and individuals with dosage received in less than 6 months and in individuals with dosage received in more than 6 months. the fitted model data’s chi-squared ratio test yielded a table 4: likelihood ratio test. effect model fitting criteria likelihood ratio tests -2 log likelihood of reduced model chi-square df sig. intercept (soi) 28.185a 0 0 . diabetes miletus 28.422 0.238 1 0 hypertension 28.29 0.105 1 0 hyperthyroidism 34.893 6.708 1 0.01 iron deficiency anemia 30.285 2.1 1 0 hematology disease 29.714 1.529 1 0.016 sle 28.185 0.06 1 0 others 28.438 0.253 1 0 the chi-square statistic is the difference in -2 log-likelihoods between the final and reduced models. the reduced model is formed by omitting an effect from the final model. the null hypothesis is that all parameters of that effect are 0. a. this reduced model is equivalent to the final model because omitting the effect does not increase the degrees of freedom. https://journals.e-palli.com/home/index.php/ajcp pa ge 36 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 32-37, 2023 score of 11.557 (p = 0.000), indicating a good model fit. furthermore, suitable outcomes for the pseudo r-squared (cox and snell: 0.907, nagelkerke: 0.726). our logistic multinomial model’s power was appropriate since it correctly recognized 93.2% of the observed data and can be relied upon to predict future guesses. the likelihood ratio tests for the model’s effects and partials are shown in table 4, and their low p-values show how highly relevant the model’s variables are. discussion this study aimed to contribute to the association between covid-19 severity in pregnancies and their outcomes. literature showed that of the 184 critically ill patients (24% of whom were female) found that 31% of them experienced thrombotic events, demonstrating that covid-19 is linked to the high incidence of thromboembolic consequences in the general population (64). the cause of this is the stimulation of coagulation pathways, which may lead to diffuse vascular coagulopathy (dic), fibrinolysis, and dynamic hypercoagulation associated with thrombocytopenia. the results showed insignificance concerning covid and gestation age(week), while the severity of the condition was significantly correlated with gestational age upon presentation. for asymptomatic patients, the median gestational age at symptom start or diagnosis was 37 weeks; for symptomatic patients, it was 33 weeks; and for patients who needed oxygen assistance, it was 36 weeks. this indicated a risk of severe covid-19 infection in late pregnancy. a modified immune system may control the extreme changing in secondary physiological aspects adapted due to pregnancy. however, the hormonal role in modifying the immune system in pregnancy was significant. according to the evidence, severe covid-19 during pregnancy is linked to iatrogenic premature births (75%), mostly due to maternal indication and during the third trimester (pierce-williams et al., 2020). the results also showed low wbcs compared with uninfected pregnant women, similar results in the observation of mohr-sasson 2020 (mohr-sasson et al., 2020). c-reactive protein was also insignificant (fig 2) and similar to the results observed by rizo-téllez in 2020 (rizo-téllez et al., 2020). the study showed that pregnant women affected by covid do not have a significant chance to transfer it. it may affect the baby during and after birth. furthermore, the modified immune system in pregnancy prevents most diseases but not viral ones. it is challenging to determine with certainty whether pregnant women are at a higher risk of covid-19’s severe repercussions based on the available research. pregnant women are nearly always excluded from more than 300 clinical trials looking into potential therapy alternatives, despite worries about their heightened sensitivity to covid-19 (whitehead & walker, 2020). pregnant participants are seldom included in clinical studies, not even those examining treatments with a wellestablished safety profile during pregnancy. in order to build a fair and knowledgeable evidence base with data from a representative population, researchers should be pushed to consider pregnant women and other groups. in several research, vaccination was seen as a prophylactic method to improve their health (taryam et al., 2021). similar beliefs about immunization being wise, necessary, or beneficial undoubtedly influenced or anticipated vaccine acceptance. other factors mentioned by tuite et al. (tuite et al., 2010) were protecting others and residing with people who could be exposed. immunization. the belief that immunization weakens one’s natural defences, is unpleasant, brings on illness, or has no bearing on one’s health was adversely connected with vaccination uptake (taryam et al., 2021). the provision of acute medical treatment has taken priority over the provision of prenatal care, resulting in a restricted capacity of clinics and a reduced capability to screen for disorders like gestational diabetes. healthcare services have had to be reorganized in many contexts due to social distancing regulations that limit direct interaction between specialists and patients to suggest identifying and treating mental health issues more effectively (brooks et al., 2020). in this situation, most women are extra susceptible to intimate partner abuse (bradbury‐jones & isham, 2020; pal & rao, 2020) and the harmful pregnancy related to it, and they are also less able to seek help. (wenham et al., 2020) the study results observed an insignificance relation between comorbid diseases with covid-infected women, similar to wang’s research in 2020. (wang et al., 2020) the vaccination and the booster dose were very helpful in reducing the intense onset of the disease that leads to death. besides all, vaccination is more important not only to stop the onset of disease but also to restrict the severity of the disease. (menni et al., 2021) social distancing and precautionary measures such as masks, gloves and hand sanitization restrict the spread. in conclusion, the overall features of individuals with covid-19 infection who appear from mid-trimester forward are comparable to those of non-pregnant people based on the clinical and scientific evidence that is currently available. there is currently no proof that covid-19 infection is more likely to affect pregnant women or that covid-19 infection increases the risk of developing severe pneumonia. conclusion the results showed insignificance with comorbid diseases when compared with covidinfected pregnant women and uninfected pregnant women, while a decrease in wbc and crp was observed in covidinfected pregnant women. acknowledgments the author thanks dha latifa hospital, dubai, for continuous support throughout the study. references bradbury-jones, c., & isham, l. (2020). the pandemic paradox: the consequences of covid-19 on https://journals.e-palli.com/home/index.php/ajcp pa ge 37 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 32-37, 2023 domestic violence. journal of clinical nursing, 29(13-14), 2047. brooks, s. k., webster, r. k., smith, l. e., woodland, l., wessely, s., greenberg, n., & rubin, g. j. (2020). the psychological impact of quarantine and how to reduce it: a rapid review of the evidence. the lancet, 395(10227), 912-920. chen, h., guo, j., wang, c., luo, f., yu, x., zhang, w., li, j., zhao, d., xu, d., & gong, q. (2020). clinical characteristics and intrauterine vertical transmission potential of covid-19 infection in nine pregnant women: a retrospective review of medical records. the lancet, 395(10226), 809-815. dorélien, a. (2019). the effects of in utero exposure to influenza on birth and infant outcomes in the us. population and development review, 45(3), 489. kamel boulos, m. n., & geraghty, e. m. (2020). geographical tracking and mapping of coronavirus disease covid-19/severe acute respiratory syndrome coronavirus 2 (sars-cov-2) epidemic and associated events around the world: how 21st century gis technologies are supporting the global fight against outbreaks and epidemics. international journal of health geographics, 19(1), 1-12. https://doi.org/10.1186/ s12942-020-00202-8 menni, c., klaser, k., may, a., polidori, l., capdevila, j., louca, p., sudre, c. h., nguyen, l. h., drew, d. a., & merino, j. (2021). vaccine side-effects and sars-cov-2 infection after vaccination in users of the covid symptom study app in the uk: a prospective observational study. the lancet infectious diseases, 21(7), 939-949. https://doi.org/10.1016/ s1473-3099(21)00224-3 mohr-sasson, a., chayo, j., bart, y., meyer, r., sivan, e., mazaki-tovi, s., & yinon, y. (2020). laboratory characteristics of pregnant compared to nonpregnant women infected with sars-cov-2. archives of gynecology and obstetrics, 302(3), 629-634. https://doi. org/10.1007/s00404-020-05655-7 mosby, l. g., rasmussen, s. a., & jamieson, d. j. (2011). 2009 pandemic influenza a (h1n1) in pregnancy: a systematic review of the literature. american journal of obstetrics and gynecology, 205(1), 10-18. https://doi. org/10.1016/j.ajog.2010.12.033 pal, a., & rao, r. (2020). intimate partner violence during pregnancy. labour room emergencies, 515-520. https:// doi.org/10.1007/978-981-10-4953-8_52 pierce-williams, r. a., burd, j., felder, l., khoury, r., bernstein, p. s., avila, k., penfield, c. a., roman, a. s., debolt, c. a., & stone, j. l. (2020). clinical course of severe and critical coronavirus disease 2019 in hospitalized pregnancies: a united states cohort study. american journal of obstetrics & gynecology mfm, 2(3), 100134. rizo-téllez, s. a., méndez-garcía, l. a., flores-rebollo, c., alba-flores, f., alcántara-suárez, r., manjarrezreyna, a. n., baltazar-lópez, n., hernándezguzmán, v. a., león-pedroza, j. i., & zapata-arenas, r. (2020). the neutrophil-to-monocyte ratio and lymphocyte-to-neutrophil ratio at admission predict in-hospital mortality in mexican patients with severe sars-cov-2 infection (covid-19). microorganisms, 8(10), 1560. sohrabi, c., alsafi, z., o’neill, n., khan, m., kerwan, a., al-jabir, a., iosifidis, c., & agha, r. (2020). world health organization declares global emergency: a review of the 2019 novel coronavirus (covid-19). international journal of surgery, 76, 71-76. taryam, m., alawadhi, d., al marzouqi, a., aburayya, a., albaqa’een, a., alfarsi, a., makki, i., rahmani, n., aljasmi, m., & mubarak, s. (2021). the impact of the covid-19 pandemic on the mental health status of healthcare providers in the primary health care sector in dubai. linguistica antverpiensia, 21(2), 2995-3015. thompson, j. l., nguyen, l. m., noble, k. n., & aronoff, d. m. (2020). covid-19-related disease severity in pregnancy. american journal of reproductive immunology, 84(5), e13339. tuite, a. r., fisman, d. n., kwong, j. c., & greer, a. l. (2010). optimal pandemic influenza vaccine allocation strategies for the canadian population. plos one, 5(5), e10520. vousden, n., bunch, k., morris, e., simpson, n., gale, c., o’brien, p., quigley, m., brocklehurst, p., kurinczuk, j. j., & knight, m. (2021). the incidence, characteristics and outcomes of pregnant women hospitalized with symptomatic and asymptomatic sars-cov-2 infection in the uk from march to september 2020: a national cohort study using the uk obstetric surveillance system (ukoss). plos one, 16(5), e0251123. https://doi.org/10.1371/journal. pone.0251123 wang, d., hu, b., hu, c., zhu, f., liu, x., zhang, j., wang, b., xiang, h., cheng, z., & xiong, y. (2020). clinical characteristics of 138 hospitalized patients with 2019 novel coronavirus–infected pneumonia in wuhan, china. jama, 323(11), 1061-1069. https://doi. org/10.1001/jama.2020.1585 wastnedge, e. a., reynolds, r. m., van boeckel, s. r., stock, s. j., denison, f. c., maybin, j. a., & critchley, h. o. (2021). pregnancy and covid-19. physiological reviews, 101(1), 303-318. wenham, c., smith, j., & morgan, r. (2020). covid-19: the gendered impacts of the outbreak. the lancet, 395(10227), 846-848. https://doi.org/10.1016/ s0140-6736(20)30526-2 whitehead, c. l., & walker, s. p. (2020). consider pregnancy in covid-19 therapeutic drug and vaccine trials. the lancet, 395(10237), e92. https://doi. org/10.1016/s0140-6736(20)31029-1 yan, j., guo, j., fan, c., juan, j., yu, x., li, j., feng, l., li, c., chen, h., & qiao, y. (2020). coronavirus disease 2019 in pregnant women: a report based on 116 cases. american journal of obstetrics and gynecology, 223(1), 111 e1. https://doi.org/10.1016/j.ajog.2020.04.014 https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 9 american journal of chemistry and pharmacy (ajcp) diabetes mellitus control and chronic kidney disease safaa mohamed matarid1* volume 2 issue 2, year 2023 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v2i2.1612 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: april 16, 2023 accepted: may 08, 2023 published: may 13, 2023 chronic kidney disease (ckd) is the rampant onset of diabetes and related complications; chronic kidney disorders and end-stage renal disease are progressing in more than 10% of the world population and mostly affect the elderly, women, minorities and patients with diabetes and hypersensitivity. the early stages of ckd are typically quiet; thus, many people are unaware they have the condition. data for this review was gathered from google scholar, scopus, pubmed, elsevier, cochrane, sage, medline, and web of science. studies were selected from 2018-2023, using keywords such as diabetes mellitus management, chronic kidney disease, microalbuminuria, target time in range, impaired fasting glucose, management ways, lifestyle modification, medication treatment, and diet control. although blood glucose levels are too unpredictable to provide a reliable evaluation, measures reflecting long-term glycemic load are used instead. the results gathered after the review suggests that optimal glycemic control along with lifestyle medication and diet control contributes to better outcomes in individuals with dm, particularly for microvascular damage. while, hba1c is the most well-known glycemic biomarker of long-term glycemic management. this suggests that effective treatments are progressing on progression.this review has discussed different ckd management parameters; the authors have discussed the disease’s treatment criteria and protocols, such as pharmacological therapies, lifestyle modifications, physical activities and insulin therapy and concluded that to avoid ckd, it’s critical to concentrate on underlying problems including hyperglycemia, hypertension, microalbuminuria, sedentary behaviour, and smoking. the occurrence of ckd can be reduced with changes in lifestyle, such as increased physical activity, nutritious food, and water consumption. keywords chronic kidney disease, end stage renal stage, albuminuria, pharmacological therapies 1 department of internal medicine, mediclinic al ain hospital, heikh khalifa bin zayed st next to choithrams supermarket, abu dhabi united arab emirates * corresponding author’s e-mail: safaamatarid12@outlook.com introduction diabetes mellitus is the most common and prevalent disease worldwide, affecting approximately 29.2% of people per year in the united states; while 537 million people around the world are reported to have diabetes, this number is expected to grow by half a billion by the year 2040 (association, 2022; idf, 2021; prevention, 2020). diabetes mellitus is a progressive metabolic disease that is implanted by the constant condition of hyperglycemia. diabetes is prompted by the action of insulin that decreases insulin production, insulin resistance, or both (goyal & jialal, 2018). hyperglycemia is a recognised marker for the onset and development of both dpn and ckd. this lethal disease causes more lethal macrovascular comorbidities such as cardiovascular diseases, diabetic peripheral neuropathy, diabetic retinopathy and ckd, leading to increased mortality rates and decreased quality of life (cole & florez, 2020). during the 1st century ad, diabetes was considered a urinary tract disease or a kidney disease due to its high urine flow. diabetes affects the kidneys primarily due to a deficiency in their retentive properties. diabetes-related urine has a sweet flavour because it contains nutrients and water that have been ingested but not broken down (eknoyan & nagy, 2005). the sweetness of diabetic urine was the initial indicator used by thomas willis in 1674 to distinguish diabetes from other types of polyuria, and he proposed that the sweet flavour initially manifests in the blood(sutherland & gruessner, 2020). a century later, matthew dobson demonstrated that the sweet flavour of urine was caused by sugar and that blood sugar had both paved and followed it. although diabetes was later linked to higher blood sugar levels, sugar in the urine was still accredited to the kidneys’ reduced ability for retentive functioning (porta, 2020). diabetic kidney disease or ckd happened by diabetes or diabetic nephropathy is the most typical onset of diabetes mellitus and also the major reason of end-stage renal disorder (ersd). diabetic kidney disease (dn) is characterised by hyperfiltration and albuminuria, followed by a loss of renal function. it can show in various ways, particularly in individuals with t2dm who may also have peripheral vascular and other glomerular/tubular diseases (sagoo & gnudi, 2020). ckd is reported to affect approximately 8 to 16 % population globally, and around 30-50 % of cases of end-stage renal disorder are considered to be preceded by diabetic kidney disease (chen et al., 2019; copur et al., 2020). the incidence of classic diabetic nephropathy in type 1 diabetes has decreased from 25% to 10-15% with blood pressure management and more use of an angiotensin converter. the overall incidence of diabetes will rise from 6% to 10% of adults in the coming years, leading to more cases of type 2 diabetes and kidney disease (winocour, 2018). ckd is recognised by a biomarker called micro-albumin, which is present in urine. the higher levels of microhttps://doi.org/10.54536/ajcp.v2i2.1612 https://journals.e-palli.com/home/index.php/ajcp pa ge 9 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 9-14, 2023 albumin in urine indicate the presence of a disorder. at the same time, albuminuria is a condition of having too much protein in the urine, resulting in kidney damage (raja et al., 2021). diabetes and ckd together increase the probability of amputations of the lower limbs by two to six times compared to diabetes alone in patients with ckd (rodrigues et al., 2022). literature review progression of micro and macroalbuminuria microalbuminuria is caused by a malfunction of the glomerular basement membrane (gbm). this enzyme is inhibited by insufficient blood sugar management, which lowers the negative charge on gbm and causes extra albumin to leak out. it is linked to type 1 and type 2 diabetic individuals (prasad & tikaria, 2022). in those with type 1 diabetes, microalbuminuria is 6% prevalent after three years but 41% common after five. type 2 diabetes affects 20% to 25% of diabetic paeople (prasad & tikaria, 2022). a decreased glomerular filtration rate, increased microalbuminuria, or both characterise diabetic nephropathy or dkd. laboratory tests are used to diagnose ckd, most frequently by estimating glomerular filtration rate (gfr) using a filtration marker like serum creatinine or cystatin c. the albumin or protein in the urine can be determined using several formulas or other methods (kovesdy, 2022). microalbuminuria, defined by urine albumin excretion of 30-300 mg/day, is thought to be a predictor of dkd. it is regarded as an adjustable problem for the progression of renal disease to its terminal stages (hussain et al., 2020). the progression of microalbuminuria can vary depending on the underlying cause and individual factors such as age, blood pressure, and blood sugar control (oshima et al., 2021). however, in general, the progression of microalbuminuria can be divided into three stages: 1. early stage: in the early stages of microalbuminuria, the albumin in the urine is slightly elevated but still within a normal range. this stage may not cause symptoms, and kidney function may still be normal (parving et al., 2015). 2. moderate stage: as microalbuminuria progresses, the albumin in the urine elevates, and kidney function may decline. this stage is often characterised by hypertension, an elevation in albuminuria, and a decline in the glomerular filtration rate (gfr) (webster et al., 2017). 3. severe stage: in the severe stage of microalbuminuria, there is a significant elevation in the amount of albumin in the urine, and kidney function is severely impaired. at this stage, patients may experience fatigue, weakness, leg swelling, and difficulty concentrating (romagnani et al., 2017). it is essential to diagnose and treat microalbuminuria in the early stages to prevent the progression of kidney disease. treatment may involve medication to control blood pressure and blood sugar, lifestyle changes such as a healthy diet and exercise, and regular monitoring of kidney function. if left untreated, microalbuminuria can progress to macroalbuminuria, which is the presence of large amounts of albumin in the urine (persson & rossing, 2018). macro albuminuria is a more advanced stage of kidney disease and is usually acknowledged with a degradation in kidney function and an increased risk of cardiovascular disease (persson & rossing, 2018). target time in range and hba1c the a1c test, blood glucose monitoring (bgm), and continuous glucose monitoring (cgm) are all used to examine glycemic control. the a1c is the commonly used biomarker for assessing blood glucose control in diabetics, as it decreases both long-term and short-term macrovascular disease and long-term microvascular consequences (wright et al., 2020). whereas time in range (tir) refers to the amount of time that a person with diabetes spends within a target range of blood glucose levels (advani, 2020). for diabetic people, maintaining blood glucose levels within a target range is important for preventing complications such as nerve damage, kidney disease, and cardiovascular disease (advani, 2020). tir is useful for monitoring blood glucose levels and assessing how well diabetes management strategies work (mayeda et al., 2020). the target range for tir can vary depending on the individual, but a common target range is between 70 and 180 mg/dl (beck et al., 2019). tir can be measured using continuous glucose monitoring (cgm) devices, which provide real-time glucose readings throughout the day and night (gabbay et al., 2020). hba1c and tir are two essential diabetes management metrics. however, they serve different functions (lu et al., 2020). tir is used to monitor short-term changes in blood glucose levels and evaluate the efficacy of diabetes management techniques, while hba1c is used to examine long-term glycemic control and assist with treatment options (yoo & kim, 2020). however, a correlation exists between hba1c and tir due to the same variables impacting blood sugar levels, such as food, exercise, and medication use (vigersky & mcmahon, 2019). a lower hba1c indicates better long-term glycemic management and is more common in patients with higher tir levels. generally, a higher tir is associated with better diabetes management and lower risk of complications (shah et al., 2021). through tir and hba1c metrics, diabetes can be managed, preventing the chances of ckd and other diabetes-related complications. prevention at the level of impaired fasting glucose and prediabetes prevention of ckd and diabetes can only be done by managing the disease at impaired fasting glucose (ifg) and prediabetes levels. impaired fasting blood sugar (ifg) is a pre-diabetic condition linked to a relatively elevated probability of developing diabetes2 (yu et al., 2020). early identification of prediabetes and individualised therapy are required to effectively manage this steadily increasing diabetic population. ifg is a significant indicator of diabetes and its comorbidities (yu et al., 2020). consistently high blood sugar levels can cause heart problems, diabetic nephropathy, retinopathy, neuropathy, and increased https://journals.e-palli.com/home/index.php/ajcp pa ge 10 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 9-14, 2023 mortality (tong & adler, 2018). impaired fasting glucose (ifg) and prediabetes are conditions that can highten the probability of developing ckd and other issues, such as cardiovascular disease (kim et al., 2020). management of these conditions typically involves lifestyle modifications and medication, as appropriate, to improve glucose control and reduce the risk of complications. 1. lifestyle modifications: a nutritious diet, frequent exercise, and weight management are examples of lifestyle changes. frequent exercises, like brisk walking or cycling, can also improve glucose control and lower the risk of comorbidities (beulens et al., 2020). 2. medication: sometimes, medication may be prescribed to improve glucose control. metformin is commonly used to treat prediabetes and ifg (beulens et al., 2020). 3. blood pressure management: high blood pressure increases ckd risk and can accelerate the development of ifg and prediabetes. periodic blood pressure assessments are recommended, and medication for hypertension can also be provided when necessary. (ohishi, 2018) 4. regular monitoring: regular blood tests can help to monitor glycemic control and kidney function. this may include a blood test called the estimated glomerular filtration rate (egfr) to assess kidney function (miller & jones, 2018). overall, early identification and management of prediabetes and ifg are important to prevent the development or progression of ckd and other complications. methodology to indicate the development of ckd into end-stage renal failure by managing diabetes mellitus, several recent studies, review articles, prospective studies, cross-sectional studies, and literature reviews, all published and peerreviewed articles, were searched and considered. the area of search was based on how effective the management of debates mellitus is in preventing chronic kidney disease, microalbuminuria and other comorbidities. data was gathered from different search engines and databases such as; google scholar, scopus, pubmed, elsevier, cochrane, sage, medline, and web of science, elsevier. numerous studies were selected from 2018-2023, using the keywords diabetes mellitus management, chronic kidney disease, microalbuminuria, target time in range, impaired fasting glucose, management ways lifestyle modification, and medication treatment, diet control. the full texts of the retrieved articles were made accessible. this article is a review. thus not all information on the prevention of ckd by management of diabetes mellitus has been provided are contained in this. we have included observational studies and all significant, pertinent big trials to highlight the overall conclusions. although we tried to incorporate the largest and most pertinent research, it is important to remember that the tiny, hopeful observational studies were likely chosen due to publication bias. discussion occurrence of ckd in t2dm for populations with type 2 diabetes or a mix of diabetes types, the yearly rate of albuminuria is typically around 8%, while for groups with type 1 diabetes, it ranges from 2% to 3%. regardless of the type of diabetes, low egfr occurs between 2% and 4% of the time. however, due to significant demographic estimate variability, combined ckd incidence rates are impractical (koye et al., 2018). microalbuminuria and albuminuria occur about 2-3% of the time in type 1 diabetes and about 8% in type 2 diabetes or mixed diabetes types. the yearly occurrence of egfr 60 ml/min/1.73 m2 is between 2 and 4 per cent. there was a relatively slight difference in prevalence rates within a single category of kidney illness, despite the considerable variation in methodologies and research design (koye et al., 2017). various studies have determined that an elevation in the occurrence rates of diabetes and hypertension increases the probability of ckd. however, according to a population-based cohort study, it was reported that diabetes and high blood pressure do not have an alliance when it comes to the incidence of chronic kidney disease (erfanpoor et al., 2021). prevalence of ckd in all phases varies globally, with 1.7% in china, 3.1% in canada, 6.7% in the us and 5.8% in australia. in europe, 2.3% in germany, 8.2% in finland, 9.2% in spain and 5.2% in england (romagnani et al., 2017). risk factors ckd is a progressive problem of diabetes, particularly in people with poorly controlled blood glucose levels. several indisputable risk factors are crucial for diagnosing, treating and managing ckd in diabetes (kazancioğlu, 2013). the risk factors for ckd in diabetes can be categorised as modifiable and non-modifiable. to stop or slow the start and advancement of ckd in patients with diabetes, early detection and management of these risk factors are crucial (hannan et al., 2021). this may include regular monitoring of blood glucose and blood pressure levels, lifestyle modifications such as healthy eating and regular physical activity, and medication as appropriate. non-modifiable factors may include genetics, male, age, table 1: modifiable and un-modifiable risk factors of ckd nonmodifiable risk factors modifiable risk factors genetics factor hypertension male sex poor glycemic control age at onset of diabetes lipid abnormalities duration of diabetes smoking family history obesity metabolic syndrome vitamin d deficiency and salt intake gestational diabetes https://journals.e-palli.com/home/index.php/ajcp pa ge 11 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 9-14, 2023 family history, the onset of diabetes etc., while modifiable factors contain hypertension, poor glycemic control, insulin resistance, smoking, metabolic syndrome, vitamin d deficiency etc. (hannan et al., 2021; koye et al., 2018; kurzhagen et al., 2020). elevated blood glucose levels can damage the kidney’s small blood vessels, leading to ckd. hypertension or high blood pressure also damages the blood vessels and contributes to the risk of causing ckd (kurzhagen et al., 2020). ckd also depends upon the duration of diabetes onset; according to the reported studies that a longer duration of diabetes can increase the risk of developing ckd in patients (hannan et al., 2021). smoking is injurious on many levels; it damages the lungs and kidneys and worsens glucose control in diabetic people (kazancioğlu, 2013). prevention of ckd ckd can be prevented or delayed by taking measures to maintain healthy kidneys. maintaining underlying conditions, quitting smoking, eating healthy foods, exercising regularly and drinking enough water can help stabilise ckd and diabetes. other than these preventions, insulin therapies play a vital role in managing ckd, while some medications are widely used to prevent mortality. some interventions to manage and prevent ckd are explained in this review. lifestyle modification healthy dietary intake and weight are vital in reducing most chronic diseases, including renal end-stage diseases. dietary and lifestyle modifications have been shown to help treat diabetes and improve glycemic control. when a patient’s egfr drops below 60 ml/min/1.73 m2, it is recommended that they seek the advice of a nutritionist to optimise their diet to improve hyperglycemia and ensure they are getting enough protein, potassium, and phosphorus to prevent the condition from worsening (williams, 2017). nutrition is essential for people with diabetic kidney disease to maintain an equilibrium of salt, potassium, phosphorus, protein, carbohydrate, and unhealthy fat intake. patients who are overweight or obese should reduce their weight and improve their exercise, as cardiac stress testing is necessary (hahr & molitch, 2015). according to the kidney disease outcomes quality initiative (kdoqi) of the national kidney foundation, following a low-protein diet (lpd) can help ckd patients live better lives and decrease the progression of esrd (alkhatib et al., 2023). it has been proven that diets high in fruits and vegetables and low in saturated fat can effectively lower blood pressure. these diets also limit salt intake (bello et al., 2005). moreover, cessation of smoking also helps in maintaining healthy kidney pharmacological management intense care of glycemic control decreases the risk of microalbuminuria and macroalbuminuria. according to a report, every increase in systolic blood pressure in the general population above 115 mm hg has been said to double the risk of cardiovascular disease (cvd). management of hypertension is essential to prevent albuminuria/proteinuria and the ensuing development of ckd in both diabetic and non-diabetic individuals. pre-hypertensive conditions, lower blood pressure levels in the general population, and more active blood pressure lowering in patients with hypertension and underlying ckd are essential (bello et al., 2005). standardising a distinct ratio for every patient is crucial for blood glucose management and ckd prevention. maintaining a blood pressure (bp) less than 130/80 in dkd patients is the typical aim advised by the national kidney foundation; nevertheless, there is considerable debate around this recommendation (williams, 2017). due to their ability to preserve the kidneys, ace inhibitors and arbs are typically regarded as first-line treatments for hypertension in diabetic people. patients with diabetes mellitus have been shown to benefit from ace inhibitors in terms of kidney, heart, and, to a lesser extent, eye and peripheral nerve function. these positive outcomes result from angiotensin ii’s hemodynamic and tissular actions being inhibited (williams, 2017). medications some treatments suggested by healthcare professionals are first-line care for the prevention of ckd, such as metformin (betônico et al., 2016; beulens et al., 2020). this medication has been advised to patients for years. the mechanism of this drug is to decrease the production of hepatic glucose, improve insulin tolerance, lowering impaired fasting glucose and plasma glucose. although metformin is the most common drug to prevent and manage ckd, it still is contradictory to its nature because metformin excretes through the renal system and the emergence of lactic acidosis, which is one of its most serious drawbacks but only occurs in about 5 cases per 100,000 patient-years of patients on average. metformin can be used to treat type 2 diabetes (t2dm) up to a gfr of 30 ml/min/1.73 m2, with a lower dose indicated at 45 ml/min/1.73 m2 (current uk guidelines) (betônico et al., 2016). sulfonylureas (sus) encourage pancreatic b cells to secrete endogenous insulin, leading to hypoglycemia, alcohol abuse, hepatic dysfunction, heart failure, malnutrition, advanced age, and interactions with other medications that displace sus from their plasma protein-binding sites (seino et al., 2016). this is due to accumulating one or more of these medications’ metabolites, increasing the risk of hypoglycemia. other medications help decrease the incidence of ckd in diabetes, such as glinides, alpha-glucosidase inhibitors, glitazones, and dipeptidyl peptidase 4 inhibitors (seino et al., 2016). insulin therapy the kidney is crucial in removing insulin from the bloodstream through two different mechanisms. in the initial stage, insulin is filtered by the glomerulus and then taken up by proximal tubular cells via endocytosis https://journals.e-palli.com/home/index.php/ajcp pa ge 12 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 9-14, 2023 (peruchetti et al., 2021). the second process includes insulin attaching to the contra luminal tubular membrane of cells and diffusing through peritubular capillaries. lysosomes transport insulin, broken down into amino acids and diffused into the peritubular arteries. insulin clearance declines when renal failure worsens, necessitating a dosage reduction to prevent hypoglycemia (peruchetti et al., 2021). this decrease in insulin clearance is initially offset by a rise in proximal tubular cells’ insulin absorption, and it is also linked to an increase in insulin resistance. muscle tissue is the predominant source of ir in ckd, occurring mostly at the extremities. in individuals with esrd, hepatic glucose production is not elevated and repressed in response to insulin, and ir may not result in a proportional rise in insulin secretion (gburek et al., 2021). lipid metabolism dysregulation of lipid metabolism leads to higher levels of triglycerides, oxidised lipoproteins, and lower levels of hdl cholesterol in chronic renal disease and esrd. advanced ckd or esrd patients have a distinctive lipid pattern that includes hypertriglyceridemia, low hdl cholesterol, and normal ldl cholesterol levels (dincer et al., 2019). in the general population, there is a clear correlation between ldl cholesterol and atherosclerotic events, but in patients with esrd, ldl cholesterol has a flat or weakly positive correlation with mortality at levels above the average and a negative correlation with these outcomes at levels below the average. reducing ldl cholesterol helps prevent serious atherosclerotic events in people with ckd and kidney transplants but not for people who need dialysis (ferro et al., 2018). limitations and strengths 1. the management and treatments discussed in this review do not guarantee the reversal of the disease. 2. several participants were not tested in this review; the review is based on factual data. 3. the most coherent risk factor of mortality due to ckd is increasing age, which has not been extensively discussed. 4. the biggest strength of this review is that very generalised terms have been used in searching strategies to combat vast unrelated data. 5. this review is based on scientifically proven facts. conclusion in conclusion of this review, diabetes mellitus is the most common and biggest risk factor for the occurrence of ckd and esrd. it is advised to maintain and focus on the underlying conditions that result in ckd over time to prevent the disease. these conditions include hyperglycemia, hypertension, microalbuminuria, sedentary lifestyle, smoking, etc. these underlying conditions may turn into symptoms and eventually convert into morbidity. changes in lifestyle, such as healthy eating, regular physical activity, and intake of advised ml of water in addition to some medications and therapies, can altogether increase improvement rates of ckd incidence. further future studies need to be done to combat the disease better. references advani, a. 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(2020). incidence and risk factors of vascular complications in people with impaired fasting glucose: a national cohort study in korea. scientific reports, 10(1), 19504. https://doi. org/10.1038/s41598-020-76661-7 https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 18 american journal of chemistry and pharmacy (ajcp) cause of land degradation and rehabilitation practices in case of amba sidist western ethiopia dessalew shitu1* volume 1 issue 1, year 2022 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v1i1.419 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: july 25, 2022 accepted: august 20, 2022 published: august 23, 2022 in ethiopia, land degradation has become a serious problem affecting all spheres of social, economic and political life of the population. it is one of the major challenges to agricultural development and food security of the country. in order to solve the problem of land degradation, a lot of efforts have been made since 1970s. however, at the end the intervention couldn’t be sustainable and able to bring the intended impact. thus, identification of challenges in relation to land rehabilitation practices is of paramount importance. the study was undertaken in amba 6 kebele. in order to achieve the objective of the study, both primary and secondary data were generated by employing qualitative and quantitative data. stratified sampling technique was used to select 76 household heads were selected from on amba 6 by using stratified random sampling as it is believed that the households are similar with respect to the characteristics that influence the research output. the process of analysis of the study was carried out using qualitative description and quantitative analysis. the quantitative data was analyzed using frequency, percentage and mean when appropriate. the qualitative data was discussed to substantiate the study. the findings of the study indicated that decrease in productivity of farm land, involvement in off-farm activities, increase in size of human population, lack of full cooperation of family members to involve in land rehabilitation practices, low assistance gained from neighbor-hoods, less access to extension service and inadequate attention from woreda agriculture and rural development office are the major challenges encountering the implementation of land rehabilitation practices in effective way. keywords land degradation, rehabilitation, amba sidist 1 college of agriculture and natural resource management, ethiopia. * corresponding author’s e-mail: dessugold@gmail.com introducton the twenty-first century is a time by which the world is getting seriously confronted by issues of sustainable use of natural resources. despite the emerging recognition of their decisiveness for the survival of humanity on the planet, these days, water and land ecosystems are being degraded at an alarming rate (hannam, 2003 cited in teketel, 2009) the case is worse in developing regions, where the majority of the population depends on these resources for its livelihould hence, the conservation and management of land and water resources for sustainable intensification and agriculture and poverty reduction in developing regions has remained one of the most challenging policy issues for a long time (bekele et al., 2007). the three inherently interrelated and interdependent land resources, namely, soil, water, and vegetation are in the immediate focus of most local, regional, and international policies, programs, initiatives, covenants, protocols, and conferences that are meant to pave the way to sustainable development. land is the most important natural resource all over the world. it is a place from which human beings are exploiting a number of resources (taffa, 2002). almost all food production for the world population is derived from land, and the need to produce more is increasing from time to time due to an increase in population. for increasing production, either area under cultivation must be expanded or its productivity needs to be increased. thus, fertility of land is decisive factor in addition to other technological input. however, land is losing its productivity due to a rising trend of land degradation (woldeamlak, 2003). land resources degradation, resulting from different causes, is threatening long-term productivity. nowadays, land degradation is reducing yield significantly and it is more acute in some parts of the world than the others. for example in central america, 75% of cropland is seriously degraded while in africa, 20% of the total land area is at risk of unrecovered (sida, 2007). in ethiopia, land degradation has become a serious problem affecting all spheres of social, economic and political life of the population. it is one of the major challenges to agricultural development and food security of the country. the rate of the country’s land degradation is very high on the other hand, like many other developing countries, ethiopia is characterized by agrarian economy and about 84 % of its total population derives means of survival from agricultural activities. moreover, the role of agriculture in the overall economy is quite significant. it contributes 50% of the total gdp and 85% of foreign exchange earnings. thus, land productivity is one of the key elements for enhancing economic development of the country. in contrast, the level of land degradation has already reached an alarming stage (moard/wb, 2007 cited in desta, 2009:2). this land degradation has been recognized to be one of the chronic problems in ethiopia and many efforts have been made against it. however, the problem of land degradation is continuing and natural resource base is deteriorating at alarming rate (yohannes, 1999; genene, 2006). the major causes of land degradation in ethiopian highlands include https://doi.org/10.54536/ajcp.v1i1.419 https://journals.e-palli.com/home/index.php/ajcp mailto:dessugold%40gmail.com?subject= pa ge 19 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 18-25, 2022 cultivation on steep slopes, deforestation, and erosive rainfall pattern, lack of fallowing, overgrazing and lack of proper conservation measures. the underlying factor for the degradation of land is population pressure (lakew et al., 2000). in response to heavy land degradation in the country, large-scale efforts for implementing natural resources conservation programs had taken place starting from the recent past. statement of the problem in ethiopia, the heavy dependence of people’s livelihoods on agriculture and inappropriate use of natural resources resulted in fast and vast land degradation (genene, 2006). on the other hand, development of agricultural sector partly depends on land productivity. however, this resource is seriously threatened by land degradation and aggravates the food insecurity problems in the country through its adverse impact on crop yield. the country could not feed its population at present and it will have difficulties doing in the near future partly due to serious land degradation (kruger et al., 1997; genene, 2006). to solve the problems of land degradation in the country, many efforts have been made since 1970s a large number of soil and water conservation activities were implemented in different parts of the ethiopian highlands in the 1970s and 1980 with a huge resource obtained from international community, particularly world food program (wfp). objective of the study the general objective of the study to assess the major cause and effect of land degradation and rehabilitation practices in case of amba 6 kebele. the specific objectives to identify major causes of land degradation in the study area to identify the social, economic and institutional factors that determines the practice of land rehabilitation to assess major challenges and shortcomings in undertaking different land rehabilitation practices research questions based on the above specific objectives, the research attempted to answer the following question: 1. what are the main causes of land degradation in the study area? 2. what are the ongoing practices of land rehabilitation? 3. what are the main socio-economic and institutional factors that affect the practice of land rehabilitation? materials and methods sampling design the researcher was adopted stratified random sampling methods to select the household this stratification method would be selected based on wealth status. the users of farming land were stratified into rich, poor and medium household head. this is because rich, poor and medium could have different attitude and perception towards effect of economic and environmental degradation this is to give equal chance for the whole target population to be selected as a respondent. sampling techniques the study area will be selected because of the existence of land degradation caused by high population growth. from the total of 319 households in the study area, 76 (7%) will be selected from the total. but, due to deficit of resources the interview hh number is declined to 10 hh. this extent of households was selected using stratified random sampling techniques. therefore, to find out representative sample for this study the researcher used the following yamane simple determination formula. n=n/1+n (e)2 (yamane, 1967) where n=sample required as a representative, n=sum of households of, e=precision level assumed to be (0.1 or 10%), n=319/1+319(0.1)2, n= 76 hh source and types of data both primary and secondary data were used in the study. the primary data were collected from the selected respondents and secondary data were collected from different written materials such as previous research, published and unpublished documents in the library. also, quantitative and qualitative data are will be used in this research this data were collected from respondents. method of data collection both primary and secondary data collected were used. primary data was collected by using: questionnaire; these questionnaires will prepare for 10 respondents. observation in this case structured and unstructured were used as different techniques of observation. we conducted interview to identify people’s experience, perception, value and opinion about the cause of deforestation, contribution and significance of forest for their living. data analysis the collected data was by analyzed using descriptive statistics technique with statically package for social scientists (spss) software to analysis data drawn house hold survey, descriptive statics like percentage, frequency, distribution was used to analyzed the data. this technique of data analysis was applied to show the number of respondents that involves in questionnaires; interview and percentage was also be used to depict responses of the sample respondent and to place them per hundred portions respectively. results and discussion sample household characteristics a total of 30 respondents were used for this study. the households were taken purposely for data collection. out of the total of 30 respondents, 60% (n=18) of them https://journals.e-palli.com/home/index.php/ajcp pa ge 20 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 18-25, 2022 were males and the remaining 40% (n=12) were females. with regards to educational status, out of the total of 20 respondents, 36.67% (n=11) of them were illiterate, 20% (n=9) were they can read and write, 23.3% (n=7) were primary school and 10% (n=3) were secondary school. as indicated in table 1; 56.67% (n=17) of the respondents were married, 23.33% (n=7) were divorced, the remaining 13.33% (n=4) of them widowed, and 6.67% (n=2) of the respondents were single. as indicated in table 2, 43.34% (n=13) were in the age group of 60 and above year followed by those in the age of group of 31 45 year 33.33% (n=10). the remaining 23.33% (n=7) of the respondents were found in the age group of 15 30 year. than, it was a question of survival. major cause of land degradation in the study area according to local community, the major cause’s land degradation in the study area overgrazing, deforestation, population growth, mismanagement of resources out of the total 30 respondents, 56.67% (n=17) of them agreed that deforestation as the major causes of land degradation, the remaining was for other rezones 23.33% (n=7), overgrazing 13.33% (n=4), and 6.67% (n=2) other services. overgrazing out of the total respondents, 56.67% (n=17) of them agreed that overgrazing as a cause of land degradation. like; teff, maize, etc were some of cereals important to generate income for local communities.the majority of local community in the study area mainly dependent on agriculture. farmers in the study area pursue mixed agriculture. both crop farming and rearing of livestock. one of the key farmer said that the reason why expansion of agriculture is due to immediate solution rather long term that means, products being obtained from agriculture takes short period of time and also it is possible to generate income in short period of time than forests which takes long period to get its outputs and also it takes long period of time to get income from it. deforestation out of the total respondents, 23.33% (n=7) of them showed that fuel deforestation as the major factor of land degradation in the study area. according to the respondents, in the study area forest was the common and major household energy source for home based activities (food cooking, water heating, etc). because of the lack of modern electric energy supply, the majority of the households are depending on fuel wood. as information obtained from the sampled households, fuel wood consumption was very exhaustive, time consuming, however, they use it as main energy source because of lack of options to use another improved energy sources table 1: marital status of sample household. marital status frequency percent married 17 56.67 divorcees 7 23.33 widows 4 13.33 single 2 6.67 source: field survey 2008 table 2: age distribution of the sample household age of the household head frequency percent 15-30 7 23.33 31-45 10 33.33 46-60 13 43.34 source: field survey 2008 communities perceptions about land degradation in the study area according to local source, forest used to support the livelihood of most rural communities the farmers used to get different functions and services from the forest; such as fire wood, herbal medicine, bee keeping, house construction materials, food, etc. however, increase land degradation in the study area was indicated as indicators of decrease in functions and services of lands. one of the community leaders said that the last few decades’ forests were depleted and evens currently the local communities supplying forest products in the market as income generation. according to the interviewee, in the study area, the local communities had no access being gained awareness creation regarding how land degradation rehabilitation mechanisms on sustainable manner. the local communities were being applied only their traditional knowledge for forests resource conservation rather than integrating their knowledge with scientific way which was being important to ensure environmental sustainability. from that local community one of the farmers also said that expanding of infrastructures for the sake of attaining erosion was being created key roles for easily occurrence of land degradation in the study area. issues which were noted earlier were some of the key factors which aggravated land degradation. according to the interviews, before two decades, land degradation was not a serious issue in the study area, but currently the problem was not only a serious issue rather table 3: current land status in the study area. current land condition frequency percent excellent 2 6 good 6 20 moderate 9 30 degraded 8 33 very much degraded 3 7 total 3 6 source: field survey, 2008 table 4: major causes of land degradation in the study area causes of land degradation frequency percentage overgrazing 17 56.67 deforestation 7 23.33 mismanagement of resources 4 13.33 population growth 2 6.67 total. 30 100 source own survey 2008 https://journals.e-palli.com/home/index.php/ajcp pa ge 21 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 18-25, 2022 mismanagement of resources out of the total of 30 respondents, 13.33% (n=4) of them agreed on urban development’s as major cause of land degradation. according to their opinions, various activates took place on the area, which replaced the area the resource was depleted. as a result, various constructions and infrastructures were built. like expansions of different building of schools, clinic, cultural offices, etc, were some of the infrastructures built on the area to expand urbanization. in the study area mismanagement of resources was being played major cause for contribution of land degradation. and to expand infrastructures huge hectares of land was being depleted in the study area population growth according to the local community due to population growth in the study area increased demand for more agricultural land, timber production, fuel wood consumption and other uses, at his time the tree cover was reduced, so the stock of tress diminishes as the forest resource was overly exploited rather than being managed on a sustainable basis. the local farmers, also perceived that over exploitation of forests resource in the study area led to land degradation and the diversion of animal dung from its traditional role as soil nutrient to direct burning for fuel, while the reduced dunging of the land and soil erosion caused by trees cover clearance or succeeded deforestation, further impairs soil fertility, this in turn, promote water runoff which takes soil from the land to produce sediment in rivers, polluting drinking water supplies, loss of productivity and aggravate formation of gullies, soil erosion and floods. major consequences of land degradation in the study area the causes and consequences of land degradation were interlinked with each other’s, that means if there was causes it also consequences and vice versa. according to most respondents the major consequences of land degradation in the area are poverty, climate change, and gully expansion, in the table 6, 63.33% (n=19) of the respondents agreed that poverty as consequences of land degradation, 26.67% (n=8) as climate change, and 10% (n=3) of the respondents said gully expansion also as a consequence of land degradation in the study area poverty poverty was the major consequences of land degradation in the study area.s to be degraded; as a result reduction in productivity was being employed for their livelihood. the majority of the community in the study area was being highly depended on agriculture. this was because, sustaining of their life as well as income generating strategy was being used mainly from mixed agriculture. as it was described earlier, one can possible to say that highly dependency of the communities on agriculture table 5: major consequences of land degradation in the study area consequences of land degradation frequency percentage poverty 20 63.33 climate change 7 26.26 gully expansion 3 10 total 30 100 source own survey 2008 was being aggravated the land in the area was happened. climate change deforestation stimulates higher accumulation of co2(carbon dioxide) on the atmosphere, these would continue depleting the ozone layers, which caused an increase of temperature on the atmosphere. the current serious issue in the study area was higher increase of temperature and climate changes, which affect the day to day activities and continued its impact on social, economic and environments. during interviews , they perceived that climate change was being taken place due to, higher increase of populations, frequent use of forests resource, urbanization, and resettlements programs which were some of the major factors highly accelerated temperature to be raised and as a result climate change in the study area. gully expansion gully expansion is the major consequence in the study area by applying high amount of water in the form of erosion to erode or deplete agricultural productivity. as a result of this people leave in the study area highly affected by gully expansion and to affect different agro forestry systems. regarding wild animals some the of the community leaders argued that nearly about 20 years ago there were a lot of wild animals, but today only few can be seen near forest fragments. the decreasing of their number was due to depletion of forests, which obligated them to go others, area having forest covers. one of the interviews of the farmer, said that the only option to mitigate loss of soil and biodiversity was that encourage the habit of protecting the environment by practicing planting of trees on a sustainable manner, practice of a forestation program, area closure and etc. according to the community whatever the case, forests land area should be increased and even should be encouraged the programs which was being described earlier, this was, because practicing of a forestation program, was being played important roles which contributed not only for environmental sustainability, but also social, and economical sustainability. soil fertility improvement practices farmers at individual level practice different land management activities mainly to increase agricultural yields and to conserve the natural environment on their plot of land. they practice a short-term benefit oriented soil fertility management and long-term benefit oriented https://journals.e-palli.com/home/index.php/ajcp pa ge 22 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 18-25, 2022 land resource management activities. for short-term effect, farmers apply chemical fertilizers to obtain high agricultural yield. inorganic chemical fertilizer application is a prerequisite in the study area as many parts of the country experience due to loss of soil fertility which is caused by erosion (see table 5.1). according to survey results, crop rotation (76.1% and 69.7% of the respondents in amba 6 kebele respectively) is one of the most important methods of improving soil fertility as well as conserving soils on cultivated fields. it is a method through which the nutrient content of the soil is improved by cultivating different crops on the same plot of land interchangeably. this method again becomes more important when leguminous crops are part of the rotation system to improve the nitrate content of the soil. according to the information obtained from amba 6 kebele and woreda experts, this system is one of the widely practiced system of soil fertility improvement in the study area which helps to obtain more outputs from the cultivated land. the rotation system mostly consists of cereals, legumes (mainly bean) and root crops like potatoes in the farm land in different seasons and years of cultivation. manure is used to be important input for promoting the fertility status of the soil. its application to farmland raises the nutrient level of the soil, increases infiltration and reduces soil erosion. about 53.7% of the respondent in amba 6 . currently, application of manure on farm lands has been decreasing from time to time as the number of livestock per household has been decreasing significantly for various reasons. participants of community indicated that fragmentation of farm plots confine the use of manure only for homestead area rather than far away from their home. in addition, the use of cattle dung as source of fuel for cooking instead of using it as organic fertilizer is another contributing factor for low application of manure for soil fertility improvement practice in the study area. regarding fallowing to improve soil fertility, the survey result indicates small contribution. only 6% and 3% of the respondents in amba 6 used fallowing to improve soil fertility respectively. participants, no fallowing that can contribute to soil fertility improvement in the study area. because of high population density, farmers have no chance to leave the land uncultivated. soil and water conservation practices in the study area, as many parts of ethiopian highlands, farmers have a pool of indigenous knowledge with which they use and manage their land resources. they make efforts to conserve their soils against erosion by applying a range of conservation techniques among soil and water conservation measures, which is widely used by farmers in the study area, in almost all villages, is contour ploughing. ninety-four percent and ninety-seven percent of the respondents in amba 6 confirmed that they protect soil from erosion through contour ploughing respectively. to establish the structure, the farm plots are ploughed horizontally; following contours so that those contour furrows are created with the help of iron plough. the furrows that are formed along contours help to hold the water until it infiltrates into the ground and hence reduces the damaging or erosive effect of surface runoff on plots of land. traditional ditches are the most extensively practiced measures used to conserve soil and water in the study area. the structures are established with the help of oxenplough deep into the ground. this kind of practice has been used by farmers since longer times. when the land is being prepared for sowing to minimize the problem of seed loss after sowing. in addition, it also minimizes water logging problems as well as soil erosion. according to information obtained from amba 6, a group of two or more farmers who possess neighbor farm plots prepare water ways at the top of the slope. this is a common practice in the study area as this area is found at the base of mount. . it helps protect crop fields from being damaged by powerful run-offs that come in such steep sloppy area. findings from the survey respondents indicated that, hillside terracing and check-dams were among the most frequently used physical structures used for soil and water conservation in the study area. however, it was noted from the amba 6 kebele information that the terracing and check-dams were of poor quality that stay only for a short period of time, in fact until they face a repeated heavy rainfall. moreover, the potential positive impacts of the conservation attempts had also been reversed by the simultaneous devastating act of the farmers themselves for cultivation, free grazing. forage strip and stone bounds (in some cases) are used as another technique for soil and water conservation practices. biological treatment of soil and water conservation activities are being practiced in combination with the physical soil and water conservation structures. the treatment of the land takes place in comprehensive way in all land use types such as in cultivated lands, grazing lands, forest areas and marginal lands of the study area. gully rehabilitation practices gullies which are intermittent stream channels longer than rills are created by concentrated rainfall runoff from the surrounding sloping land. usually, gullies follow sheet erosion or result from neglect of rills. most of the reasons behind gully formation in the study area include deforestation, cultivation of steep slopes, lack of fallow, less vegetative cover, overgrazing and insufficient soil conservation measures. according to table 6: mechanisms of land rehabilitation practice in the study area mechanisms of land rehabilitation practices frequency percentage fertility management 19 63.33 soil and water conservation 8 26.67 gully rehabilitation 4 12 total 30 100 https://journals.e-palli.com/home/index.php/ajcp pa ge 23 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 18-25, 2022 the survey respondents, most of the aforementioned factors behind gully formation are testified to be existent and to be the major causes of land degradation in the study sites. in order to curb gullies, check-dam was widely constructed along water ways from tree shrubs or stones. this eventually helps raise the floor up to the level of surrounding and original ground. cut off drains and planting trees and grasses along water ways are additional gully rehabilitation practices in the study conclusion this study has attempted to investigate the challenges and prospects of land rehabilitation practices in amba 6. the findings of the study indicated that the livelihood of the farmers in the study area depends on subsistence agriculture. the major economic activity for all sampled households is based on farming (about 92%). the average size of farm land owned by farmers is less than one hectare which limits the amount of production in the study area. on the other hand, the findings indicate has serious land degradation problems as observed in other part on highland ethiopia. this problem appeared to be one of the major challenges for crop production. sheet and rill erosion are commonly observed in farmland whereas gullies are common in communal lands and edges of farmlands. limited use of conservation structures, lack of fallowing, cutting trees for fuel and construction purposes, ploughing steep slopes and overstocking are the major immediate root causes of land degradation. the underlying causes of land degradation as identified by the study are population pressure, steep slope nature of the area and erratic rain fall pattern. more than half of the farmers in the study area are practicing crop production in erosion prone areas and still there is an expansion of cultivation in to marginal lands due to population pressure. this expansion of farming towards marginal area resulted in shortage of grazing land, deforestation and increasing trend in land degradation. the livelihood of the community in the study area is affected by these land degradation problems. the productive lands become less productive and subsequently left uncultivated. about 76% of the sampled households recognized that there is land degradation in their locality and about 30% of the sampled households agreed that leaving lands due to loss of quality is the main cause of decreasing trend in land holding in the study area. regarding response to land degradation problem, there have been a range of land rehabilitation practices underway in the study area by local communities, government and non-government organizations. the activities practiced in the study area include soil fertility improvement activities, soil and water conservation structures and rehabilitating degraded lands by agro forestry, afforestation and area closure system. however as compared to the magnitude of the problem, these land management and rehabilitation practices are not enough to curb land degradation problem. there are also challenges that affect land rehabilitation practices in the study area. the study revealed that different socio-economic and institutional factors determine land rehabilitation practices in the study area. among the socioeconomic factors increase in human population is the major challenge. with the increased population, there is subsequent increase in the size of cultivated area which in turn resulted in the shrinking of grazing lands and expansion of the cultivation into areas formerly considered as marginal and extremely fragile. continuous search for new cultivable land also challenges the construction and maintenance of soil and water conservation practices and closing extremely degraded area for rehabilitation. regarding to the involvement of family members and neighborhood in land rehabilitation practices, the finding of the study revealed that slightly more than half percent of the population have willingness and cooperation to involve in land rehabilitation practices. this indicates the increase of awareness of the community about the negative impact of land degradation. but the population which accounts nearly half percent did not show any willing to involve inland management and rehabilitation works. the reasons identified during the survey, exhaustion of the fertility of land and subsequent reduction in farm yields, deterioration of social values of helping each other and subsequent focus on personal gains, and decrease in productivity of land which discourage the need for additional labor from outside are the major one. economically, the decreased productivity of farm land and involvement in offfarm activities pushes away people from participating in land rehabilitation practices. in addition, people who engaged in off-farm activities cannot contribute labor supply for any land rehabilitation practices as these activities require large number of labor. thus, from this, it can be concluded that the economic factors have played their own role in land rehabilitation practices. regarding the institutional support related factors, though efforts being made were reported by the government and non-government officials in the study area, none of the activities are enough to halt the problem of land degradation. from this, it is possible to conclude that inadequate integration of the government officials with local communities during planning and introduction of different soil and water conservation activities and the intention of government to fulfill the immediate need of the people are some of the problems observed in the study area. in general, among the considered socio-economic and table 7: the extent of land degradation in the study area extent of land degradation frequency percentage very high 20 66.6 medium 7 23.33 low 3 10 no land degradation -----https://journals.e-palli.com/home/index.php/ajcp pa ge 24 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 18-25, 2022 institutional factors that influence land rehabilitation practices, the increasing number of population, involvement in off farm activities, decline in farm productivity, and inadequate attention paid by government toward securing alternate means of livelihood to ease pressure on land, the need to obtain short term benefits rather than long-term from land are the very important ones in the study area. recommendations the findings of the research indicate that there is increasing trend in land degradation problem in the study area. different land management activities are being practiced in order to conserve resource base and ngos, none of the interventions can efficiently curb the problem. therefore, based on the finding of the research, the following actions that are believed to play significant role in improving land rehabilitation practices and solving, at least minimizing problems of land degradation are recommended. in association with the increased population pressure a number of adverse effects on the environmental conservation of the study area and land rehabilitation practices have been identified. easing population pressure on natural resource dependence needs due attention. this can be achieved by resettlement program, continuous training and awareness creation on family planning, technological improvements in agriculture and development of other sectors of the economy to minimize burdens on natural resource. in making intervention in land management practices, there should be active participation of local people primarily farmers. this helps to integrate indigenous land management practices with the new ones and enhance easy adoption and sustainable use of effective introduced practices. in addition, it is also essential to provide benefits to the local communities from enclosed area in sustainable manner which, in turn, increased the sense of one’s resource references aklilu, a. 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(1986). soil conservation revised print. bt batsford limited, london. hussen haycho (2006). land use change and challenges of land degradation in adaba area, bale zone, a thesis submitted to school of graduate studies, institute of regional and local development studies, irwin b (2001) managing forest as common property: collective forest management in ethiopia. in zenebework tadesse (ed). environment and development in ethiopia. proceedings of the symposium of the forum for social studies, aa, september 15-16, 2000, for social studies, aa ethiopia kebede tato (1989). development forum of reclaimed land, report of the second departmental workshop community forestry and civil conservation development department (cfsc dd) ministry of agriculture (moa) addis abab, ethiopia. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 1 american journal of chemistry and pharmacy (ajcp) subclinical hypothyroidism: a review safaa mohamed matarid1* volume 2 issue 2, year 2023 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v2i2.1594 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: april 21, 2023 accepted: may 11, 2023 published: may 12, 2023 both the mother and the fetus are physiologically stressed out during pregnancy. the second most prevalent endocrine illness among pregnant women, behind diabetes mellitus, is thyroid disease. several differences in the mother’s thyroid function are seen in the complete life cycle of pregnancy, and thyroid dysfunction occurs from the mother’s inability to adjust to these physiological changes. the possibility for maternal and fetal outcomes can be extremely high when endocrine abnormalities like hypothyroidism are present during pregnancy. the fetal development and perinatal outcomes can be significantly impacted by thyroid illness, which is frequent in women of reproductive age. for the first part of pregnancy, the fetus depends on the mother’s thyroid hormone, which is essential for optimal fetal neurodevelopment. there is authentication of an association between overt maternal hypothyroidism and overt maternal hyperthyroidism. with higher obstetrical risks and negative impacts on the offspring’s cerebrospinal nervous system expansion, grey matter”, and neurocognitive potential. treatment for overt thyroid disorders improves results. subclinical maternal hypothyroidism can have a negative impact on a baby’s neurocognitive and obstetrical outcomes, despite conflicting findings. subclinical hypothyroidism has not yet been successfully treated. pregnancy-related subclinical hyperthyroidism is easily tolerated. however, new research has revealed no improvement with levothyroxine treatment, indicating that thyroid autoantibodies alone might also affect foetal and neurodevelopmental outcomes.. the fetus may be impacted by several uncommon maternal genetic thyroid conditions, such as a tsh receptor mutation that causes hcg hypersensitivity or“thyroid hormone contrary. the thyroid”perform a key function. for the best care, it is crucial to understand fetal health. data was gathered from different search engines and databases such as; google scholar, scopus, pubmed, elsevier, cochrane, sage, medline, and web of science. numerous studies were selected from 2017-2022, using the keywords subclinical hypothyroidism, hypothyroidism, pregnancy, fetal damage, risk factors, current challenges, thyroid hormone, and american thyroid association. the full texts of the retrieved articles were made accessible. keywords subclinical hypothyroidism, age group, infertility, pregnancy, prevalence, fetal health, morbidity 1 department of internal medicine, mediclinic al ain hospital, heikh khalifa bin zayed st next to choithrams supermarket, abu dhabi united arab emirates * corresponding author’s e-mail: safaamatarid12@outlook.com introduction the thyroid gland disease known as subclinical hypothyroidism (sch) is distinguished by increased tsh and normal ft3 and ft4 levels. the only approach to detect this illness is through biochemical testing due to the wide variety of clinical presentations. the causes are similar to those of overt hypothyroidism; the most common is chronic autoimmune thyroiditis (hashimoto’s thyroiditis), characterised by anti-thyroid peroxidase antibodies. other causes include previous hyperthyroidism, postpartum thyroiditis, subacute thyroiditis, thyroid injury and inflammation from radiation, surgery, medication, and thyroid infiltration. (khan et al., 2017). pregnant women frequently have hypothyroidism. the detection frequency, particularly in emerging india, for example, has not kept up with the severity of the issue. during pregnancy, the mother’s thyroid function changes. there are several causes for these alterations. similarly inflation in thyroglobulin due to high-rise estrogen and human chorionic gonadotrophin, inflation in renal losses of iodine due to inflation of glomerular filtration rate, during pregnancy, the need for iodine and the production of thyroid hormone both increase by 50%. this is due to alterations in the peripheral metabolism of maternal thyroid hormone and changes in iodine transport to the placenta (pokhanna et al., 2017). pregnancy is a stress test for the thyroid that can lead to hypothyroidism in women with a low thyroidal reserve or iodine shortage. subclinical hypothyroidism affects 2-7% of pregnant women, and overt hypothyroidism affects 0.22 to 2.5% (sakr & sakr, 2020) (mandel, 2004), (syamala et al.). retardation in neurodevelopment is a result of maternal hypothyroidism, particularly in the first trimester, and hinders cognitive growth (wade & mandel, 2018), ((aravelli et al., 2022; malti). making a clinical diagnosis of hypothyroidism during pregnancy is difficult due to nonspecific features that may be masked by already present obstetric symptoms. thyroid function testing is therefore required for the diagnosis of subclinical hypothyroidism. since hypothyroidism is easily managed by early diagnosis and medication, this could lessen the burden of unfavorable maternal and fetal outcomes, which are extremely frequently seen. screening should ideally be done during prenatal testing or after a pregnancy is confirmed. there aren’t any national https://doi.org/10.54536/ajcp.v2i1.1594 https://journals.e-palli.com/home/index.php/ajcp pa ge 2 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 1-8, 2023 guidelines for treating thyroid dysfunction, and there are few statistics on its prevalence during pregnancy in india. the study aims to assess the prevalence and consequences of thyroid malfunction, particularly hypothyroidism in pregnancy. according to reports, the prevalence of sch is 4–10% in adults. however, this varies according to the population, with more instances occurring in regions with enough iodine (canaris et al., 2000). even more patients who take thyroid drugs are affected (abdurazzakova, 2021). like other thyroid conditions, sch is more prevalent in women than males and worsens with age. every year, it’s likely that 2-5% of sch patients will develop overt hypothyroidism (abdurazzakova, 2021). according to the increase in serum tsh levels, there are typically two types of sch: mildly raised tsh levels (4.0–10.0 m iu/l) and severely increased tsh values (>10 m iu/l). however, the lower limit of tsh that should be utilised is still debatable, with numerous studies employing different cutoffs. most sch patients (4–10 m iu/l) have lesser levels of elevated tsh (kim et al., 2017). a collection of conditions known as thyroid diseases damage the thyroid gland. the thyroid is a tiny, butterflyshaped gland that produces thyroid hormones on the front of your neck (cooper & biondi, 2012). nearly every organ in your body, including your heartbeat, is impacted by thyroid hormones because they regulate how your body uses energy (farling, 2000). sometimes the thyroid produces these hormones in excess or insufficiently. hyperthyroidism, a condition in which your body produces too much thyroid hormone, can make numerous bodily processes faster. the term hyper denotes an overactive thyroid. learn more about pregnancy-related hyperthyroidism. hypothyroidism, or having too little thyroid hormone, can slow down many bodily processes. hypo denotes the under activity of the thyroid. learn more about pregnancy-related hypothyroidism (mullur et al., 2014). thyroid hormones part in central nervous system development (morreale de escobar et al., 2000). since the fetus does not begin manufacturing thyroid hormone until weeks 16 to 20, the maternal thyroid is the only source of thyroid hormone in the early stages of pregnancy (morreale de escobar et al., 2000). by having frequent thyroid function testing and taking any medications your doctor prescribes, you can still have a safe pregnancy and preserve the health of your unborn child if you have thyroid issues (shahid et al., 2018). pregnancy is crucial for women since it involves significant physiological changes. today, it has been discovered that fetal programming has consequences beyond fetal development; as a result, poor prenatal programming significantly impacts the development of most chronic diseases that threaten human life (huang et al., 2017). one of the most frequent endocrine abnormalities during pregnancy is thyroid disease. the relationship between maternal thyroid dysfunction during pregnancy and unfavorable pregnancy outcomes and long-term health effects has drawn much attention over the past two decades (korevaar et al., 2017). understanding variations in thyroid function and the effects of thyroid disease during pregnancy is extremely important, given the significance of thyroid hormones for a healthy pregnancy and fetal development. numerous shortand long-term negative effects on the health of the mother and fetus are associated with overt thyroid problems. when detected and addressed at an early enough stage, the majority of these complications can be avoided. uncertainty exists over whether maternal subclinical hypothyroidism affects pregnancy or the subsequent cognitive growth of their offspring. practical issues figure 1: shows tsh levels and their symptoms figure 2: shows the key practical issues with thyroid hormones and with no thyroid hormones methodology to indicate the treatment outcomes of subclinical hypothyroidism in different age groups and in during pregnancy, several recent studies, review articles, prospective studies, cross-sectional studies, and literature reviews, all published and peer-reviewed, were searched and considered. the area of search was based on how effective is treatment with thyroxine in preventing morbidity in different age groups, treatment of infertility due to subclinical hypothyroidism, prevention of fetal loss, and fetal brain damage. this article is a review. thus not all information on thyroid https://journals.e-palli.com/home/index.php/ajcp pa ge 3 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 1-8, 2023 illness and its implications on fetuses has been provided are contained in this. we have included observational studies and all significant, pertinent big trials to highlight the overall conclusions. although we tried to incorporate the largest and most pertinent research, it is important to keep in mind that the tiny, hopeful observational studies were likely chosen due to publication bias. physiology of thyroid during pregnancy due to the increased metabolic demands during pregnancy, the thyroid gland undergoes considerable physiologic changes that affect its form and function. depending on baseline iodine supplies, the thyroid expands by 0–30% in size when iodine requirements rise. (berghout & wiersinga, 1998), (vannucchi et al., 2017). thyroid binding globulins (tbg), in particular, rise 2to 2.5-fold in response to increased serum estrogen (sorrenti et al., 2021). the thyroid gland must produce more thyroid hormone to resist standard extent of available thyroid hormone in response. additionally, human chorionic gonadotropin (hcg) is released prematurely in pregnancy. it has an alpha subunit one is comparable to thyroid stimulating hormone’s (tsh) alpha subunit, this causes the thyroid gland to be directly stimulated, a rise in free thyroid hormone, and a reduction of tsh in the first trimester of pregnancy(erdoğan et al., 2022; glinoer et al., 1993). this is assumed to happen so the fetus receives enough available thyroid hormone to by pass the placenta. inlarged tbg, hcg stimulation, renal excretion, and transplacental transit of available thyroxine (ft4) cause a nearly 50% rise in thyroid hormone production during the first trimester (lebeau & mandel, 2006). iodine and conception iodine is a crucial element of thyroid hormones, and pregnant women need more of it. iodine shortage is linked to thyroid malfunction, affecting fetal development (zimmermann, 2016). today, it is generally acknowledged that severe maternal iodine deficiency can have negative effects on both the mother and the baby, including miscarriage and conditions including hypothyroidism and goiter and stillbirth; for the newborn, including neonatal mortality; and for the child, including stunted growth, cretinism, and poor brain development (delange, 2007). in societies with severe iodine deficiency, iodine supplementation is advised as a treatment for maternal hypothyroidism, and there is strong evidence that it improves clinical outcomes, such as rates of cretinism and infant mortality (li et al., 2016). concerns have lately been voiced over the uk and other affluent nations iodine intake requirements for pregnant and childbearing women. some formerly believed to be iodine-rich regions were mild to moderately iodine deficient (zimmermann, 2007). additionally, recent research from the large longitudinal avon project in the uk has indicated a linear relationship between mild-to-moderate maternal iodine shortage and worse cognition in children between the ages of eight and nine (bath et al., 2013). despite the positive evidence, it is debatable whether pregnant women from areas with mild to moderate iodine deficiency should take iodine supplements. concerning supplements, not all of the research used rigorous techniques (bath & rayman, 2013). these studies have suggested that iodine supplementation in populations with mild-tomoderate iodine deficiency may have some positive effects on maternal newborn serum thyroglobulin and thyroid volume (i.e., a smaller increase), though data on thyroid function are inconsistent, and there is a lack of hard evidence regarding long-term effects like pregnancy outcomes, childhood neurodevelopment, and growth (zimmermann, 2007). some research suggests that starting iodine supplementation sooner can lead to better results (taylor et al., 2014). the american thyroid association’s guidelines for diagnosing and treating thyroid disorders during pregnancy and postpartum served as the reference ranges for the test values used in this study. regulation 14.2 of the ata recommendations, states that the following normal reference ranges should be used in the laboratory in the absence of trimester-specific tsh reference ranges in the laboratory. “1st trimester0.1 to 2.5 m iu/l”, “2nd trimester0.2 to 3m iu/l”, “3rd trimester0.3 to 3m iu/l”. standard free t4 volume is 0.7 to 1.8ng/ml the free t3 volume is 1.7 to 4.2 pg/ml. thyroid physiology in fetus in her opening remarks, dr. morreale de escobar discussed the various neurodevelopmental consequences in infants born in regions with endemic iodine shortages and children born with inherited hypothyroidism. infants born to mothers severely deficient in iodine risk developing neurologic cretinism, which can cause psychological issues, deafness, mutism, squinting quadriplegia. if these mental defective infants acquire enough iodine, their thyroid function will be completely normal at birth. on the other hand, infants with inherited hypothyroidism are hypothyroid at birth and are thought to have been so while still in utero. however, babies born to moms who are iodine insufficient will suffer acute, irreparable brain damage originating by an event that occurred during the 1st half of pregnancy, but neonates born with inherited hypothyroidism, if discovered and medicate in advance, will have a positive result for their neurodevelopment. understanding fetal thyroid development and possible maternal thyroid rise to the fetus is necessary to explain these disparities, particularly in the first half of pregnancy, before the foetus develops any significant thyroid hormone (eng & lam, 2020). first trimester fetal thyroid hormone levels are determined by fetal coelomic fluid and serum. the existence of thyroid hormone in the fetus during the 1st trimester has been shown in fetal coelomic fluid at https://journals.e-palli.com/home/index.php/ajcp pa ge 4 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 1-8, 2023 six weeks and fetal serums at twelve weeks of gestation, which are probably of maternal genesis (contempré et al., 1993). thyroid hormone levels in the circulation of the fetus continue to rise throughout pregnancy, as sum-up here: 1. at 12 weeks gestation, the average foetal serum t4 level is about 26 nmol/l, and at term, it is 128 nmol/l. foetal serum-free t4 (ft4) levels climb over the course of pregnancy, with a mean value of around 1.3 pmol/l at 12 weeks and 25.7 pmol/l at duration. (jansen et al., 2019). 2. the total triiodothyronine (t3) and free t3 concentrations in fetal serum remain low until 30 weeks of gestation due to the placental inner deiodination process of t4 to reverse triiodothyronine (rt3) and increase slowly from roughly 0.09 nmol/l at the 12th gestational week to 0.68 nmol/l at birth. 3. tbg concentrations also increase noticeably, rising from 5 mg/l during 12 weeks gestation to about 25 mg/l at delivery, with higher concentrations in babies than in adults. the research team led by dr. morreale de escobar concluded that the levels of thyroid hormone-binding proteins in fetal fluids and serum is extremely low and independent of the maternal thyroid state. before midgestation, most maternal t4 and t3 enter these compartments in the free form, causing levels to rise to physiologically important levels. they suspect there is proof that these maternal hormones play a significant role in fetal evolution, notably in the neurological structure, as is explained below. they also emphasize the importance of placental barrier to stop free t4 and particularly t3 from entering the fetal tissues at extent, can be hazardous. iodothyronine deiodinases in fetuses physiology of the fetal thyroid thyroid hormones (th) is produced by the mother during early pregnancy. despite being first noticed at 10–12 weeks, fetal th and hypothalamic-pituitary maturation do not start to rise until 18–20 weeks. the main fetal deiodinase that activates thyroid function is deiodinase type 2, or d2, which becomes more active by the end of the first trimester and enhances thyroid receptor occupancy with t3. apo-trs: thyroid hormone receptors that are not occupied. second and third trimester thyroid hormone levels in serum around 18 to 20 weeks of gestation, the pituitaryportal vascular system fully develops, and fetal thyroid hormone secretion starts. foetal serum t4 rises without involvement of maternal-fetal vascular connections from a mean of around 2 g/dl (26 nmol/l) at 12 weeks to 10 g/dl (138 nmol/l) at term in samples obtained via cordocentesis. (ziegler et al., 2022). the rise in hepatic fabrication of serum tbg and, to a minor extent, the stimulation of fetal thyroidal t4 production by tsh are both responsible for the rise in serum t4. from a mean of roughly 0.1 ng/dl (1.3 pmol/l) at 12 weeks to 2.0 ng/dl (25.7 pmol/l) at term, fetal serum due to placental type 3 or 5-deiodinase 3 (5-d3) venture, which changes t4 to reverse triiodothyronine (rt3) and t3 to reverse t2, the increase in fetal serum and free t3 levels is significantly less. foetal serum t3 levels range from 6 ng/dl (0.09 nmol/l) at 12 weeks to 45 ng/dl (0.68 nmol/l) at term. adult values of euthyroid persons serum tsh levels are particularly execessive than in the maternal circulation, rising progressively from 4 mu/l at 12 weeks to 8 mu/l at term. it is well thought out that the genesis and evolution purpose of this foetal tsh remain unknown. because of intrauterine levels of free t4, it appears that the development of the hypothalamicpituitary-thyroid compact and negative response is not finished prior a few months after birth. of free t4 and tsh are favourably related up to delivery. the fact that intrauterine ft4 levels are significantly greater than the one found in age-paired preterm babies leads to the perspective that maternal transfer of t4 continues to donate remarkably to the hormones accessible to foetal tissues (ares et al., 1997), (morreale de escobar, 1998). hypothyroidism primary maternal hypothyroidism is characterised as a high tsh level throughout pregnancy without uncommon exceptions such as thyroid hormone resistance, pituitary tumors that secrete tsh, and a few cases of central hypothyroidism with physiologically inactive tsh (zimmermann, 2007). hypothyroidism and the results of pregnancy (overt hypothyroidism) overt hypothyroidism has been associated with preterm birth, prenatal hypertension, placental abruption, low birth weight, postpartum hemorrhage, perinatal morbidity, and mortality. (wasserstrum & ananla, 1995). in overt hypothyroidism, the probability of preterm birth was higher (or 1.19, p 0.00001), according to a synopsis of 14th cohort studies and one case-control study (sheehan et al., 2015). the risk of problems was greatly reduced by earlier treatment of overt hypothyroidism and normalisation of thyroid function during pregnancy (leung et al., 1993). subclinical hypothyroidism poor perinatal outcomes could also be made more likely by subclinical hypothyroidism (maraka et al., 2016). according to a synopsis of 18 cohort studies, women’s with subclinical hypothyroidism had a inflated risk of miscarriage (rr 2.01), placental abruption (rr 2.14), preterm membrane breach (rr 1.43), and infant death (rr 2.58) (maraka et al., 2016). contrarily, a study of 10,990 patients found no increase in unfavorable pregnancy outcomes in the 483 (4.4%) women who had subclinical hypothyroidism (tsh >97.5% , ft4 2.5-97.5%) (rosario et al., 2018). the variations in these studies findings could be attributed to the various https://journals.e-palli.com/home/index.php/ajcp pa ge 5 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 1-8, 2023 ways subclinical hypothyroidism has been defined and the presence or absence of tpo antibodies. the subclinical hypothyroidism definition used in the metaanalysis varied between research studies; some adjusted for tpo status while others did not (maraka et al., 2016). positive antibodies raise the likelihood of complications for women with subclinical hypothyroidism”, and 3060% of pregnant women with high tsh have greater levels of tpo antibodies (mansouri et al., 2017). as was previously mentioned, a meta-analysis of three randomised trials that examined the influence of levothyroxine medication for subclinical hypothyroidism did’nt find any differences in obstetrical consequences after 3, 5, or 9 years of persue. to summarise, gestational hypertension, squat birth weight, and premature delivery are all increased by overt maternal hypothyroidism. treatment, particularly in the early stages of pregnancy, helps reduce these risks. although the evidence is mixed, maternal subclinical hy additionally, the treatment’s results have not yet been observed. this may not come as a surprise given the biological improbability that even a very slight thyroid dysfunction would cause like a wide variety of unfavourable pregnancy outcomes.pothyroidism may also negatively impact obstetrical consequences. there are conflicting recommendations about when to start treating subclinical hypothyroidism. currently, the american thyroid association advises starting treatment. when a pregnant woman has subclinical hypothyroidism, her tsh should be less than 4.0 miu/l when the pregnancy-certain reference value is not accessible to their community, 2.5 miu/l if she has negative tpo antibodies”, and more than that if she has positive tpo antibodies (anagnostis et al., 2017). the american college of obstetricians and gynaecologists states that there is no evidence that subclinical hypothyroidism can be identified or treated and makes no recommendations about its management; pregnant women with subclinical hypothyroidism have better baby outcomes (obstetricians aco, 2015). the royal australian and new zealand college of obstetricians and gynaecologists advises against testing for tpo antibodies or subclinical hypothyroidism and against using levothyroxine to treat subclinical hypothyroidism during pregnancy (james-mcalpine, 2019). the treatment of subclinical hypothyroidism in the course of pregnancy can not be necessary, according to the findings of more recent investigations. discussion one of the most prevalent endocrine conditions in pregnant women is thyroid disease, which negatively affects both the mother and the fetus. the initial antenatal visit’s universal screening aids in identifying thyroid problems in pregnancy. thyroid dysfunction should ideally be screened for before conception because any hypothyroid status can be treated before trying for a baby. this clinical study’s primary goal was to ascertain the frequency of thyroid problems in pregnancy. geographical differences in the prevalence of hypothyroidism during pregnancy are significant. according to data from western nations, subclinical hypothyroidism affects an estimated 2.5% of people, and overt hypothyroidism complicates up to 0.3% of pregnancies (akbaş & çarlioğlu, 2020). in india, pregnancy-related hypothyroidism is significantly more common than in western nations. we still have iodine deficiency in many areas of india. therefore prevalence varies greatly between different states. iodine deficiency is the most typical cause of hypothyroidism in pregnancy in underdeveloped nations like india. according to the study, 6.8% of people had hypothyroidism, with 0.7% having overt hypothyroidism and 6.1% having subclinical hypothyroidism. subclinical hypothyroidism was shown to be prevalent in 633 people in the indian population in a prior study, which was carried out in 2010 (sahu et al., 2010). our data is on par with recent research on prevalence. according to the statistics gathered, pregnant women have a high frequency of hypothyroidism, which necessitates rapid intervention. this high prevalence may be caused by multiple pregnancies, adolescent pregnancies, the low nutritional content of local foods, malnutrition and iodine deficiency, poor socioeconomic conditions of the local population, and high physiological demand during the growing years. in addition to iodine shortage, environmental factors may contribute to the prevalence of hypothyroidism in the area. compared to the west, asian countries were found to have a higher prevalence of hypothyroidism. the prevalence of hypothyroidism was considerably higher in the high-risk group than in the non-high-risk group in a huge chinese research study that collectively 2899 pregnant women (10.9 vs. 7.0%, p = 0.008) (ayyar, 2011). according to a chinese study, goitrogens are present in the food, as evidance from india, and micronutrient deficiencies, like those in selenium or iron, might result in hypothyroidism and goiter (marwaha et al., 2003) (teng et al., 2011), (das et al., 2011). therefore, it is hypothesized that india and asia have greater rates of hypothyroidism during pregnancy. in india, hypothyroidism is also not consistently prevalent (gayathri et al., 2009). bandela et al. 10% of people in andhra pradesh reported having sch. 2.8% of people had sch, according to gayathri et al. tsh’s varying upper limit cutoffs could contribute to this discrepancy (unnikruishnan et al., 2013). rate of live births per cycle “using data from nine studies involving 4396 women, busnelli et al. discovered that women with positive tai had lower lbr than women with negative tai (or 0.73, 95% ci [0.54-0.99], p14.04). the pooled effect estimate was also obtained using a random effect (notmod0.64, 950.64,90.42-0.99], p 14.05).werhofer et al. discovered that low tsh levels were linked to a higher likelihood of getting pregnant (13.9%) as opposed to https://journals.e-palli.com/home/index.php/ajcp pa ge 6 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 1-8, 2023 high tsh levels, which were linked to a lower likelihood of getting pregnant (5%); however, the authors note that statistical analysis of the link between baseline tsh levels and pregnancy potential was not possible in their study because of the small sample size (2-0.99], p. 14.05).“werhofer et al.’s”observation towards upgraded pregnancy potential in the presence of low-normal tsh levels (13.9%) compared to the high-normal tsh group (5%), but the authors note that statistical analysis of the effect of baseline tsh levels on pregnancy potential was not possible in their research study due to the least number of pregnancies’. conclusion to prevent harmful results for the mother and fetus, thyroid dysfunction during pregnancy must be identified and treated early. both overt hypothyroidism need to be properly managed. levothyroxine is frequently used to treat subclinical hypothyroidism; however, its ability to improve maternal or fetal outcomes has not been conclusively demonstrated. treatment for subclinical hyperthyroidism is typically unnecessary, although it should be remembered that nonthyroidal disease or pregnant thyrotoxicosis are both possibilities. although monitoring studies with euthyroid women who had thyroid autoantibodies showed worse maternal and fetal outcomes, there is currently insufficient evidence to show that these outcomes are improved by thyroxine supplementation. more rcts are required to examine the effects of treating pregnant euthyroid women with autoimmune thyroid disease, subclinical hypothyroidism, and isolated hypothyroxinemia. a secure basis for practise is provided by current practise guidelines, and they are strikingly similar and differing in significant ways that generally point to gaps in our present understanding of thyroid problems in pregnancy. references abdurazzakova, d. 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(2016). the importance of adequate iodine during pregnancy and infancy. hidden hunger, 115, 118-124. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 1 american journal of chemistry and pharmacy (ajcp) the relationship between the type of dimensions (3d, 2d) and mental interaction (high, low) in the augmented reality environment and its effects on learning samy abdelwahab safaan1* volume 2 issue 1, year 2023 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v2i1.1075 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: december 15, 2022 accepted: january 09, 2023 published: january 15, 2023 the research aims to reveal the relationship between the type of dimensions (3d, 2d) and the type of mental interaction (high and low) and its impact on students’ knowledge of the types of wireless networks and their components in the course “wireless communications and mobile security” based on their satisfaction with this learning, and their academic achievement. the research sample is 144 students in the second year, fourth level of the cybersecurity diploma, and they were divided into four experimental groups. the first experimental group studies with books augmented with three-dimensional models with high mental interaction, and the second experimental group studies with books augmented with three-dimensional models with low mental interaction. the third experimental group is taught with books augmented with two-dimensional models with high mental interaction, and finally, the fourth experimental group is taught with virtual books with two-dimensional models with low mental interaction. the research results that there is an interaction between the type of dimensions in augmented reality books and the mental interaction (high, low) in identifying the types of wireless networks and their components, where the high mental interaction groups excelled, regardless of the type of dimensions (3d, 2d) in the achievement test, and there was clear satisfaction for the students of the groups that study with 3d models compared to the 2d groups. keywords ar technology, augmented reality, digital world, qassim university, virtual reality 1 department of natural and applied sciences, community college of buraydah, qassim university, buraydah, 52571, saudi arabia * corresponding author’s e-mail: samyabdelwahabsafaan@outlook.com introduction augmented reality lets you unite the real world with the virtual world, in additional words you can do whatever you can do things that you cannot in the real world with the help of virtual objects and digital convenience. ar technology is based on three key factors: alignment of virtual and real objects, actual time relations, and a combination of digital and real environments. the phenomenon of ar involves adding a real-world environment into a device with additional virtual elements, according to milgrim’s mixed reality study, for example, if a map is considered through augmented reality, it will portray a modified version of the area with an easy glance to all the checkpoints and destinations in that area. in terms of entertainment, augmented reality provides a huge variety of video games and virtual entertainment allowing a person to explore the digital world (see figure 1) (alqifari et al., 2021; iordache et al., 2012; khan et al., 2019; küçük et al., 2014; miller et al., 2019). figure 1 shows a taxonomy related to the studies through which the actual human world and virtual fundamentals may be interlinked. the continuum varies from a whole different perspective in a real environment in contrast to a digital/virtual environment. according to this continuum, interlinked reality can be explained as a platform where the virtual world meets the real world. ar enables one to interact in the depths of the digital world and be a part of it. in consideration of augmented reality in smartphones, the features come through the camera that connects both worlds on a technical level and helps explore. a very well-known and easily understandable example is the virtual map application which allows accessing different locations appearing in the digital view mocking the real view of the person while augmented reality gps drive/ navigation enables an ar-based tracking and navigation system. such a combination of both worlds can discover a vast aspect of possibilities and opportunities for things that were not possible before. the attractive fact about ar is also the fact that it is portable and does not require any sort of specific instruments or equipment and can be accessed through a digital device with ease and minimum effort. an augmented reality mimics the real world through small segments of information in order to interpret the data on digital screens, the amount of accessed data and information decides the degrees of immersion of tracking and displaying technical results. figure 1: milligram’s mixed reality continuum [2] https://doi.org/10.54536/ajcp.v2i1.1075 https://journals.e-palli.com/home/index.php/ajcp pa ge 2 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 1-7, 2023 literature review digital growth takes its course to a further level each day as easy access enables the average people to possess access to ar through their devices which also plays its part in learning and educational purposes. many students face difficulties in learning and remembering the information they need as the cognitive load has its certain capacity, therefore, it is not wise to overload the mental capacity with information one cannot process. hence, augmented reality devices are introduced into the learning and educational systems that allow the students to develop an interest and pay attention to the knowledge that is being provided to them. the commonly used devices grant experience to graphics hardware, fast processors, and multiple onboard sensors. it also works on platforms like military and marketing. many researchers acknowledged augmented reality as a potential source of learning (alqifari et al., 2021; iordache et al., 2012; khan et al., 2019; küçük et al., 2014; miller et al., 2019) the research intends to examine the progress of the students of the qassim university in qassim, saudi arabia, when they were provided an ar-based learning system. impact of ar in diverse departments apart from the educational department, augmented reality is now gaining a reputation in other departments as well, especially in the medical department. techniques like vr exposure therapy help heal patients with anxiety claustrophobia, acrophobia, and social anxiety. it uses the mechanism of safely exposing them to a virtual situation designed according to their condition in order to overcome their mental illness (thwaites, 2021). another technique is the autism glass projects which lets kids with autistic issues understand their emotions without using google glass and maintain socially healthy relationships with people in their lives (van krevelen & poelman, 2010). ar phantom limb pain treatment in this situation the patients with missing limbs or parts of limbs are given the experience of physical feelings in those parts of their body using ar technology, which lets the amputee comprehend the virtual arm that is shown on the screen while the patient functions the amputated arm, the virtual arm on the monitor will also be displayed in the same action through the interfaces to stimulate and permit the patient to regulate the originally amputated limb with their mind, in directive to experience a satisfying effect. vr surgery simulation in addition to the opportunities ar provides, helps in ordinary surgical training and surgeries by medical experts as it reduces the chances of errors. numerous medical organizations are now adopting this technique for multiple varieties of surgical performances like knee arthroscopy where physicians practice injection of anesthesia during knee cap replacement surgery, this makes the procedure a lot simpler and more accurate as surgeons are able to understand the sensitivity of the situation accordingly. other than the risk-free factor it also enables the interns a chance to learn from their errors. following technologies should be encouraged and implied in clinical procedures rather than the hurdles that are put in their way like construction cost, the addition of system software for the record, and the reliability of quality (alqifari et al., 2021; khan et al., 2019) augmented reality has set its course and remains on track as it enters different domains of working organizations such as a virtual toolkit for robotic manufacture or fusion of physical/digital robotic covering workflows to validate the probable of augmented and diversified actuality for computerization and support in human-appliance original interface workflows, while fresh cases of ar use in manufacture locations show the upcoming potentials of the equipment (alqifari et al., 2021; hussein, 2017; krüger et al., 2022) the unique usage of virtual reality and augmented reality methods permits a continuous modification of the spray at dissimilar levels throughout the duplication development and benefits in rectifying or stopping the procedure quick if a certain part of the assemblies is exposed to be unattainable, or if they are predictable to unanticipated risky circumstances and struggles like if some curves are distorting a lot, then the spraying has to instantaneously be halted and assortments of suitable distortion are to be arranged after repetitive physical trials and detailed plotting of the adequate variations (alqifari et al., 2021; diegmann et al., 2015; hussein, 2017; krüger & bodemer, 2020; krüger et al., 2022). this way it can permit the instant re-arrangement of serious limitations, such as the approach of deposition, swiftness, the compression of the spray, pathways, remoteness to the arrangement in progress, and variations in the material physiognomies despite the fact being applied, for instance, the level of moisture, thickness, the number of threads, size of stones, amongst others. augmented reality has previously a noteworthy amount of presentation in the construction field, as a regulator device. it is applied in construction positions for the constructors to have an improved acceptance of wherever mistakes can have some precarious damaging inferences in the constructions. augmented reality still has not been meaningfully utilized so far as a project and optimizing means for the duration of the manufacturing course (alqifari et al., 2021; diegmann et al., 2015; kesim & ozarslan, 2012; krüger & bodemer, 2020; krüger et al., 2022). material and methods problem-based learning (pbl) is a model methodology in educational sectors where actual-world problems facilitate students’ learning instead of the forthright knowledge of facts and conceptions of conventional teaching. the research considers the learning implementation hypothesis that is based on observing the behavior of the https://journals.e-palli.com/home/index.php/ajcp pa ge 3 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 1-7, 2023 students after opting out of a learning approach involving 2d and 3d dimensional types and observing the changes in their cognitive load and attention driven towards the learning procedure. participants a study was conducted based on a sample of 144 students in the second year, the fourth level of the cybersecurity diploma, and they were divided into four experimental groups, for mapping the scientific research skills of students enrolled in wireless communications and mobile security curricula by using ar tools and applications, students from al-qassim university were selected, and enrolled in the course of computer science research for the year 2018/2019. the teaching of educational courses using 2d and 3d dimensional type was considered the independent variable while mental interaction i.e. the cognitive load and learning skills were considered the dependent variable. in order to judge the credibility of the hypothesis and its results two tools were designed, an achievement test to inspect the influence on students’ learning skills and a track of the amount of attention and pleasure ar learning provided, using the likert scale. the students were tested for the reliability coefficient based on an academic test taken from 15 students per hour (table 1) (alqifari et al., 2021; hussein, 2017; iordache et al., 2012; khan et al., 2019; küçük et al., 2014; miller et al., 2019). table 1: total test variance no of students total test variance reliability coefficient = 0.75 15 14.75 design the attentiveness and learning indexes of the test queries were determined through the calculations of results of the investigational sample which comprised (15) students. a two-by-two between-subject design was used in the investigation. mental interaction, which might be low (m–) or high (m+) using 2d dimensional type, was one influence. the mental interaction, which might be low (m–) or high (m+) measured using 3d dimensional type, was the other element. the values of attentiveness and learning index ranged between (33% 67%). the cognitive load reached (67%), which is statistically accepted (table 2). the outcomes of the study showed that the technology has been well acknowledged by students and facility members. they claimed that it helped them in teaching and evaluating their abilities in learning. furthermore, it was time-saving and required less amount of effort. it was also fast and easy to use and it provided excellence of data, according to the participants. the research approached the data implementation technique on different themes based on pre and post-test trials where the data of 34 students were investigated who were provided the opportunity of ar learning. the mean score of the pre-test for this theme observed was 23.1, whereas the post-test equaled 50.9. this indicated the escalation of the statistically substantial changes between the mean scores of students in the theme of data collection expertise for the pre and post-test of the digital learning skills (table 3). the former conclusion shows that there was a positive impression of using ar research for the progressive learning skills among students of the course. materials and apparatus the study also concluded different independent and dependent variable results based on the practice of dimensional type (2d and 3d) in educational departments, keeping a view of the mental interactions as variables for the research (alqifari et al., 2021; hussein, 2017; iordache et al., 2012; khan et al., 2019; krüger & bodemer, 2020; küçük et al., 2014; miller et al., 2019)[1-7]. the research embraced a 2x2 design, intersecting levels of difficulty and struggles in the tasks. while applicants finalized an anagram task, the dimensional framework was operated (i.e., the use of 2d and 3d dimensional type), and the task struggle was set to two conditions, either hard or easy. there were four experimental situations, mentally-easy using 2d dimensional type, mentally-hard using 2d dimensional type, mentally-easy using 3d dimensional type, and, mentally-hard using 3d dimensional type. members were unsystematically allocated into one of each potential order of the four conditions. each situation was observed individually at the four sequential positions correspondingly crossways participants, permitting for both a subject-based inquiry concluding all four trials and an among subject analysis only by means of the participants’ initial anagram trials. procedure the participants were greeted and given an explanation of the study’s protocol and content at the start of the study. after all of their questions were answered, they signed an informed consent form. participants individually assessed their capability principles, expectations for accomplishment, and subjective assignment standards regarding the topic and task on a mutual feedback form that was launched on a computer screen. participants were then directed to the research room, where they finalized two short preparation tests. first was an anagram-deciphering task with the sample anagram and a couple of training anagrams. this confirmed that the participants were able to see the anagrams and implicit the anagram task. then, the research provided the ar headset to the participant in order to carry out the second task which was a navigation trial using ar objects. the experimenter requested participants to make sure if they observed a virtual ball and then to move in its direction until the ball changed color. this procedure was implied again with a digital cube. the number of anagrams answered was recognized by evaluating the footage of every single participant. to accelerate examination, a program was introduced by means of https://journals.e-palli.com/home/index.php/ajcp pa ge 4 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 1-7, 2023 python (version 2.7) to figure out the audio and cut large segments of stillness from each of the demos. a human coder then attended to the clipped recordings and made it clear what words were communicated by participants. each participant was awarded a point for each anagram deciphered appropriately (alqifari et al., 2021; hussein, 2017; iordache et al., 2012; khan et al., 2019; krüger & bodemer, 2020; küçük et al., 2014; miller et al., 2019). statistical tables the groups’ individual ratings of their familiarity with the matter, their expectation of how well they would unravel the tasks, and their apparent worth, significance, and awareness of information were equated to ensure that the groups did not fluctuate in their pre-study data about and attentiveness in the matter. table 2: cognitive load and attentiveness positive impression cognitive load attention span 67% 33% table 3: test score test credit dimensional type using 2d dimensional type using 2d dimensional type mental interaction low (m) 22.1% 24.4% high (m) 49.6% 52.4% table 4: mean score data collection significance 0.01 pre-test mean score 23.2 post-test mean score 50.9 figure 3: interactions using ar results participants deciphered multiple queries in the easy situations than in the tough situations representing that the management of exertion in the middle of situations was effective. social reserve indicates that contributors can solve harder cyber security assessments when they are alone than socially. the chief result of striving (easy vs. hard) on the mark was noteworthy, with easy (2d &3d) as the dependent variable = 7.68, while hard (2d &3d) was the independent variable = 4.16 based on interactions in the anagram test. the results showed major differences when the bar chart was designed according to the results of assessment trials, mental interactions using 2d dimensional type ranged from 3.24%-2.11% while mental interactions using 3d dimensional type ranged from 5.70%-9.70%. the subjects with a high level of accomplishment gave forth a positive attitude toward augmented reality applications. the developing statistic showed that the participants who were given ar opportunities in terms of knowledge possessed a high level of attainment, which made a constructive approach obvious towards this technology and put forth a low extent of energy during the application procedure. moreover, the outcome of the examination showed that the subjects who have used the applications of augmented reality, had a future intent to use these techniques in the future, as they were pleased with the practice of using virtual models, and had a reduced level of nervousness while functioning through this technology. these circumstances of results can be enlightened by the detail that the ar applications fascinate the responsiveness of the users, also enables an operative learning atmosphere, and encourage them for focusing better on the learning (alqifari et al., 2021; diegmann et al., 2015; hussein, 2017; kesim & ozarslan, 2012; krüger & bodemer, 2020; krüger et al., 2022; mason, 2020; miller et al., 2019). discussion the fundamental object that turned the attitude of the participants into affirmative could probably be the fact that the learners discovered a diverse learning objective other than depending on the old-style learning techniques. the prominent situation was that using augmented reality applications escalates enthusiasm and the users can also have fun working with them, resulting in a positive outlook towards these applications. it was also witnessed that the participants could raise the capability of their understanding, reading, speaking, and learning using ar techniques, more in comparison to the ones who followed the customary ways of learning and research. as it was proven from the collected data that the participants developed better expertise and talents like spatial learning capability, technology efficiency, computer science https://journals.e-palli.com/home/index.php/ajcp pa ge 5 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 1-7, 2023 approximation, finding solutions for their problems, and collaboration in order to avoid any difficulties (bartosh & anzalone, 2019; fernando et al., 2017). metzler and shepard (1974), r. n. shepard and cooper (1982), and corballis (1986) all describe a similar process of mental rotation (corballis & blackman, 1990; metzler & shepard, 1974; shepard & cooper, 1986). how much can this approach take use of the fact that 3-d figures have steeper slopes than 2-d ones? if one considers the “dimensionality” of a stimulus to be equivalent to its “complexity,” then one may say that the “effect” of dimensionality (as in the case of complexity) results from the higher amount of information that must be maintained in the case of 3-d stimuli or more complex stimuli. this argument is supported by the fact that it is possible to make it. folk and luce (1987) demonstrated this for 2-d polygons by finding that mental rotation rates were slower for polygons with more vertices than for polygons with fewer vertices. this was true for simple and complicated polygons alike (folk & luce, 1987). folk and luce claimed that they were able to produce the complexity effect because they did not depend on reflected or perturbed distractors, but cooper (1975, 1976) was unable to do so because they used distractors that were more similar to their standard polygons. folk and luce were able to achieve the complexity effect for this reason. participants were required to encode and modify a more comprehensive representation of the stimulus so order to avoid being fooled by a distractor that resembled it too closely. this was done to prevent being fooled by a distractor that resembled it too closely. slower rotation rates were detected as a result of the increased information load brought about by the more realistic presentation of more complex pictures (cooper, 1975, 1976). since it is thought that students may learn accurate mental models of a topic by envisioning and engaging with the portrayal of the phenomenon, much study has been conducted on the influence of virtual reality technology on the teaching of chemistry (antonoglou et al., 2011; chiu & wu, 2009; halpern & collaer, 2005; phillips et al., 2010). this was the most comprehensive research conducted to date on how chemistry students engage with desktop 3d virtual reality learning environments. it examined the students’ perceptions of the surroundings, sensation of presence in the environment, spatial orienting abilities, and sentiments of self-efficacy. throughout the length of the experiment, students will be able to zoom in and out of the 3d virtual reality environment to observe molecules and the angles at which they are bonded from a variety of perspectives. in addition, they may spin and control a molecule in order to explore how the atoms contained inside it are bonded to one another by using the many opportunities presented by the surrounding environment. as part of this kind of chemistry education exercise, students are tasked with mentally manipulating or changing an object into a mental configuration. students with a high level of spatial intelligence may be able to perform the mental adjustments required for molecular organization more quickly. this is a reasonable conclusion. researchers have shown, however, that students often lack this mental capacity for seeing and manipulating three-dimensional chemical groupings (halpern & collaer, 2005; wu & shah, 2004). on the basis of this examination, applying the ar implicational technology in computer laboratories supervised by the expert’s leadership results in having no trouble while using the opportunities provided by the application and dealing with reduced levels of unease among the learners (bartosh & anzalone, 2019; fernando et al., 2017). the purpose of students to use ar applications in the forthcoming can be clarified by the basis of the matters that go further for revolution to catch the attention of users and give growth to the learning impulse of students. conclusion the research concludes that the use of ar can either increase or decrease the cognitive load and attention span of an individual as provides various possibilities and learning techniques for the users. learning techniques that involve augmented reality will allow educational departments to give forth promising results and develop a better reputation through their unique teaching styles and technicalities. understanding such occurrences needs the construction of a mental model based on knowledge about the separate stages and connecting these phases according to the basic rules of causation (lowe & boucheix, 2008; narayanan & hegarty, 2002). both lowe and boucheix (2008) and narayanan and hegarty (2002) discovered the same thing. both 2.5d and 3d provide a third dimension, which might be important for constructing accurate mental representations. this is especially true in instances when spatial expansion is necessary for understanding. since monoscopic presentations already communicate a variety of spatial signals, the move from monoscopic 2.5d to stereoscopic 3d is a less important one. however, shifting from two dimensions to two and a half dimensions may provide crucial information. recent research has shown that students can only profit from stereopsis in a restricted number of situation. it is obvious that for the purpose of reducing the cognitive load of the users and making certain of efficient learning, the fundamentals of multimedia and learning concepts have the tendency to work when designed in an augmented reality application. hence, the study recommended techniques that can help improve the quality in accordance with dependent and independent variables of augmented reality, firstly, planning ar applications that can be adopted in order to decrease the cognitive load and increase the achievement levels in students to enhance the learning abilities of the learners. secondly, for the purpose of attention increasing and inspirational increase, an updated learning environment should be created that provides ar application access. courses that involve ar applications should be put in https://journals.e-palli.com/home/index.php/ajcp pa ge 6 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 1-7, 2023 the curriculum so that the learners can have access to opportunities at their homes. reasonable studies should be planned by evaluating the insolence, success, and perceptive loads of learners for augmented reality’s uses in dissimilar grounds of learning (alqifari et al., 2021; corballis & blackman, 1990; fernando et al., 2017; hussein, 2017; metzler & shepard, 1974; miller et al., 2019; shepard & cooper, 1986). there are certain drawbacks and limitations to this technology specifically when it comes to the cost and maintenance of augmented reality and applications, it also requires expertise and an appropriate crew to keep the technology functional and advanced, speaking of advanced, it takes specific understanding to update and keep track of the advancements that the technology requires and the technical issues that ar devices face at certain times. there is also a need for a further vigorous discussion about virtual knowledge. there remains the issue of a compressed understanding of the discriminations of countless simulated spaces, stages, and technologies. another issue is that access to the gadgets with ar tech, the admittance to the internet, a computer or laptop, a nintendo switch domain, or even the intellectual space to get involved in the virtual world is not appropriately circulated. the capability to observe the situation and provide current reports, information and projections is reflected as an unachievable luxury in lots of parts of the world. also, many regions are still unaware of this technology and the possibilities they are missing out on that is why there should be certain sourcing to promote awareness of the opportunities of the virtual world (alqifari et al., 2021; corballis & blackman, 1990; folk & luce, 1987; hussein, 2017; krüger & bodemer, 2020; metzler & shepard, 1974; miller et al., 2019; shepard & cooper, 1986). even though augmented reality has no limit to the informational opportunities it provides, there still are certain limitations that get in the way of evolution because much of this data likely possesses doubtful consistency when associated with outmoded bases. the innovative generation is familiarized with constructing virtual existence in the digital world of their own according to their own requirements and desires. as a result, their aims and objects have taken a different course by communal worldwide connections as a substitute to textbooks and certain other customary written sources of knowledge and learning, on the other hand, exist to be the predictable and steadfast basin of understanding. in concern to that, there are certain boundaries and hurdles that are uncrossable, for example, the complication that rises while arranging overlaid means of information (cooper, 1975, 1976). many students face malfunction problems while using augmented reality applications and the case situations where it gets rather troublesome for the students to make use of the device or technical markers to get access to the augmented information or the virtual world. it is often believed that the possible explanation for this boundary is that application designers need to advance the set of rules in the pursuit and processing of representative departments. there is another suggested possibility to be considered, that is for the common future scientist to discover further into the realm of augmented reality and its virtual world in order to understand the aspects of this domain further and provide answers to the complications so that those solutions can be considered and tally in the future ar application designing in the academic contexts (antonoglou et al., 2011; chiu & wu, 2009). augmented reality sometimes also proves itself to be a distraction for the users when they divert their attention towards the charm of the virtual world being a novelty rather than the information that they are supposed to be absorbing it also has imaged itself as a disadvantage in terms of learning and getting tasks done using the virtual assets. it is also stated that augmented reality can be measured as a “disturbing technology,” as augmented reality can be a source of interference for users trying to learn and develop reading abilities while they use tools such as head-mounted displays (hdm) in terms coaching and educational session. some researchers declared in their analogies that the device that transports virtual figures can disorder the original commitment among the user with other users and related folks in relation to communication. an additional way of intrusiveness is the circumstance that whenever to completely make use of this opportunity in the learning session, a discussion requires to be entirely equipped in the computer hardware division and partake unwavering internet connections so the session stays on its flow while in commencement (antonoglou et al., 2011; chiu & wu, 2009; halpern & collaer, 2005; lowe & boucheix, 2008; mason, 2020; narayanan & hegarty, 2002; phillips et al., 2010; wu & shah, 2004). acknowledgments the writer is thankful to qassim university. conflict of interest there is no conflict of interest. consent for publication the author agrees to the final version submitted to the journal. funding none references alqifari, a., alghidani, m., almazyad, r., alotaibi, a., alharbi, w. a., aljumail, e., alqefari, g., alkamees, a., & alqifari, h. (2021). burnout in medical undergraduate students in qassim, saudi arabia. middle east current psychiatry, 28(1), 1-6. antonoglou, l., charistos, n., & sigalas, m. (2011). design, development and implementation of a technology enhanced hybrid course on molecular symmetry: students’ outcomes and attitudes. chemistry https://journals.e-palli.com/home/index.php/ajcp pa ge 7 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 1-7, 2023 education research and practice, 12(4), 454-468. bartosh, a., & anzalone, p. (2019). experimental applications of virtual reality in design education. chiu, m.-h., & wu, h.-k. (2009). the roles of multimedia in the teaching and learning of the triplet relationship in chemistry. in multiple representations in chemical education. springer, 251-283 cooper, l. a. (1975). mental rotation of random twodimensional shapes. cognitive psychology, 7(1), 20-43. cooper, l. a. (1976). demonstration of a mental analog of an external rotation. perception & psychophysics, 19(4), 296-302. corballis, m. c., & blackman, a. r. (1990). the effect of apparent movement on mental rotation. memory & cognition, 18(5), 551-555. diegmann, p., schmidt-kraepelin, m., eynden, s., & basten, d. (2015). benefits of augmented reality in educational environments-a systematic literature review. fernando, s., reinhardt, d., & weir, s. (2017). simulating self supporting structures. sharing of computable knowledge!, 177. folk, m. d., & luce, r. d. (1987). effects of stimulus complexity on mental rotation rate of polygons. journal of experimental psychology: human perception and performance, 13(3), 395. halpern, d. f., & collaer, m. l. (2005). sex differences in visuospatial abilities: more than meets the eye. cambridge university press. hussein, l. m. (2017). the effectiveness of teaching educational research course on the development of scientific research skills, academic and personal integrity among female students of al-qassim university. international journal of asian social science, 7(5), 392-409. iordache, d. d., pribeanu, c., & balog, a. (2012). influence of specific ar capabilities on the learning effectiveness and efficiency. studies in informatics and control, 21(3), 233-240. kesim, m., & ozarslan, y. (2012). augmented reality in education: current technologies and the potential for education. procedia-social and behavioral sciences, 47, 297302. khan, t., johnston, k., & ophoff, j. (2019). the impact of an augmented reality application on learning motivation of students. advances in humancomputer interaction, 2019. krüger, j. m., & bodemer, d. (2020). different types of interaction with augmented reality learning material. 2020 6th international conference of the immersive learning research network (ilrn), krüger, j. m., palzer, k., & bodemer, d. (2022). learning with augmented reality: impact of dimensionality and spatial abilities. computers and education open, 3, 100065. küçük, s., yýlmaz, r. m., & göktaþ, y. (2014). augmented reality for learning english: achievement, attitude and cognitive load levels of students. education & science/ egitim ve bilim, 39(176). lowe, r., & boucheix, j.-m. (2008). learning from animated diagrams: how are mental models built? international conference on theory and application of diagrams, mason, m. (2020). the elements of visitor experience in post-digital museum design. design principles and practices, 14(1), 1-14. metzler, j., & shepard, r. n. (1974). transformational studies of the internal representation of threedimensional objects. miller, m. r., jun, h., herrera, f., yu villa, j., welch, g., & bailenson, j. n. (2019). social interaction in augmented reality. plos one, 14(5), 0216290. narayanan, n. h., & hegarty, m. (2002). multimedia design for communication of dynamic information. international journal of human-computer studies, 57(4), 279315. phillips, l. m., norris, s. p., & macnab, j. s. (2010). the concept of visualization. in visualization in mathematics, reading and science education (pp. 1934). springer. shepard, r. n., & cooper, l. a. (1986). mental images and their transformations. the mit press. thwaites, h. (2021). heritage preservation in the postdigital era: how much information is enough? virtual creativity, 11(1), 9-31. van krevelen, d., & poelman, r. (2010). a survey of augmented reality technologies, applications and limitations. international journal of virtual reality, 9(2), 1-20. wu, h. k., & shah, p. (2004). exploring visuospatial thinking in chemistry learning. science education, 88(3), 465-492. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 37 american journal of chemistry and pharmacy (ajcp) phytochemical and biological screening of catholic vegetable (jatropha tanjorensisjatropha tanjorensis) aqueous extract in bali, taraba state, nigeria o. imohiosen1* volume 2 issue 2, year 2023 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v2i2.1602 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: april 18, 2023 accepted: may 23, 2023 published: may 25, 2023 jatropha tanjorensis has been used as a medicinal herb in treatment and management of disease because they are relatively safer, more affordable and sometimes offers better therapeutic value than synthetic drugs. the study investigated the phytochemical and biological screening of catholic vegetable (jatropha tanjorensis) aqueous extract with the objectives of evaluating some secondary metabolites in the plant using standard qualitative methods and ascertaining the antimicrobial effect of jatropha tanjorensis aqueous leave extract on bacteria of medical importance such as pseudomonas aeruginosa, klebsiella pneumonia, staphylococcus aureus, salmonella typhi and escherichia coli using the agar well diffusion method. the results revealed that the extract of the leaf of j. tanjorensis contain some important bioactive components (phytochemicals) in a high amount such as saponins followed by flavonoids, alkaloid and tannins in moderate amount whereas phenol, phlobatannins and glycosides are in low amount. it also shows that the extract was active against e. coli, p. aeruginosa, s. aureus, k. pneumonia and s. typhi measuring clear zones of inhibition 30.0mm, 22.0mm, 19.0mm, 18.0mm and 17.0mm respectively. thus, the exhibition of strong antibacterial activity and phytochemical profiles obtained from the analyses of j. tanjorensis leaf indicated that it might be recommended for human consumption and adequate amount of consumption could contribute greatly towards meeting human nutritional needs for normal body growth and adequate protection and curative measures to common diseases related to the test organisms. keywords biological screening, catholic vegetable, phytochemical, aqueous extract 1 department of science laboratory technology, federal polytechnic bali, taraba state, nigeria * corresponding author’s e-mail: ojeaga2003@yahoo.com introduction plants constitute a pool and storehouse of herbal medicines (oyewole et al., 2007). in addition, herbal medicines have received greater attention as an alternative to clinical therapy and the demand for these remedies has currently increased (usda, 2007). their use is highly dependent on experimental screening to ascertain active components, safety, and efficacy of the plant products (oyewole et al., 2007; igbinaduwa et al., 2011). jatropha tanjorensisjatropha tanjorensis plant description of jatropha tanjorensisjatropha tanjorensis jatropha tanjorensis is a multipurpose perennial shrub/small tree of 3-6 m height. the word ‘jatropha’ is derived from greek words ‘jatros’ and ‘trope’ (food/nutrition) which implies medicinal uses. the genus jatropha belongs to family euphorbiaceae and subfamily acalyphoideae and includes about 175 species. it may be evergreen or deciduous, depending on climate. it has a short tap root, robust laterals and many fine tertiary roots. the stem is woody, erect, cylindrical, solid and branched. branches are stout, green and semi woody. leaves are palmate and have 5-7 shallow lobes and are arranged in alternate with spiral phyllotaxis. length and widths of leaves varies from 16-21 and 14-18 cm and are cauline and ramel, exstipulate, petiolate. petioles are 12-19 cm long (akhilesh & tewari, 2015). figure 1: picture of jatropha tanjorensis (source: fieldwork 2023) taxonomy/classification of jatropha tanjorensisjatropha tanjorensis kingdom plantae phylum tracheophyta class magnoliopsida order malpighiales family euphorbiaceae genus jatropha species jatropha tanjorensis common names: catholic vegetable, lapalapa or iyanaipaja (yoruba), ugu-oyibo (igbo), “hospital too far” (pidgin english) (ochulor, njoku, uroko & egba, 2018). https://doi.org/10.54536/ajcp.v2i2.1602 https://journals.e-palli.com/home/index.php/ajcp pa ge 38 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 37-40, 2023 binomial name: jatropha tanjorensis j.l. ellis and soroja (gbif secretariat, 2021; usda plants, 2007-2010). importance and uses of jatropha tanjorensisatropha tanjorensis jatropha tanjorensis ellis & saroja is among these screened plants, which belongs to genus of flowering plants in the spurge family, euphorbiaceae (carlasabandar, 2010). it is a multipurpose plant, nurtured for medicinal applications and used as food. almost every fragment of the plant is useful and nutritional in several ways. its many benefits depend on which part of the plant is being used. the leaves are generally cooked and eaten like vegetables such as fluted pumpkin leaves (ugu) and spinach (nwachukwu, 2018). j. tanjorensis mainly use is for fencing, and as a source of edible leafy vegetables and natural medicinal therapy against ailments which include diabetes in many parts of southern nigeria (iwalewa et al., 2005). jatropha tanjorensis possesses important pesticidial, anticancer and hepatoprotective activity. the sap from stem stops itching, scratches and bleeding of cuts. the root decoction is used as a mouth wash for treating bleeding gums, toothache, ringworm, eczema and scabies and also treat dysentery and venereal diseases (nwachukwu, 2018). the leaf aqueous extract is consumed as a blood tonic with the claim that it increases blood volume. it can also be used as alternate to lettuce in salad preparation amongst other very vital usages as reported by many authors (igbinaduwa et al., 2011). the oxidant potential of the plant extract has been examined and to have antioxidative potential against reactive oxygen species produced in protein energy malnutrition. the seed oil of j. tanjorensis is used for the treatment of paralytic and rheumatism affections. traditional medicine practitioners use the root in the treatment of urinary tract infections and sexually transmitted infections. it is also used for irregular periods, to treat menstrual pains, and to ensure a strong foetus during pregnancy (nwachukwu, 2018). notwithstanding the countless potentials of jatropha tanjorensis as a therapeutic plant has not found a wide application in pharmaceutical systems. its usage has mostly been restricted due to insufficient scientific information on the plant. hence, this study aimed to evaluate the phytochemical and biological screening of catholic vegetable (jatropha tanjorensis) aqueous extract. materials and methods study area the study was carried out in federal polytechnic bali, bali local government area (lga) of taraba state, nigeria (see fig.2). the local government area lies between latitude 7046 n and 7054 n of the equator and longitude 10 030 e and 110 00 e of the prime meridian (topographic sheet, 1968). this falls within the dry guinea savannah with an estimated land area of 11,540 km2. it has some mountains like gazabu, dakka, maihula, bagoni, among others. based on the 2006 national population census, bali had a population of about 211,024 persons (npc, 2006). figure 2: map of bali local government area showing the study area (source: bureau for land and survey jalingo, taraba state) thermostated water bath, electronic balance (model: 3003h),hot plate (model: orl 2080), oven (thermcool), desiccators, cotton wool, filter paper, pestle and mortar, glass wares (volumetric/conical flask, beakers, funnel), jatropha tanjorersis leaves reagents/chemicals methanol, diethyl ether, ammonia solution, mayer’s regent, wagner’s regent, iron (iii) chloride, hydrochloric acid, ethanol, acetic acid, tetraoxosulphate (vi) (h2so4) acid, sodium hydroxide (naoh) collection and identification of jatropha tanjorersis fresh jatropha tanjorersis leaves were harvested from rest house residence in bali l.g.a of taraba state, in the in the month of april, 2023 and identified using pertinent taxonomic literatures (gbif secretariat, 2021; usda plants, 2007-2010). the harvested leaves were sorted and then washed with clean water to remove dirt and unwanted materials that may be adhering on the leaves and after washing. the samples were air dried at room temperature on the laboratory table of science laboratory technology department, federal polytechnic bali and after drying, it was then milled using pestle and mortar and the powder obtained were used for analyses. aqueous extraction preparation of jatropha tanjorersis leaves 100 gm of fine powdered sample of jatropha tanjorersis was weighed and mixed with 500 ml of distilled water in a round bottom flask and then the reflux condenser apparatus was setup. a reflux condenser was attached to the flask and inserted with a heating mantle. the mixture was refluxed for one hour and filtered with watman filter paper no. 1. the reflux was done again using new distilled water at each stage. the filtrate was heated to dryness by https://journals.e-palli.com/home/index.php/ajcp pa ge 39 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 37-40, 2023 evaporation. part of the dry filtrate (extract) was used for phytochemical and antimicrobial screening. analyses determination of phytochemicals alkaloid, phenol, phlobatannins, flavonoids, saponins (froth test), volatile oils and tannin were determined using qualitative methods described by trease & evans (2009). microorganisms the bacteria used are known to be potentially pathogenic to humans. they include salmonella spp., escherichia coli, staphylococcus aureus, pseudomonas aeruginosa and klebsiella spp. all the organisms’ strains were obtained as clinical isolates from sancta maria clinic, bali, taraba state and optimum laboratory, jimeta-yola, adamawa state, nigeria. antimicrobial susceptibility testing the antibacterial activity of jatropha tanjorersis leaf extracts (100mg/ml) in vitro on the isolates was determined by the agar well diffusion method as described by (olakunle et al., 2013: osho & bello, 2010). this was done using pour plate method in which small colonies from each clinical isolates of the test organisms were made into suspension with 1ml of sterile distilled water in test tubes. 0.1ml of each suspension was dispensed into sterile petri dishes after which melted and sterilized nutrient agar maintained at 450c was poured (15 aliquot) into the respective plates. the plates were allowed to set, four equidistant wells of 6mm in diameter were punched in each plate using a sterile cork borer. 0.2ml of extract was introduced into each of the wells. a well filled with sterile water served as control and the plates were allowed to stay for15 minutes for pre-diffusion to take place followed by incubation for 24-48 hrs at 370c. the zones of inhibition were measured with the use of a metric rule and were recorded in centimeters (millimeters). table 1: phytochemical screening of jatropha tanjorensis leaves aqueous extract chemical constituent confirmation phlobatannins + saponins +++ glycosides + alkaloids ++ phenols + flavonoids ++ tannins ++ (+++= highly detected, ++= moderately detected, += lowly detected -= not detected) table 2: antimicrobial activity of jatropha tanjorensis leaves aqueous extract against some bacteria isolates organisms dose zone of inhibition (bacteria) (ml) (mm) staphylococcus aureus 0.2 19 salmonella typhi 0.2 17 escherichia coli 0.2 30 klebsiella pneumonia 0.2 18 pseudomonas aeruginosa 0.2 22 results and discussion discussion the result of the phytochemical screening of the jatropha tanjorensis leaf aqueous extract is presented in table 1. the tannin content of the j. tanjorensis leaf sample was moderately detected. according to ujonwundu et al. (2010) and who (2002), tannins have antioxidant effects which help in prevention of cancer and a level below 5mg/100g in foods are safe for human consumption. flavonoids are free radical scavengers and potent watersoluble super antioxidants (onimawo & akubor, 2012). flavonoid was moderately detected in the leaf extract of j. tanjorensis. though, moderate consumption of flavonoid is essential for normal heart beat, metabolic and neuromuscular activities. it also protect against all stages of carcinogenesis, prevents oxidative cell damage and have strong anticancer activity (bello et al., 2008). the alkaloid content of the jatropha tanjorensis leaf sample was moderately detected. alkaloids are medicinal and important secondary metabolites. alkaloids isolated in pure form and their synthetic derivatives are used as basic medicinal agents for their analgesic and bactericidal effects (morton, 2001). saponin was highly detected in the aqueous leaf extract of j. tanjorensis. some properties of saponins include cholesterol binding properties, formation of foams in aqueous solution, bitterness and hemolytic activity (sodipo et al., 2000). however, saponins’ anti-microbial activities make them good for treating yeast and fungal infections (okwu & ndu, 2006). though, high level of saponin has been associated with gastroenteritis manifested by diarrhoea and dysentery (gernah et al., 2007), acute poisoning is relatively rare (awe & sodipo, 2001). however, these toxic principles exhibit useful medicinal properties (afiukwa & igwe, 2015). thus, the presence of some important phytochemicals highly detected (saponin), moderately detected (tannin, flavonoid and alkaloid) and lowly detected (phenols, glycosides and phlobatannins) in the jatropha tanjorensis leaf is an indicator to the medicinal value of the plant. table 2. show the result of the antimicrobial activity of j. tanjorensis leaves aqueous extract against five isolated bacteria identified to be pathogenic to humans. the extract exhibited effective antibacterial action against the isolated bacteria by their clear zones of inhibition identified as pathogenic to humans. e. coli displayed greater level of susceptibility 3.0cm (30mm) than other tested bacteria. followed by p. aeruginosa with level of susceptibility 2.2cm (22mm). s. aureus, k. pneumonia and s. typhi indicated the least zone of inhibition 1.9cm, https://journals.e-palli.com/home/index.php/ajcp pa ge 40 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 37-40, 2023 1.8cm and 1.7cm (19mm, 18mm and 17mm) respectively. the most resistant isolate tested was s. typhi with the least zone of inhibition of 1.7cm (17mm). conclusion the study revealed that the aqueous extract of the leaf of j. tanjorensis contain saponins in high amount followed by flavonoids, alkaloid and tannins in moderate amount. in contrast, phenol, phlobatannins and glycosides is in low amount. it also shows that the extract was active against e. coli, p. aeruginosa, s. aureus, k. pneumonia and s. typhi measuring clear zones of inhibition 30.0mm, 22.0mm, 19.0mm, 18.0mm and 17.0mm respectively. thus, the display of strong antibacterial activity and phytochemical profiles of j. tanjorensis leaf showed that it may be recommended for human consumption and adequate amount consumption could contribute greatly towards meeting human nutritional need for normal body growth and adequate protection and curative measures to common diseases related to the test organisms. acknowledgements the researcher wishes to acknowledge the management of federal polytechnic bali for sponsoring this study via tet fund. reference afiukwa, a. c. & igwe, o. d. 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(2002). the world health report 2002 – reducing risks, promoting healthy life. geneva, who. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 11 american journal of chemistry and pharmacy (ajcp) adsorption of some heavy metals from wastewater using fine sand and zeolite benjamin. i.1*, rose. e. kukwa1, teghtegh. f. samoh2, john. o. ajegi3, terhile. m. iortile1 , tor p. ngunoon4, solomom. d. igbawase1, christopher. n. abah.1 volume 1 issue 1, year 2022 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v1i1.385 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: july 11, 2022 accepted: july 18, 2022 published: july 20, 2022 in this study, successive columns containing different types of solid state materials were used to investigate the treatment efficiency of soya bean oil mill wastewater (soap stock) using heavy metals analysis. soap stock is a liquid waste obtained from soya bean oil industry from the neutralization process which has several environmental issues due to its high amount of organic and inorganic pollutants. soap stock emerges from the refining process when oil is treated with a dilute alkali solution separating the ffas as soaps. in this present study, an attempt was made at treating the wastewater using successive columns containing different types of solid state materials. the wastewater was passed through successive columns of fine sand, zeolite and zeolite/fine sand composite.. this treatment method decreased the toxic concentration of heavy metals assessed which include pb, cu, zn, ni and cd by mean percentages of 96.60%, 100%, 62.0%, 16.10% and 96.60% respectively. most contaminants were removed in the zeolite/sand composite column possibly as a result of ion exchange capacity in addition to high sorption affinity of zeolite on its active sites. this simple zeolite based method enable us to obtain environmentally friendly treated soya bean oil water that can be safely used for irrigation purposes. keywords zeolite, heavy metals, soap stock, physical precipitation, fine sand, column 1 department of chemistry, benue state university, makurdi, nigeria. 2 department of chemistry, university of ilorin, ilorin, kwara state, nigeria. 3 college of education oju, benue state, nigeria. 4 department of chemistry, joseph sarwuan tarka university, makurdi, nigeria. * corresponding author’s e-mail: ishwahlizer@gmail.com introduction wastewater obtained from edible refined oil processing industries is a major issue of environmental concern particularly in under-developed or developing countries. the waste streams that come out from such oil refinery create serious environmental problem such as great threat to aquatic life due to its high organic content and among other issues. hence its treatment is essential prior to its disposal. the choice of effluent treatment method depends on the organic content present in the effluent and its discharge conditions (rose et al, 2020). vegetable oil effluent entering the sewer system consists mainly of fats, oils, greases (fog), sulphate and phosphates resulting in both high inorganic and organic loading for the receiving wastewater works (rose et al, 2020). edible oil industrial wastewaters usually come from the degumming, deacidification, deodorization and neutralization steps. in the neutralization step sodium salts of free fatty acid (soap stocks) are produced whose splitting through the use of sulphuric acid generates highly acidic and oily wastewaters. its characteristics depend largely on the type of oil processed and on the process implemented that are high in chemical oxygen demand(cod), oil and grease, sulphate and phosphate content, resulting in both high inorganic as well as organic loading of the relevant wastewater treatment (rose et al, 2020). moreover, vegetable oil refinery wastewater is known to contain chemical compounds like phenol, heavy metals from bleaching earth, catalysts used in the hydrogenation process, oxidizable substances and fats and oils. because of quantity and characteristic variations and complexity, wastewater treatment to meet the desired effluent standards is complicated, and the choice of methods of wastewater treatment depends on many local conditions and therefore, cannot be standardized (rose et al, 2020). different methods of treating edible oil mill wastewater have been reported, such methods include aerobic treatment, anaerobic digestion, reverse osmosis, electro dialysis, ultrafiltration and composting (ahmaruzzaman, 2008) however, a cost effective and environmentally friendly method for the treatment of oil processing wastewater is not yet found. building an effective treatment technology for oil processing wastewaters is not economically feasible due to the cost implications. recently, a zeolite based treatment method was developed for the treatment of olive mill wastewater which seemed to be a promising technology in the treatment of oil processing wastewaters (ahmaruzzaman, 2008). this study adopts a similar zeolite based method in the treatment of soya bean oil mill wastewater known as soap stock. adsorption has many advantages over the other processes of edible oil waste water because it has the characteristics of convenience, easy operation and simplicity in design. it has a wide application for removal of different pollutants. other important advantages of this process include: low operation cost, high flexibility, simple design and operation, easy automation, lack of sensitivity to toxic pollutants and the capability of operation at very low concentration, environmentally friendly, less investment in terms of initial cost (anagnostopoulos et al, 2013) the most important criteria in adsorption processes is to find a low cost adsorbent that is widely available, having high adsorption capacity, possess rapid rate of removal and having low adverse effect on the treated https://doi.org/10.54536/ajcp.v1i1.385 https://journals.e-palli.com/home/index.php/ajcp http://ishwahlizer@gmail.com pa ge 12 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 11-17, 2022 water. many adsorbents have been investigated reported for the treatment of edible oil wastewater but zeolite has been proved to be more effective (anagnostopoulos et al, 2012) literature review edible vegetable oils are derived from the seed of such plants as soya bean, corn, palms, groundnut, olive tree, rapeseed/canola, linseed, cottonseed, castor, rice bran and sunflower. the oils are fatty acid esters of glycerol, commonly called triglycerides. the production of edible vegetable oils can be separated into two phases: extraction and refining (anagnostopoulos et al, 2012) .the oils are extracted from the seeds by mechanical or hydraulic pressing or solvent extraction. the crude oil that results from the extraction process must be refined to make the oil acceptable for edible purposes. there fining process removes components such as free (unesterified) fatty acids, phosphatides, unsaponifiable components such as sterols and tocopherols, carotenoids, chlorophyll (color), and metals (anari al, 2010) some by-products of low commercial value are obtained from vegetable oil refining processes. important amounts of by-products such as soap stocks (ss), deodorizer fatty acid distillates (fad) and acid oil (ao) are produced from the oil refining processes. these by-products are harmful to the environment if they cannot be used for any beneficial or industrial activity (anari al, 2010). soap stock emerges from the refining process when oil is treated with a dilute alkali solution separating the ffas as soaps. this wet lipid mixture is separated from the crude oil by centrifugation. it is generated at a rate of about 6 % of the input of oil entering the refining operation and its cost represents 1/10 of the refined oil cost. soap stock is quite acidic, with ph values between3.505.50. this residual is also referred to as residual oleins (auta et al, 2013). neutralization process result in a byproduct known as stock soap consist of the sodium soaps of free fatty acids (ffas) present within the oil. soap stocks always require subsequent treatment. the traditional method of treating soap stock is known as soap splitting. the soaps tock is split into fatty acids and water by acidification with strong acids usually h2s04. this is often considered an attractive solution because it is relatively simple (auta & bansal et al, 2009). soap stock is the least valuable by-product from oil processing, and it is generated at a rate of about 6% of the volume of crude soya bean oil refined, amounting to as much as 0.8 million mt in the united states annually. typically, soya bean oil mill wastewater is composed of the water from the tissue of the fruit, the water used for the various stages of oil production, soya pulp, mucilage, pectin, and oil, among other things, suspended in a relatively stable emulsion. moreover, it contains toxic organic and inorganic compounds. in general, soya bean oil mill wastewater is characterized by an intensive violet dark brown to black color, strong specific soya oil smell, high degree of organic pollution (chemical oxygen demand (cod) of 220 g/l and biochemical oxygen demand (bod) of 110 g/l), ph of 4.5-5.6, 45 g/l of organic compounds in total, high electrical conductivity (ec), high content of polyphenols (24 g/l), reduced sugars (60% of the dry weight), and high solid matter content (bhatnagar & borrega et al, 2013). soya bean oil mill is an acidic effluent with a high nutrient content that can be used to fertilize soil; however, it is very rich in toxic phenolic compounds. in recent years, there has been increased attention directed toward finding the best methods to treat vegetable oil wastewater and toward recycling both the organic matter and nutritive elements in the crop production system. some vegetable oil mill wastewater characteristics are favorable for agriculture, because this effluent is rich in water, organic matter, n, p, k, and ca; however, other characteristics are unfavorable for agriculture, including the presence of phenolic compounds (carro, l et al, 2010) the atomic structures of zeolites are based on threedimensional frameworks of silica and alumina tetrahedral that is, silicon or aluminum ions surrounded by four oxygen ions in a tetrahedral configuration. each oxygen atom is bonded to two adjacent silicon or aluminum ions, linking them together, clusters of tetrahedral form boxlike polyhedral units that are further linked to build up the entire framework. in different zeolites the polyhedral units may be equidimensional, sheetlike, or chainlike. the aluminosilicate framework of a zeolite has a negative charge, which is balanced by the cations housed in the cagelike cavities (chakrabarti, s, et al, 2008). zeolites have much more open, less dense structures than other silicates; between 20 and 50 percent of the volume of a zeolite structure is voids. .zeolites are safe, naturally occurring crystalline aluminosilicate that have a threedimensional structure; aluminum, silicon and oxygen which are arranged in a regular structure of [sio4]and [alo4]-tetrahedral units that form a framework with small pores (also called tunnels, channels or cavities) of about 0.1 2 nm diameter running through the material. in these small channels, solid, liquid and gaseous substances can be trapped. high ion exchange capacity, the molecular sieve properties and the relatively high surface area make zeolite a promising adsorbent media for treating effluent with different suspended solids (dang et al, 2009) zeolites have wide application as gas and odor filter, as a part of animal feed, and as ammonia removers from different wastewaters. the metallic ions sorbent behavior of natural zeolite has been also studied by several researches, and it has been recognized as a promising sorbent for heavy metals (galá, j et al, 2013). another characteristics of zeolite is their large surface area (20-50 m2/g by natural species, however above 1000 m2/g by synthetic ones). both physisorption and chemisorption bounding may occur within the zeolite voids during the removal of pollutants. zeolites, nowa-days can be used in almost all pollutants removal processes such as atmospheric, municipal or industrial wastewaters treatment and purification (hansen, c.l et https://journals.e-palli.com/home/index.php/ajcp pa ge 13 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 11-17, 2022 al, 2007) heavy metals are also present in the soya bean oil mill wastewater. heavy metals are elements having atomic weights between 63.5 and 200.6, and a specific gravity greater than 5.0. most of the heavy metals are dangerous to health or to the environment. heavy metals in industrial wastewater include lead, chromium, mercury, uranium, selenium, zinc, arsenic, cadmium, silver, gold, and nickel (ismail, b et al, 2013). the main threats to human health from heavy metals are associated with exposure to lead, cadmium, mercury and arsenic. these metals have been extensively studied and their effects on human health regularly reviewed by international bodies such as the world health organization and food and agricultural organization. acute heavy metal intoxications may damage central nervous function, the cardiovascular and gastrointestinal (gi) systems, lungs, kidneys, liver, endocrine glands, and bones. non-biodegradable contaminants pose a serious health and environmental hazard and removal of these wastes cannot be achieved using secondary methods (jambulingam et al, 2007) hence, tertiary/advanced wastewater treatment methods such as ion exchange, precipitation, membrane separation, electrolysis and adsorption can be used to remove these recalcitrant wastes. so far, a number of efficient methods have been developed for heavy metal removal. some researchers has reviewed various methods for the removal of heavy metals such as chemical precipitation, ion exchange, reverse osmosis, electrodialysis, ultrafiltration, nanofiltration, coagulation, flocculation and floatation. generally, these processes are efficient in removing the bulk of metals from solution at high or moderate concentrations. however, chemical processes produce a large amount of metallic sludge, making metal recovery difficult (jayarajan, m et al, 2011 & johar, n et al, 2012). for these reasons, adsorption technology has gained a wider application due to its inherent low cost, simplicity, versatility and robustness. adsorption is commonly used technique for the removal of metal ions from various industrial effluents (khan, n.a et al, 2004 & diez, v et al, 2012) materials and methods study area this research work was conducted in makurdi town, the headquarters of benue state. makurdi town is the headquarters of makurdi local government area and capital of benue state. the town is located between latitude 7°38’n 7°50’n, and longitude 8°24’e and 8°38’e and 104 meters elevation. it is situated in the benue valley in the north central region of nigeria. it is traversed by the second largest river in the country, the river benue. collection and preservation of soap stock exactly 5 liters of fresh soap stock was collected (using plastic container) at seraph oil mills, nigeria limited, which is located at km 7 gboko road makurdi, benue state. the soap stock was then transported immediately to chemistry laboratory at benue state university and was stored at room temperature prior to analysis. sample digestion the digestion method described by anwar e tal (2014) was adopted with slight modification. 50ml of the sample was measured and transferred into beaker containing 10ml of concentrated hno3. the sample was boiled slowly on a hot plate to 20ml. another 5ml of conc. hno3 was t hen added and heated until digestion was completed. the sample was filtered and transferred into 100ml volumetric flask and made to mark with distilled water. the digests was used for flame photometry and heavy metals analysis of the wastewater (anwar, a et al, 2014) physical precipitation of soap stock physical separation was carried out using glass cylinders with diameter 10 cm and length 20 cm. after 5 h of gravity separation, two kinds of suspended solids were observed: a floating part and a precipitated part. the floating part was decanted and the precipitate filtered using filter paper and a less turbid sample was obtained (anwar, a et al, 2014). soya bean oil mill wastewater treatment using successive steps the soya bean oil mill wastewater (soap stock) was passed through three designed treatment steps, following physical separation. each treatment consists of successive column containing different solid-state materials. the columns were made of transparent glass material with an internal diameter of 2 cm and a length of 20 cm. the columns were sealed from the bottom using two pieces of gauze fabric firmly held by strings and tape. the columns were packed with solid state materials to 15 cm height. the packing process was done in 5 cm increments to avoid segregation of particles. the columns were mounted vertically on a wooden holder in the laboratory. each treatment system was replicated three times. the contact times between the soya bean oil mill wastewater and solid-state materials were 1 h for fine sand, 2 h for zeolite and 4h for the fine zeolite/sand composite. the physicochemical parameters used to evaluate the treatment efficiency for each method were ph, phosphate, turbidity, sulphate, nitrate and metal ions concentration (anwar, a et al, 2014). characterization of zeolite y zeolite y was synthesized and characterized using pxrd, eds and ftir. the pxrd used was a bruker d8 advance diffractometer operating with a copper x-ray tube, a monochromator with a linx eye detector. the pxrd data were collected using cu kα1 (1.5406a) radiation, over the 2θ range between 5 60° using a step size of 0.022° for 43 minutes. the eds spectrum was produced on an edax pheonix, edx with a carl zeiss 1530 vp spectrometer. the samples were sprinkled onto https://journals.e-palli.com/home/index.php/ajcp pa ge 14 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 11-17, 2022 12mm aluminium stubs using “carbon sticky tabs”. these were then gold coated using an emitech sc 7640 gold/ palladium sputter coater to reduce the static charges during the analysis. the perkinelmer paragon 1000 ftir spectrophotometer was used to collect ftir data for the sample. the sample was prepared by making discs of a small amount of the sample (t 3 mg) in kbr and measurements were carried out over ir region of 1200-400cm-1 for the zeolite. a background spectrum was measured before the sample to compensate for atmospheric conditions around the ftir instrument determination of heavy metals in soap stock using atomic absorption spectrometry the concentrations of heavy metals in the wastewater (soap stock) were analyzed before and after treatment to assess the decrease in concentrations of the pollutants using atomic absorption spectrometry zinc approximately11.24g of zinc oxide was dissolved in deionised water and diluted to 1l in a volumetric flask. 1 ppm, 2 ppm, 3ppm, 4 ppm and 5ppm solutions were made from the stock solution. the aas was the calibrated at a wavelength of 213.9 nm under lamp current of 5 ma using fuel acetylene (diez, v. et al, 2012). lead about 1.5g of lead nitrate pb(no3)2was dissolved in deionized water and diluted to 1l in a volumetric flask . 1 ppm, 2 ppm, 3 ppm 4 ppm and 5 ppm were prepared from the stock. the aas was calibrated at 217.0 nm wavelength, spectra bond of 1 nm and current of 5 ma using acetylene and air as fuel (diez, v. et al, 2012). nickel about 4.0g of ni (no3)2.6h2o was dissolved in deionised water and diluted to 1l in a volumetric.1 ppm, 2 ppm, 3 ppm 4 ppm and 5ppm were prepared from the stock solution. the aas was calibrated at 213.9 nm wavelength, spectra bond of 1 nm and current of 5 ma using acetylene and air as fuel (diez, v. et al, 2012). cadmium exactly 2. g of cadmium nitrate was dissolved in 250 ml of de-ionized water and diluted to 1 l in a volumetric flask. 1 ppm, 2 ppm, 3 ppm, 4 ppm and 5 ppm were prepared from the stock solution. the aas was calibrated table 1: characterization data of fine sand using x-ray fluorescence spectrometer compounds/elements % weight sio2 92.49 al2o3 7.44 fe2o3 nd cao 0.06 mgo 0.09 k2o nd key nd: not detected table 2: elemental composition of the synthesized zeolite y (eds) elements % weight 0 48.7±0.07 na 9.2±0.04 al 11.5±0.0 si 30.7±0.1 si/al 2.7 figure 1: pxrd pattern for zeolite y (experimental pattern) matched with the reference pattern 01-070-4285 (vertical tick marks), anhydrous formula (na54.91al56si136o384). at a wavelength of 228.8 nm at a spectra bond of 0.4 nm and lamp current of 3.5 ma using fuel acetylene. copper 1g of copper was dissolved in 25 ml of hcl. few drops of nitric acid was added and diluted to 1 l in a volumetric flask with de-ionzed water. 1 ppm, 2 ppm, 3 ppm, 4 ppm and 5 ppm were prepared from the stock solution. the aas was calibrated at a wavelength of 309.3 nm and spectra band pass of 0.4 nm, lamp current of 10 ma using fuel acetylene and nitrous oxide. https://journals.e-palli.com/home/index.php/ajcp pa ge 15 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 11-17, 2022 figure 2: ftir spectrum for the synthesised zeolite y table 3: heavy metals concentration in the soap stock soya bean oil mill before and after treatment heavy metals (mg/l) untreated wastewater physical pre-cipitation fine sand treatment zeolite treatment zeolite/sand treatment fao (2010) epa (2011) pb 0.7555±0.00 0.1475±0.00 0.103±0.00 0.0760±0.00 0.0260±0.00 0.5 0.01 cu 0.2850±0.00 0.0350±0.00 0.0260±0.00 0.0050±0.00 nd 0.1 0.02 zn 1.3505±0.00 1.1505±0.00 1.0725±0.00 0.5975±0.00 0.5135±0.00 1.0 0.5 ni 0.0465±0.00 0.0355±0.00 0.0580±0.00 0.0400±0.00 0.0390±0.00 0.01 0.02 cd 0.1455±0.00 0.0815±0.00 0.0195±0.00 0.0620±0.00 0.0270±0.00 0.01 0.02 key: nd= not detected results and discussion chemical composition of fine sand the chemical composition of the fine sand used in this present study was determined by x-ray fluorescence spectrometer (arl 9900 oasis model) as shown in (table 1). silicon dioxide was found to be the predominant oxide, 92.49%. other oxides present were al2o37.44%, cao 0.06% and mgo 0.09%. the silicon to aluminum ratio of the fine sand used was 12.40. this characterization data obtained is in line with earlier investigation reported (alluri et al, 2007). pxrd the pxrd data compared against the icdd database for the theoretical phases showed good agreement that the targeted zeolite y was formed as clearly seen from the matched patterns (fig. 1).the pxrd patterns for the zeolite y was a single crystalline phase with high degree of crystallinity. synthesis of zeolite y sem-eds the elemental composition of zeolite y was determined using energy dispersive spectroscopy (edax pheonix model) as shown in (table 2). the result revealed that zeolite y consist of 48 7% oxygen, 9.2% sodium, 11.2% aluminum and 30.7% silicon. the si/al ratio (2.7%) calculated was found to be in good agreement with the data obtained from pxrd (2.4%) as calculated from the molecular formula of the zeolite y (na54.91al56si136o384). ftir a broad band was observed in the region 1019 cm-1 which was attributed to the asymmetric vibrations of si-o bridging and si-o non-bridging bonds. the band in the range 717 cm-1 is due to the symmetric stretching of internal vibrations while the internal vibrations due to the bending of the t-o tetrahedra occurred between 455 cm1. vibrations of the double six rings (d6r) connecting the sodalite cages occurred between 579 cm-1 while that around 626 cm-1 is assigned to the symmetric stretching of external t-o linkages in the zeolite. the band at 1150 cm-1 was attributed to the asymmetric stretching of external t-o linkages in the zeolite respectively. adsorption of heavy metals the results presented in table 3 showed a decrease in the concentration of heavy metals using the zeolite-based technique. lead for the raw wastewater sample 0.7555±0.00mg/l was obtained, 0.1475±0.00mg/l for physical precipitation, 0.103±0.00mg/l for sand filter, 0.0760±0.00mg/l for zeolite filter and 0.026±0.00mg/l for zeolite/sand filter respectively. the decrease in the values could be attributed to the ion exchange and absorption affinity of the zeolite material used. the values obtained were within that of maximum allowable concentration (mac) limits by fao (2011) and epa (2016) for irrigation water with values ranging from 0.5 to 0.01mg/l (babel, s al, 2003) https://journals.e-palli.com/home/index.php/ajcp pa ge 16 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 11-17, 2022 copper for the raw wastewater sample 0.285±0.000mg/l was obtained, 0.035±0.00mg/l for physical precipitation, 0.026±0.00mg/l for sand filter, 0.005mg/l for zeolite filter and was less than the detectable limit using the zeolite/sand filter. the decrease in the values could be attributed to the ion exchange and adsorption properties of zeolite. the values obtained were within the maximum allowable concentration limits by fao (2011) for irrigation water (babel, s al, 2003) nickel about 1.04465±0.00mg/l was obtained for the raw soya bean oil mill wastewater sample, 0.0355±0.00mg/l for physical precipitation, 0.058±0.00mg/l for sand filter, 0.040±0.00mg/l for zeolite filter and 0.039±0.00mg/l for zeolite/sand filter. the result shows a gradual decrease in the pollutant load from the raw wastewater to the final zeolite/sand filter treatment, reducing the concentration within the maximum allowable limit of 0.2mg/l for agricultural water. similar results were also reported for oil refinery wastewater treatment using the zeolite and carbonaceous adsorbents (kwon j.s et al & muthukumaran et al, 2010) zinc about 1.3505±0.00mg/l was obtained for raw soya bean oil mill wastewater sample, 1.1505±0.00mg/l for physical precipitation, 1.0725±0.00mg/l for sand filter, 0.5975±0.00mg/l for zeolite and 0.5135±0.00mg/l for zeolite/sand filter. these values are within the maximum allowable concentration limit of 1.002.00mg/l for irrigation water. similar works were also reported by some researchers on adsorption of heavy metals from water and wastewater using low cost adsorbents from agricultural by-products (pitakpoolsil et al & varga et al 2013) cadmium for the raw wastewater sample, 0.1455±0.000mg/l was obtained 0.0815±0.00mg/l for physical precipitation, 0.0195±0.00mg/l for sand filter, 0.062±0.00mg/l for zeolite filter and 0.027±0.00mg/l for zeolite/sand filter. the value obtained at the zeolite/sand composite filter is in accordance with the recommended values of 0.01-0.002mg/l by fao and epa (2011). similar results were also reported on heavy metal removal of edible oil content in wastewater by advanced oxidative process (aslan, s. et al 2001 & chipasa et al 2009). conclusion the treatment of soap stock was using successive steps that are raw wastewater, physical precipitation, sand treatment, zeolite treatment and zeolite/sand composite using solid state materials and testing the treatment efficiency using heavy metals analysis proved to be effective for edible oil effluent. this study found that the filtering abilities of zeolite/sand composite works better for soya bean oil mill wastewater treatment. the simple zeolite-based method can be used for the soya bean oil mill wastewater treatment as an environmentally friendly and cost effective method. the treatment efficiency on the removal of heavy metals such as zn, cu, pb, ni and cd using aas gave a good heavy metal reduction from the wastewater. references ahmaruzzaman, m. 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(2013). comparative study of sorption kinetics and equilibrium of chromium (vi) on charcoals prepared from different low-cost materials. journal of micro chemical science, 107, 25–30 11(1), 70-81. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 21 american journal of chemistry and pharmacy (ajcp) synthesis and characterization of alpinia calcarata loaded nanoparticles to control hyperglycemia b.s. wanniarachchi1, p.g.u. chathuranga1, p.h.u.w. de silva1, b.m. jayawardena1* volume 2 issue 1, year 2023 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v2i1.1293 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: february 12, 2023 accepted: march 02, 2023 published: march 10, 2023 bovine serum albumin (bsa) nanoparticles loaded with the bioactive compounds of a. calcarata, which is known to exert its antidiabetic activity through the inhibition of pancreatic enzymes, are a good form of an antidiabetic nutraceutical as they have reduced side effects, protection of active compounds from environmental agents, specific delivery to target sites and prolonged shelf-life. the objective of the present study was to synthesize and characterize a. calcarata loaded nanoparticles (alnp) to be used as a powder form nutraceutical with higher antidiabetic activity. in this study an aqueous a. calcarata extract (4.00 ml) was added to bsa (20 mg/ml, 4.00 ml, ph 9) in the presence of citric acid as the cross-linking agent. the alnp gave an ic50 value of 147 µg/ml, a glucose (5 mm) uptake percentage of 73.09% at a 0.5 mg/ml concentration, a solubility value of 64%, a. calcarata loading percentage of 6.66% and a. calcarata entrapment efficiency of 87.71%. they had a spherical morphology and uniform size with a particle size of 1030.70 nm, pdi of 0.199 and a zeta potential of 2.57 mv. the uv-visible absorbance spectra and ft-ir spectra showed that citric acid had caused conformational changes in the protein structure of bsa and that the active compounds were successfully loaded into the synthesized nanoparticles which interacted with the protein matrix via covalent bonds. therefore, it can be concluded that the synthesized nanoparticles have an antidiabetic effect and the antidiabetic activity of bioactive compounds of the aqueous a. calcarata extract become enhanced when loaded onto the nanocarriers. keywords a.calcarata, antidiabetic, bsa nanoparticles, citric acid, nutraceutical 1 department of chemistry, university of kelaniya, sri lanka * corresponding author’s e-mail: bimali@kln.ac.lk introduction diabetes mellitus, the most common non-communicable disease in the world is coined from the greek word diabetes, meaning to pass through a large discharge of urine and the latin word mellitus, meaning sweet. it is a general term used to describe a variety of metabolic disturbances, the primary cause of which is chronic hyperglycemia. (sapra & bhandari, 2022; kerner & brückel, 2014) type 1 diabetes mellitus (t1dm), type 2 diabetes mellitus (t2dm), ‘other’ and gestational diabetes mellitus (gdm) are the classifications of diabetes based on etiology and pathology. (guthrie & guthrie, 2004) according to the world health organization (who), in 2019, diabetes was the ninth leading cause of death and is also responsible for the largest rise in male deaths with an 80% increase since 2000. it is the no.1 cause of kidney failure, adult blindness and lower-limb amputations. also in 2019, among the us population, 283,000 children and adolescents younger than 20 years suffered from diagnosed diabetes. the 2022 national diabetes statistics report states that currently about 382 million (8.3%) people are affected by diabetes and this number is projected to increase to 552 million (53%) by 2035. (kharroubi & darwish, 2015) oral antidiabetic drugs that are used in the treatment of diabetes have serious side effects such as hypoglycemia, weight gain, anemia and congestive heart failure. also, due to their high cost and these side effects, scientists are searching for more effective and safer antidiabetic drugs. therefore, recently attention has been focused on natural products including food plants, as possible sources of more potent and safer antidiabetic therapy. (kazeem & davies, 2016) alpinia calcarata roscoe which belongs to the family zingiberaceae is a rhizomatous plant that is widely used as a medicinal source in sri lanka. rhizomes of this plant are known as heen araththa or katu kikiriya in sinhala and snap ginger in english. the mature rhizomes are dense and branched with a light to dark brown colour and they are the most important part of this plant, as they are a major part of indigenous medicinal formulation for the treatment of blood impurities, indigestion, throat inflammation, voice improvement and to marinate youthful vigor. the decoction of alpinia calcarata rhizome is widely used to treat respiratory ailments, cough, bronchitis, asthma and arthritis. the ethanolic as well as aqueous extracts of alpinia calcarata rhizomes show antibacterial, antifungal, anthelminthic, anti-inflammatory, antioxidant, anticancer, antinociceptive, gastroprotective, aphrodisiac and antidiabetic effects. this herb is also used as a traditional medicine for stomachache, fever and rheumatism. (rahman & islam, 2015) analysis of a.calcarata rosc. grown in sri lanka has revealed the presence of quercetin, protocatechuic acid, 1,8-cineole, β-pinene, vanillic acid, 4-o-methyl-syringic acid and methyl cinnamate as well as several terpenes and diterpenes as constituents. novel bis-labdanic diterpenoids such as calcaratarin d and calcaratarin https://doi.org/10.54536/ajcp.v2i1.1293 https://journals.e-palli.com/home/index.php/ajcp pa ge 22 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 21-31, 2023 e have been isolated and identified from its rhizomes, which are known to be a pair of stereoisomers according to spectral evidence. qualitative phytochemical analysis, of the hot water extract and hot ethanol extract, also revealed the presence of alkaloids, flavonoids, steroid glycosides, polyphenols and tannins in its rhizomes. the relative percentages of water extractable matter and ethanol extractable matter are 18.6-20.5 and 22.6-24.8, respectively. (rahman & islam, 2015) the essential oils of rhizomes, roots and leaves when analyzed using capillary gc and gc/ms showed the presence of 18 compounds. the major compound in the leaf and rhizome oils was 1,8-cineole and in the root oil it was α-fenchyl acetate. a. calcarata, having such a huge range of phytochemical diversity, has been ensured non-toxic and safe in animal studies. (arambewela et al., 2005) scientific studies carried out with the oral administration of the hot ethanolic and hot water extracts of the rhizomes of a. calcarata to normoglycemic and streptozotocin-induced diabetic rats have shown hypoglycemic and antihyperglycemic effects, respectively. both hot water and hot ethanol extracts significantly reduce the blood glucose level, body weight gain, plasma triglyceride and total cholesterol levels as well as improve the glucose tolerance. alpinia calcarata extract exerts its potent antidiabetic activity through the inhibition of intestinal glucose absorption by inhibiting the activity of pancreatic α-amylase and α-glucosidase enzymes. however, the hypoglycemic effect of hot ethanol extract is usually higher than that of hot water extract. phenolic compounds such as flavonoids, polyphenols and tannins that are present in its rhizomes are known to be responsible for its antidiabetic activity. (rahman & islam, 2015; wasana et al., 2021) extraction procedures are carried out to separate the medicinally active portions of plant or animal tissues from the inactive or inert components by using selective solvents. (handa et al., 2008) some of the extraction techniques that can be used to extract the active compounds of the bark of a.calcarata are microwave digestion, pressurized water extraction, solvent extraction, decoction water extraction and infusion water extraction. out of these techniques, the pressurized water extraction has been proven to result in an aqueous extract that is more active than the other. (wariyapperuma et al., 2018) unlike the traditional extraction methods that require large volumes of non-environmental friendly organic solvents and is time consuming with low extraction efficiency, pressurized water extraction, which is a green solvent extraction method, is environmentally friendly as it uses minimum or no organic solvent, cheap with shorter extraction time and high extraction efficiency. the analytes extracted using this method are also safe for human consumption, testing and processing as organic solvents are not involved. as water is easily available, non-toxic and can be recycled or disposed with minimal environmental problems, pressurized water extraction has steadily become an efficient and low-cost method of extraction for less-polar organic components. (teo et al., 2010; jayawardena & smith, 2010) “pressurized hot water” is used to denote the region of condensed phase of water from 100 ℃ to 374 ℃, which is the critical point of water. the density of water remains almost constant over this temperature range so that the pressure effect on the properties of water is minimal. during extraction, moderate pressures such as 15 bar at 200 ℃ and 85 bar at 300 ℃ are needed to keep a condensed phase of water. under these conditions the dielectric constant of water decreases, and it starts behaving as an organic solvent such as ethanol and methanol, which can then extract organic and non-polar compounds from numerous kinds of matrices. however, if water was used at room temperature and atmospheric pressure, it will not be suitable for the extraction of organic and non-polar compounds, because of its high polarity and high dielectric constant due to its hydrogen bonded structure. (teo et al., 2010) with the advance of nanotechnology, scientists have developed nanoencapsulation techniques for the targeted release and protection of pharmaceuticals and food bioactive components, so that they can be safely added into formulations and result in maximum bioavailability. conventional microencapsulation technologies include physical and chemical processes such as spray drying, freeze drying, extrusion, coacervation, liposomes and conventional emulsions. unlike these methods, nanocarriers could provide more bioavailability through increased surface-to-volume ratios and therefore higher muco-adhesive possibility within the small intestine and higher feasibility of interacting with enzymes and metabolic factors, and also these tiny particles could easily pass through the cell membranes and penetrate into the target cells and release their encapsulated material. (assadpour & mahdi jafari, 2019) in addition, these nanocarriers can also improve the solubility of hydrophobic compounds, such as the active compounds of cinnamon, protect the chemical structure of nutraceuticals from environmental agents such as light, temperature, ph, radicals and oxygen, allow specific delivery to target sites, allow a controlled release of the encapsulated compound, result in reduction of side effects and prolonged shelf-life, and also, they have minimum influence on the appearance of final food products. (assadpour & mahdi jafari, 2019; paolino et al., 2021) a nanocarrier is a nano-sized system and is scientifically defined in the pharmaceutical area as a particle with a size of a few nm to just below 1000 nm. (assadpour & mahdi jafari, 2019) nanoparticles fall into the category of colloidal drug delivery system as they behave as a whole unit with respect to its properties and transport mechanism. several types of nanoparticle systems have been identified such as polymeric nanoparticles, polymeric micelles, solid nanoparticles, lipid-based nanoparticles for example, solid lipid nanoparticles (sln), nanostructured lipid carriers (nlc) and lipid drug conjugate (ldc), liposomes, inorganic nanoparticles, dendrimers, magnetic https://journals.e-palli.com/home/index.php/ajcp pa ge 23 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 21-31, 2023 nanoparticles, nanocrystals and nanotubes. in recent years, biopolymer-based nanoparticles including protein nanoparticles are actively used in pharmaceuticals due to their low toxicity and biodegradability. they also have better biocompatibilities and the possibility for surface modification. these nanocarriers can be synthesized using proteins like albumin, gelatin, whey protein, gliadin, legumin, elastin, zein, soy protein and milk protein. these protein nanoparticles are an ideal material for delivery of bioactive compounds as their amphiphilic nature allows them to interact with both the bioactive compounds and the solvent. (verma et al., 2018) albumin is an attractive macromolecule carrier due to its high binding capacity and is obtained from a variety of sources such as bsa, egg white (ovalbumin) and human serum albumin (hsa). it has a molecular weight of 66.5 kda and is a water-soluble protein which is therefore used in maintaining the osmotic pressure, binding and transport of nutrients to the cells. albumin also dissolves in ethanol (40% w/v), it is stable in the ph range of 4-9 and can be heated at 60 ℃ for 10 hours without showing any kind of denaturation effects. it is widely used in the preparation of nanospheres and nanocapsules as these albumin nanocarriers are non-toxic, biocompatible, biodegradable, easy to prepare, non-immunogenic, have well defined sizes and also carry some reactive groups such as thiols, amines and carboxyl. these groups can work as surface modifiers during the cross-linking process. also, the bioactive compounds encapsulating albumin nanoparticles can be easily digested by the enzyme protease, releasing the entrapped material. achieving the required particle size of albumin nanoparticles is easy and reproducible. (verma et al., 2018; amighi et al., 2020) albumin nanoparticles can be prepared by various techniques such as desolvation, emulsification, nanoparticle albumin-bound (nab) technology and nano-spray drying. (amighi, et al., 2020). among these methods, the most commonly used methods are desolvation and emulsification. (niknejad & mahmoudzadeh, 2015) under the desolvation method, a desolvation agent which is an organic solvent such as ethanol or acetone or even a natural salt is added into the aqueous solution of albumin. by adding desolvation agents, albumin starts to change its tertiary structure slowly. then at a certain level, protein clumps (aggregates) are made and finally these unstable aggregates are hardened by the cross-linking agent. (niknejad & mahmoudzadeh, 2015) in order to separate the particles, the turbidity of the system should be increased. in the emulsification method, an aqueous phase of albumin is prepared with distilled water, which is added to an organic phase plant oil such as cotton seed oil under mechanical homogenizer until an oil-water emulsion is prepared. this emulsion is then added into preheated oil over 120 ∘c drop by drop. this results in evaporation of water and irreversible destruction of albumin which leads to the formation of nanoparticles. the resulting particles are then suspended in an ice-cold bath. (verma et al., 2018) bsa nanoparticles can be synthesized using the desolvation method, where ethanol can be used as the desolvation agent to obtain nanoparticles with narrower size distribution and controllable particle size. crosslinking is an essential step in nanoparticle synthesis, as it influences bio-decomposability of the loaded bioactive compounds and their release from the nanocarrier system. a cross-linker is added to stabilize the newly formed nanoparticles. the addition of a cross-linking agent changes the surface charge of nanoparticles. also, it modifies the colloidal stability and electrostatic potential of the bsa nanoparticles in solution. recently, natural origin cross-linkers such as citric acid which guarantees human health have started to become very popular. during the cross-linking process of citric acid, more than one carboxyl group reacts with proteins. also, citric acid starts its cross-linking process in alkaline ph values. at higher ph values, amino groups are deprotonated, so the free amino groups attack the partially positively charged carbonyl carbons of citric acid resulting in a nucleophilic substitution. (amighi et al., 2020; aniesrani delfiya et al., 2016)) the objective of the present study was to synthesize and characterize alnp, so that it could be used as an antidiabetic nutraceutical to treat hyperglycemia. materials and methods raw materials alpinia calcarata rhizomes chemicals bsa, ethanol, citric acid, sodium hydroxide, alpha amylase, starch, dinitrosallicylic acid reagent (dns), sodium hydrogen phosphate, sodium dihydrogen phosphate, dimethyl sulfoxide (dmso), anhydrous kbr, baker’s yeast and anhydrous dextrose. instruments electric grinder (sumeet, india), analytical balance (kern alj 120-4 germany), pressure cooker (prestige india), centrifuge machine, magnetic stirrer, ph meter, thermometer, electric oven, microplate reader (spectra max m5, molecular devices, ca, usa), malvern zetasizer nano zs apparatus (malvern instruments ltd., malvern, uk), field emission scanning electron microscope (hitachi su6600 fe-sem), uv-vis spectrophotometer (agilent technologies, germany. cary 60), ft-ir (fourier transform infrared) spectrometer (perkinelmer, l 1600300 spectrum two lita, liantrisant, uk). preparation of alpinia calcarata powder dried alpinia calcarata rhizomes grown in sri lanka were obtained and ground into fine powder using an electric grinder and refrigerated at -10 ℃ until further use. preparation of aqueous alpinia calcarata extract a. calcarata powder (10.00 g, < 0.5 mm) was digested using https://journals.e-palli.com/home/index.php/ajcp pa ge 24 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 21-31, 2023 distilled water (100.00 ml) under a pressure of 15 psi (121 ℃) for 15 minutes in medium pressure. the extract was filtered using a muslin cloth and the filtrate was centrifuged at 3000 rpm for 15 minutes. the supernatant obtained was then stored at -20 ℃ until further analysis. preparation of alnp for the synthesis of alnp, an aqueous extract of a. calcarata (4.00 ml) was mixed with ethanol (16.00 ml) and added at a rate of 1.00 ml/min to a bsa solution (20 mg/ml, 4.00 ml, ph 9) while the solution was constantly stirring at 600 rpm at 4 ℃. then citric acid (8% w/v, 230 µl) was added as the cross-linking agent, and the mixture was allowed to stir for another 2-3 hours, at 4 ℃. the mixture was then stored at 4 ℃ for 24 hours, for the formation of stable nanoparticles, by facilitating the cross-linking process. the nanoparticle bearing solution was centrifuged at 3000 rpm for 30 minutes and the pellet containing the nanoparticles was purified by five cycles of centrifugation (15000 g, 10 min) using milli-q water. the obtained particles were then dried at 50 ℃ until a constant weight was observed and then stored at 4 ℃ until further analysis. determining the yield of alnp weight of the obtained product was measured. total concentration of all compounds in a.calcarata extract was determined by evaporating a.calcarata extract (1.00 ml) on an evaporating glass and weighing the remaining. using the obtained data, the yield was calculated according to the following equation. yield = (weight of the product (g) /weight of used bsa + weight of a.calcarata extract)×100 determining antidiabetic activity of alnp antidiabetic activity of the aqueous alpinia calcarata extract and alnp were determined by carrying out invitro alpha amylase inhibition assay. alpha amylase inhibition assay the alpha amylase inhibitory activity of aqueous a.calarata extract and alnp were determined by following the method specified in oyedemi, et al., 2017 with slight modifications. a concentration series (1 – 8 mg/ml) of the aqueous a.calarata extract was prepared. each solution (500 µl) was placed in a separate eppendorf tube. alpha amylase enzyme (0.05 mg/ml, 250 µl) in sodium phosphate buffer (0.02 m, ph 6.9) was added to each eppendorf tube. the mixtures were then incubated at room temperature (28 ± 2 ℃) for 15 minutes. starch solution (1%, 250 µl) in sodium phosphate buffer (0.02 m, ph 6.9) was added. the reaction mixtures were incubated at room temperature (28 ± 2 ℃) for 15 minutes. dns reagent (250 µl) was added to each eppendorf tube, and all the tubes were boiled for 5 minutes. absorbance was measured at 540 nm using the microplate reader. this procedure was triplicated. three more separate series were carried out, one where the enzyme was replaced with sodium phosphate buffer (0.02 m, ph 6.9, 250 µl), another where the aqueous a.calcarata extract was replaced with sodium phosphate buffer (0.02 m, ph 6.9, 500 µl) and another series where both the aqueous a.calcarata extract and the enzyme were replaced by sodium phosphate buffer (0.02 m, ph 6.9, 750 µl). each of these series were also performed in triplicates. percentage inhibition of the alpha amylase enzyme was calculated using the following formula. a = absorbance at 540 nm without inhibitor and with enzyme b = absorbance at 540 nm without inhibitor and enzyme c = absorbance at 540 nm with inhibitor and enzyme d = absorbance at 540 nm with inhibitor and without enzyme based on the percentage inhibition values, the ic50 value of the aqueous a.calcarata extract on alpha amylase enzyme was calculated using the graphpad prism 9.2.0 software. concentration series of alnp (0.13 – 1.00 mg/ml) was prepared using dmso as the solvent. the percentage inhibition and ic50 value of alnp on alpha amylase enzyme was calculated using the same procedure that was followed for the aqueous a.calcarata extract. yeast glucose uptake assay glucose uptake by yeast cells in the presence of a. calcarata aqueous extract and alnp were determined by following the method described by cirillo, et al., 1962 with slight modifications. a 1% yeast solution was prepared by dissolving commercial baker’s yeast (0.25 g) in distilled water (0.25 ml) and was left overnight at room temperature. the following day, the solution was centrifuged at 4200 rpm for 5 min repeatedly until a clear supernatant was obtained. 10% (v/v) suspension was prepared in distilled water. glucose solution (25 mm) was prepared by dissolving anhydrous glucose (0.45 g) in distilled water (100.0 ml). then using the prepared glucose solution, 10 mm and 5 mm glucose concentrations were prepared by diluting with distilled water. a concentration series of a. calcarata aqueous extract (0.48 – 7.60 mg/ml) as well as a concentration series of alnp (0.03 – 0.54 mg/ml) were prepared by using distilled water and dmso as solvents, respectively. the extract (1.00 ml) and nanoparticle suspension (1.00 ml) were added separately to glucose solution (5 mm, 10 mm, and 25 mm, 1.00 ml) and incubated at 37 °c for 10 min. then the yeast suspension (100 μl) was added to start the reaction, and the mixtures were vortexed and further incubated at 37 ℃ for 60 min. the tubes were then centrifuged (2500 g, 5 min) and glucose in the supernatant was estimated by adding dns reagent (250 μl) to supernatant (1.00 ml) and the mixtures were boiled for 5 min. absorbance was measured at 540 nm by microplate reader. blank was prepared by adding distilled water (300 μl) instead of the sample solution. https://journals.e-palli.com/home/index.php/ajcp pa ge 25 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 21-31, 2023 the procedure was triplicated. the percentage increase in glucose uptake by yeast cells was calculated using the following formula. characterization of alnp solubility alnp (5.0 mg) was added to distilled water (5.00 ml) and stirred for 30 minutes at 600 rpm at room temperature. after that the mixture was centrifuged at 10 000 rpm for 15 minutes. pellet was dried at 100 ◦c in an oven until a constant weight was observed. the procedure was triplicated. the solubility was calculated according to the following formula. a.calcarata loading and entrapment efficiency a concentration series from the pressured water extract of a.calcarata was prepared. absorbance of the concentration series was measured at 310 nm. a graph of absorbance at 310 nm vs concentration of a.calcarata extract was plotted (attieh, et al., 2015). absorbance of the supernatants which remained after the separation of the product containing pellet, was measured at 310 nm. by comparing the absorbance value of the supernatant with the standard curve, the concentration of remaining a.calcarata compounds in the supernatant was found. the procedure was triplicated. a.calcarata loading and entrapment efficiencies were determined by using following formulae. weight of a.calcarata loaded = weight of a.calcarata added weight of a.calcarata in supernatant determination of particle size and zeta potential the particle size, pdi and zeta potential of the alnp were determined with the malvern zetasizer nano zs apparatus. an aqueous suspension of alnp was diluted 1:100 with ultrapure water and the solution was placed in a disposable polystyrene cuvette and the particle size measurement was obtained. the solution was also placed in a folded capillary zeta cell and the zeta potential measurement was obtained. both procedures were triplicated. morphological observations field emission scanning electron microscope was used to visualize the morphology and shape of the synthesized alnp. the sample was mounted onto the sample stub using carbon tapes and the images were taken after gold sputter coating for 15 seconds. uv-visible absorbance spectra the uv-visible absorption spectra were analyzed using a uv-visible spectrophotometer from 200 to 500 nm within a 1 cm quartz cell. a 0.8 mg/ml synthesized alnp sample was prepared after dissolving it in dmso and its absorbance spectrum was obtained. it was then compared with the absorbance spectra of 1 mg/ml pure bsa, 0.07 mg/ml aqueous a.calcarata extract and 0.8% w/v citric acid. the absorbance spectrum of distilled water was subtracted from all sample spectra. ft-ir spectroscopy the molecular characteristics of the alnp were examined and compared with that of pure bsa, oven dried aqueous a.calcarata extract and pure cross-linking agent, citric acid using an ft-ir spectrometer. each sample was mixed with anhydrous kbr in a 1:10 ratio and ground using a motor and pestle until a fine powder was obtained. a small portion of the powder was placed in the pellet forming mold and pressed under pressure. then the pellet was placed in the ft-ir spectrometer and scanned in the wavenumber range of 750-4000 cm-1. statistical analysis the obtained data were statistically analyzed by one-way analysis of variance (anova) using minitab software package. the results were expressed in the form of mean ± standard deviation of triplicate determinants. the level of significance was taken at 5% confidence interval (p < 0.05). results physical appearance of alnp table 1: yield and morphology of alnp yield (%) morphology 94.11 light brown coloured powder table 2: ic50 values of alpha amylase enzyme sample ic50 value on alpha amylase enzyme (µg/ml) aqueous a.calcarata extract 152.10 (±0.10) a alnp 147.00 (±0.97) b means followed by different letters are significantly different (p < 0.05) antidiabetic activity alpha amylase inhibition assay https://journals.e-palli.com/home/index.php/ajcp pa ge 26 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 21-31, 2023 figure 1 : the graph of percentage inhibition (%) of aqueous a.calcarata extract on alpha amylase enzyme vs. concentration figure 2 : the graph of percentage inhibition (%) of alnp on alpha amylase enzyme vs. concentration table 3 : percentage increase in glucose uptake by yeast cells at different glucose concentrations in the presence of aqueous a.calcarata extract concentration(mg/ml) glucose uptake percentage (%) at different glucose concentrations 5 mm 10 mm 25 mm 7.6 68.66 (±0.02) a 43.57 (±0.26) a 24.20 (±2.02) a 3.8 64.61 (±0.29) b 32.28 (±0.26) b 22.22 (±0.13) ab 1.9 58.45 (±0.34) c 20.68 (±0.91) c 20.20 (±0.39) b 0.95 50.78 (±0.07) d 17.14 (±0.47) d 16.69 (±0.16) c 0.475 45.30 (±0.97) e 15.23 (±0.40) e 14.16 (±0.30) d means followed by different letters are significantly different (p < 0.05) table 4 : percentage increase in glucose uptake by yeast cells at different glucose concentrations in the presence of alnp concentration(mg/ml) glucose uptake percentage (%) at different glucose concentrations 0.54 73.09 (±0.06) a 49.81 (±0.09) a 28.89 (±0.56) a 0.27 71.28 (±0.52) b 38.41 (±0.28) b 26.82 (±0.88) b 0.135 68.97 (±0.80) c 31.66 (±0.40) c 24.82 (±0.76) c 0.0675 64.17 (±0.16) d 25.38 (±0.72) d 20.97 (±0.87) d 0.03375 60.80 (±0.70) e 18.03 (±0.82) e 16.63 (±0.35) e means followed by different letters are significantly different (p < 0.05) yeast glucose uptake assay characterization of alnp table 5 : solubility, a.calcarata loading percentage and a.calcarata entrapment efficiency of alnp solubility in water (%) a.calcarata loading percentage (%) a.calcarata entrapment efficiency (%) 64.00 (±1.00) 6.66 (±0.01) 87.71 (±0.07) table 6 : particle size, pdi and zeta potential of alnp parameter mean ± standard deviation particle size 1030.70 (±75.3) nm pdi 0.199 (± 0.003) zeta potential 2.57 (±0.32) mv https://journals.e-palli.com/home/index.php/ajcp pa ge 27 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 21-31, 2023 figure 3: sem image of alnp figure 4 : uv-visible absorbance spectra of pure bsa, aqueous a.calcarata extract, pure citric acid and alnp figure 5 : ft-ir spectra of pure bsa, aqueous a.calcarata extract, pure citric acid and alnp https://journals.e-palli.com/home/index.php/ajcp pa ge 28 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 21-31, 2023 discussion as the rhizome extract of sri lankan alpinia calcarata has shown potential antidiabetic effects, its pressured water extract was used to synthesize nanoparticles, in order to be used as an antidiabetic nutraceutical. (samarasinghe, et al., 2020) nanoparticles were synthesized using its extract in order to increase the solubility of its bioactive compounds while allowing specific delivery and controlled release to target sites with reduced side effects and prolonged shelflife. (assadpour & mahdi jafari, 2019; paolino et al., 2021) the synthesized alnp appeared in powder form (table 1). when compared to the conventional microencapsulated products obtained from techniques such as spray drying and freeze drying, these powder form nanocarriers provide more bioavailability through increased surface-to-volume ratio providing them the ability to easily pass through the cell membranes and penetrate into target cells releasing their encapsulated compounds. (assadpour & mahdi jafari, 2019) the obtained colour of the alnp should be due to the entrapment of the bioactive compounds of the extract within the synthesized nanocarriers. as a high concentration of bsa was used during the nanoparticle synthesis, a high yield of 94.11% of alnp was recorded. (aniesrani et al., 2016) ic50 values of aqueous a .calcarata extract and alnp are given in in table 2. when comparing the ic50 values, it can be seen that the synthesized alnp showed a higher alpha amylase inhibitory activity than the crude a. calcarata extract. this proves that the antidiabetic activity of bioactive compounds becomes much higher when loaded onto nanocarriers probably due to the increased surface area to volume ratio resulting in a higher feasibility of interacting with the alpha amylase enzyme. the percentage increase in glucose uptake by yeast cells in the presence of aqueous a.calcarata extract and alnp are given in table 3 and table 4 respectively. the glucose uptake by yeast cells is indicated by the change in glucose concentration in the medium after a particular time interval. the aqueous a.calcarata extract as well as alnp have promoted the uptake of glucose across the plasma membrane of yeast cells according to tables 3 and table 4 respectively. however, the glucose uptake capacity at approximately 0.5 mg/ml aqueous extract was 45.30 (±0.97) %, and for that of synthesized alnp was 73.09 (±0.06) %, for the 5mm glucose concentration. similarly, the alnp exhibited a higher activity than the aqueous extract at all glucose concentrations used in the study. therefore, this once again proves that the antidiabetic activity of the aqueous extract becomes further enhanced when going into the nanoscale. moreover, the glucose uptake capacity at 0.03 mg/ml of alnp was 60.80 (±0.70) % and that has reached up to 73.09 (±0.06) % when 0.5 mg/ml of alnp was used, in the presence of a 5 mm glucose concentration. this means that by increasing the nanoparticle concentration, the capability of yeast cells to uptake more glucose from the environment can be increased. a similar increment in the glucose uptake capacity was observed when 10 mm and 25 mm glucose concentrations were used. however, an inverse relationship to the molar concentration of glucose was observed with the percent increase in glucose uptake by yeast cells, when 5 mm, 10 mm and 25 mm glucose concentrations were compared for the same amount of alnp. from the results it is clear that lower the glucose concentration, higher the uptake by yeast cells. this observation is in good agreement with the earlier work of rehman, et al., 2018 and bhutkar, et al., 2016. the solubility of alnp in water is given in table 5. solubility of the alnp is determined by the ph of the bsa solution as it affects the size of the nanoparticles formed. the isoelectric point of bsa is around 4.7. hence, at ph 9 the bsa protein molecules are ionized and therefore they tend to repulse each other. this causes the synthesized alnp to become smaller and smaller, increasing their surface area and thereby resulting in greater solubility in water. table 5 also shows the a. calcarata loading percentage and a. calcarata entrapment efficiency of the alnp. during nanoparticle synthesis, as the bsa concentration that was used was high (20 mg/ ml), it may have increased the number of nanoparticles that were formed. when the quantity of nanoparticles is higher, it will decrease the amount of a. calcarata that gets loaded with constant addition. also, the high ph of the bsa solution (ph 9) will also decrease the amount of a. calcarata that gets loaded due to the high electrostatic repulsion between bsa and a.calcarata. this explains the small value for the a. calcarata loading percentage of the synthesized alnp. also, a high bsa concentration will increase the entrapment efficiency as larger particles have larger volumes which increases the quantity of a. calcarata that can be held by the synthesized alnp. this explains the high value obtained for the a.calcarata entrapment efficiency of alnp. (aniesrani et al., 2016; ranasinghe, et al., 2013) alnp showed a mean diameter of 1030.70 (±75.3) nm (table 6). according to amighi, et al., 2020, the nanoparticle size that is obtained under the conditions that were used in alnp synthesis with citric acid as the cross-linking agent is 1201.00 (±58.4) nm. as the mean particle size obtained for alnp is very close to this value, it is clear that these nanoparticles have been formed effectively. according to danaei, et al., 2018, the pdi of a nanoparticle sample with effective particle size distribution should be between 0.05-0.7. as the pdi value of alnp falls within this range, it can be stated that a homogenous population of nanocarriers have been synthesized, which can act as safe, stable and efficient nanocarriers of a. calcarata. therefore, the tendency of this antidiabetic nutraceutical to accumulate in the target tissue, which depends on the particle size distribution will be minimal. nanoparticles with a zeta potential between -10 and +10 mv are considered neutral, while nanoparticles with zeta potentials greater than +30 mv are considered strongly cationic and those with zeta potentials less than -30 mv are considered strongly anionic. since most cell membranes https://journals.e-palli.com/home/index.php/ajcp pa ge 29 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 21-31, 2023 are negatively charged, zeta potential can strongly affect a nanoparticle’s tendency to penetrate membranes, with cationic particles displaying toxicity due to cell membrane disruption. (clogston & patri, 2011) as the zeta potential of alnp is 2.57 (±0.32) mv, these nanoparticles are neutral and will be able to easily penetrate the cell membranes without causing any toxicological effects. this further ensures that the synthesized alnp can be effectively used as an antidiabetic nutraceutical. the morphological analysis of alnp was carried out with fe-sem, and the obtained image is shown in figure 3. the sem micrograph obtained revealed morphological aspects of alnp with a spherical shape and uniform size. when comparing the uv-visible absorbance spectrum of alnp with that of the aqueous a.calcarata extract, peaks at 210 nm and 280 nm were observed in both spectra (figure 4). this means that the active compounds of a. calcarata have been successfully loaded into the bsa nanoparticles. therefore, this proves that the active compounds of the aqueous alpinia calcarata extract have been successfully loaded into the bsa nanoparticles. however, a peak corresponding to citric acid was not observed in the uv absorbance spectrum of the a. calcarata loaded nanoparticles. this means that citric acid was completely washed away during the washing step performed in the purification of nanoparticles. the pure bsa spectrum showed a characteristic peak at 280 nm. therefore, the peak observed at 280 nm in the alnp spectrum may be due to both bsa and the active compounds of a. calcarata. when considering the ft-ir spectrum of alnp, it too showed peaks that could also be observed in the ftir spectrum of the aqueous alpinia extract (figure 5). characteristic peaks including =c-h stretch at 3300 cm1, c=c benzene stretch of two peaks between 1400-1600 cm-1 and oh stretch by a broad peak between 30003600 cm-1 were observed in both spectra. therefore, this gives further evidence for the successful entrapment of the a.calcarata compounds within the synthesized alnp. according to xu, et al., 2015, one to two carboxyl groups of one citric acid molecule can react with bsa, resulting in an increase in the total amount of carboxyl groups and a decrease in the total amount of amine groups in the cross-linked nanoparticles. this explains the presence of a peak corresponding to the oh stretch and decrease in intensity of the peak at 3400 cm-1 corresponding to the n-h stretch in the spectrum of alnp. this means that interactions between bsa and citric acid have caused conformational changes in the protein structure of the synthesized nanoparticles. a similar observation has been made by amighi, et al., 2020 during the synthesis of bsa nanoparticles using citric acid as the cross-linking agent. furthermore, two peaks at 1665 cm-1 and 1537 cm-1 were observed in the pure bsa spectrum corresponding to the amide i and amide ii stretches respectively. however, in the alnp spectrum the peak corresponding to the amide i stretch was absent and a noticeable shift was seen in the peak corresponding to the amide ii stretch. hence, loading of a. calcarata compounds have also induced conformational changes in the structure of the bsa protein. this means that the active compounds of a. calcarata have interacted with the protein matrix of the synthesized nanoparticles via covalent bonds. this result is similar to the results obtained by rani, 2016. conclusion the present study has proven that the antidiabetic activity of bioactive compounds of alpinia calcarata becomes enhanced when loaded onto nanocarriers. a high yield of 94.11% of alnp were synthesized which appeared in powder form, had a spherical morphology, uniform size, with effective particle size distribution and a neutral surface charge. as the bsa solution used had a ph value of 9, which was further away from its isoelectric point, alnp reported a solubility percentage of 64%. a high a. calcarata entrapment efficiency of 87.71% and an a.calcarata loading percentage of 6.66% were also recorded for the synthesized nanoparticles. the mean particle size of the alnp was 1030.70 nm. the ft-ir spectrum of nanoparticles showed that the cross-linking agent, citric acid had caused conformational changes in the protein structure of bsa and that the active compounds were successfully loaded into the synthesized nanoparticles which interacted with the protein matrix via covalent bonds. the uv-visible absorbance spectrum of alnp further proved the successful entrapment of the active a. calcarata compounds within the nanoparticles and also showed that citric acid was completely washed away during the washing step. therefore, it can be concluded that alnp have been synthesized effectively, which can be used as a powder form antidiabetic nutraceutical. a limitation of this study was the high particle size obtained for the synthesized alnp and therefore future work can be carried out to reduce it by testing various other desolvation agents and cross-linking agents. also, in-vivo antidiabetic analysis can be carried out to test the effect of the synthesized alnp within living organisms in the future. acknowledgements this study was funded by the university grant rp/03/02/06/01/2021 which is gratefully acknowledged. conflict of interest there is no conflict of interest. references amighi, f., emam-djomeh, z., & labbafi-mazraehshahi, m. 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(2015). controlled delivery of hollow corn protein nanoparticles via non-toxic crosslinking: in vivo and drug loading study. biomedical microdevices, 17(1), 8. https://doi. org/10.1007/s10544-014-9926-5 https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 13 american journal of chemistry and pharmacy (ajcp) computer forensics education: the applications of forensic tools in modern education system samy abdelwahab safaan1* volume 1 issue 2, year 2022 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v1i2.1077 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: december 15, 2022 accepted: december 23, 2022 published: december 29, 2022 cybercrime and computer-related incidents are becoming more prevalent and common, resulting in billions of dollars in damages. to address these crimes and scams, there is an urgent need to build digital forensics training programme, that will equip relevant professionals to investigate computer crime and events efficiently. there is presently no standard that governs the development of an academic program’s digital forensics curriculum. this study extensively investigates prior work on curriculum design in digital forensics as well as existing computer forensics course offered at the first is the community college in buraidah, and the second is the community college in unaizah namely, “cyber security diploma” the cyber security diploma is offered in both colleges, the community college in buraidah and unaizah, and the diploma is at the third level of the course “computer forensic analysis,” which is the focus of the research topic. interviews were conducted with both digital forensic trainers and practitioners, and the findings were examined in order to identify and assess the skills and knowledge required by the industry and law enforcement agencies for their digital forensic examiners. hands-on training may be delivered by constructing and managing a student laboratory specifically designed for digital forensic examination instruction. a laboratory can be outfitted with a wide range of software solutions, ranging from commercial investigation suites to free command line tools. similarly, the forensic workstations that execute the software might be supplied by the vendor as stand-alone devices or assembled from separate components in-house. building and administering a student lab successfully can be a daunting task, but it can also be done on a little budget as long as the focus stays on student achievement. keywords computer forensics education, cyber security, digital forensics, digital forensic tools, encase, ftk 1 department of natural and applied sciences, community college of buraydah, qassim university, buraydah, 52571, saudi arabia * corresponding author’s e-mail: samyabdelwahabsafaan@outlook.com introduction the use of digital items has grown engrained in our professional and personal lives as computer and internet technologies have advanced. email and online chat, for example, have become commonplace modes of communication. computer systems and the internet are used by organizations and businesses for e-commerce, corporate communication, and internal administration (sindhu & meshram, 2012)society is so reliant on computers and internet technology that the internet infrastructure has become the backbone of communications, education, healthcare, transportation, and warfare, among many other things (tsai, 2015). because of its importance in our society, technology has become a target for cybercriminals, scammers, and terrorists (casey, 2020). cybercrime and device incidents continue to play a role and common, resulting in hundreds of billions in damages, highlighting the need to train a skilled workforce to limit, prevent, and prosecute these crimes, frauds, and attacks through efficient digital forensics. however, because digital technology systems are so complicated and dynamic, digital forensic investigators must have appropriate understanding as well as a diverse range of skills. computer forensics is used to acquire digital evidence form a wide range of crimes such as child pornography, financial fraud, identity theft, cyberstalking, homicide, kidnapping, and rape (harvey, 2019). during the investigation of digital devices, computer forensics investigators employ a number of software tools. these technologies are essential for gathering and assessing digital evidence (tilekar). forensic investigation in general, and especially hard disks examination, is difficult for an investigator because of the requisite technical background, such disk examinations typically have a pretty high learning curve (garfinkel, 2013). the high learning curve is partially caused by the wide spectrum and accessibility of forensic investigative tools. there are numerous tools to consider, both commercial and open source. increasingly popular tool, particularly free source, are becoming accessible on a regular basis. to varied degrees, these tools offer layers of information that assist forensic experts to locate and safely preserve digital evidence, as well as undertake routine examinations (sikos, 2021). however, depending on the sort of examination, investigators are always required to know how to operate and configure/parameterize multiple tools, particularly open source types. the existence of a large number of such tools involves the expertise to address the following research questions: “how do i apply these tools properly?” and “where/when can i apply them effectively?” in fact, forensic examiners have various degrees of it background and technical proficiency, ranging from computer security specialists to investigators with basic computer skills (mattijssen et al., 2020). as a consequence, irrespective of their computer and it skills, investigators want practical tools that could assist them in obtaining findings quickly and with https://doi.org/10.54536/ajcp.v1i2.1077 https://journals.e-palli.com/home/index.php/ajcp mailto:samyabdelwahabsafaan@outlook.com pa ge 14 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(2) 13-20, 2022 minimal usage complexity (case et al., 2008). whenever we consider human participation, especially for the investigation of today’s normally vast amounts of data, such tools that can reduce the investigation burden on the human investigator irrespective of their technical expertise are becoming increasingly important. when it comes to open source tools, it is usual for one software product being unable to gather all of the necessary data. as a result, the examiner must adopt a variety of methods in order to obtain useful data from the target. due to the obvious expertise demanded by the tools, all this needs advanced training and adds to the learning curve. these tools are also not usually compatible with one another. users of today’s technologies must appropriately examine the outcomes of the tools in order to decide the next steps in performing a deeper study. related work according to the united states government accountability office (gao), preventing computer crimes and cyberattacks entails several obstacles (fellows). some examples include a lack of cybercrime detection and reporting technologies, a lack of education or training standards that may equip law enforcement with sufficient analytical and practical skills, and a lack of structure to enhance cybersecurity and raise public awareness. the digital forensics community is profoundly concerned about the industry’s lacking of expertise and skills standards. so far, only a few efforts have been made to define computer forensics programme standards. the digital forensics education programs accreditation commission (fepac) published guidance for computer forensics education and skills in 2008 (ruffell, 2010), which were bestowed upon them by the american academy of forensic science (aafs). these papers simply provide basic requirements for computer forensics training and education, such as the amount of credits required, the key forensics subjects that should be covered, and so on. the west virginia university forensic science initiative established recommendations for computer forensics training and education, which were issued by the national institute of standards and technology (nist) (cunha et al., 2017). this work may be used as a fantastic guide for developing an educational programme. however, building an educational programme based primarily on these standards would be prohibitively expensive for educational and training institutes; 24 courses is a substantial number in an academic programme. in reality, no previous educational or training programme have included such a diverse range of computer forensics courses (yasinsac et al., 2003). policy maker for network forensics, computer and network forensics professional, and computer and network forensics researcher are examples of topics taught in an education programme that differ significantly from those offered in a training programme. a training programme emphasizes practical skills and application, whereas an education curriculum emphasizes theory and principles. according to the designers of the approach, an undergraduate programme can ideally include themes present in both training and teaching programmers (gottschalk et al., 2005). the undergraduate course introduces students to the core tools and approaches of the subject. the undergraduate course mentioned above is necessary for the graduate course, which covers advanced issues related to evidence analysis and presentation, as well as the modification and integration of available technology into standard operating procedures. it does not give comprehensive instruction on several issues, notably the practical use of equipment and talents necessary for a forensics education programme (digabriele, 2008). the “high tech crime consortium “(htcc) set up an online certificate program that displays the viewpoints or competence of a computer forensics degree graduate (lee & pagliaro, 2013). security principles, system administration, web publishing, and two computer forensics courses were all suggested. its central emphasis was on security and network topics, and students were not involved in the development of practical skills and technology (gottschalk et al., 2005). erbacher and swar stressed the need of including training and education themes in computer forensics education programme. however, their primary concentration, however, is on the management or administrative aspects of digital forensics (fakouri & teimouri, 2019). literature review computer forensics is a forensic science subject with varied meanings depending on who you ask. however, it is best defined as “the application of scientifically derived and proven methods to the preservation, collection, validation, identification, analysis, interpretation, documentation, and presentation of digital evidence derived from digital sources for the purpose of facilitating or furthering the reconstruction of criminal events, or assisting in the anticipation of unauthorized activities shown to be disruptive to planned operations” (taylor et al., 2007). sub-disciplines of digital forensics include computer forensics, mobile device forensics, network forensics, and database forensics. that analyze different equipment, figure 1: the investigational method of digital forensics https://journals.e-palli.com/home/index.php/ajcp pa ge 15 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(2) 13-20, 2022 media, or evidence (raval, 2020b). our study focuses on computer forensics in particular, as well as the tools utilized for training. as a result, the vast majority of the examples and explanations in this study will be provided with this goal in mind. the above description of computer forensics encompasses a wide variety of operations, from device seizure through evidence presented to judicial authorities. however, we would want to reduce the process phases to more basic methods, as indicated in the us department of justice’s “a guide for first responders” (brinson et al., 2006). the suggested paradigm is divided into four stages: data collection, investigation, analysis, and reporting (see figure. 1). phases of digital forensics during this step, the examiner must guarantee that the copy of the media that will be used in the examination and analysis stages is correct, as well as that the original media’s integrity is protected. otherwise, the entire inquiry may be deemed illegitimate by the court (branham, 2019). the examination phase is the stage at which detectives conduct a thorough search for evidence connected to the alleged crime. this focuses on finding and locating possible evidence, which may be in unusual areas (mellars, 2004). this procedure should fulfil a number of goals. first and foremost, the status of the evidence must be meticulously documented. examiners especially explore unorthodox areas for the existence of hidden or obscured data throughout the whole procedure. because the final outcome of this procedure may be big, data reduction can be conducted after all of the information is revealed (mellars, 2004). an investigator retrieves evidence using a variety of techniques and tools. some of these tools will be briefly described later in this chapter. during this phase, the investigator searches for information that will help him or her answer questions about the case. an investigator, for example, has to know: “given the reviewed files/data, might this data confirm or deny my hypothesis?” “how so?” because they have little help from the inquiry tools, much of their work is cognitive (casey, 2020) depending on the investigator’s expertise and experience. there may be a need to gather and examine additional data that has never been covered previously. as a result, the investigative process may return from the analytical phase to the collection and inspection stages. it may also require multiple iterations of investigation and analysis to back up a criminal theory (casey, 2020). the reporting step is the final stage of the digital forensic inquiry. a written report typically includes and summarizes the examination procedure, the pertinent data retrieved with their associated hash values, and the conclusions derived from the analysis phase (casey, 2020). furthermore, the tools utilized in earlier stages, as well as the reasons for their usage, are indicated in the reporting phase. examination notes must be kept for the purposes of discovery or testifying (casey, 2020). some automated (particularly commercial) tools provide reports for investigators. however, nearly no single purpose tool generates such a report, hence it is important for the investigator to collect information for every activity and tool used in all of the above-mentioned phases for reporting purposes. digital forensic tools the results of a survey conducted between both computer forensics practitioners and colleges or institutions offering a computer forensics programme (see table 1). to establish the technical skills needed by computer forensics practitioners, as well as the tools that should be taught in forensic analysis courses, professionals in computer forensics both from public and private sectors were surveyed, for each group asked a variety of questions (kessler, 2010). teachers of digital forensics were asked what analytical tools they used in their curriculum and if they were willing to engage with digital forensics practitioners for teaching purposes. they were also polled on why they did not cooperate with digital forensics practitioners for instructional purposes. the study also inquired about their thoughts on how to increase education in digital forensics. these survey questions were sent to universities and colleges that provide computer forensic degrees. this poll includes seventeen volunteers from various institutions and universities, as well as nine volunteers from the united states’ digital forensics practitioner association. sixtyseven percent of digital forensics practitioner responders had fewer than ten years of expertise in the field. the digital forensics practitioner’s category had the most responses, with 44.4 percent coming from corporations or private companies. the furthermore share of respondents, 22.2 percent, came from law enforcement and non-governmental organizations. moreover, 11.1 percent of digital forensics professionals worked for government agencies, with little or no responses from of the private investigation sector. table.1 highlights the use of main digital forensics technologies by both professionals and digital forensics professors encase is the most widely used tool for both instructors and educators, with 94.1 percent of digital forensics teachers and it is used as the primary tool for computer forensics collection and analysis by 66.7 % of computer forensics professionals. ftk is the second most popular tool, with 70.6 percent of digital forensic professors and 56.6 percent of digital forensic professionals using it. winhex, helix, md5sum, and mobiledit are other handy tools. computer forensics experts frequently employ forensics; yet, it appears that professors seldom use it. other tools used by computer forensics professionals but not by digital forensics instructors ptk, celldek, videofocus, dtective, clearid, dvelepor, and magnifi are a few examples. similarly, foremost, pyflag, and outguess are tools used by teachers rather than specialists in digital forensics. the willingness to collaborate for the two components (e.g., the survey results are shown in figure.2 was analyzed to establish how closely industry and allied organizations https://journals.e-palli.com/home/index.php/ajcp pa ge 16 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(2) 13-20, 2022 table 1: use of digital forensics tools (30). computer forensics tools results from universities and colleges results from computer forensics practitioners encase 94.1% 66.7% access data forensics toolkit (ftk) 70.6% 57.6 ftk imager 70.5% 66.7% win hex 64.6% 55.3% autopsy forensics tool 23.5% 11.1% helix 64.7% 55.7% smart 0.00% 33.3% ptk, 0.00% 11.1% celldek 0.00 11,1% video focus, 0.00 0.00% dtective 0.00 11.1% clear id 0.00 11.1% dvelepor 0.00 11.1% magnifi 0.00 11.1% md5sum 29.5% 55.7% py flag 5.9% 0.00% foremost 17,6% 0.00% mobil edit 11.8% 55.6% figure 1: digital forensics instructors and practitioners’ desire to contribute in the progress of digital forensics (31). can engage with universities for digital forensics training. according to the poll results, a digital forensics professional should be proficient in the use of the most often used tools, such as ftk and encase. as a result, graduates of forensic institutions receive extensive training in these technologies. furthermore, forensic training should include a comprehensive lecture on forensic tools, focusing on ftk and encase as well as helix, winhex, and other open-source tools. the survey shows that the how the training of these tools helps the graduates to enter into the practical aspect of the computer forensics and how they both the educational institutes and the professionals can work together to bring a suitable learning and training environment for the students of computer forensics. therefore, it is very important to understand that combined efforts towards the development of the computer forensics can help educational institutions and students to understand the practical aspect of the computer forensics. proposed models for digital forensics the most common commercial test software packages nowadays are accessdata’s forensic toolkit (ftk) and guidance software’s encase. both commercial offers are employed by law enforcement agencies, have advantages and disadvantages, and are frequently compared and contrasted. ftk is a digital investigative tool that has been used in court. it is built for speed, stability, and ease of use, it outperforms all other products in terms of substantial pre-processing and indexing, filtering, and searching (cantrell & dampier, 2012). ftk is wellknown for its use of front-end indexing, which results in near-instant search results. a standalone imager, registry viewer, and password recovery programme are also included in the test suite. guidance software describes encase as the global standard in digital investigative technology for forensic specialists that want rapid, forensically sound data gathering and investigations with a repeatable and defensible methodology (cantrell https://journals.e-palli.com/home/index.php/ajcp pa ge 17 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(2) 13-20, 2022 & dampier, 2012). it also includes extra tools like an independent imager and decryption solutions in the form of modules. both suites are also noted for their distributed processing capacity, which allows the programme to handle one case at a time using up to eight workstations. this dramatically accelerates case processing and reduces case processing durations from days to hours. while this commercial software might be rather pricey, both provide academic programme that significantly lower the cost of educational reasons. both provide substantial training programme that may be utilized to teach new programme trainers. these advantages can help to offset some of the expense of commercial software. another advantage of commercial suites such as ftk and encase is that they both provide industry-recognized certifications for their products’ use and performance. accessdata certified examiner (ace) eligibility is issued by ftk, and encase offers a comparable qualification, encase certified examiner (ence). both certifications may be obtained by students at diploma level and after finishing a degree and can aid in their career search. there are several free digital forensic investigation tools available. these tools are portable and may be set up in a matter of minutes. they are frequently used in the field to gather information on the go or to conduct investigations when time is of the essence. that isn’t to suggest they aren’t as productive as commercial suites; in fact, commercial tools are frequently easier to use for novices. both sorts of tools evaluate data in the same way, and a whole digital forensics programme could and is developed around free toolkits (cantrell & dampier, 2012). current challenges for educational institutes cybersecurity”, often known as information security, seems to be a rapidly expanding job sector as a result of rising and costly cyberattacks. graduates of cybersecurity schools are often prepared for entry-level roles which includes “cybersecurity analysis” at “buraydah community college, qassim university” and “unaizah community college, qassim university”. as per the “u. s. department of labor, labor statistics (bls)”, workforce of cybersecurity professionals is expected to grow from 2012 to 2022, much faster than the average growth rates for all fields of work of 11 percent and 18 percent for computer-related professions (gonzález jaimes, 2016). there seems to be a nationwide lack of cybersecurity specialists with the necessary knowledge and expertise, and education is anticipated to become the vital solution (tu et al., 2012). as a response, there seems to be an increase in the demand for and availability of cybersecurity education and skills. meanwhile, cybersecurity education faces significant obstacles. to excel academically and professionally in cybersecurity, students must have a solid foundation and preparedness in computer and information science and technology. however, there has been a perpetuated failure of education in the middle east to prepare a strong and world-leading workforce in computing professions. the following are the major aspects of this catastrophe in middle eastern undergraduate computer programmes: the primary source of the gap among students’ education as well as the real abilities required in the labor market may be outdated curricula and course material, as well as a lack of understanding practical experience. other issues include a decline in enrolment and a failure to take advantage of service learning programmes that develop application skills. this study presents a projectbased curriculum educational outreach paradigm for cybersecurity education to assist reduce the gap between college education courses and work training necessary in the real life. to that aim, saudi arabia has launched two major standards that serve as principles for enhance the effectiveness of cybersecurity learning and skills for both education institutes and employment. the first framework is (scywf) (liu & tu, 2020), which categorized 40 job positions in cybersecurity and described the required responsibilities, expertise, skills, and competencies for each position. the second framework (scyber edu) (gamlo & bamasak, 2011), describes the minimal standards and academic units which should be provided by college cybersecurity programmes in saudi arabia, ranging from certificate to doctoral levels. some institutions have steadily incorporated one or two cybersecurity courses into academic curriculum in past few years, including “buraydah community college” and unaizah community college” qassim university” however, introducing such courses frequently finds obstacles in teaching the practical aspect of the diplomas namely, forensic computer analysis” which is one of the courses taught at the third level of the study plan for the “diploma in cyber security”, and the “diploma in cyber security” is a two-year diploma, after secondary school, divided into four levels and teaching staff finds it difficult to teach the practical side of this course, as well as the students in understanding it. the paper will address the teaching methods to implement and make things easy for the students and teachers to understand the practical side of these courses. the technical working group on education and training in digital forensics (tu et al., 2012), suggests creating a computer forensics laboratory to supply students with equipment. and software to train them in practical skills, particularly with the popular digital forensic tools featured in our findings. individual machines are involved in the majority of white-collar crime in the public sector. the corresponding counter-investigative powers are limited to the average end-user. according to the study results, there is a considerable increase in the number of occurrences on networks, protocols/devices, and internet apps, and many of these incidents involve adversaries with skills much beyond those of typical end users (raval, 2020b). to examine these criminal cases and their perpetrators effectively and efficiently, as well as discover relevant evidence, digital forensics specialists require more broad knowledge and abilities that introduce the discipline of https://journals.e-palli.com/home/index.php/ajcp pa ge 18 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(2) 13-20, 2022 network / internet forensics. there are currently relatively few educational programme that provide such training, and there is no agreement on what tools and subjects should be included in educational courses covering network / internet forensics. there are about three key methods to understand cybersecurity: (i) incorporating one or two classroom learning into the overall (ii) curriculum. incorporating cybersecurity themes into other network and computer courses. (iii) and using less formal approaches, such as training courses. cybersecurity modules are frequently provided throughout the latter semesters of a student’s programme. in certain circumstances, a security course is an elective, which has the unintended consequence of students avoiding it during the last semester. some students are found to fear that somehow this security course will be challenging and will damage their ability to complete their education. under the second technique, cybersecurity topic is introduced in other information and communication technology (ict) curriculum such as, “operating systems”, “computer networks”, “databases, and “software application programming”. it was frequently suggested that cybersecurity should be covered in a range of learning courses. this addition is contingent on both the teachers’ understanding of the subject and their effort in addressing such topics in the curriculum. when a talented professor quits the college, this might radically alter. cybersecurity material differs over institutions and departments including both techniques. we therefore presented participants with a list of cybersecurity topics, such as computer security, network security, it system security, security management, and incident response, to assist them better grasp overall competencies of colleges. the majority of interviewees felt that teaching the first three categories is more practical. the majority of interviewers felt that teaching the first three topics is more doable, and they’re less convinced regarding teaching “incident response” in the course. the third strategy is the use of informal activities to enhance understanding about cybersecurity. some universities host lectures, conferences, and other events to strengthen students’ understanding of cybersecurity by inviting guest speakers. such activities are said to be warmly appreciated by students and to pique their interest in the topic. finally, certain cybersecurity topic is clearly included in professional certification material. to meet the demand for qualified investigators, educational institutions have implemented a variety of degree and certificate programme in digital forensics. training a student in digital forensics, or any investigative discipline, presents unique obstacles in that educators are faced with teaching a student how to analyze a case and identify evidence through discovery in the absence of an absolute definitive blueprint. every case is unique, and there is no “recipe” or “by the numbers” way for conducting investigations. a digital forensics investigation cannot be completed by simply following a set of procedures on a reference card. a successful curriculum necessitates activities other than theoretical lectures and multiple-choice tests. a digital forensics programme must include hands-on lab work in which students execute exercises and tests on actual equipment in a lab setting. when theory can be implemented or applied, it becomes more understandable (sánchez et al., 2011). this teaching technique promotes kinesthetic learning, in which students learn via physical action rather than through listening to lectures or reading powerpoint presentations. kinaesthetic learning happens when students actively participate in learning through hands-on activities and other ways, and students learn more as a result (pollitt et al., 2008). modern approach towards digital forensics education an investigator in digital forensics must be well-versed in a wide range of operating systems. according to practitioners’ experience, windows pcs are the most prevalent in inquiry situations, whereas unix / linux accounts for around 20% of total systems (tu et al., 2012). this implies that the digital forensics curriculum should cover a range of operating systems, with a primary concentration on windows and a secondary focus on unix / linux and macintosh. unfortunately, while it is theoretically ideal to teach as many operating systems as possible, educational programmes have limited resources, including time, equipment, and staff resources. the difficulty for students seeking hands-on experience is to create and maintain a proper laboratory environment for instruction. the many considerations that must be taken to account for space needs, software, hardware, network concerns, and the many other varied peripherals and devices necessary for such a programme can make building and maintaining such a laboratory a daunting job. there appears to be considerable worry about how students can be prepared to fulfil the expectations of both industry and law enforcement authorities (cummins flory, 2016). there are numerous options to tackling this problem; one recommended strategy is to collaborate with digital forensics specialists from both industry and law enforcement. it is impossible for digital forensics specialists to commit a significant amount of time to designing teaching programme, especially given budget and timeline restrictions. it is critical that digital forensics curriculum combine industry and law enforcement agency experiences and ideas. specialty training for academic staff that don’t have prior experience in certain fields of cybersecurity is an important component of establishing stronger curriculum. similarly, suitable training for students in practical cybersecurity topics must be improved through the introduction of laboratories and skills acquired even outside the college. additional actions that should be considered include, • incentives for local industry to assist education programs, such as paid internships and the availability of https://journals.e-palli.com/home/index.php/ajcp pa ge 19 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(2) 13-20, 2022 trainers. • promoting inter-temporal professional relationship among academics and governmental entities in order to boost growth. • obtaining assistance from global counterparts (organizations or private businesses), including the organization of american states (in uruguay), ibm (in costa rica), and microsoft (in india). • creating educational programs and services, such as forensic labs. • “virtual training environments” being adopted at collage level. • envisioning and promoting training courses to provide students with cybersecurity relevant work experience. • attention on detailed practical learning and training courses. numerous governments have recognized the usefulness of retraining programs in enhancing practical–technical cybersecurity training. in the united kingdom, cybersecurity training programs helped country major industries and are funded by the government (thompson, 2019). in the united states, community college cyber job training programs also begun to appear (thompson, 2019). industrial cooperation help fund these and other related activities. the cybersecurity field is incredibly dynamic and everchanging. as the need for skilled individuals in this industry grows, education institutions are engaging in a number of ways. advanced nations, at both the governmental and corporate levels, have begun initiatives to assist educational institutions in developing adequate cybersecurity curricula in order to meet the growing need for cybersecurity specialists. moreover, a number of scholars have presented cybersecurity curriculum design approaches and suggestions. this research, i trust, will provide educators with a better understanding of the entire efforts undertaken in the cybersecurity field, as well as assist them in developing increasingly successful cybersecurity curriculum and instruction in future work. professional project, internship, and/or court experience are examples of suitable courses that can be classed in this category. further study should be conducted to investigate the association between students who complete professional projects and internships and their competitiveness in the labor market following graduation. anecdotal evidence suggests that students who conduct internships in this profession are more likely to obtain related work within six months of graduation than those who did not do an internship. students must comprehend the basic principles, techniques, and procedures that skilled criminals use to perpetrate such crimes, as well as the various countermeasures that organizations and businesses may take to defend themselves, in order to successfully solve cybercrime. according to the findings made above, network forensics courses should include a wide range of subjects, including: operating systems, network and internet protocols, malware, devices, applications, network hacking methods and tactics, as well as countermeasures and security mechanisms. to meet the demand for qualified investigators, educational institutions have implemented a variety of degree and certificate programme in digital forensics. training a student in digital forensics, or any investigative discipline, presents unique obstacles in that educators are faced with teaching a student how to analyze a case and identify evidence through discovery in the absence of an absolute definitive blueprint. every case is unique, and there is no “recipe” or “by the numbers” way for conducting investigations. conclusion this study looked at the design of digital forensics curriculum and current educational programme that offered a generic list of computer forensics courses but provided no recommendations on which topics to include and which tools to teach. both digital forensics instructors and practitioners were interviewed, and the data was evaluated in order to determine the information, methodology, and abilities required by the industry and law enforcement groups. the most often used commercial tools are encase and ftk, and the majority deal with windows operating systems, followed by unix /linux and macintosh. the increase in cybercrime implies the continued need for well-trained detectives. universities may respond by offering basic and advanced digital forensics courses. based on our findings, we propose some next research directions. we would want to first study the problems and methods for building online security and forensics courses in order to assure flexibility and cost effectiveness, as well as to enhance participation. all commercial and open source technologies should be available for use in the online courses as much as the on-campus learning environment, and the solution should be properly scalable and flexible enough to adapt to quickly developing computer and forensic technology second, the design of digital forensics programme for both elementary and secondary school graduates should be studied to determine how they might be integrated into existing computer and network security programs. this paper has provided a high-level overview of a typical student laboratory setting. to better prepare students for a future in investigation, an emphasis on evolving technology should be maintained on a continuous basis. of course, the focus of any programme should always be on student learning, which may be more successfully accomplished by constructing and employing a purpose-built student digital forensics laboratory. acknowledgments the writer is thankful to qassim university. conflict of interest there is no conflict of interest. https://journals.e-palli.com/home/index.php/ajcp pa ge 20 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(2) 13-20, 2022 consent for publication the author agrees to the final version submitted to the journal. references branham, r. a. 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(2003). computer forensics education. ieee security & privacy, 1(4), 15-23. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 8 american journal of chemistry and pharmacy (ajcp) a study on the therapeutic effect of 5-azacytidine to attenuate the ramifying repercussions of ischemia reperfusion injury on mitochondrial molecular machinery vasisht yegneshwaran1*, priyanka n prem1, sri rahavi boovarahan1, gino a kurian2 volume 1 issue 2, year 2022 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v1i2.1169 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: january 05, 2023 accepted: january 29, 2023 published: february 05, 2023 5-azacytidine is a hypomethylating agent that has for long been used in cancer therapy due to its ability to inhibit the protein dna methyltransferase responsible for hyper-methylating dna strands. recently, studies involving in vitro, ex vivo, and in vivo experiments have assessed the cardioprotective effects of 5-azacytidine during myocardial ischemia-reperfusion injury (iri). however, the effect of this compound in restoring the damage induced to mitochondrial molecular machinery during iri has not yet been explored. understanding this would help us analyze the ways through which mito-targeted therapeutics can be used. the purpose of this study is to investigate the therapeutic impact of 5-azacytidine, as dna methylation is a very common epigenetic modification observed during iri. furthermore, the protective effect of the compound in alleviating the damage induced to mitochondria during iri can be identified, as dna methylation can leave a direct impact on the mitochondrial genes as well. an isolated mitochondria model will be used to determine the effects of 5-azacydine on mitochondrial molecular machinery as the capacity to generate dna, rna, and proteins are preserved in isolated mitochondria. in this study, we focus on the mechanisms of mitochondrial replication, and translation to understand the effect of 5-azacytidine on the iri affected mitochondrial system. mitochondrial dysfunction is also another key turn of events that happens during iri. the role of 5-azacyidine in preserving the functionality is also being assessed in our research. the findings of these experiments would help us determine the plasticity the compound imparts on mitochondrial molecular mechanism’s integrity and function post-induced iri. keywords mitochondria, ischemiareperfusion injury, in-vitro replication, mitochondrial dysfunction 1 vascular biology lab, 117, anusandhan kendra, sastra deemed university, tirumalaisamudram, tamil nadu, india 2 school of chemical and biotechnology, sastra deemed university, tirumalaisamudram, tamil nadu, india * corresponding author’s e-mail: vasisht31@gmail.com introduction ischemia, which is caused by the blockage of blood vessels, is a condition that leads to infarction of tissue. the best possible treatment identified till date is reperfusion, which involves removing the obstruction to blood flow using drugs and partially or completely invasive surgery depending on the severity of the infarction. reperfusion of tissues affected by ischemia is contraindicated in its ability to cause ischemia reperfusion injury, which is identified by certain unique pathophysiological hallmarks such as the generation of reactive oxygen species (ros) due to the reestablishment of circulation, elevated inflammation caused by the excessive ros, calcium overload and mitochondrial dysfunction, which includes the abnormal opening of the mptps. (frank et al., 2012; hausenloy and yellon, 2013; sánchez-hernández et al., 2020) researchers have been trying to understand the mechanisms and develop therapeutic measures against this condition. recent studies implicated epigenetic modifications in regulating the expression of genes involved in pathways, contributing to iri. a complete understanding of these epigenetic mechanisms is essential to identify an appropriate target to curb the damage caused by iri, as a prophylactic or as a treatment option. since they partially control gene expression patterns, these epigenetic mechanisms go beyond genetics. to better understand how ischemia reperfusion damage may be treated or prevented, the exact definition of epigenetics has been employed and controlled with the aid of inhibitors to alter gene expression patterns. epigenetics is the mechanism that influences heritable changes in gene expression and function without altering the genome’s sequence. these epigenetic pathways are influenced by external environmental elements as well. these epigenetic mechanisms control various mediators which results in ischemia reperfusion injury. some of mediators of reperfusion injury are oxygen free radicals (ros), endothelial dysfunction and microvascular injury, alterations in calcium level and altered myocardial metabolism. yet these concepts, key events and the complete order of how these mechanisms can be understood is still a work in progress. many studies believe that targeting abnormal dna methylation in ischemia reperfusion injury is a key technique for the prevention and treatment of the disease. methylation of dna at cytosine phosphate-guanine (cpg) dinucleotides is a typical epigenetic alteration that serves as a link between the genotype and the environment. the methyl group (-ch3) is added to the 5th position of cytosine residues in cytosine-phosphoguanine (cpg) dinucleotides during dna methylation, resulting in chromatin condensation and gene expression changes. enzymes known as dna methyltransferases (dnmts) catalyse this process, which is reversed by enzymes known as ten-eleven-translocation protein https://doi.org/10.54536/ajcp.v1i2.1169 https://journals.e-palli.com/home/index.php/ajcp http://vasisht31@gmail.com pa ge 9 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 8-20, 2023 1 (tet1), which converts 5-methyl cytosine (5mc) to 5-hydroxymethyl cytosine (5hmc). (moore et al., 2012) dna methyltransferase inhibitors (dnmti) have also been demonstrated to be useful in the treatment of ischemia-related illnesses. 5-azacytidine, a dnmti is a medication licensed by the us food and drug administration (fda) for the treatment of acute myeloid leukaemia (aml) and myelodysplastic syndrome (mds), with an inhibitory impact on dna methylation as the underlying mechanism. 5-azacytidine has a long history of clinical use in cancer therapy. moreover, several studies have been conducted to demonstrate the role of 5-azacytidine as a cardioprotective drug in the treatment of ischemia/reperfusion (i/r) damage. it is done in the animal model of rat. (boovarahan & kurian, 2021) the discovery that 5-azacytidine was integrated into dna and that it blocked dna methylation when present in dna led to its widespread use to show the relationship between the loss of methylation in particular gene areas and the activation of the related genes. (christman, 2002) since methylation works on dna directly, it may work on nuclear and mitochondrial dna. it also allows for the observation of both direct and indirect effects on epigenetic modulations on various components involved in iri. an extensive, broad-spectrum target like methylation pattern may provide for the development of therapeutics for ischemic conditioning in the future. the main focus of this paper, will therefore be with respect to 5-azacytidine and its therapeutic effects on ischemia reperfusion injury. materials and methods animals the guidelines from the committee for the purpose of control and supervision of experiments on animals (cpcsea), government of india were strictly adhered to throughout all the animal experimental procedures involved in the study. all the rights and terms has been approved by the institutional animal ethical committee (iaec) at sastra deemed to be university, thanjavur, india for the conduct of experiments. male wistar rats of weight between 250-300 grams inbred in the central animal facility at sastra deemed to be university, thanjavur, india was used in the study. the rats were all housed in a well-ventilated polycarbonate cage in a temperature-controlled room at (22 ± 2°c) with a relative humidity of (60 ± 5%). the animals were exposed to 12 hours of light and dark cycle with ad libitum supply of water and food. isolation of mitochondria the organelle mitochondria was isolated using density gradient differential centrifugation of the organ homogenate following the guidelines from palmer, et al. a 10% organ homogenate was prepared momentarily before the process of density gradient differential centrifugation using isolation buffer (220 mm mannitol, 70 mm sucrose, 5 mm mops, 2 mm edta, and 0.2% bsa) with a ph of 7.4. the organ homogenate was then centrifuged at lower speed of 800g for 10 minutes at 4°c to pellet the nuclear fraction. the supernatant was then transferred into a new eppendorf tube. the supernatant was then centrifuged at 8000g for 10 minutes at 4°c to remove any cellular debris. the supernatant was removed and the pellet was briefly resuspended in isolation buffer. this suspension was subjected to high centrifugation of 12,000g for 10 minutes at 4°c to pellet a pure fraction of mitochondria (graham j. m. et al. 2002). the mitochondria was resuspended in storage buffer (100mm kcl, 100mm tris-hcl, 75mm sorbitol, 25mm sucrose, 10mm k2hpo4 , 5mm mgcl2 , 0.05mm edta, 0.2% bsa) at a ph of 7.4 and their protein concentration was determined with the use of bradfords reagents (biorad). a bovine serum albumin standard was used to determine the concentration. after the process of protein estimation, the isolated mitochondria were randomly divided in to groups and subjected to normoxia and hypoxia reperfusion using respiratory buffer (300mm mannitol, 100mm kcl, 20mm hepes, 10mm kh2po4, 5mm mgcl2 , 1mm egta, 0.2% bsa) at ph 7.1 and hypoxia buffer purged with n2 (75mm nacl, 25mm hepes, 20mm lactate, 16mm kcl, 10mm nahco3 , 5mm deoxy-d-glucose, 1.2mm mgcl2 , 1.2mm cacl2, 1mm kh2po4) at ph6.8. experimental groups isolated mitochondria from rat hearts after normalisation were randomly divided into six groups and details of each experimental groups are as follows: 1. normal after the process of mitochondria isolation, the organelle was subjected to equilibration using respiratory buffer for 1 hour 15 minutes to maintain normal respiration. 2. normal + 5-azacytidine after the process of mitochondria isolation, the organelle was subjected to equilibration using respiratory buffer for 15 minutes followed by 0.5 μm 5-azacytidine pre-treatment for another 15 minutes. at the end of drug pre-treatment, the mitochondria were centrifuged at high speed of 12,000g for 10 minutes at 4°c to pellet a pure fraction of mitochondria. then, the mitochondrial pellet was resuspended in respiration buffer for 45 minutes to maintain normal respiration. 3. normal + dmso after the process of mitochondria isolation, the organelle was subjected to equilibration using respiratory buffer for 15 minutes followed by 0.5 μm dmso pre-treatment for another 15 minutes. at the end of drug pre-treatment, the mitochondria were centrifuged at high speed of 12,000g for 10 minutes at 4°c to pellet a pure fraction of mitochondria. then, the mitochondrial pellet was resuspended in respiration buffer for 45 minutes to maintain normal respiration. 4. ir after the process of mitochondria isolation, the organelle was subjected to equilibration using respiratory buffer for 30 minutes followed by highspeed centrifugation at 12,000g for 10 minutes at 4°c to pellet a pure fraction of mitochondria. then, the mitochondrial https://journals.e-palli.com/home/index.php/ajcp pa ge 10 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 8-20, 2023 pellet was resuspended in hypoxia buffer for 15 minutes to hypoxia followed by a 30 minutes incubation in respiration buffer to facilitate reperfusion. 5. ir + 5-azacytidine after the process of mitochondria isolation, the organelle was subjected to equilibration using respiratory buffer for 15 minutes followed by 0.5 μm 5-azacytidine pre-treatment for another 15 minutes. at the end of drug pre-treatment, the mitochondria were centrifuged at high speed of 12,000g for 10 minutes at 4°c to pellet a pure fraction of mitochondria. then, the mitochondrial pellet was resuspended in hypoxia buffer for 15 minutes to hypoxia followed by a 30 minutes incubation in respiration buffer to facilitate reperfusion. 6. ir + dmso after the process of mitochondria isolation, the organelle was subjected to equilibration using respiratory buffer for 15 minutes followed by 0.5 μm dmso pre-treatment for another 15 minutes. at the end of drug pre-treatment, the mitochondria were centrifuged at high speed of 12,000g for 10 minutes at 4°c to pellet a pure fraction of mitochondria. then, the mitochondrial pellet was resuspended in hypoxia buffer for 15 minutes to hypoxia followed by a 30 minutes incubation in respiration buffer to facilitate reperfusion. evaluation of mitochondrial function using mitochondrial electron transport chain complex activity after the estimation of protein and normalisation of mitochondria, the mitochondrial electron transport chain activity was measured spectrophotometrically by employing a specific donor-acceptor oxidoreductase in a 0.1m phosphate buffer (frazier et al. 2012). for determination of mitochondrial complex1 activity a rotenone sensitive nadh oxidoreductase was used. a succinate decyl ubiquinone 2,6-dichlorophenolindophenol (dcpip) reductase was used determine the mitochondrial complex ii activity. to determine the mitochondrial complex iii and iv activity cytochrome c reductase and cytochrome c oxidase was used as previously described by ansari et al. oxidative stress assessment the antioxidant profile of the isolated mitochondria homogenate was evaluated using catalase activity, sod activity and gsh:gssg ratio. all the experiments used a multimode spectrophotometric plate reader to measure kinetic absorbance as well as endpoint absorbance. a reaction buffer containing 0.1 m sodium phosphate buffer, ph 7.2, 4 mm h2o2 , and 5 n h2so4 was added to the isolated mitochondrial samples. the reaction was initiated by the addition of 0.005m kmno4 to the reaction buffer containing samples. the change in optical density was measured kinetically measured at 515 nm to assess the catalase activity (goldblith et al. 1950). a reaction buffer containing 45mm tris, 1mm edta was added to the isolated mitochondria samples. the reaction was initiated by the addition of 2.5mm pyrogallol to the reaction buffer containing samples. the change in optical density was measured kinetically at 420 nm to assess the superoxide dismutase activity (nandi et al. 1988). a reaction buffer containing 0.25m sodium phosphate and 5% trichloroacetic acid was added to the isolated mitochondria samples. the reaction was initiated by the addition of ellman’s reagent (5,5’-dithiobis-2nitro-benzoic acid) to the reaction buffer containing samples. the change in colour due to formation of thionitrobenzoate was measured at 412 nm to assess the gsh activity (sedlak et al. 1968). a reaction buffer containing 0.25m sodium phosphate buffer, 0.5mm edta, 4mm oxidised glutathione and 0.2mm nadph was added to the isolated mitochondria samples. the reaction was initiated by the addition of ellman’s reagent (5,5’-dithiobis-2-nitro-benzoic acid) to the reaction buffer containing samples. the change in optical density due to oxidation of nadph was measured at 340 nm to assess the gssg activity (sedlak et al. 1968). in vitro mitochondrial protein synthesis the experiment in vitro mitochondrial protein synthesis was carried out according to the procedures earlier by fernandez-silva et al. the isolated mitochondria were briefly suspended in maite buffer (75mm sorbitol, 25mm sucrose, 10mm kcl, 10mm k2hpo4 , 0.05mm tris-hcl) at ph 7.4 containing 10mm glutamate, 10mm succinate, 2.5mm malate, 1mm adp and 1mg/ml of bsa. the process of translation was initiated by adding 100 μg/ml emetine, 100 μg/ml cycloheximide, and 10 μm of the 20 l-amino acids to the medium followed by an incubation for 25 minutes in gentle shaker. at the end of incubation process, mitochondria were pelleted by a high-speed centrifugation at 12000g for 10mins at 4°c. the pelleted mitochondria were then suspended in lysis buffer (137mm nacl, 20mm tris-hcl, 50mm edta, 1% np40) with protease inhibitors (2mm na2vo4, 2mm naf, 0.1 pmsf[phenylmethylsulfonylfluoride]) followed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (sds page) (garrido et al., 2008). a 12% resolving gel was used for sds page. after the process of electrophoresis, the gels were stained using coomassie brilliant blue stain for a period of 4 hours. at the end of 4 hours, a de-staining solution was used to remove the excess stain for a period of 2 hours. the gels were then visualised using quantity one software (biorad, california, usa). the band intensity was quantified using imagej software. in vitro mitochondrial dna synthesis the experiment in vitro mitochondrial dna synthesis was carried out according to the procedures earlier by fernandez-silva et al. the isolated mitochondria were briefly suspended in maite buffer (75mm sorbitol, 25mm sucrose, 10mm kcl, 10mm k2hpo4, 0.05mm tris-hcl) at ph 7.4 containing 10mm glutamate, 10mm succinate, 2.5mm malate, 1mm adp and 1mg/ ml of bsa. the process of translation was initiated by adding 50 μm of each dntp to the medium followed by an incubation for 5 hours in gentle shaker. at the https://journals.e-palli.com/home/index.php/ajcp pa ge 11 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 8-20, 2023 end of incubation process, mitochondria were pelleted by a high-speed centrifugation at 12000g for 10mins at 4°c. the pelleted mitochondria were then suspended in lysis buffer (150mm nacl, 20mm tris-hcl, 20mm edta, 1%sds) at ph-8.75 in the presence of 10 μm proteinase k and 10 μm rnase a followed by agarose gel electrophoresis (garrido et al., 2008). mitochondrial dna isolation and dna quantification intact mitochondrial dna was isolated by following the exact experimentation procedure mentioned by martia, et al. phenol, cholorform and isoamyl alcohol was used for biphase separation followed by precipitation of dna by 100% ethanol. the precipitated was washed thrice using 75% ethanol and the dried dna pellet was dissolved in elution buffer ae (5mm tris-hcl) at ph 8.5. the dna sample were then quantified using a nanodrop spectrophotometer by thermo fisher scientific (nanodrop 2000 spectrophotometer) (enriquez et al., 1996). statistical analysis all the statistical analysis involved through the study was carried out using prism version 8 (graph pad software inc., san diego, ca, usa). the data analysis was carried out using two-way analysis of variance (anova), followed by dunnet’s post-test. the experimental results were expressed as mean ± sd, and a p < 0.05 was considered statistically significant. results and discussion 5‐azacytidine preserves the mitochondrial function during ir in an isolated mitochondrial system mitochondrial dysfunction is one of the key events to occur during ischemia reperfusion injury. this has an irreversible impact over the mitochondria even after various therapeutic intervention. so, an ideal therapeutic should overcome permanent mitochondrial dysfunction. mitochondrial integrity with 5-azacytidine pre-treatment was evaluated. from the evaluation we were able to identify that 0.5 μm 5-azacytidine was protective against ir. so, an optimal dose 0.5 μm was fixed as standard throughout all the experimentation procedures involving isolated mitochondria. we further evaluated the electron transport chain enzyme activity to asses the mitochondrial function. the ir groups pre-treated with 5-azacytidine has improved levels of electron transport chain activity. 5-azacytidine pre-treatment in ir preserved mitochondrial electron transport chain complex activity of complex i, ii, iii and iv by %, %, % & % in the mitochondria isolated from heart, when compared to mitochondrial groups subjected to ir. 5-azacytidine pre-treatment in ir preserved mitochondrial electron transport chain complex activity of complex i, ii, iii and iv by %, %, % & % in the mitochondria isolated from kidney, when compared to mitochondrial groups subjected to ir. figure 1: dosage determination: optimal concentration of 5-azacytidine drug was estimated to be 0.5 µm using nqr assay for an isolated mitochondrial system subjected to ir figure 2: effect of 5-azacytidine on mitochondrial electron transport chain complex activity in isolated rat heart mitochondria https://journals.e-palli.com/home/index.php/ajcp pa ge 12 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 8-20, 2023 effect of 5-azacytidine on mitochondrial electron transport chain complex activity in isolated rat heart mitochondria. (a) complex i activity (nqrnadh dehydrogenase) was measured as μmol nadh oxidized/min/mg protein, (b) complex ii activity (sqrsuccinate dehydrogenase) was measured in nmol dcpip reduced/min/mg protein, (c) complex iii activity (qcrcytochrome bc1) was measured in nmol cytochrome c reduced/min/mg protein, and (d) complex iv activity (coxcytochrome c oxidase) was measured in nmol cytochrome c oxidized/ min/mg protein.(5-aza,5-azacytidine; dmso, dimethyl sulfoxide; ir, ischemia-reperfusion.) data were represented as mean ± sd. ( n=6 per group). *p < 0.05 versus ir. figure 3: effect of 5-azacytidine on mitochondrial electron transport chain complex activity in isolated rat kidney mitochondria effect of 5-azacytidine on mitochondrial electron transport chain complex activity in isolated rat kidney mitochondria. (a) complex i activity (nqrnadh dehydrogenase) was measured as μmol nadh oxidized/min/mg protein, (b) complex ii activity (sqrsuccinate dehydrogenase) was measured in nmol dcpip reduced/min/mg protein, (c) complex iii activity (qcrcytochrome bc1) was measured in nmol cytochrome c reduced/min/mg protein, and (d) complex iv activity (coxcytochrome c oxidase) was measured in nmol cytochrome c oxidized/min/mg protein.(5-aza,5-azacytidine; dmso, dimethyl sulfoxide; ir, ischemia-reperfusion.) data were represented as mean ± sd. ( n=6 per group). *p < 0.05 versus ir. effect of 5-azacytidine in alleviating damage induced to mitochondria due to oxidative stress during iri a robust release of free radicals is one of the most critical events involved in ir injury pathology. this is also one of the key contributors to mitochondrial dysfunction. the therapeutic effect of 5-azcytidine in overcoming and altering the damages due oxidative stress induced by ir is vital. mitochondrial oxidative stress parameters were assessed with 5-azacytidine pre-treatment. from the evaluation, the ir groups pre-treated with 5-azacytidine has improved levels of catalase and superoxide dismutase activity by % and % in the mitochondria when compared to mitochondrial groups subjected to ir, isolated from heart. 5-azacytidine showed improved levels of catalase and superoxide dismutase activity by % and % in the mitochondria when compared to mitochondrial groups subjected to ir, isolated from kidney. further experimental evaluation revealed that the ir groups pre-treated with 5-azacytidine has improved levels of gsh:gssg ratio by % in the mitochondria when compared to mitochondrial groups subjected to ir, isolated from heart. 5-azacytidine showed improved levels of gsh:gssg ratio by % in the mitochondria when compared to mitochondrial groups subjected to ir, isolated from kidney. assessment of oxidative stress damage in cardiac mitochondria subjected to 5-azacytidine pretreatment. (a) catalase enzyme activity, (b) superoxide dismutase (sod) enzyme activity, and (c) gsh:gssg activity; all measured in isolated mitochondrial lysates of rat kidneys. values are represented as mean ± sd of six individual animals per group. *p < 0.05 versus ir. abbreviation:5aza,5-azacytidine; dmso, dimethyl sulfoxide; ir, ischemia-reperfusion. https://journals.e-palli.com/home/index.php/ajcp pa ge 13 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 8-20, 2023 assessment of oxidative stress damage in renal mitochondria subjected to 5-azacytidine pre-treatment. (a) catalase enzyme activity, (b) superoxide dismutase (sod) enzyme activity, and (c) gsh:gssg activity; all measured in isolated mitochondrial lysates of rat kidneys.. values are represented as mean ± sd of six individual animals per group. *p < 0.05 versus ir. abbreviation:5aza,5-azacytidine; dmso, dimethyl sulfoxide; ir, ischemia-reperfusion 5-azacytidine pre-treatment overcomes impaired protein translational ability in an isolated mitochondrial system in vitro protein translation ability of mitochondria in an isolated mitochondria system is assessed and visualised in the experiment. epigenetic modification during iri is one of the most reasons for translational inaccuracy. hampered protein synthesis is one of the main reasons behind irreversible recovery from iri. the ameliorative effect of 5-azacytidine in overcoming translational inaccuracy when subjected is explored in our study. the significant improvement in ir groups treated with 5-azacytidine compared to ir groups can be clearly visualised through the electrograms of the sds page gels. mitochondrial translational ability was assessed after 5-azacytidine pre-treatment. before subjecting the mitochondria from heart to in vitro protein translation, the mitochondrial protein was normalised to mg/ml across all groups. after the process of in vitro translation, the ir groups pre-treated with 5-azacytidine showed an increase in mitochondrial protein by % when compared to mitochondrial groups subjected to ir in heart. before subjecting the mitochondria from kidney to in vitro protein translation, the mitochondrial protein was normalised to mg/ml across all groups. after the process of in vitro translation, the ir groups pre-treated with 5-azacytidine showed an increase in mitochondrial protein by % when compared to mitochondrial groups subjected to ir in kidney. figure 4: assessment of oxidative stress damage in cardiac mitochondria subjected to 5-azacytidine pre-treatmen figure 5: assessment of oxidative stress damage in renal mitochondria subjected to 5-azacytidine pre-treatment table 1: quantitative mitochondrial protein estimation by bradfords method in heart tissue before in vitro protein translation s.no group name protein unit 1. normal 0.67333333 mg/ml 2. normal + 5-aza 0.67333333 mg/ml 3. normal + dmso 0.67333333 mg/ml 4. ir 0.67333333 mg/ml 5. ir + 5–aza 0.67333333 mg/ml 6. ir+dmso 0.67333333 mg/ml table 2: quantitative mitochondrial protein estimation by bradfords method in heart tissue after in vitro protein translation s.no group name protein unit 1. normal 0.909 mg/ml 2. normal + 5-aza 0.97 mg/ml 3. normal + dmso 0.612 mg/ml 4. ir 0.42 mg/ml 5. ir + 5–aza 0.873 mg/ml 6. ir+dmso 0.359 mg/ml https://journals.e-palli.com/home/index.php/ajcp pa ge 14 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 8-20, 2023 in-vitro cardiac mitochondrial translated proteins analysis using sds-page gel electrophoresis. (a) sds page gel electrogram, (b) protein estimation, (c) total band intensity, and (d) representations for gel electrogram lanes. values are represented as mean ± sd of six individual animals per group. *p < 0.05 versus ir. (5-aza,5-azacytidine; dmso, dimethyl sulfoxide; ir, ischemia-reperfusion.) figure 6: in-vitro cardiac mitochondrial translated proteins analysis using sds-page gel electrophoresis table 3: quantitative mitochondrial protein estimation by bradfords method in kidney tissue before in vitro protein translation s.no group name protein unit 1. normal 0.94833333 mg/ml 2. normal + 5-aza 0.94833333 mg/ml 3. normal + dmso 0.94833333 mg/ml 4. ir 0.94833333 mg/ml 5. ir + 5–aza 0.94833333 mg/ml 6. ir+dmso 0.94833333 mg/ml table 4: quantitative mitochondrial protein estimation by bradfords method in kidney tissue after in vitro protein translation s.no group name protein unit 1. normal 1.1505 mg/ml 2. normal + 5-aza 1.2545 mg/ml 3. normal + dmso 0.9008 mg/ml 4. ir 0.69 mg/ml 5. ir + 5–aza 1.1896 mg/ml 6. ir+dmso 0.67087 mg/ml in-vitro renal mitochondrial translated proteins analysis using sds-page gel electrophoresis. (a) sds page gel electrogram, (b) protein estimation, (c) total band intensity, and (d) representations for gel electrogram lanes. values are represented as mean ± sd of six individual animals per group. *p < 0.05 versus ir. (5-aza,5-azacytidine; dmso, dimethyl sulfoxide; ir, ischemia-reperfusion.). https://journals.e-palli.com/home/index.php/ajcp pa ge 15 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 8-20, 2023 figure 7: in-vitro renal mitochondrial translated proteins analysis using sds-page gel electrophoresis 5-azacytidine enhances dna replication machinery in an isolated mitochondrial system in vitro dna replication ability of mitochondria in an isolated mitochondria system is assessed and visualised in the experiment. dna hypermethylation during iri is one of the most reasons for transcriptional impairment and translational inaccuracy. upregulation of dnmt1 during ir is one of the major reasons behind extensive and farfetched damage. the therapeutic effect of 5-azacytidine in overcoming impaired dna replication machinery when subjected is explored in our study. the significant enhancement in mitochondrial dna copy number in ir groups treated with 5-azacytidine compared to ir groups can be clearly visualised through the electrograms of the agarose gels. mitochondrial dna replication ability was assessed after 5-azacytidine pre-treatment. before subjecting the mitochondria from heart to in vitro dna replication, the mitochondrial protein was normalised to mg/ml across all groups. after the process of in vitro dna replication, the ir groups pre-treated with 5-azacytidine showed an increase in mitochondrial dna by % when compared to mitochondrial groups subjected to ir in heart. before subjecting the mitochondria from kidney to in vitro dna replication, the mitochondrial protein was normalised to mg/ml across all groups. after the process of in vitro dna replication, the ir groups pre-treated with 5-azacytidine showed an increase in mitochondrial dna by % when compared to mitochondrial groups subjected to ir in kidney. table 5: quantitative mitochondrial protein estimation by bradfords method in heart tissue before in vitro dna replication s.no group name protein unit 1. normal 0.763 mg/ml 2. normal + 5-aza 0.763 mg/ml 3. normal + dmso 0.763 mg/ml 4. ir 0.763 mg/ml 5. ir + 5–aza 0.763 mg/ml 6. ir+dmso 0.763 mg/ml table 6: mitochondrial dna quantification in heart using nanodrop spectrophotometer after in vitro dna replication s.no group name nucleic acid unit 260/280 260/230 sample type 1. normal 578.25 ng/µl 1.81 2.12 dna 2. normal + 5-aza 583.14 ng/µl 1.82 2.14 dna 3. normal + dmso 462.69 ng/µl 1.9 2.11 dna 4. ir 258.18 ng/µl 1.9 2.12 dna 5. ir + 5–aza 565.82 ng/µl 1.89 2.17 dna 6. ir+dmso 264.28 ng/µl 1.83 2.0 dna https://journals.e-palli.com/home/index.php/ajcp pa ge 16 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 8-20, 2023 figure 8: agarose gel electrophoresis analysis of in-vitro cardiac mitochondrial dna replication agarose gel electrophoresis analysis of in-vitro cardiac mitochondrial dna replication. (a) agarose gel electrogram, (b) total band intensity, and (c) representations for gel electrogram lanes. data were represented as mean ± sd. ( n=6 per group). *p < 0.05 versus ir. abbreviation:5-aza,5-azacytidine; dmso, dimethyl sulfoxide; ir, ischemia-reperfusion. agarose gel electrophoresis analysis of in-vitro renal mitochondrial dna replication. (a) agarose gel electrogram, (b) total band intensity, table 7: quantitative mitochondrial protein estimation by bradfords method in kidney tissue before in vitro dna replication s.no group name protein unit 1. normal 0.984 mg/ml 2. normal + 5-aza 0.984 mg/ml 3. normal + dmso 0.984 mg/ml 4. ir 0.984 mg/ml 5. ir + 5–aza 0.984 mg/ml 6. ir+dmso 0.984 mg/ml table 8: mitochondrial dna quantification in kidney using nanodrop spectrophotometer after in vitro dna replication s.no group name nucleic acid unit 260/280 260/230 sample type 1. normal 578.25 ng/µl 1.89 2.0 dna 2. normal + 5-aza 583.14 ng/µl 1.82 2.1 dna 3. normal + dmso 462.69 ng/µl 1.91 2.1 dna 4. ir 258.18 ng/µl 1.8 2.12 dna 5. ir + 5–aza 565.82 ng/µl 1.83 2.17 dna 6. ir+dmso 264.28 ng/µl 1.9 2.0 dna figure 9: agarose gel electrophoresis analysis of in-vitro renal mitochondrial dna replication https://journals.e-palli.com/home/index.php/ajcp pa ge 17 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 8-20, 2023 and (c) representations for gel electrogram lanes. data were represented as mean ± sd. ( n=6 per group). *p < 0.05 versus ir. abbreviation:5-aza,5-azacytidine; dmso, dimethyl sulfoxide; ir, ischemia-reperfusion discussion ischemia reperfusion injury is one of the most common and unavoidable forms of injury to occur during a revascularisation procedure or because of therapeutic interventions to an atherosclerotic plaque or an ischemic stroke (piper et al., 1998). the injury might seem inevitable because of huge number of variable complications arising after interventional therapy. till date, a lot of different therapeutics have been tested, repurposed and trailed for the treatment of iri but a lot of them fails in one or many concern. in previous study by rahavi, et al., has reported the ameliorative effect of 5-azacytidine against iri preliminarily due to its inherent ability to combat the dnmt1 by inhibiting this action (a.a.mangoni et al. 2018). the role of 5-azacytinde in combating cancer is also well explored. but, the role of 5-azacytidine in alleviating mitochondrial damage due to iri is not explored. as mitochondria is one of the key players in regulation and resuscitating cell survival during iri, it is of immense importance to have a deep understanding of the mechanism through which 5-azacytidine helps alleviating iri. the mechanistic action on drug on mitochondria is not well documented and is unexplored. so, having a better understanding of the way through which mitochondria mediates ir protection would help enable a complete overlay (d.jain et al. 2017). even though there are multiple drugs to treat iri, such multifaceted view on the action of drug is absent thereby most drugs fail to transition even into clinical trials. therefore, phrenological manipulative ability without enough substantial evidence is not fruitful yield (i.andreadou et al. 2020). many drugs like methotrexate, hydroxychloroquine which are used for the treatment of various diseases like cancer, malaria have been repurposed to treat cardiovascular diseases as well (a.daiver et al. 2021). 5-azacytidine mediated epigenetic reprogramming is used extensively in cancer research. the mechanism of action and epigenetic modulation mediated by 5-azacytidine is clearly established and well documented (e.hervout et al. 2013). also, many studies have reported the cardioprotective, nephroprotective and vasculo-protective nature of the compound. but the effect of this compound on cardiac iri and renal iri are not well established (s.sou et al, 2016). therefore, a proper evidence-based study would help us repurpose the drug effectively to manage many cardiovascular and renal complications in a clinical scenario as a potent pharmacological agent (l.badimon et al. 2019). from our study, 5-azacytidne has been to shown to provide an ameliorative effect against iri in an isolated mitochondrial system thereby, providing a promising role as a pharmacological intervention. the organs heart and kidney have been shown to exhibit an elevated levels of mitochondrial dysfunction because of the free radical stack and extensive calcium overload (h.ma et al. 2011). this immediately results in the change in functionality of the mitochondria leading to loss of mitochondrial bioenergetics (k.yang et al. 2017). based on these evidences, we identified a significantly decreased mitochondrial electron transport chain activity due to deterioration in function of mitochondrial complexes i, ii, iii, iv. there was also decreased tendency to scavenge the free radical resulting increased oxidative stress. our studies have shown that pre-treatment with 5-azacytidine resulted in improved mitochondrial electron transport chain activity. the results have also indicated that the scavenging effect of 5-azacytidine but it was not that considerable when compared to its other therapeutic effect. however, the compound enhanced the survival key for mitochondria when subjected to iri by considerably enhancing the mitochondrial functionality and improved resistance to oxidative stress induced mitochondrial damage. translational inaccuracy and reduced mitochondrial copy number are another key deteriorative feature of iri. so, a pharmacological agent that has the ability to modify the epigenetics of a cellular system is vital (p.e.nikolaou et al. 2019). often times we fail to realise that mitochondria is a system of its own and it has a trivial role to play in cellular epigenetics as well (b.a.hemmings et al. 2015). when subjected to iri, the levels of mitochondrial protein translation were significantly reduced and it also resulted in decreased mitochondrial copy number. however, once the groups were pre-treated with 5-azacytidine and when subjected to iri hey showed improved to resistance to translational inaccuracy by enhanced protein synthesis. the epigenetic modification that 5-azacytidine imparts on the dna replication machinery also resulted improved mitochondrial copy number even when to iri. based on the results obtained from these independent experiments throughout the long run we could demonstrate the therapeutic effect of 5-azacytidine to attenuate the ramifying repercussions of ischemia-reperfusion injury on mitochondrial molecular machinery. conclusion ischemia reperfusion injury mediated mitochondrial damage is inevitable and without proper pharmacological intervention it can be irreversible. this study shows the therapeutic effect of 5-azacytidine in attenuating iri mediated mitochondrial dysfunction. 5-azacytidine pre-treated groups showed improved mitochondrial translational accuracy and increased mitochondrial copy number. these evidences indicated the rooted ability of 5-azacytidine to act on mitochondria to ameliorate iri induced mitochondrial damage and thereby contributing towards cellular homeostasis. future prospects from our findings pre-treatment with 5azacytidine has enhanced the ability of protein translation, increased https://journals.e-palli.com/home/index.php/ajcp pa ge 18 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(1) 8-20, 2023 mitochondrial copy number and have restored the damage caused to mitochondrial function when subjected to ischemia reperfusion injury in an isolated mitochondrial system. an isolated mitochondrial system acts as great tool in understanding the specificity, targeted action and therapeutic potential of 5-azacytidine. however, this system doesn’t exactly replicate environment and interactions of an organ system, whereas it replicates the environment of the cell to sustain a healthy mitochondria population. so, extensive animal studies consisting animals pre-treated with 5-azacytidine are required to further evaluate the effect of 5-azacytidine on mitochondria wherein a lot of other factors can interplay to contribute, impair or enhance the therapeutic effect. also, the effect of 5-azacytinde on mitochondrial gene expression associated with ischemia reperfusion and need to be studied. furthermore, studies can also be extended to analyse the efficacy of 5-azacytidine in attenuating any repercussions on distant organ due to ischemia reperfusion injury. acknowledgements funding: we would like to acknowledge the department of science and technology (dst), government of india, new delhi, for funding the instrumentation facility required for research through grand‐in‐aid (emr/2017/000669). references andreadou, i., schulz, r., papapetropoulos, a., turan, b., ytrehus, k., ferdinandy, p., daiber, a., & di lisa, f. 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(2017). the key roles of gsk-3β in regulating mitochondrial activity. cellular physiology and biochemistry. international journal of experimental cellular physiology, biochemistry, and pharmacology, 44(4), 1445–1459. https://doi. org/10.1159/000485580 https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 26 american journal of chemistry and pharmacy (ajcp) magnitude and associated factors for sexually transmitted infections among hawassa industrial park workers, southern ethiopia solomon tesfaye doelaso1* volume 1 issue 1, year 2022 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v1i1.431 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: july 11, 2022 accepted: july 18, 2022 published: july 20, 2022 sexually transmitted infections (sti) are a group of infectious diseases spread through unprotected sexual intercourses, resulting in curable and incurable diseases. the syndrome is a variety of clinical signs and symptoms caused by pathogens that can be acquired and transmitted through sexual activity. around the world estimated about 376 million people become infected each year with one of four common curable stis where about 86 million new cases of curable stis occurred in the african region. the objective of this study to assess the magnitude and associated factors for sexually transmitted infections in 2020 among hawassa industrial park workers, southern ethiopia. the multistage sampling technique was used to select a total of 663 study participants. the data were entered into the epi-data version 4.4 and analyzed by spss version 21. both bivariate and multivariable logistic regression analyses were employ p-values of <0.05 and aor with 95%ci was used to determine the presence of association between covariates and dependent variable.. the self-reported sti magnitude in the last 12 months was 18.7% (18.54-18.91) among the hawassa industry park workers. hometown residence [aor=2.29; 95%ci: 1.34-3.92], drinking alcohol (aor=3.26; 95%ci: 1.74-6.09), view/read pornography (aor=4.38; 95%ci: 2.67-7.18) and poor knowledge (aor=2.69; 95%ci: 1.65-4.420 were significantly associated with the magnitude of stis. the self-reported magnitude of stis among hawassa industrial park was found to be high. keywords stis, associated factors, syndromes, risky sexual practices, hip workers 1 sidama regional state health department, ethiopia. * corresponding author’s e-mail: gersam2016@gmail.com introduction sexually transmitted infections (stis) are a group of infectious diseases transmitted through unsafe sexual intercourse as the primary mode of transmission. (workowski k, 2015). please follow the referencing style of the journal (apa 6/7th edition). the organisms causing stis can also be spread through other routes such as blood transfusion, tissue transfer, skin-to-skin sexual contact, mother-to-child, and blood or blood products (who, 2014) (fmoh, 2015). to date more than 30 pathogens; bacterial, viral, fungus, protozoa and ectoparasites have been identified that can be transmitted through sexual intercourse. commonly known curable sis is syphilis, gonorrhea, chlamydia, trichomoniasis, chancroid, lymph granuloma venereum and donovan’s. the stis that are preventable, but not curable are the viral stis which includes hiv, human papilloma virus, hepatitis b virus and herpes simplex virus (de schryver a ma, 1990). conventionally, a supposed stis has been diagnosed by either clinical appearance alone, which is often inaccurate and incomplete or a laboratory-based test, which is complex, very expensive, and commonly delay treatment (fmoh, 2015). also, clinical diagnosis of several stis is problematic due to broad-based signs and symptoms. etiologic-confirmed diagnosis is scientific, but the service is often unreachable in many developing countries or may be located in urban centers (sahu l mp, 2005). another strategy, the syndromic approach, offers an alternative approach, which depends on the ability to identify and treat the syndromes caused by an sti, with minimal or no requirement for laboratory diagnostic support (tesfaye f et al., 2000) (mehul t et al., 2013). instead, it is based on the identification of a group of signs and symptoms associated with a number of well-known etiologies approach that can be attained and spread through sexual intercourse. treatment is provided for the majority of the etiologies locally responsible for the syndrome (de schryver a ma, 1990). commonly known sti syndromes are: urethral discharge in men, genital ulcer/sores, vaginal discharge, inguinal bubo, scrotal swelling, lower abdominal pain in women and neonatal conjunctivitis (fmoh, 2015) (csa, 2017). also, it is inclusive approach because in addition to the provision of treatment, it includes: patient education, condom supply, counseling, partner notification and management, and hiv testing and counseling (fmoh, 2015). according to the who 2019 estimate around the world, about 376 million people become ill each year with one of four common stis: syphilis, gonorrhoea, chlamydia and trichomoniasis. it is projected about 86 million new cases of curable stis occurred in africa regions (who, 2019). in developing countries prevalence and their complications are amongst the top five disease groups for which adults seek health care (mehul t et al., 2013). their burden in sub-saharan african countries is very high. it is estimated that 80-90% of the global burden of stis found in the developing countries because there is inadequate or inaccessible diagnosis and treatment facilities (who, 2007). however, large scale up of health care investments and strategies applied for the prevention and treatment of stis in ethiopia the issue continues https://doi.org/10.54536/ajcp.v1i1.431 https://journals.e-palli.com/home/index.php/ajcp mailto:gersam2016%40gmail.com?subject= pa ge 27 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 26-36, 2022 to be a young adults’ health problem. the prevalence of stis rises from 1.15% in 2005 to 4% in 2016 among 1549 years old (csa, 2017). its magnitude, health and socio-economic impacts are still not well known due to a combination of social stigma, under-reporting, asymptomatic nature of infections and lack of diagnostic facilities (who, 2013). the reason for high prevalence is risky sexual practices (rsp) and low use of preventive services among young age groupies. the health of these group is a key element for social and economic development of the country, overlooking the srh of these groupies can lead to high social and economic costs, both immediately and in the years ahead (who, 2007). majority of stis are asymptomatic and only part of the symptomatic population seeks health care and even a smaller number of cases are reported. which can lead to the development of serious complications with severe consequences for the individuals and community (mehul t et al., 2013). about 70-80% of infected women are asymptomatic and so do not seek treatment. they are risky for complications and possibly infecting others (fmoh, 2015). it may increase the risk of hiv acquisition, motherto-child transmission of stis can result in adverse pregnancy outcomes. also, late treatment or untreated infection can lead to pelvic inflammatory disease, ectopic pregnancy, infertility and cervical cancer (who, 2014). the association between stis and hiv has an epidemiologic interaction and share the same risk factors. so, it is very critical to strengthen sti prevention and control program not only to improve quality of life and control their complications, but also to prevent the spread of hiv infection (fmoh, 2015) (mehul t et al., 2013). counselling can improve people’s ability to recognize the syndromes of stis, and increase the likelihood they will seek care early. as well, people seeking treatment for stis also face several challenges including limited resources, social stigma, poor quality services, and no followup of sexual partners (who, 2014). although sexual abstinence is the preferred method for sti prevention among young age group, it is not practical for many of the individuals. therefore, consistent and correct condom use is a rational method of preventing the transmission stis (who, 2007). in hawassa city, urbanization and rural-urban migration is a growing phenomenon for a job opportunity that set for younger age groups to employ at the newly established hawassa industrial park (hip), that predisposes them to high risk for stis. report from symptomic presentation of stis among people living in rural areas of lucknow shows that about 11.18% were diagnosed stis based on syndromic approaches (mishra s et al., 2016). another community-based survey on stis associated symptoms and health-seeking behaviors among iranian adults shows that, 39.9% of women and 17.6% of men reported having at least one sti associated syndromes under consideration at the time of study (nasirian m et al., 2015). according to ethiopian demographic and health survey 2016, overall 4% of women and men age 15-49 reported having syndromes of sti in the last 12 months (csa, 2017). other studies conducted among wolaita sodo university and university of gonder students shows that 12 month period self-reported prevalence of stis to be 19.5% and 18.2% respectively (yohannes b et al., 2013) (ayanaw b et al., 2019) . similar studies conducted among high school students at bahir-dar and hiv patients in ayder referral hospital, northern ethiopia reported that the prevalence of stis is 13.1% and 8.5% reported to have syndromes of stis respectively (gebremichael h et al., 2017) (gebrelibanos a et al., 2015). study conducted among young women in northern ethiopia shows that self-reported prevalence of stis in the last 12 months was 21.3% (fisseha g ae, 2015). surveillance conducted in ethiopia in 8 health facilities located in amhara, oromia and addis ababa reported that commonest syndrome was vaginal discharge 50%, urethral discharge 31%, genital ulcer/sores 9%, lower abdominal pain 7.3%, and two syndrome were present in few patients 3% (fmoh, 2015). national survey in new caledonia among youths shows that hometown residence showed significant association with the prevalence of the stis (corsenac p et al., 2012). another study at debre birhan high school students shows that, residence being urban had positive association with knowledge of syndromes of stis (adera a, et al., 2015). similarly, living with families and relatives helps the parent/relatives to monitor their children and improve decision-making capacity on rsps (bettinger j et al., 2004). another study conducted among young people in south africa shows that, study subjects aged 20-24 years and women had more than twice the odds of having a curable sti compared to study participants aged 15-19 years and men, respectively (francis s et al., 2018). also, divorced individuals were about 5 times more likely to have had risky sexual activities than married individuals (kassa m et al., 2013). one study among seasonal migrant workers in metema, northwest ethiopia shows that study subjects who got a daily income above usd 5.00 were 2.2 times more likely to have rsps during the prior 6 months than those having a daily income lower than usd 5.00 (tiruneh k et al., 2015). migrants were frequently forced into physically demanding works with poor living conditions and little benefits, while having to live apart from their partners and families, may develop new sexual partners and involve in high rsps that may increase the chance of stis (tiruneh k et al., 2015). study among people living in rural area of lucknow shows that significant association between educational status and prevalence of stis (mishra s et al., 2016). respondents whose educational level less than fourth grade were 12 times more likely to be engaged in rsps than whose educational level tenth grade or more (kassa m et al., 2013). similarly occupation and social classes were also marked as significant socio-economic factors https://journals.e-palli.com/home/index.php/ajcp pa ge 28 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 26-36, 2022 associated with the prevalence of the stis (mishra s et al., 2016). having good knowledge on modes of transmission, prevention methods and complications of stis helps to protect themselves from stis and its complication (ayanaw b et al., 2019). also, many studies shows that those who have poor knowledge of sti were more likely to have stis than students with good knowledge (yohannes b et al., 2013) (ayanaw b et al., 2019) . adverse impacts associated with substance use include, increase the chance of unprotected sex, decrease the selection of sexual partners, increase the number of sexual partners and careless sexual activity. such combinations significantly increase their vulnerability to the stis (winters k., 1999). study shows that overall, 53.8% of them used at least one substance in lifetime. commonly used substances are: alcohol 41.7%, khat 30.3%, cigarette 11.3% and illicit substances 3.9% (derese a et al., 2014). alcohol use found significantly associated with rsps including unprotected sex, multiple sexual partners (msps), paying for sex and selling sex that increases likelihoods of stis (weiser s et al., 2006). those using alcohol having about 3 times higher chances of rsps compared to those not using it (alemu a et al., 2015). khat chewer were 3.4 times more likely to have rsps than those who didn’t chew khat. moreover, those who smoke shisha were 3.44 times more likely to have risky sexual behaviour as compared to those who didn’t smoke shisha (gizaw a et al., 2014). different reasons were stated for the substance use such as, to get personal pleasure, increase work performance, peer pressure, to get relief from tension, to be sociable and for other reasons (alemu a et al., 2015). view/read pornographic materials could change the normal sexual desire and care taking of exposing to rsps (ayanaw b et al., 2019). those who attend sex films, movies frequently were about 2 times more likely to be involved in rsps than those who didn’t view/read pornographic materials (henok a et al., 2015). risky sexual practice is any human sexual practices which put individual’s physical, social and psychological health at the risk (malhotra s et al., 2008). it also includes early sexual practices, unprotected sexual intercourse, and msps occur in a wider context. although rsps doesn’t always shows a high-risk lifestyle, it often clusters with other risky behaviors such as substance use and violence participation (blum r mk, 2005). earl age sexual initiation was significantly associated with increased risk of stis (upchurch d et al., 2004). those who first sexual start from 15-19 years were about 3 times more likely to have had rsps than from 20-24 years (kassa m et al., 2013). and those who have no sexual partner currently was riskier sexually practices than those who had current partner currently (kassa m et al., 2013). studies conducted among wolaita sodo and madawulabu university students revealed that those who had msps in a lifetime were more likely to have stis than those who had one sexual partner in the life time (yohannes b et al., 2013) (setegn t et al., 2013). similar studies conducted among school youth at bahirdar and young women in northern ethiopia revealed that those who had msps in the last 12 months had higher chances of experiencing stis compared to those with one partner (gebremichael h et al., 2017) (fisseha g, 2015). one study conducted among migrant daily laborers in metema district show that isolated work sites lead to a lack of social cohesion and social norms governing behavior of workers, which may lead to engagement in rsps, about 68% of sexually active respondents reported non-marital sexual intercourse in the last 6 months (tiruneh k et al., 2015). studies conducted in wolaita sodo university students and young women in northern ethiopia, those who had never used condom during intercourse had higher odds of experiencing stis than those who had ever used condom in their lifetime (yohannes b et al., 2013) (fisseha g, 2015). the most common reasons stated for not using at all or inconsistently using condom are partners refusal, condom inaccessibility, trust of partner, forgetfulness after alcohol intake, religious prohibition, hurry to have sex and additional reasons for not using condoms (alemu a et al., 2015) (abeje a aa, 2017). also, those who reported themselves to have had extra-marital sexual contacts in the last year had higher prevalence of sti (rostami f et al., 2017). people who experienced a syndrome of sti may delay to seek care timely or do not seek care despite the available service. health-seeking behavior affects people’s actions when they suspect an infection (voeten h et al., 2004). perceived severity of sti was also one of the predictor factors for early health care-seeking behavior (tsadik m et al., 2019). according to edhs 2016 report only 32% of those who had syndromes sti sought advice or treatment (csa, 2017). the most important reasons for not receiving treatment by study subjects were feeling guilty of telling problem to the health worker, thinking symptom as incurable, thinking symptom not serious, thinking symptom as curable without treatment, lack of money, not knowing where to get treatment, and others including lack of time (yohannes b et al., 2013). methods and materials study design and period institution based cross-sectional study was conducted among hawassa industrial park workers from july 26 to august 26, 2020. self-reported syndromic approach was used to assess magnitude and associated factors for stis among hip workers. source and study population source population was hip workers and workers in randomly selected factory sheds during the study period were study population. inclusion and exclusion criteria all hip workers were included, and hip workers who are severely ill during study period and recruitment time less https://journals.e-palli.com/home/index.php/ajcp pa ge 29 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 26-36, 2022 than 30 days were excluded. operational definitions early sexual initiation: having sexual intercourse before the age of 18 years (csa, 2017). risky sexual practices: workers who have at least one of the following: inconsistent condom use with non-regular partner, having multiple sexual partners, starting sexual intercourse before age of 18 years and sexual intercourse with csws. knowledge about sti: it was measured using a series of 36 knowledge questions about stis. those who scored mean and above were taken as good knowledgeable. sti cases for male/female: they were considered stis cases if he/she reported at least one of history of urethral discharge, genital ulcer/sores, scrotal swelling, inguinal bubo, abnormal vaginal discharge, or lower abdominal pain syndromes in the last 12 months. sexually active: workers who had sexual intercourse at least once prior to the study. sample size determination the sample size was determined using single population proportion formula by taking the prevalence of stis (50%) because absence of similar studies among industry workers at national level. n= minimum sample size z = standardized normal distribution value for the 95%cl, which is (1.96). p= prevalence of stis (50%). d= margin of error 5% (0.05) design effect of 1.5 non-response rate of 15% nf = final sample size using population proportion formula: n= z2 a/2 p(1-p)/d2 n= (1.96)2 (0.5) (0.5)/(0.05)2 n =3.84×0.25/0.0025 =384 n= 384, multiplied by design effect of 1.5 and nonresponse rate (15%) was added final sample size (nf) = [(384×1.5) + 15%] =663 sampling technique and procedures there are 52 sheds in hip and about 30,000 regular workers within it. multistage sampling technique was used to select representative study subjects. simple random sampling technique (lottery method) was used to select the factory shed from the total of 52 sheds by taking 29% (15 sheds) of the total factory sheds. to assure the representativeness of the data, the sample size was proportionally allocated to all (15 sheds) proportional to their number of workers. finally, the study participants were selected from factory sheds using a simple random sampling technique by using attendance sheet. data collection procedures data was collected using structured self-administered questionnaires. the questionnaire was first prepared in english, and translated into amharic then, translated back into english, to check the consistency. data was collected using a structured questionnaire with open and closed end questions. data quality management one week prior to data collection a pre-test was conducted on 5% (setegn t et al., 2013) of the sample size at moha soft drink factory in hawassa. depending on the result of pretest, correction and modification were done on questionnaire before applying on the study population. data processing and analysis data was cleaned, coded and entered in spss version 21.0 software for further analysis. frequencies and cross tabulations were used to summarize descriptive statistics of the data. tables and texts were used for data presentation. bivariable logistic regression analysis was used to identify candidate variables for multivariable logistic regression at p-value of ≤0.25. the strength of association was determined using multivariable logistic regression at p-value <0.05, and 95% ci of adjusted odd ratio (aor). results and discussion socio-demographic and economic characteristics in this study, 657 study participants were involved in the study making a response rate of (99 %). the mean age of study participants was 27 years (sd ± 3.2 years). among study participants 117 (17.8%) were male and 540 (82.2%) were females, 530 (80.7%) were single in marital status, and 363 (55.3%) were protestant. about 452 (68.8%) respondents stayed for more than one year at hip. the mean income of the respondents was 2350 etb (sd ±1183) and 405 (61.6%) were from rural residential. and about 298 (45.4%) of the respondents have secondary educational attainment. knowledge of respondents about stis the overall knowledge of study participants shows that, about 323(49.2%) of the study participants had good knowledge of stis. about 651(99.1%) respondents have heard about stis. about 641 (97.6%) of the respondents know at least one mode of sti transmission. among the total of 657 study participants 640 (97.4%) were known at least one preventive methods of sti. regarding the syndromes of stis, 640 (97.4%) of the study participants know how stis manifest. about 216 (32.9%) respondents know asymptomatic case transmits stis. among total respondents about 54 (90.4%) study subjects reported that early treatment has benefit, and 490 (74.6%) study participants know that sti have complication. non sexual behavioral characteristics respondents were asked their experience of non-sexual risky practices to assess their exposure to substances and pornographic materials; and the findings shows that, about 96 (14.6%) drink alcohol, 67 (10.2%) chew khat, https://journals.e-palli.com/home/index.php/ajcp pa ge 30 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 26-36, 2022 table 1: socio-demographic characteristics of hip workers, southern ethiopia, 2020. variables frequency percent sex male 117 17.8 female 540 82.2 age 18-22 59 9.0 23-27 306 46.6 28-32 268 40.8 33-37 24 3.6 length of stay at hip less than 6 months 37 5.6 6-12 months 168 25.6 greater than 12 months 452 68.8 religion protestant 363 55.3 orthodox 184 28.0 muslim 67 10.2 catholic 43 6.5 ethnic groups sidama 350 53.3 oromo 96 14.6 amhara 85 12.9 wolayita 62 9.4 kambata 35 5.3 other* 29 4.5 marital status single 530 80.7 married 112 17.0 divorced 15 2.3 living arrangement living alone 234 35.6 friends 199 30.3 wife or husband 108 16.4 family 76 11.6 relatives 40 6.1 income of respondents 450-1000 25 3.8 1001-1500 208 31.7 1501-5000 379 57.7 5001-6500 45 6.8 hometown residence urban 252 38.4 rural 405 61.6 education status read and write 20 3.0 primary 92 14.0 secondary 298 45.4 247 37.6 *others: hadiya, silte, gurage, halaba table 2: knowledge of sti among hip workers, southern ethiopia, 2020 (n=657) frequency percent heard about sti yes 651 99.1 no 6 0.9 common curable sti syphilis yes 620 94.4 no 37 5.6 gonorrhea yes 536 81.6 no 121 18.4 chlamydia yes 155 23.6 no 502 76.4 trichomoniasis yes 105 16.0 no 552 84.0 transmission routes of sti yes 641 97.6 no 16 2.4 routes of sti transmission sexual intercourse yes 641 97.6 no 16 2.4 blood/blood product yes 343 52.2 no 314 47.8 https://journals.e-palli.com/home/index.php/ajcp pa ge 31 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 26-36, 2022 sharp materials yes 284 43.2 no 373 56.8 mother to child yes 396 60.3 no 261 39.7 sti prevention methods yes 640 97.4 no 17 2.6 sti prevention methods abstinence yes 631 96.0 no 26 4.0 condom use yes 487 74.1 no 170 25.9 faithful partner yes 380 57.8 no 277 42.2 sti syndromes yes 640 97.4 no 17 2.6 common syndromes of sti genital ulcer or sores yes 460 70.0 no 197 30.0 urethral discharge in male yes 589 89.6 no 68 10.4 vaginal discharge yes 608 92.5 no 49 7.5 lower abdominal pain in women yes 357 54.3 no 300 45.7 scrotal swelling yes 317 48.2 no 340 51.8 inguinal bubo yes 145 22.1 no 512 77.9 neonatal conjunctivitis yes 153 23.3 no 504 76.7 asymptomatic case transmits sti yes 216 32.9 no 441 67.1 early treatment has benefit for sti yes 594 90.4 no 63 9.6 sti has complications yes 490 74.6 no 167 25.4 complications of sti still birth yes 375 57.1 no 282 42.9 abortion yes 438 66.7 no 219 33.3 infertility yes 281 42.8 no 376 57.2 ectopic pregnancy yes 229 34.9 no 428 65.1 increase hiv transmission yes 240 36.5 no 417 63.5 some kind of cancer yes 165 25.1 492 74.9 absolute treatment for sti yes 619 94.2 no 38 5.8 place of absolute treatment for sti available hospital yes 556 84.6 no 101 15.4 health center yes 432 65.8 no 225 34.2 family guidance association yes 281 42.8 no 376 57.2 private health centers yes 233 35.5 424 64.5 overall knowledge of sti poor 334 50.8 good 323 49.2 https://journals.e-palli.com/home/index.php/ajcp pa ge 32 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 26-36, 2022 table 3: non-sexual behavioral characteristics of hip workers, southern ethiopia, 2020 variables frequency percent chew khat yes 67 10.2 no 590 89.8 khat chewing frequency daily 6 9.0 more than once in a week 23 34.3 weekly 31 46.3 monthly and above 7 10.4 drink alcohol yes 96 14.6 no 561 85.4 alcohol drinking frequency daily 25 26.0 more than once in a week 18 18.8 weekly 21 21.9 monthly and above 32 33.3 shisha smoke yes 16 2.4 no 641 97.6 shisha smoking frequency more than once in a week 3 18.8 weekly 3 18.8 monthly and above 10 62.5 over all substance use yes 126 19.2 no 531 80.8 reason for substance use (n=126) satisfaction 76 60.3 work hard 13 10.3 peer pressure 20 15.9 relief tension 17 13.5 view/read pornographic materials yes 185 28.2 no 472 71.8 pornography types mobile video 99 53.5 internet 51 27.6 movies or television 31 16.8 reading materials 4 2.2 and 16 (2.4%) smoke shisha in the last 12 months. overall, 126 (19.2%) of them used at least one substance in the last 12 months. about 185 (28.2%) of study participants view/read pornographic materials in the last 12 months. sexual behavior of the respondents about 647 (98.5%) respondents had a history of sexual intercourse in life time. the mean age reported at first sexual intercourse was 18.8 (± 2.7) years. about 577 (89.2%) of the study participants has msps in lifetime. only 119 (28.4%) sexually active study participants used a condom for every sexual intercourse in lifetime. during the last 12 months, 453 (73.2%) of study participants reported to have msps, and 398 (64.3%) didn’t used a condom during sexual intercourse. about 29 (4.7%) had sexual intercourse for the benefit/gift in the last 12 months. self-reported sti syndromes and health-seeking behavior the number of respondents who have a syndrome of sti in a lifetime was 182 (28.1%) with (95%ci; 28.0328.72) and among them 121 (18.7%) with (95%ci; 18.54 18.91) were within the last 12 months among sexually active study participants. out of these 15 (12.4%) were males and 106 (87.6%) were females. factors associated with sexually transmitted infections bivariate and multivariable analysis were applied to identify the factors significantly associated with the magnitude of stis. the bivariate analysis shows that, sex of respondents, marital status, hometown residence, monthly income, education status, drinking alcohol, view/read pornographic materials, age at first sex, didn’t use condom ever, msps in the last 12 months, didn’t use condom in the last 12 months, no current sexual partner, sex for the benefit/gift and poor knowledge of stis found to have a statistically significant association with the magnitude of stis. variables such as; sex of respondents, marital status, hometown residence, monthly income, education status, drinking alcohol, view/read pornographic materials, current sexual partner and knowledge of stis were considered in multivariate logistic regression analysis. this study shows that 121 (18.7%) with (95%ci; 18.5418.91) of sexually active hip workers had self-reported stis syndromes in the last 12 months. the finding of this study is comparable with studies conducted among wolaita sodo university and university of gondar students, 19.5% and 18.2% respectively (yohannes b et al., 2013) (ayanaw b et al., 2019). however, it is somewhat lower than studies conducted among young women in northern ethiopia 21.3% (fisseha g, 2015). this difference could be due to the study subjects of northern https://journals.e-palli.com/home/index.php/ajcp pa ge 33 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 26-36, 2022 table 4: sexual practices among hip workers, southern ethiopia, 2020 (n=657) variables frequency percent ever had sexual intercourse(n=657) yes 647 98.5 no 10 1.5 extra-marital sex(n=112) yes 5 4.5 no 107 95.5 age at first sex(n=647) < 18 324 50.1 ≥18 323 49.9 no. of life time sexual partners(n=647) 1 70 10.8 ≥2 577 89.2 condom ever used(n=647) yes 419 64.8 no 228 35.2 condom use frequency(n=419) always 119 28.4 mostly 141 33.7 sometimes 159 37.9 reason for not use condom always (n=528) trust partner 222 42.0 partner refuse 127 24.1 i didn't get it 73 13.8 i dislike it 70 13.3 ashamed to buy 36 6.8 sexual intercourse in the last 12 months(n=647) yes 619 95.7 no 28 4.3 no. of sexual partners in the last 12months(n=619) 1 166 26.8 ≥2 453 73.2 condom use in the last 12 months(n=619) yes 221 35.7 no 398 64.3 having current sexual partners(n=657) yes 254 38.7 no 403 61.3 having sex for the benefit/gift(n=619) yes 29 4.7 no 590 95.3 sex after substance use(n=619) yes 13 2.1 no 606 97.9 sex with csws(n=117) yes 12 10.3 no 105 89.7 condom use with csws(n=12) yes 8 66.7 no 4 33.3 ethiopia were only women and selection was from health facilities there is a high chance to find suspected cases. while, the finding from this study is higher when compared with the edhs 2016 national report 4% (csa, 2017) people living in rural areas of lucknow 11.18% (mishra s et al., 2016), population based survey in the city of são paulo, brazil 6.3% (monteiro v et al., 2016) and school youths at bahir-dar 13.1% (gebremichael h et al., 2017). this could be due to edhs survey, people living in rural areas of lucknow and population-based survey in brazil was community-based, in which most study subjects could be all age groups and also may be differences in data collection method. the difference from study conducted among school youth in bahir dar may be due to differences in age group, living condition and work habit of the hip workers. this study shows that those workers with poor knowledge of stis were 2.7 times more likely to develop stis than workers with good knowledge of stis. the finding consistent with other studies conducted at the wolaita sodo university 4.8 times and university of gondar students 3.3 times more risky than good knowledge of sti (yohannes b et al., 2013) (ayanaw b et al., 2019). overall, 334 (50.8%) of the study subjects have poor knowledge on sti. this finding was consistent with a studies conducted among university of gondar students 55.3%, young women in northern ethiopia 40.4% and madawalabu university students 57.5% have poor knowledge of sti (ayanaw b et al., 2019) (fisseha g ae, 2015) (setegn t et al., 2013). this study indicated that having drunk alcohol statistically significant association with risks of stis. those workers who had drunk alcohol were about 3.2 times more likely to have risky for stis when compared to those who didn’t drink alcohol. the finding of this studies was consistent with study conducted among the female partners of inmates in brazil, alcohol drinkers were 1.7 times more likely to have a sti than non-drinkers (martins c et al., 2018). exposing to pornographic materials could alter the normal sexual desire and care taking of exposure to stis. this study shows that about 28.2% of respondents were view/read pornography materials in the last 12 months. those who view/read pornographic materials were 4.4 https://journals.e-palli.com/home/index.php/ajcp pa ge 34 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 26-36, 2022 table 5: sti syndromes in the last 12 months among hip workers, southern ethiopia, 2020 variables frequency percent self-reported stis(n=121) male 15 12.4 female 106 87.6 sti syndromes among males(n=15) genital ulcer or sore 4 26.6 urethral discharge 5 33.3 inguinal bubo 2 13.3 scrotal swelling 4 26.6 sti syndromes among females (n=106) genital ulcer or sore 23 21.7 vaginal discharge 63 59.4 inguinal bubo 4 3.8 lower abdominal pain 16 15.1 treatment history(n=121) yes 92 76.0 no 29 24.0 place of treatment(n=92) hospital 27 29.3 health center 33 35.9 pharmacy 6 6.5 family guidance 4 4.3 private clinic 22 23.9 reason for not taking treatment (n=29) thought syndrome is incurable 4 13.8 syndromes are not serious 5 17.2 well without treatment 8 27.6 don't know where it can be treated 8 27.6 lack of money 2 6.9 ashamed of health professionals 2 6.9 hiv status of respondents (n=657) negative 361 54.9 unknown 282 42.9 positive 14 2.1 times more likely to have sti than when compared with those who didn’t view/read pornographic materials. the finding was comparable with studies conducted among university of gondar students, 1.5 times more likely to have an sti than counterpart (ayanaw b et al., 2019). this study shows that those who comes from rural areas where 2.3 times more likely to have the risks for stis when compared to those who are urban areas. this finding was consistent with community based survey in adami tullu, which shows that rural residents where 2.3 times more likely to have stis than urban residents [6]. this difference may be poor knowledge on transmission and prevention methods of stis in rural areas. most of workers came from different rural areas having different cultures and values. they are vulnerable to rsps in new environment, living away from the family, may limit workers to protect from peer pressure and could also be easily deceived with monetary incentives from persons who seek out sex because low monthly payment in hip. limitations since sexual behavior and practice is a private, intimate and sensitive issue, respondents may feel embarrassed to report syndromes (may subject to bias). so, selfadministered questionnaires were used to keep privacy, and study participants were informed the purpose and confidentiality of the study before data collection. in this study sti was assessed only through the self-report of the workers, no physical and laboratory examination was done and since we are using a syndromic approach, we may miss asymptomatic workers and we may misdiagnose signs and symptoms due to other health problems as similar manifestations with sti syndromes. so under/ over reporting of stis may be possible. conclusion the self-reported prevalence of stis among hawassa industry park workers was high. hometown residence, drinking alcohol, view/read pornographic materials and poor knowledge of stis were factors significantly associated with the magnitude of sexually transmitted infections. the findings in this study suggest that sti magnitude among hip workers found to be higher than the general population, thus advances in sti prevention and control hard work targeting among hip workers were required. large scale studies including all other industry institutions and use of better diagnostic modalities were recommended. references adera a, wudu m, yimam y, tamirie m, aregah m, dessie g, et al., (2015) assessment of knowledge, attitude and practice of students towards sti in hailemariam mamo preparatory school debre birhan, ethiopia. science journal of public health. 3(4), 563-71. ayanaw b, yenus h, berhe r, e. a. 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(1999). treatment of adolescents with substance use disorder. in: us department of health and human services phs, substance abuse and mental health services administration., editor. center for substance abuse treatment. usa world health organization. (2005). risk and protective factors affecting adolescent reproductive health in developing countries. yohannes b, gelibo t, m. t. (2013). prevalence and associated factors of stis among students of wolaita sodo university, southern ethiopia. international journal of scientific and technology research, 2(2), 86-94. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 1 american journal of chemistry and pharmacy (ajcp) nutritional and phytochemical screening of raw and boiled hypocotyls of african fan palm (borassus aethiopum) benjamin. i.1*, rose. e. kukwa1, teghtegh. f. samoh2, terhile. m. iortile1, tor. p. ngunoon3, john. o. ajegi4, solomon d. igbawase1 volume 1 issue 1, year 2022 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v1i1.351 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: june 25, 2022 accepted: july 03, 2022 published: july 06, 2022 the proximate analysis, phytochemical screening and mineral properties of palmyra palm (borassus aethiopum) were evaluated on the dried powder of the raw and boiled hypocotyl. the results obtained showed that the moisture, ash, fat/lipid content, crude fibre, protein and carbohydrates of the hypocotyl had values at (7.17, 3.00, 3.35, 2.63, 11.25, 72.60 and 9.33, 2.67, 2.80, 4.03, 10.00 and 72.57) %. the phytochemical analysis performed were alkaloids (76.00 and 43.47) %, cyanogenic derivative (78.84% and 48.60%), tannins (19.97 and 19.96)%, and phytate (1.19 and 0.07). the mineral composition; potassium, calcium, zinc and iron was determined using atomic absorption spectrometry with values 633.70 and 452.27 mg/100 g, 148.65 and 120.78 mg/100g, 1.36 and 2.13 mg/100 g, 2.05 and 3.24 mg/100 g respectively. furthermore, the result of the t-test analysis carried out showed that there is a significant difference between the raw and boiled sample of palmyra palm. the quantitative phytochemical analysis of the borassus aethiopum hypocotyls revealed a good percentage of phytochemicals such as alkaloids, saponin, flavonoids, steroids, cyanogenic derivatives, tannins and phytate and hence can be used in pharmaceutical and medical science to produce drugs and supplements that can prevent diseases. keywords anti-nutrients, borassus aethiopum, mineral, phytochemicals, proximate 1 department of chemistry, benue state university, makurdi, nigeria. 2 departments of chemistry, university of ilorin, ilorin, kwara state, nigeria. 3 departments of chemistry, joseph sarwuan tarka university, makurdi, nigeria. 4 college of education oju, benue state, nigeria. * corresponding author’s e-mail: ishwahlizer@gmail.com introduction the significance of plants cannot be overemphasized in our day-to-day lives as they are the major source of oxygen needed by humans and other animals for breath and existence, they serve as a means of livelihood to many house hoods, provide raw materials for many industrial products such as pulp which is used to produce paper and other raw materials. other organic extracts such as dyes, pesticides and drugs are gotten from plants (benjamin et al, 2022). food is no doubt the most basic necessity for one to effectively function in his ecosystem. it is a substance that often composed of carbohydrates, lipids, proteins, vitamins and water which are eaten or drunk by animals or humans for nutrition (benjamin et al, 2022). the constituent in food contains important chemical substances known as nutrients. these are ingested, digested, absorbed and circulated in the blood streams to feed the cells which constitute the body building blocks and consequently, the increase in body resistance to diseases and faster recovery of illnesses is witnessed (anhwange et al, 2020). most of the food consumed by humans are sourced from plants and animals, the former has been grouped into; leafy vegetables, seeds, tubers and fruit (karou et al, 2006). there are over 30,000 known edible plants, from which only 300 were domesticated accounting for more than 95% of the required human plant food (kamanzy et al, 2002). the part of plant responsible for bearing of seeds is known as fruit and is considered a healthy food supplement because it composed of an appreciable amount of water, carbohydrates, lipids, proteins, vitamins and minerals such as calcium, magnesium, potassium, sodium, zinc, copper and iron (edeoga et al, 2005). the food which humans depend on wholly for survival is derived from primary metabolites. these foods when eaten raw may however contain secondary metabolites such as tannins, terpenes, alkaloids, flavonoids just to mention but a few which may be deleterious to health. african fan palm shoots are beneficial to man as they contain carbohydrates for energy, proteins for buildup of body tissues, vitamin a which improves night vision, vitamin b1[thiamine] which converts sugars and starches into energy, vitamin b2[riboflavin] which enables utilization of fats, proteins and sugars ,vitamin b6 which aid the metabolism of protein, carbohydrates and fats and as well controls the body cholesterol level, vitamin e which protects the body store of vitamin a, tissues and fats from destructive oxidation, they also contain folate and pantothemic acids. minerals found in african fan palm shoots include calcium, magnesium, phosphorus, potassium, sodium, zinc, copper, manganese, selenium and iron. however, the amount of carbohydrate present in these shoots do not amount to more than 3-4 grams per 100grams serving which if exceeded would have led to a high calorie content detrimental to health. apart from primitively consuming the shoots as food, they help boost the immune system, they are anti-cancer fighting, have anti-inflammatory properties, they are heart friendly and help in developing a healthy weight loss and contain a negligible amount of fat. shoots also contain a significant amount of dietary fiber (amos, s et al, 2001 & adamu, b. et al, 2012) literature review palmyra palm (borassus aethiopum) is a non timber forest product (ntfp). it belongs to the family arecaceae. it is https://doi.org/10.54536/ajcp.v1i1.351 https://journals.e-palli.com/home/index.php/ajcp mailto:ishwahlizer%40gmail.com%20?subject= pa ge 2 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 1-10, 2022 a multipurpose species native to the semiarid zones and sub humid tropical regions of africa, in the south of asia, islands of the pacific and the indian ocean. palmyrapalm (borassus aethiopum) is a tropical plant with multiple uses and the hypocotyl is appreciated by the population in food and in traditional medicine. in addition all parts of this plant such as the stem, roots, fruits, leaves, hypocotyl, petioles, seeds and sap are a source of richness for rural populations (dewole, et al, 2013). ethno botanical survey revealed the aphrodisiac properties of borassus aethiopum as well as anti-oxidant, analgesic and anti-inflammatory properties of this plant which contribute immensely to the treatment of various ailments (dewole, et al, 2013). borassus aethiopum is an evergreen tree growing to 20 m (65 ft) by 5 m (16 ft) at a slow rate. it is hardy to zone. the flowers are pollinated by insects. the plant is not self-fertile and it is noted for attracting wildlife. it is also suitable for: light (sandy), medium (loamy) and heavy (clay) soils, it prefers well-drained soil and can grow in nutritionally poor soil. it requires a suitable ph of acid, neutral and basic soils growing in very acidic and alkaline soils. borassus aethiopum does not grow in shade hence it prefers moist soil and can tolerate drought. boiled hypocotyl of borassus aethiopum (palmyra palm) are widely used in most areas in benue state; this is carried around by vendors who use it as a source of income in their daily lives. as a result of its high rate of consumption mostly by the old and little by the young in the benue valley, due to its medicinal value and health implications it is of utmost importance scientifically to investigate and identify the bioactive constituents present in this plant (kolawole, s.e. & obueh, h.o. (2013).). the borassus aethiopum plant in the benue communities is recognized and kept within the ecosystem for not only its social importance but also for its economic value. the borassus aethiopum mart is commonly known as the african fan palm, palmyra palm, toddy palm in english, akuu (kuugh) in tiv, odoo in idoma, muruchi in hausa and kolabula by the benin people. the different part of this plant such as the roots, petioles, sap, fruits, seeds, final buds, shoots, wood and the resin are important sources of income for the rural communities especially for the women (gbesso f et al, 2013). numerous ways of the fruit consumption are known. the fruit can be directly boiled for eating or germinated before cooking. however, the hypocotyls obtained after fruit germination is the best-selling derivative product consumed in various cities in benue state and nigeria as a whole, and also in many countries like benin, senegal, and burkina faso etc. in benin the hypocotyl is mostly appreciated by the rural and urban populations especially by the department of zou-colliness and still remains the centre of production and distribution (kolawole, gbesso f et al, 2013 & waziri et al, 2010). research has revealed that about 90% of the households in central benin consume the products of borassus aethiopum with the hypocotyl inclusive. the high level of consumption explains the interest of the hypocotyl in the diet of the local population. besides its use in the diet, the hypocotyl is also renowned for its medicinal properties. an ethno botanic investigation revealed the aphrodisiac properties of the hypocotyls which is use in the treatment of erectile dysfunction in men (waziri et al, 2010). hypocotyls are most often consumed boiled or sometimes smoked. the hypocotyls are mostly vended in streets of benue state, benin and also certain streets in various states in nigeria. ornamentally, the leaves of the palm have served the basketry and mat industries. the trunk has been used in constructing bridges, and telegraphic poles due to its tough and termite resistant nature. the roots, leaves, flowers and fruits are used for multiple purposes such as nutrition agents, treatment for sexually transmitted diseases (e.g., beign herpes), cutaneous fungal infections, and viral infections particularly measles (waziri et al, 2010, gbesso et al, 2016). the flowers are used to treat impetigo, whereas the roots are used for asthma treatment. the sap of borassus aethiopum is usually boiled immediately after extraction to make sugar or fermented to produce an alcoholic beverage. also, the mature hard nuts are grounded and used in porridge. other studies have revealed that the young shoot of the germinating fruit of borassus aethiopum extract contain an anabolic effect of androgens; therefore, supporting its local use as an aphrodisiac. furthermore, the methanolic seed coat of borassus aethiopum has been shown to possess free radical scavenging action and its leaves have an effective anthelmintic activity against indian adult earth worms. the young shoots of this plant are rich in starch and fibre, which aids in the control of various ailments especially diabetes. it is believed that the regular consumption of the flour of this plant increase body strength, reduce hunger and the incorporation of it in other foods would positively reduce malnutrition. however, due to the presence of bitter compounds (steroid and saponin) has limited its consumption rate. owing to the presence of bitterness and inadequate supplies of starch and fibre, research to the food properties and the possible use of starch and fibre from economic, under-utilized fibre rich food plants based has become of expected attention(sakande. j et al, 2012). materials and methods study area the research was conducted in makurdi, town the benue state capital. the town is located at latitude 7o 38’n 7o 50’n and longitude 8o 24’e 8o 38’n. it is situated in the benue valley in the north central nigeria sample collection and identification fresh samples of raw and boiled palmyra palm hypocotyls were obtained from ugba market in logo local government area of benue state and were taken to department of biology science of the benue state university makurdi for identification and authentification. https://journals.e-palli.com/home/index.php/ajcp pa ge 3 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 1-10, 2022 sample preparation dirt’s and other extraneous materials were removed from the hypocotyls with a stainless steel knife. the hypocotyls were washed, chopped into pieces and dried under shade and thereafter reduced to powdered form using mortar and pestle and blended with an electronic blender. sample digestion 5 g of the hypocotyl powder was weighed into a 250 ml beaker. the sample was then digested with 200 ml of distilled water with 1 ml of hno3and 2 ml of hcl added. this was then heated to boil after which it was filtered. the digested sample was then stored in a 100 ml volumetric flask prior to analysis with aas. determination of mineral contents potassium, calcium, magnesium, zinc, iron, copper and manganese were analyzed after digestion with hno3 and hcl. this was done by transferring the digested samples into a 60 ml sample bottle after which the atomic absorption spectrophotometer (model pg 990) with appropriate hollow cathode lamps was used for the elemental analysis. determination of proximate composition moisture content empty clean crucible dishes were dried in the oven at a temperature of 105ºc for 30 minutes and cooled for 10 minutes in a desiccator. 2 g of the samples (raw and boiled hypocotyl) were weighed and put in the dishes and heated for 3 hours at a temperature of 105ºc (sakande. j et al, 2012). the dishes were then removed from the oven, cooled in a desiccator and weighed. the moisture content was calculated using the formula below: ash content the ash content was estimated by complete incineration. 5 g of both raw and boiled hypocotyl samples were weighed in to a pre-heated and cooled crucible and was incinerated in a muffle furnace at 600°c for 3 hours (sakande. j et al, 2012). the ash was then cooled in a desiccator and weighed. the ash content was determined using; crude fibre exactly 5 g of the powdered sample were weighed and placed in 500ml conical flask containing 200 cm3 of 1.25% h2so4 which was boiled gently for 30 minutes. the content was filtered and the residue was scrapped back into the flask with a spatula. 200 cm3of 1.25% naoh was added and allowed to boil gently for 30 minutes. the content was then filtered and washed thoroughly with hot distilled water. the precipitate was rinsed once with 10% hcl and twice with ethanol. the content was then allowed to dry and the residue scraped into a weighed crucible and dried overnight at 105ºc in hot oven. it was then cooled in a desiccator. the sample was heated again at 500ºc for 2 hours in a furnace. it was finally cooled in a desiccator and weighed (sakande. j et al, 2012). the percentage of crude fibre was calculated using the equation below: crude fat round bottom flask for each sample was washed and oven dried and their respective weights taken with that of the boiling chips. the lipid content was then determined according to the soxhlet method using hexane as a solvent (ahmed. a, el al, 2010)..10g of flour from each sample was weighed and inserted into a soxhlet extraction cartridge (thimble). the assembly was then placed in the extractor, where 150 ml of hexane was added. it was then allowed to heat for 3 hrs after which the hexane was collected. the fat extracted was then put in a water bath at 68oc to enable complete escape of hexane. this was then followed with oven drying for 30 minutes and the weight of the round bottom flask with the fats extracted measured. the amount of lipid extracted was obtained from the difference between the weights of the flask with the antibumping agents before extraction and the weight after extraction. the crude fat content was calculated using the equation below; crude proteins about1 g of the sample was weighed and 7.5 g of kjeldahl catalyst was weighed into a kjeldahl digestion flask and 20 ml of sulphuric acid was added. the content in the flask was then heated in the kjeldahl digestion flask until a clear blue coloration was formed, which showed that digestion was complete. the flask was then cooled and the content diluted with 250ml of distilled water and 70 ml of 50% naoh was then added. the content was then distilled using kjeldahl distilled apparatus. the distillate was then received into a flask containing 50 ml of 4% boric acid solution and bromcresol green solution indicators after distillation, 150 ml of the distillate collected was titrated against 0.1 mhcl to the end point (ahmed. a, el al, 2010). this is calculated with the equation given below; % n = percentage of nitrogen s = titre value of sample b = titre value of blank m = molarity of hcl crudeprotein=% n×6.25 carbohydrate the carbohydrate content was determined using the differential method (ahmed. a, el al, 2010). according to https://journals.e-palli.com/home/index.php/ajcp pa ge 4 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 1-10, 2022 the principle of this method, the sample consists essentially of water, minerals, proteins, fats, and carbohydrates. the content of carbohydrates is determined by reduction according to the following formula: carbohydrate (%) = 100 – (protein + fat + fibre + ash + moisture)...……….3.5 quantification of phytochemicals these hypocotyls were pulverized and used for the preparation of the other extracts, methanol and aqueous were obtained by successive extractions with solvents, according to the polarity level. determination of alkaloids exactly 2 g of the was weighed and added into a 250 ml beaker and 100 ml of 10% acetic acid in methanol was added and covered with aluminum foil and allowed to stand for 4 hours. after which the solution was filtered using a whatman filter paper (no: 125). 12 ml of ammonium hydroxide solution was then added to the filtrate and allowed to cool. the precipitate was dried in the oven at 60ºc and reweighed to determine the weight of the alkaloid (ahmed. a, el al, 2010). determination of steroids this was determined according to (akinniyi. j.a et al, 2010) without modification. 2 g of the methanol extract of the samples were macerated with 50 ml of chloroform for 24 hours. it was then filtered and evaporated. the dried masses were then combined to give the chloroform extract. determination of saponin about 1.40 g of the powdered sample was added into a conical flask and 150ml of 20% aqueous ethanol added. the samples were heated over a hot water bath for 5 hours with continuous stirring at 50ºc. the solution was filtered and the residue re-extracted with 200 ml of20% ethanol. the combined extract was reduced to 40 ml over water bath at 90ºc. the concentrate was transferred into a 250 ml separatory funnel and 20 ml of diethyl ether added and shaken vigorously. the aqueous layer was recovered while the ether layer discarded. the purification process was repeated. furthermore, 60 ml of n-butanol was added and washed thrice with 10 ml of 5% aqueous sodium chloride. the remaining solution was then heated in a water bath. after evaporation the sample was dried in the oven to a constant weight in a measured crucible. the saponin content was calculated using standard formulae. volume of 3 ml of the filtrate, (sample) standard tannic acid solution and distilled water was added into the test tubes, labeled sample standard and blank respectively. 1.0 of folin-denis reagent was added to all the test tubes followed by 2.5 ml of saturated sodium bicarbonate solution which was then added and allowed to incubate at room temperature for 120 minutes. the absorbance of the sample and the standard was read against the blank at 490nm. the percentage of tannin is calculated thus: where: at = absorbance of the test sample as = absorbance of the standard solution c = concentration of standard solution w = weight of the sample used vf = total volume of the extract va = volume of the extract analyzed determination of flavonoids exactly2.5 g of the samples were weighed into a 250 ml beaker and 50 ml of 80 % aqueous methanol was added, covered with a filter paper and allowed to stand for 24 h, at ambient conditions. the supernatant was discarded so that the residue at the bottom remains. 50 ml of ethanol was added to the residue; it was covered with a filter paper and allowed to stand for 24 h, at ambient condition. the mixture was filtered into a conical flask using a whatman filter paper (no 12, 125 mm). the procedure was repeated two times with the same sample and the filtrate combined. empty crucibles were weighed and their weight recorded. the filtrates were transferred into the crucible and evaporated to dryness over a water bath. the crucible was cooled in a desiccator to a constant weight. the percentage of flavonoids was estimated as: determination of tannins about 2 g of the sample was weighed and added into test tubes. 15 ml of distilled water was added and stirred at 10 minute interval for 1 hour and then filtered. a total determination of cyanogenic derivatives exactly 1.0 g of the sample was weighed into a 250 ml round bottom flask and 200 ml of distilled water was added and the mixture allowed to stand for 2 h. 1 ml of tannic acid was added into the mixture and distilled into a 250 ml conical flask containing 20 ml of 2.5 % naoh. 100 ml of the distillate was measured into a 250 ml conical flask and 8 ml of 6 m nh4oh and 2 ml of 5 % ki was added. the mixture was titrated with 0.02 m agno3from a micro-burette against a black background to the end-point marked by permanent turbidity. the amount of cyanogenic glycoside was estimated as: where titre value = volume of agno3 used, extract volume = 100 ml, aliquot volume = 110 ml, weight of sample = 1 g and conversion factor = 1.08. phytate determination https://journals.e-palli.com/home/index.php/ajcp pa ge 5 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 1-10, 2022 table 1: mineral composition of raw and boiled hypocotyl of borassus aethiopum mineral raw (mg/100 g) boiled (mg/100 g) potassium 633.70 ± 0.021 452.27 ± 0.021 sodium 43.54 ± 0.021 40.16 ± 0.000 zinc 1.36 ± 0.007 2.13 ± 0.028 magnesium 71.54 ± 0.021 63.25 ± 0.007 manganese 63.31 ± 0.078 61.49 ± 0.014 calcium 148.65 ± 0.000 120.78 ± 0.007 iron 2.05 ± 0.000 3.24 ± 0.021 values are mean duplicate ± s.d determination. table 2: quantitative phytochemical contents of raw and boiled hypocotyl of borassus aethiopum parameter raw (%) boiled (%) flavonoid 14.40 ± 0.000 9.00 ± 0.848 alkaloids 76.00 ± 1.131 43.47 ± 0.000 tannins 19.97 ± 0.000 19.96 ± 0.000 saponins 17.85 ± 3.040 6.00 ± 0.000 steroids 6.00 ± 0.353 5.25 ± 0.707 cyanogenicderivatives 78.84 ± 0.901 48.60 ± 0.708 phytates 1.19 ± 0.077 0.07 ± 0.021 values are mean duplicate of ± s.d determination table 3: proximate composition of raw and boiled hypocotyl of borassus aethiopum component raw (%) boiled (%) moisture 7.17 ± 0.763 9.33 ± 2.254 ash 3.00 ± 0.800 2.67 ± 2.193 crude fat 3.35 ± 1.62 2.80 ± 2.828 protein 4.03 ± 0.247 2.63 ± 0.247 crude fibre 11.25 ± 1.060 10.00 ± 1.414 carbohydrate 71.20± 0.000 72.57 ± 0.000 values are average duplicate of s.d determination. the method used was that of (ahmed. a, el al, 2010, & akinniyi. j.a et al, 2010) with slight modification. 0.2 g of the samples was weighed into 250 ml conical flask. this was then soaked in 100 ml of 20% hcl for 3 hours, the samples were then filtered and 50 ml of the filtrates was placed in a 250 ml beaker and 100 ml distilled added to the samples. 10 ml of 0.3% ammonium thiocyanate solution was added as indicator and titrated with standard iron (iii) chloride solution which contained 0.00195 g iron per 1 ml. it was then calculated as; statistical analysis data were analyzed using t-test analysis which was carried out on 95% confidence level and the probability of t= 0.05 obtained values at 0.509, 0.007 and 2.64 showed that there is a significant difference between the raw and boiled sample of the palmyra palm hypocotyl. results and discussion the results of the mineral, proximate and phytochemical screening of palmyra palm hypocotyls (borassus aethiopum) is presented table 1, 2 and 3 respectively. discussion mineral content potassium the values of potassium obtained in this study ranges from 633.70 ± 0.021 mg/ 100g and 452.27 ± 0.021 mg/ 100 g this is greater than (akinniyi. j.a et al, 2010) which show no concentration of potassium at all. the values reported were greater than that of (sastry. n.y et al, 2012). at 236.7 mg. it could be seen from table 1 that, all the minerals analyzed potassium have the highest concentration than all the other minerals. potassium is both a mineral and an electrolyte. it aids the muscle to work including the muscles that control breathing and heart beat. potassium is mostly gotten from the food we eat and the excess potassium that is not required by the body is removed from the blood by the kidney. potassium has a daily dietary intake of 3500-4700 mg(sastry. n.y et al, 2012). sodium the values of sodium obtained for the raw and boiled sample of palmyra palm as shown in table 1 ranges from 43.54 ± 0.021 and 40.16 ± 0.00 mg/100 g. this result shows that there is no much variation in the sodium content of the raw hypocotyls from the boiled hypocotyls with a difference of 3.38 mg/ 100 g. this shows a higher yield of the sodium mineral as compared to (jamkhande et al, 2014) which shows an average value of 1.45 ± 0.80 mg/ 100 g. these differences could be as a result of the composition of the soil whereby the hypocotyls are produced (sastry. n.y et al, 2012 & jamkhande et al, 2014). the world health organization recommended intake of calcium in adults is 1000 mg in male and female, which shows that both the raw and boiled hypocotyls contain lesser amount of the world health organization recommended daily standard. zinc and iron the value for zinc and iron obtained from this study were 1.36 ± 0.007 mg/ 100 g for raw , 2.13 ± 0.028 mg/ 100 g for boiled, and 2.05 ± 0.00 mg/ 100 g , 3.24 ± 0.021 mg/ 100 g of iron. these values are different to those obtained by [18] at 2.08 mg/100 g for boiled and 0.06 mg/100 g for the raw which shows a significant difference from the result of this study. the values ofiron obtained by[19] were 1.41 mg/100 g for boiled and 0.05 mg/100 g. the values of the zinc and iron content obtain from the study is less than that of [20, 21, 22] which were 12.74 mg/100 g for zinc and 11.51 mg/100 g for iron. the daily recommended nutritional intake of zinc for adults is 12 mg/day and 18 mg/day for iron. cell growth regulations, gene expression, are important roles played by zinc in the body. iron is use for the formation of melanin and transport of oxygen to the body tissues. this is a vital part of dieting in infants, pregnant women, breast feeding mothers, and it is also use to prevent anemia in the elderly. the significant difference between the zinc and iron content could be due to the mineral composition https://journals.e-palli.com/home/index.php/ajcp pa ge 6 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 1-10, 2022 of the soil and climatic conditions. this could also be attributed to the anti-nutritional factor such as phytate that can bind to essential dietary minerals such as zinc and iron (oumarou. z. nadège w.n et al, 2019) magnesium the values of magnesium in this study indicate that magnesium has a concentration of 71.54 ± 0.021 mg/100 g for raw hypocotyls and 63.25 ± 0.007 mg/100 g for boiled hypocotyls which shows a good deal of magnesium in the samples. the results obtained from this study are higher than those of and (sobert s.j. & struwig, m., 2019) which were 8.42 mg/100 g for boiled and 9.11 mg/100 g for the raw hypocotyls. magnesium is the second most intracellular cation right after potassium and has a dietary daily intake of between 50-400 mg. the results of this study show that the raw and boiled hypocotyls of palmyra palm falls within the range of the daily dietary intake of magnesium. magnesium plays a vital role in the physiological functions of the body such as maintaining normal nerve and muscle function, it also supports a healthy immune system, maintains heart beat and also helps to regulate glucose level in the blood. manganese the values of manganese gotten from this study showed its concentration at 63.31 ± 0.077 mg/100 g and 61.49 ± 0.014 mg/ 100 g for raw and boiled sample. the values obtained in this study are greater than those reported on the previous work on the evaluation of the nutritional and anti-nutritional composition of the african palmyra palm by (onwuka, g. i. al, 2005, & oduwaye. o. f) at 12.85 mg/ 100 g and 11.89 mg/100 g. manganese is one of the micro elements analyzed and from the results of the research work carried out it had more concentration than the other micro elements such as iron and zinc analyzed. the dietary intake of manganese is 2.3 mg for men older than 19 years of age, this shows that the amount of manganese in the palmyra palm hypocotyls is very high and above recommended dietary daily intake. calcium calcium is one of the essential macro elements it plays an important role in bones building and also blood clotting. about 99% of calcium in our body is in our teeth and bones. table 1 shows the concentration of calcium for the raw and boiled samples at 148.65 ± 0.00 mg/100 g and 120.78 ± 0.007 mg/100 g this values were higher than those reported by [28,29] which were 14.16 mg/100 g for boiled and 19.32 mg/100 g for raw. this difference could be attributed to the mineral composition of the soil and climatic condition. phytochemical screening flavonoids the ranges of values obtained for this research work are 14.40 ± 1.13 and 9.00 ± 0.85 for raw and boiled samples as shown in table 4.2. this shows that there is more flavonoid content in the raw sample than the boiled. the results obtained is higher than that of (morton j.f. 1992., & akinpelu. d. a et al, 2008) on the preliminary phytochemical screening of borassus flabellifer which were 9.80 ± 0.03 and 8.80 ± 0.02. comparatively, the study of (temitope. o. o et al, 2016) on proximate analysis, phytochemical and mineral composition of boiled borassus aethiopum hypocotyl reported flavonoid value at 1.42 ± 0.07. flavonoids have been reported to exhibit antibacterial, antifungal and antiviral effects. they also perform antioxidant, protective effects and inhibit the initiation, promotion and progression of tumors (temitope. o. o et al, 2016, ajai, a.i, 2012) alkaloids alkaloids are the most efficient therapeutically significant plant substances. pure isolated alkaloids and their synthetic derivatives are used as basic medicinal agents because of their analgesic, antispasmodic and antibacterial properties. the value obtained from this study shows the mean value of the raw and boiled palmyra palm hypocotyls at 76.00 ± 0.00 and 12.80 ± 0.00% respectively. these values are above those reported by (ajai, a.i, et al, 2012) in his study on the phytochemical constituents and nutrient evaluation of black rhun palm with values of alkaloid ranging from 24.16±0.034 to 22.80 ± 0.769. similarly some authors had reported a value of 0.76 ± 0.01% for alkaloid in their previous study on the minerals and nutritional profile of borassus heineanus which is lower to the value obtained by the present study. these differences in values could be attributed to the difference in chemical composition of the soil and the hypocotyl used (rignero. r. al, 1997). tannins the range of values for tannins obtained from the present study for raw and boiled samples are 19.97 ± 0.00 and 19.96 ± 0.00. the values obtained show a minor difference between the raw and boiled sample of the palmyra palm hypocotyls. the value obtained from this research are lower than those reported from the previous research on the proximate, mineral and anti nutritional composition of borassus akeassii with values ranging from22.80 ± 0.02 and 22.78 ± 0.02 [35] and also lower than that of [36] which was 279.36 ± 30.79 g/100 g. another researcher reported tannin value at 31.77 ± 1.67 mg/100 g on his study on the nutritional and antioxidant properties of some african edible plants (aoac, al, 1990). the presence of tannin in the plant indicates the astringent properties of palmyra palm hypocotyls and in addition, aid in the healing of wounds and burns (aoac, al, 1990) saponin the value of saponin obtained from this study is as shown in table 2 for the raw and boiled hypocotyls. the values show that there is more saponin value in the raw sample than the boiled sample 17.85 ± 3.040 and 6.00 https://journals.e-palli.com/home/index.php/ajcp pa ge 7 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 1-10, 2022 ± 0.000 respectively. previous studies reported the antinutritional values of saponin at 643.03 ± 33.20 mg/100g and 2.18 ± 0.28 mg/100 g in their work on the nutritional and anti-nutritional composition of the african palmyra palm. the values obtained in the present study are lower than those reported by but higher that reported by (aoac (2002) on the physicochemical characterization of date palm. some authors also reported the saponin value of borassus aethiopum mart at 0.25 ± 0.01, these value is also lesser as compared to that obtained from this study. the variation could be partly attributed to the method of analysis used and as well as the climatic condition of the hypocotyls harvested region. the difference could also be attributed to the storage time before the analysis was carried out. steroids table 2 shows the result of steroids obtained from this study with values at 6.00 ± 0.00 and 5.25 ± 0.35 which is not consistent with those obtained by previous works which are 11.00 ± 0.87 and 9.80 ± 0.65. these values are similar to those reported by (aoac al, 2002) with 5.85 ± 0.001 and 5.92 ± 0.003. the difference in the values could be due to the composition of the hypocotyl and mineral composition of the soil from which there are harvested as well as the climatic conditions. cyanogenic derivatives hydrogen cyanide is of high detriment for human consumption. small amount of it in the human body can cause harm to one’s health. table 2 of this study showed the values of the cyanogenic derivative at 78.84 ± 0.901 % and 48.60 ± 0.708 %. previous research work on the phytochemical screening of borassus akeassii reported the cyanide concentration to be 0.35 g. the value varies and is lower with that obtained from the present study. it can be deduced from the study that the cyanide concentration reduces on boiling, this is because during boiling, disruption of tissue and parenchyma occur and this facilitates the release of the free cyanide into the boiling water and also the enzyme, beta glycosidase is destroyed. also, evaporation of free cyanide is volatile to heat. it is also worthy of note that the disruption of tissue and parenchyma during boiling leads to softening of the shoots which makes it palatable for consumption (aoac 2002 & alhooti, j.s. al, 1998). phytate the ranges of values obtained from this research work on phytate are shown in table 2. the value obtained shows a little significant difference between the two samples. phytate are responsible for the inhibition of absorption of minerals such as iron, zinc and calcium to the body. some authors in their work on the physico chemical properties of borassus aethiopum reported phytate concentration in the hypocotyls at 87.88 ± 19.59 mg/ 100 g. another study on the aphrodisiac properties of hypocotyls extracts of borassus aethiopum mart collected in central of benin republic reported its value at 275.75 ± 53.54 mg/100 g. the result obtained from this study are lesser than those reported by previous works. the present study revealed phytate value to be 27.61 ± 0.59 mg/100 g. this variation may be as a result of the hypocotyl composition and also due to the method of analysis employed. proximate analysis moisture content the mean value of the moisture content obtained from this study were 7.17% for raw and 9.33% for boiled as presented in table 3. these values are higher than those obtained by (camara, f. & amaro, c.a. 2003 & fergusin, e.l et al, 1993) which were 0.98% for raw and 2.16% for boiled. these show a significant difference between the samples under study with respect to moisture. from literature it has been reported that high moisture content leads to difficulty in storage and hence spoilage. the significant difference between these two flours could be partly attributed to the drying time of the samples before usage for analysis. ash content the mean values of ash content for this research work ranges from 3.00% and 2.67% for raw and boiled samples, the results of this study are similar to those obtained by on the physicochemical properties of palmyra palm (borassus aethiopum) fruits from northern cameroon which were 2.88% for raw and 2.53% for boiled samples. these results are slightly above from those obtained by which are 1.17% and 1.18% respectively. the difference could be attributed to the effects of climate and the composition of the sample. fat the ranges of values obtained for fat in this study are 3.35% and 2.80% for raw and boiled samples. this trend shows a variation in the results obtained by (ahmed. a, et al, 2010] in his study of the proximate analysis, phytochemical and mineral composition of boiled borassus aethiopum which shows the results at 0.23% for raw and 0.17% for boiled. the result obtained from this study shows that the lipid is higher than those obtained by (al-samarai et al, 2016 & aina d.o, al, 2018) which are 0.01%, 1.49% and 1.11%. this result is lower than that obtained by (alli. g al, 2017) with 10.73%. the low fat content of borassus aethiopum allows it better storage stability while avoiding rancidity and it is also an ideal food for weight control protein the values of protein obtained from this study are 4.03% and 2.63%as shown in table 3 for the raw and boiled samples. this shows a high protein content in the raw sample. the protein content obtained from this study was higher than those reported by (jatau, d.f. al, 2008).) 3.06% for the raw and 2.71 for the boiled. the result obtained was similar to (ojha,v, 2013) with a value https://journals.e-palli.com/home/index.php/ajcp pa ge 8 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 1-10, 2022 of 4.90%. studies have shown that boiling denatures proteins. from literature it has been reviewed that plants with a protein content of greater than 12% are considered a good protein source (sharma, l al, 2004). hence, the hypocotyls of palmyra palm shoots studied are a good source of protein if consumed in the right proportions crude fibre the value of crude fibre obtained from this study ranges from 11.25% for the raw and 10.00% for the boiled. the results obtained from this study work were lower than those obtained by (sharma, l al, 2004 & bhatt,p., al, 2005). previous work on optimization of phenolics and dietary fibre extraction from date palm seeds reported higher values at 23.92% and 28.20% respectively (kumbhare.v al, 1996). the presence of crude fibre in food or plant is an indication of the level of non-digestible carbohydrate and lignin. carbohydrate the carbohydrate values of the raw and boiled sample of borassus aethiopum obtained from this study were 72.57% and 71.20% as reported in table 3. this results show that hypocotyls of borassus aethiopum are rich in carbohydrate. the results obtained from this study are lower than those of (kumbhare.v al, 1996 & giri, s al, 2000) which reported values of 86.75% for raw and 87.19% for boiled. from the present study it can be drawn that carbohydrates are the major macronutrients of palmyra palm hypocotyls (shoots). the results obtained are also lower than those obtained in ivory coast by (zhang,j., 2008) which averaged 83.79 ± 1.0 and higher than those reported in benin by (wan-you al, 2005) in his work on the valorization of the palmyra palm hypocotyl with value at 43.50%. the major function of carbohydrates is to produce energy to the body. palmyra palm shoots are comparable to most starchy foods such as yam, cassava, with little protein and fat. conclusion borassus aethiopum hypocotyl is a nonconventional food resource with potentially exploitable attributes. the flour of these hypocotyls contains an appreciable amount of physico-chemical properties. this study evaluated the physico-chemical composition of borassus aethiopum and revealed that it is rich in carbohydrates, with low lipid (fat) content, also with an appreciable amount of crude fibre. the hypocotyls also contain certain amount of protein; moisture and low ash content the hypocotyls also contains appreciable amount of minerals that are vital for metabolic activities in the body.the mineral composition revealed all the minerals analyzed with appreciable amounts in potassium, calcium, magnesium, manganese, and sodium. the study also reports a low amount of minerals with respect to zinc and iron which is due to the presence of phytate that inhibits its adsorption. the quantitative phytochemical analysis of the borassus aethiopum hypocotyls revealed a good percentage of phytochemicals such as alkaloids, saponin, flavonoids, steroids, cyanogenic derivatives, tannins and phytate and hence can be used in pharmaceutical and medical science to produce drugs and supplements that can prevent diseases. references aaina d.o (2018).proximate analysis, phytochemical and mineral composition 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(1993).the zinc, calcium, copper, magnesium, non-starch polysaccharide and phytate content of seventy eight locally grown prepared african foods. journal of food analysis. 6: 337-342. gbesso. f., adjatin a., dansi, a.a. & akoegninou.,a. (2016), “aphrodisiac properties of hypocotyls extracts of borassus aethiopum mart (arecaceae) collected in central of benin republic,” international journal of current microbiology and applied sciences.5(3), 802-814. gbesso f, akouehon, g, tente, b, & akoegninou. a. (2013). “aspects technico-economiques de la transformation de borassus aethiopum mart (arecaceae) au centre-benin,” journal afrique science, 9(1),59-173. house, w.a. (1999). trace element bioavailability as exemplified by iron and zinc. international journal of field crops research. 60: 115-141. jatau, d.f. (2008). profitability assessment of borassus jamkhande. p. g, suiyawanshi. v. a, wattamwar. a.s, barde. s. r. 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(2002). evaluation of the antibacterial activities of medicinal plants from ivory coast. journal of phytoether research, 16(5), 479-50 mahan, m.r., konan, n’g.y., sidibe, d., coulibaly, a., ezoua, p., chatigre, k.o. & biego, g.h.m. (2016). nutritive compounds from leaves of moringa oleifera l and beans of vigna unguiculata w for improvement of the meal deriving with new shoots of borassus aethiopum m in côte d’ivoire. international journal of environmental and agriculture research. 2:1-11. morton j.f. (1992). widespread tannin intake via stimulants and masticatories, especially guarana, kola nut, betel vine and accessories. journal of basic life science. 2: 739-765 nethagia. s, sivasamya. a, thennarasua. g, saravanan. g.(2010). adsorption of malachite green dye onto activated carbon derived from borassus aethiopum flower biomass. journal of hazardous materials 181: 271-280. oumarou. z. nadège w.n, souleymane k, cissé h, adama s, bassibila a.z, fulbert n, yves t, aly. s. 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(1997). isolating bioactive compounds from marine organism journal of marine biotechnology. 5(4), 187-193. sakande. j, nikiema .a, kabie. e, lompo. m, bassene. e. (2012). invitro assay of potential antifungal and antibacterial activities of extracts of borassus aethiopum mart. international journal of nigerian society. 24:48-51. sastry. n.y, padmaja . j, rao.r.p, kirani. krls, kaladhar d, (2012). invitro dose dependent study of anti-human pathogenic bacteria and free radical scavenging activities of methanolic seed coat extract of borassus flabellifer. asian journal of pharmaceutical and clinical research.5:83-86. sakande. j, kabre. e, lompo. m, pale. e, nikiema j.b. (2013). anti-inflammatory and antioxidant activities of a fraction 11 of male inflorescences of borassus aethiopum mart (arecaceae). american journal of biochemical and molecular biology. 13: 101-109. sobert s.j. & struwig, m., (2019), ‘borassus aethiopum mart. (arecaceae) in limpopo province with a key to south african palms, bothalia. international journal of tropical agriculture. 49(1), 2374-2376. temitope. o. o., fasusi. o. a., ogunmodede. a. f., thonda. a. o., oladejo b. o., yusufbabatunde. a. m., & ige. o. o. (2016). phytochemical composition and antimicrobial activity of daniellaoliveri extracts on selected clinical micro-organisms. international journal of biochemical research. 14(1), 1-13. waziri, m,. akinniyi. j. & salako. a. (2010). “ latoxicite de l’extrait acetonique muruchi, shoot de borassus aethiopum mart,” european journal of scientific research, 41(1), 6-12. zaharaddeen. n.g, afidah. a.r, saadath. a.h. (2014). potential of borassus aethiopum shells as precursor for activated carbon preparation by physicochemical activation; optimization, equilibrium and kinetic studies. journal of environmental and chemical engineering. 2: 1423-1433 https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 8 american journal of chemistry and pharmacy (ajcp) seroma formation after mastectomy with or without thoracoabdominal binder – a randomized control trial maryam essa alfard1*, shereya devendra2 volume 2 issue 2, year 2022 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v2i2.1540 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: april 10, 2023 accepted: may 01, 2023 published: may 16, 2023 the study’s main objective is to reduce seroma production post-mastectomy with the help of using thoracoabdominal binders. the randomized controlled trial (rct) included patients (no=40) that received breast surgery. they were divided into two groups; the control group (no=20) was the one that did not use the binders, and the study group (no=20), complied with adorning the thoracoabdominal binder for two weeks post-breast surgery. the drainage of both breasts and axillary drains was examined during this duration. the amount of drainage was noted, and charts were created on that basis. a total of 40 patients initially became a part of this trial, equally divided into two groups of 20 patients each. all of the participants of this group had undergone breast surgery (either mastectomy or mrm) and had to appear for the follow-up examination 2-3 weeks post-surgery. in these follow-up meetings in the outpatient clinic, the drainage charts were maintained, including the levels of drain output and compliance rate of the thoracoabdominal binder group; some patients were excluded from the study due to exclusion criteria.there was no difference between both groups in the drainage output, thus imposing no major impact of thoracoabdominal binders on seroma reduction. other implications of binders in breast care post-surgery were explored to add to the positive effects of binders. this suggests that more studies need to be done to find the best management methods for postoperative seroma formation. keywords breast care, drainage, mastectomy, seroma, thoracoabdominal binders 1 specialized surgeon, dubai hospital, united arab emirates 2 222, al khaleej st deira, dubai, united arab emirates * corresponding author’s e-mail: maryamessa23alfard@outlook.com introduction breast cancer these days addresses the most widely recognized cancer among ladies, with more than 2 million new cases in 2018. its careful treatment depends on oncoplastic medical procedure, a combination of those two disciplines with the last objective of a result adjusted between the best oncologic and therapeutic outcomes. without a doubt, since its origination, breast remaking has assisted patients with looking “typical” when dressed; all the more, as of late, progressions in careful procedures and clinical advancements have increased present expectations so patients can feel tastefully satisfied and likewise unclothed (salgarello, 2012). during the many previous years, the straightforward way to deal with breast cancer has developed from extremist mastectomy to the improvement of breast‐conserving medical procedures and reconstructive strategies. after an areola or skin‐ sparing mastectomy, the chance of having played out a prompt remaking addresses an extraordinary benefit for patients, inferable from its critical psychosocial benefits. as of now, quick implant‐based breast remaking (ibr) addresses 81.9% of all breast reconstructive strategies (masià et al., 2020). axillary lymphadenectomy (aln) remains the standard choice in many cases of breast tumors. likewise, to agony, hematoma and contamination, the fundamental early unfavorable impact of aln is the post-usable creation of liquid in the axillary bowl. seroma development is a consistent component; however, the volume has a wide variety. subsequently, the pace of symptomatic seromas that require needle yearning goes from 20 to 60% (kopelman et al., 1999). seroma is defined as the clear fluid deposited under the primary layer of the skin, which is usually found near the incision site after surgery, typically of reconstructive type (pogson et al., 2003). this occurs because space is created in such surgical procedures due to tissue removal. seroma is a typical post-surgery complication in breast cancer. it is considered a common cause of discomfort and irritation in patients, which might result in extending the hospital stay. it can cause problems like an infection on the site of surgery, slow laceration healing, and mortification of the skin flap. substantial research is being carried out to comprehend the etiology to uncover different means of reduction of seroma formation (de rooij et al., 2020). literature review countless rcts are being established to reduce complications created by seroma formation. many studies give a mixed opinion over the insertion of drainage postsurgery as a method of seroma reduction. florian et al. (ebner et al., 2014) explain that the most common reasons for the introduction of a drain in breast cancer surgery are: (i) reducing the risk of a possible hematoma, (ii) drainage of the seroma of the incision, or (iii) controlling the infections occurring at the surgical site. it has also been stated that not including a drain might have the following side effects: a larger amount of seroma-related complications. other risk factors like the size of the breasts, age, number of lymph nodes with tumors and blood pressure have also been examined in the past years (droeser et al., 2009; srivastava et al., 2012). https://doi.org/10.54536/ajcp.v2i2.1540 https://journals.e-palli.com/home/index.php/ajcp mailto:maryamessa23alfard@outlook.com pa ge 9 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 8-19, 2023 drain using a drain has been a typical practice to demolish the dead space made after a medical procedure. the utilization of closed suction drainage in patients who went through mastectomy speeds up injury recuperating and is likewise connected with a lower occurrence of wound contamination, necrosis, and breakdown (srivastava et al., 2012). research by bourke et al (bourke et al., 1976) found no distinction between utilizing closed suction and creased wound drainage in 51 patients who underwent straightforward mastectomy (thoren, 1964). in a concentrate by whitfield and rainsbury (whitfield & rainsbury, 1994), no huge contrast was seen between suctioned and closed siphon drainage on seroma development. the amount of quantity of drain tubes used has been researched. two randomized trials reported that using different drainage tubes presents no critical benefit on the sum or length of seroma seepage (petrek et al., 1992; terrell & singer, 1992). thoracoabdominal binders a thoracoabdominal binder is a wide belt encompassing the thorax and abdomen and supporting the incision. binders have been around for quite a while, yet their significance has fluctuated over the long run; there has forever been disagreement concerning whether they were great for patients or useless. accordingly, we led the current randomized controlled trial to explore the clinical impacts of thoracoabdominal binders. we plan to give proofbased ideas to its expected in postoperative consideration for breast surgeries (jiang et al., 2021). stomach versatile folios (trusses, girdles, stomach belts, longuette, and so forth) (cheifetz et al., 2010; fagevik olsén et al., 2009; christopher m larson et al., 2009) are often utilized regularly after laparotomy and ventral hernia fix. the beneficial impacts incorporate, among others, relief from discomfort, decreased chance of seroma development, worked on respiratory capability and postural strength. in any case, using abdominal binders depends on unclear or no proof. besides, the clinical impacts of stomach covers have been tested because of an expected gamble of postusable pneumonic inconveniences (rothman et al., 1966). on the positive side, documentation from a couple of studies infers that stomach fasteners reduce postoperative torment, seroma development, mental distress, and postoperative uneasiness (p. chowbey et al., 2000; jin et al., 2009; christopher m larson et al., 2009; k. leblanc, 2004). abdominal binders have likewise been shown to upgrade activation, safeguard the patient’s injury, help in coughing, and ease deep breathing (rothman et al., 1966). lymphedema is a well-known condition occurring right after breast surgery, but no agreement has been reached about the best treatment course. careful methodologies, despite, are getting expanded consideration. different microsurgical reconstructive procedures plan to reestablish life structures and capability of the lymphatic framework in upper limb breast cancer-related lymphedema (bcrl) (masia et al., 2016). since the lymphatic system is a complex vascular system, we concentrate on its life systems and usefulness in every individual patient utilizing symptomatic imaging strategies: lymphoscintigraphy, figured tomography angiography (cta), indocyanine green (icg) backhanded lymphography, and magnetic resonance (mr)lymphography. a preoperative limb lymphoscintigraphy is performed for every patient to evaluate the lymphatic capability of the limb. materials & methods study design this is a randomized controlled trial (rct) based on exploring the effectiveness of using thoracoabdominal binders in seroma reduction post-mastectomy. the dubai hospital ethical committee examined and approved the study procedure. all participants provided their consent in writing after being fully informed. the participants’ privacy was protected throughout the study, and the confidentiality of the data acquired was guaranteed.the patients who consented to participate in this study were allotted to one of the two groups: the thoracoabdominal group, where patients adorned binders post-surgery. at the same time, the other was the control group (without binders). both groups were carefully observed to compare the seroma drainage amount and other complications (if any). setting and patient selection patients were selected from three different saudia arabia regions: central, northern and southern. patients who had received breast surgeries such as mastectomy or modified radical mastectomy (mrm) because of invasive breast cancer or ductal carcinoma in situ (dcis) were qualified for consideration in the present trial. patients undergoing a breast-enlargement procedure or primary breast reconstructive treatments were barred. interim analysis the interim analysis for this seroma reduction after mrm or mastectomy trial was initially conducted to identify the effectiveness of thoracoabdominal binders. none of the postoperative methods seemed superior to discontinuing the rest of the practice at that particular interval. randomization according to a pc-created randomization plan, patients will be arbitrarily allocated to one of the two bunches with a 1:1 distribution. randomization will occur upon the arrival of the medical procedure, half an hour preceding incision closure. blinding blinding of the procedure was impossible as the patients were physically wearing thoracoabdominal binders, and they could not hide the main element of the study to make the trial non-biased. pre-op medications patients were given kefzol (antibiotic) once before the https://journals.e-palli.com/home/index.php/ajcp pa ge 10 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 8-19, 2023 surgical procedure. these medications were administered to reduce the surgical site infections (ssi) rate. surgical site infections (ssi) influence the oncologic consideration of breast cancer patients because of postpones in different treatments, expansions in care expenses, failed recreations, and possibly development in recurring disease rates. the rate of surgical site infections has decisively increased in literature as per the reported cases. the increment range is dramatic as if it skyrocketed from 0.8% to being raised to 26% (edwards et al., 2009; neumayer et al., 2007; prospero et al., 2006). surgical procedure in simple mastectomy, the whole breast tissue is eliminated; axillary tissues are undisturbed. at times, the “sentinel lymph node” — the central axillary lymph node that the metastasizing cancer cells would be supposed to deplete into — is taken out. individuals who go through a simple mastectomy can typically leave the clinic after a brief stay. regularly, a drainage tube is embedded during a medical procedure in their chest and joined to a little suction device to eliminate subcutaneous liquid. these are normally eliminated a few days after the medical practice as drainage declines to less than 20-30 ml daily. when this procedure is completed on a tumourous breast, it is also performed on a healthy breast to prevent cancer recurrence. the decision of this “contralateral prophylactic” choice has become more run of the mill as of late in california, most outstanding in individuals more youthful than 40, moving from only 4% to 33% from 1998 to 2011. nonetheless, the potential advantages give off an impression of being negligible, best case scenario, without any genetic markers, as per a huge scope study distributed in 2014 (kurian et al., 2014; newman, 2014; tanner, 2014). customary incision closure the incision was then closed using conventional, absorbable skin sutures, and none of the modern skin flaps were utilized after the mastectomy. medications after surgery most patients were given 500mg of kefzol every six hours for two weeks. some patients were advised to use kefzol only once for seven days, and only a few were given augmentin for 7 days. hospital stay the patients have been advised of the duration of their hospital stay as per their post-op condition. some were even kept in the hospital for about ten days. post-op complications several complications occur post-mastectomy or mrm, including lymphedema, shoulder morbidity and seroma aspiration. the one we encountered during this rct was a hematoma. hematoma is a pool of mostly coagulated blood that structures in an organ, tissue, or body space. a hematoma is typically brought about by a messed up vein that was harmed by a medical procedure or a physical injury (hematoma, toenail, gross, 2013). it can accumulate at any place in the body, including the brain. not at all like cancer, breast hematomas are mostly harmless. treatment of a hematoma relies upon its seriousness. small hematomas might resorb following a couple of days. more extreme hematomas that keep expanding might expect a medical procedure to deplete the gathered blood or potentially control any draining vessels and reclose the surgical site (hoda & cheng, 2017). follow up the patients were told to show up for the follow-up 2-3 weeks before mastectomy or mrm. evaluation the assessment was tentatively directed in the two groups, with the same philosophy and absence of pain convention. the principal factors explored were seroma production, pain before and after the medical procedure, post-op complications, the number of days the drain was attached, etc. symptomatic seroma was characterized as an axillary liquid collection initiating inconvenience or pain with clinical consultation. exclusion criteria the exclusion criteria were based on the following points: (1) patients unwilling to share their details publicly. (2) patients who were unable to comprehend the reason behind the study and its extent and, for that reason, table 1: basic history collection of the trial. n o. c on tr ol / tr ea tm en t m r n g en de r a ge n at io na lit y r eg io n t yp e of s ur ge ry m ed ic al h is to ry 1 pr ev io us s ur ge ry at th e sa m e si te t yp e of c an ce r b lo od l os s n eo ad ju va nt c he m ot he ra py t um ou r s iz e 1 c on tro l 51 19 06 7 fe m al e 30 -3 9 sa ud i c en tra l m rm fi br o c y st ic d is e a se n o id c < 50 m l ye s 25 cm https://journals.e-palli.com/home/index.php/ajcp pa ge 11 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 8-19, 2023 2 c on tro l 51 24 13 9 fe m al e 30 -3 9 sa ud i c en tra l m rm o be sit y n o id c < 50 m l ye s < 2 cm 3 c on tro l 51 34 04 6 fe m al e 60 -6 9 sa ud i c en tra l m rm h tn , o lf c va , c k d , a n e m ia n o id c 50 -1 00 m l n o 25 cm 4 c on tro l 51 26 83 6 fe m al e 50 -5 9 sa ud i c en tra l m as te ct om y po st m en op au sa l n o id c + d c is < 50 m l n o > 5 cm 5 c on tro l 51 15 88 7 fe m al e 40 -4 9 sa ud i c en tra l m rm 0 n o id c < 50 m l ye s > 5 cm 6 c on tro l 51 21 76 5 fe m al e 40 -4 9 sa ud i c en tra l m rm d m , h tn n o il c g ra de 2, e rp r+ , h e r2 < 50 m l ye s > 5 cm 7 c on tro l 51 17 08 0 fe m al e 60 -6 9 sa ud i n or th m rm d m n o id c < 50 m l ye s > 5 cm 8 c on tro l 51 19 00 7 fe m al e 50 -5 9 sa ud i n or th m rm d m , b a n o id c 50 -1 00 m l ye s > 5 cm 9 c on tro l 51 22 17 7 fe m al e 50 -5 9 sa ud i n or th m rm d m , h tn n o id c > 10 0 m l ye s > 5 cm 10 c on tro l 51 15 11 9 fe m al e 40 -4 9 sa ud i c en tra l m as te ct om y h tn , e sr d n o id c < 50 m l ye s 25 cm 11 c on tro l 94 94 54 fe m al e 40 -4 9 o th er c en tra l m rm 0 n o id c < 50 m l ye s 25 cm 12 c on tro l 83 25 01 fe m al e 50 -5 9 sa ud i c en tra l m as te ct om y d m , h tn ye s d c is < 50 m l ye s+ ra di at io n < 2 cm 13 c on tro l 51 21 97 1 fe m al e 50 -5 9 sa ud i c en tra l m as te ct om y 0 n o id c r ig ht br ea st < 50 m l n o 25 cm 14 tr ea tm en t 51 30 88 8 fe m al e 50 -5 9 sa ud i c en tra l m rm 0 n o id c < 50 m l n o > 5 cm https://journals.e-palli.com/home/index.php/ajcp pa ge 12 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 8-19, 2023 15 tr ea tm en t 51 12 13 0 fe m al e 50 -5 9 sa ud i c en tra l m rm d m , h tn , fa m il y h is to ry o f u te ri n e c a rc in om a n o po or ly d iff er en tia te d c ar ci n om a 50 -1 00 m l ye s > 5 cm 16 tr ea tm en t 51 24 09 1 fe m al e 50 -5 9 sa ud i so ut h m rm ba n o id c g ra de 3 < 50 m l ye s 25 cm 17 tr ea tm en t 51 17 94 8 fe m al e 50 -5 9 sa ud i n or th m as te ct om y 0 n o id c g ra de 3 < 50 m l ye s 25 cm 18 tr ea tm en t 50 22 25 7 fe m al e 30 -3 9 sa ud i c en tra l m rm 0 n o h yp er pl as ia a ty pi ca l w it h ic d < 50 m l n o 25 cm 19 tr ea tm en t 51 30 82 0 fe m al e 40 -4 9 sa ud i n or th m as te ct om y 0 ye s id c g ra de 1 < 50 m l n o 25 cm 20 tr ea tm en t 48 01 30 fe m al e 30 -3 9 sa ud i c en tra l m rm o st eo sa rc om a ri gh t f em ur , le ft b re a st c a n c e r n o id c 50 -1 00 m l ye s 25 cm 21 tr ea tm en t 99 54 56 fe m al e 70 -7 9 sa ud i ce nt ra l m as te ct om y d m , h tn , le ft br e a st c a n c e r, b a n o d c is < 50 m l n o < 2c m 22 tr ea tm en t 41 43 5 fe m al e 70 -7 9 sa ud i n or th m rm d m , h tn n o id c < 50 m l ye s 25 cm 23 tr ea tm en t 51 24 61 5 fe m al e 50 -5 9 sa ud i ce nt ra l m as te ct om y d m , h tn n o id c < 50 m l ye s < 2c m 24 tr ea tm en t 51 23 21 8 fe m al e 40 -4 9 ot he r c en tra l m rm 0 n o id c < 50 m l ye s 25 cm 25 tr ea tm en t 51 20 82 4 fe m al e 50 -5 9 sa ud i c en tra l m rm 0 n o id c < 50 m l ye s 25 cm https://journals.e-palli.com/home/index.php/ajcp pa ge 13 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 8-19, 2023 table 2: data of drain collection and post-op condition. n o. m et s to l n s antibiotics coverage h ei gh t w ei gh t b m i d ra in 1 d ra in 2 r em ov al o f d ra in o n po st -o p d ay n o. ph ys io th er ap y e xe rc is es pa tie nt c om pl ia nc e w ith w ea rin g th e b el t po st -o p co m pl ic at io ns 1 n1 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 17 5 79 .3 25 .9 111ml to 5ml 120ml to 25ml d ay 1 4 ye s c on tro l g ro up n o 2 n0 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 16 3 89 .6 33 .7 200ml to 20ml 130ml to 10ml d ay 1 4 ye s c on tro l g ro up n o 3 n2 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 14 8. 5 72 .9 33 160ml to 30ml 130ml to 10ml d ay 1 4 ye s c on tro l g ro up n o 4 n0 kefzol once pre-op, postop augmentin 7 days 16 4 62 .7 23 .3 132ml to 20ml 0 d ay 9 n ot n ee de d c on tro l g ro up n o 5 n2 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 15 0 97 .1 43 .2 210ml to 10 ml 115ml to 10ml d ay 3 3 ye s c on tro l g ro up n o 6 n2 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 15 8 87 .3 35 150ml to 65 ml 100ml to 10ml d ay 1 8 ye s c on tro l g ro up n o 7 n2 kefzol once pre-op 15 0 54 .4 24 .2 167ml to 15ml 110ml to 5ml d ay 1 4 ye s c on tro l g ro up n o 8 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 16 6 85 .5 31 140ml to 23ml 120ml to 5ml d ay 1 4 ye s c on tro l g ro up n o https://journals.e-palli.com/home/index.php/ajcp pa ge 14 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 8-19, 2023 9 n3 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 15 9 67 .5 26 .7 150ml to 30ml 120ml to 10ml d ay 1 4 ye s c on tro l g ro up h em at om a po st -o p d ay 3 10 n0 kefzol once pre-op 16 4 76 .9 28 .6 100ml to 5ml 0 d ay 10 n ot n ee de d c on tro l g ro up n o 11 n1 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 15 5 69 .7 29 150ml to 40ml 110ml to 10ml d ay 1 4 ye s c on tro l g ro up n o 12 n0 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 15 5 64 26 .6 170ml to 10ml 0 d ay 1 4 n ot n ee de d c on tro l g ro up n o 13 n0 kefzol once pre-op and post-op for 7 days 15 6 56 23 .7 130ml to 10ml 0 d ay 1 4 n ot n ee de d c on tro l g ro up n o 14 n1 kefzol once pre-op and post-op for 7 days 15 3 76 .9 32 .9 140ml to 10ml 124ml to 5ml d ay 9 ye s ye s n o 15 n3 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 16 4 82 .1 30 .5 122 ml to 10ml 120ml to 5ml d ay 1 4 ye s ye s n o 16 n1 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 16 8 73 25 .9 220ml to 10ml 160ml to 5ml d ay 1 4 ye s ye s n o 17 n1 kefzol once pre-op, postop augmentin 7 days 15 8 71 .4 28 .6 150ml to 30ml 0 d ay 1 4 n ot n ee de d n o n o https://journals.e-palli.com/home/index.php/ajcp pa ge 15 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 8-19, 2023 18 n1 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 15 0 58 .4 26 120ml to 20ml 50ml to 5 ml d ay 1 4 ye s ye s n o 19 n0 kefzol once pre-op and post-op for 7 days 16 0 86 .6 34 .5 190ml to 10ml 0 d ay 1 4 n ot n ee de d ye s n o 20 n3 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 16 3 90 53 200ml to 10ml 170ml to 10ml d ay 1 4 ye s ye s n o 21 n0 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 15 6 73 .2 30 .1 150ml to 15ml 0 d ay 1 4 n ot n ee de d ye s n o 22 n1 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 14 2. 5 77 .2 38 144 ml to 10ml 110ml to 10ml d ay 1 4 ye s ye s n o 23 n0 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 14 7 80 .4 37 .2 110 to 5ml 0 d ay 1 4 n ot n ee de d ye s n o 24 n1 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 15 8 72 28 .8 130ml to 20ml 120ml to 5ml d ay 1 4 ye s ye s n o 25 n1 kefzol once pre-op, keflex 500 mg q6hrs 2 weeks 16 2 70 .7 26 .9 110ml to 25ml 150ml to 10ml d ay 1 4 ye s ye s n o did not sign the consent form. (3) patients who never showed up for a follow-up. (4) those whose drainage charts were not managed were also excluded. results and discussion as per the research, no recognizable difference was found in the group of patients wearing the thoracoabdominal binders compared to the patients not wearing the binder in the quantity of seroma production. statistical analysis when the values of the drain removal of the control group (g1) and treatment group (g2) are compared statistically, the standard deviation of drain 1 of the control group (s=16.753) and that of the treatment group (s=7.5252) vary greatly as it gives us the p-value of 0.01249. this shows a distinguishable difference, but the sd values of drain 2 (g1= 6.95 & g2 = 3.964) did not show a distinct difference with the p-value of 0.07272, implementing the non-significant difference in the sd of both groups. this https://journals.e-palli.com/home/index.php/ajcp pa ge 16 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 8-19, 2023 nullifies the difference in the first drain and thus brings us back to the conclusion that there was no remarkable difference in the seroma production of the binder and non-binder groups. (all the calculations were performed using spss (ibm spss statistics for windows, version 25, new york, usa). primary outcome the quantity of drain extracted the first time ranged from 5ml to 65ml in the control group, while in the treatment group, the range was from 5ml to 30 ml. in the second drain removal, the field dropped from 0ml to 25ml in the control group, while the treatment group’s range fell from 0ml to 10ml (table no.2). however, since only 25 patients were left for the conclusive results, the result’s significance declined. also, the statistical analysis suggests otherwise. the sizes of the tumors extracted (table no. 1), bmi, body weight (table no. 2) etc., are also mentioned in the tables. the size of the tumor barely affects seroma production, but the increased body weight and bmi may increase the levels of seroma production (srivastava et al., 2012). secondary outcomes although there was no major difference in the seroma production of the patients undergoing mastectomy or mrm, many other positive changes were detected in this rct. pain binders worked as a pain-relief mechanism as they applied enough pressure on the thoracoabdominal region to relieve the patients of postoperative pain after the surgery. in another study (daniel & matheson, 1969), it was also evident that abdominal binders acted as painrelieving equipment. it was a short study following in a logical order, where patients were their control group (n=16). this study was conducted on patients who had to undergo major abdominal surgeries and wore binders for only about 10 minutes. the patients brought to attention that they felt a minor difference in the feeling of pain while wearing the binder. they felt less pain than those without the abdominal binder. pulmonary stress the patient reported that the stress in breathing was significantly reduced for those wearing the binders compared to those not. to support this statement, one rct (c. m. larson et al., 2009) (n=23) reported the aid of abdominal binders in the pulmonary function stating that the vital capacity of the lungs increased recognizably in the binder group compared to the control group. physical activity as it is common knowledge, shoulder immobility is also frequent post-breast surgery. the patients wearing a binder showed less discomfort while moving their hands than the non-binder group. still, the hand movement remained restricted. an approach supported by most breast surgeons is to suggest that patients limit signs at the shoulder to move away from the midline to no more prominent than 90 and that active upper extremity physiotherapy is postponed until drainage catheters have been eliminated (vitug & newman, 2007). also, a study (rothman et al., 1966) suggested that abdominal binders greatly help with the patient’s physical functions. (where physical function means a six-minute walk) (n=75). the two groups were examined for the distance they walked in the given time before the surgery and five days following the surgery. it was concluded that the binder group walked a significantly longer distance on day 5 than the non-binder group, which supports our statement claiming improvement in hand and shoulder movement post-surgery with the help of a thoracoabdominal binder. nevertheless, still, the results are ambiguous when compared to other research. it is safe to say that binders may help in improving physical function. still, more studies need to implement thoracoabdominal binders as safe and beneficial post-op care equipment. discomfort discomfort is one of the main reasons many patients disagree with using a binder. the feeling of uneasiness and constantly being compressed may be a difficulty most people do not want to endure. the pressure binder applied to the incision site may be uncomfortable to some, but it benefits as a pain-relieving compression for others. in one rct, 21 out of 28 patients (in the abdominal binder group) claimed no discomfort due to the adherence to the binder, while only 7 reported pain (jiang et al., 2021). seroma formation is considered one of the most recurrent complications post-breast surgery, be it a mastectomy, modified radical mastectomy, or even reconstructive surgery (agrawal et al., 2006; carless & henry, 2006; kumar et al., 1995; woodworth et al., 2000). needle aspiration for seroma removal is a common cause of post-op clinical visits (hashemi et al., 2004). this is why seroma reduction using different measures has become the goal of breast surgeons to make breast surgery possible with lesser pain and complications. seroma accumulation causes the skin to stretch and causes it to sag as well, creating an even more discomforting and unpleasing appearance. suppose the seroma formation continues for an extended period. in that case, it causes a delay in the healing of the wound, may cause skin flap necrosis, and be associated with surgical site infections and arm lymphedema (kuroi et al., 2006). the pathophysiology or the cause of seroma is still vague (ebner et al., 2014), but the leading cause, as per many rcts and meta-analyses, is considered dead space (agrawal et al., 2006; de rooij et al., 2020; kuroi et al., 2006; van bastelaar et al., 2016; van bemmel et al., 2011). one of the main reasons for this rct is to study the results of applying pressure with the help of a thoracoabdominal https://journals.e-palli.com/home/index.php/ajcp pa ge 17 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 8-19, 2023 binder to compress the dead space and reduce seroma production. although the findings of this rct are unclear and do not present extraordinary evidence in support of the usage of binders for the reduction of seroma formation, a couple of other larger studies do give positive outcomes analyzing the use of binders in seroma reduction, pain management and improvement in physical activity (lesser shoulder morbidities) (rothman et al., 1966). one of the studies (kontos et al., 2008) shows that pressure dressings (which can be modified into thoracoabdominal binders) make a significant difference in seroma production post-mastectomy. two hundred patients were randomized into two groups; one of them was the pd (pressure dressing) group, while the other was non-pd. all the different surgical techniques and pre and postoperative supervision were kept similar. their discoveries are in favor of pd as a viable, economical and simple to-apply technique for the decrease (a) of the time with drains in situ after mrm, (b) the number of patients producing seromas and (c) of the seroma aspirations. this might diminish further complexities, require clinical consideration, and cut expenses (kontos et al., 2008). the use of such pressure dressings, binders and compressions of the wound has been discovered widely in other horizons of innovative surgery (p. k. chowbey et al., 2000; k. a. leblanc, 2004). however, it is still in the early stages of breast surgery. more widespread, larger and more organized studies need to be executed to accumulate more and more verification and evidence to make the use of binders a common practice in the field of surgery (rothman et al., 1966). pain management post-surgery is one of the extensively recognized uses of binders in other fields of study. once again, in breast surgery, the use is limited. thus, the findings are ambiguous on whether it is beneficial or not. however, the mechanical support it provides to the wound decreases patients’ discomfort due to incisional pain. the pressure the binder applies supports the incision and even helps in wound healing to a small extent (rettenmaier et al., 2017). arm morbidity is one of the most inconvenient longlasting complexities of breast cancer therapy and essentially affects the regular routines of breast cancer survivors (bcs). despite its significance, persistent arm morbidity is moderately under-investigated (kwan et al., 2002). arm and shoulder morbidity reduces the quality of life of (bcs) greatly. binders have shown slight improvement in shoulder movement in a few patients. this is also a reason to consider more research devoted to thoracoabdominal binder applications to determine their effectiveness, as references have been provided for inappreciable improvement in physical activity with the help of binders in this rct. cancer is fatal, so most patients also have mental health issues. surgeries also bring out different levels of anxiety in most patients. this psychological stress worsens postsurgery due to the thought of any negative symptoms related to the operation (rhodes et al., 2000). an rct (cheifetz et al., 2010) compared the pre and postoperative stress levels in patients divided into binder and nobinder groups to study the use of binders. the outcome suggested that the stress level remained the same in the binder group and rose significantly in the non-binder group. similarly, a prospective controlled study (daniel & matheson, 1969) was held for a similar cause; it was detected that the psychological distress caused by coughing was exponentially reduced in the binder group compared to the others. the main reason behind the better emotional response in the binder group is the feeling of being held by the pressure of the binder wrapping around the incision. the pain of coughing post-surgery acts like trauma; thus, the compression provided by the binder eases that pain and helps reduce stress levels. it also facilitates the breathing process by covering the pain caused by the wound at the surgical site, and easy breathing also aids in the help of relieving depressive thoughts. as per the fore mentioned studies, binders help manage postoperative psychological distress; some patients feel extremely uncomfortable in the tightness of a thoracoabdominal binder. this discomfort is personal and non-scientific (rothman et al., 1966). this study concludes that using binders causes no compromise to pulmonary function. conclusion for the sake of actual implementation, the current investigation is uncertain as to the impact of thoracoabdominal binders on seroma development. although there is faint evidence in favor of a slight reduction in the production of free fluid, the lack of concrete evidence and other factors like increased bmi etc., masking the impacts of compression by the binders make its effectiveness ambiguous. insubstantial evidence is also mentioned in the binder groups’ positive pain management effects, emotional and physical distress, and moving capability. thus, more large-scale studies and comparative meta-analyses need to be taken to justify using thoracoabdominal binders in patients post-mastectomy and mrm. b binders can help make breast surgery care post-surgery less painful and more economical for the patients, resulting in fewer clinic visits. limitation one restriction of this study was puzzling due to neoadjuvant chemotherapy and axillary clearance. axillary clearance and neoadjuvant chemotherapy have been demonstrated to expand the chances of patients undergoing seroma production. another limitation may be the small size of the groups. most trials require more cases to ensure the results are not constricted to a small environment or group of people. also, the lack of versatility, as the entire trial is based in one country, may restrict the results to a certain environment. the pathophysiological reason for seroma formation is https://journals.e-palli.com/home/index.php/ajcp pa ge 18 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 8-19, 2023 still ambiguous. still, some surgeons might suggest that the bmi, other medical conditions and surgery at the same site before breast surgery may be a reason behind it. moreover, in our data [table no.1], it can be seen that most patients suffer from conditions like diabetes mellitus and hypertension. in table no. 2, we can see the raised bmis of some patients, which may cause hindrances in evaluating the accurate results. reference agrawal, a., ayantunde, a. a., & cheung, k. l. 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(2002). chronic arm morbidity after curative breast cancer treatment: prevalence and impact on quality of life. j clin oncol, 20(20), 42424248. https://doi.org/10.1200/jco.2002.09.018 larson, c. m., ratzer, e. r., davis-merritt, d., & clark, j. r. (2009). the effect of abdominal binders on postoperative pulmonary function. am surg, 75(2), 169-171. larson, c. m., ratzer, e. r., davis-merritt, d., & clark, j. r. (2009). the effect of abdominal binders on https://journals.e-palli.com/home/index.php/ajcp pa ge 19 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 8-19, 2023 postoperative pulmonary function. the american surgeon, 75(2), 169-171. leblanc, k. (2004). laparoscopic incisional and ventral hernia repair: complications—how to avoid and handle. hernia, 8(4), 323-331. leblanc, k. a. 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(2014). double mastectomy doesn’t boost survival for most. https://web.archive.org/ web/20140914001755/; http://bigstory.ap.org/article/ double-mastectomy-doesnt-boost-survival-most terrell, g., & singer, j. (1992). axillary versus combined axillary and pectoral drainage after modified radical mastectomy. surgery, gynecology & obstetrics, 175(5), 437440. thoren, l. (1964). z-plasty and suction drainage in radical mastectomy. acta chirurgica scandinavica, 128, 75-84. van bastelaar, j., beckers, a., snoeijs, m., beets, g., & vissers, y. (2016). flap fixation reduces seroma in patients undergoing mastectomy: a significant implication for clinical practice. world journal of surgical oncology, 14(1), 1-5. van bemmel, a. j., van de velde, c. j., schmitz, r. f., & liefers, g. j. (2011). prevention of seroma formation after axillary dissection in breast cancer: a systematic review. eur j surg oncol, 37(10), 829-835. https://doi. org/10.1016/j.ejso.2011.04.012 vitug, a. f., & newman, l. a. (2007). complications in breast surgery. surgical clinics of north america, 87(2), 431-451. whitfield, p., & rainsbury, r. (1994). suction versus siphon drainage after axillary surgery for breast cancer: a prospective randomized trial. journal of british surgery, 81(4), 547-547. woodworth, p. a., mcboyle, m. f., helmer, s. d., & beamer, r. l. (2000). seroma formation after breast cancer surgery: incidence and predicting factors. the american surgeon, 66(5), 444-451. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 27 american journal of chemistry and pharmacy (ajcp) documenting pharmacist intervention: a technology-driven solution to overcome challenges murooj shukri1, amna mukhtar2, sahar alsharif3, abdilahi mohamed4 volume 2 issue 2, year 2023 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v2i2.1630 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: april 20, 2023 accepted: may 22, 2023 published: may 25, 2023 clinical pharmacist conducts multiple activities governed by the best evidence for disease management and safe medication practices. the ashp highlighted the importance of documenting and recording clinical pharmacy interventions to prevent adverse drug events and reduce hospital length of stay. our institution utilizes an in-house electronic documentation system (e-ds) to document clinical interventions. we aim to study the impact of enhancing the e-ds. this study is a retrospective single-center descriptive review post-implementation of in-house e-ds enhancements. data were collected between september 2017 and december 2019 for documented interventions by clinical pharmacists for admitted patients. fields for analysis include clinical interventions, expected outcomes of interventions, cycle time for documentation, and drug cost. outcomes measures include several documented interventions, preventing medication errors/adverse drug events, timesaving and cost avoidance. descriptive analysis will be performed for categorical variables. the study showed an increased number of documented interventions for preventing medication errors/adverse drug events, time-saving, and cost avoidance, with an average time-saving of 20 minutes per 10 interventions and estimated cost avoidance of 20 000 sar per month. study limitations include a lack of interface with ehr and non-feasibility to estimate indirect cost. investment in documentation system optimization to reduce medication errors/adverse drug events, time-saving and cost avoidance. keywords clinical interventions, documentation system, clinical pharmacy 1 medication safety officer, king faisal specialist hospital and research center, jeddah, saudi arabia 2 medication safety/clinical support pharmacy, king faisal specialist hospital and research center, jeddah, saudi arabia 3 pharmacy automation and support, king faisal specialist hospital and research center, jeddah, saudi arabia 4 clinical pharmacy specialist, king faisal specialist hospital and research center, madina, saudi arabia * corresponding author’s e-mail: mshukry@kfshrc.edu.sa introduction clinical pharmacist conducts multiple activities governed by the best evidence for disease management and safe medication practices with other providers to optimize patient outcomes (pharmacy, 2014). their interventions must be documented directly in the patient’s medical record or equivalent retrievable documentation system. according to kim y. et al., a pharmacist intervention may be defined as an act or action that prevents medication therapy problems and optimize drug therapy for individual patients in cooperation with other healthcare professionals (kim & schepers, 2003). documentation of clinical activities is an integral part of efficient healthcare delivery, particularly to the clinical pharmacist (pharmacists, 1996). this documentation can also be used as a metric system to justify clinical pharmacists’ productivity and demonstrate their integral role in multidisciplinary healthcare teams. literature review the ashp highlights the importance of documenting and recording clinical pharmacist interventions (pharmacists, 1996). interventions by clinical pharmacists in a hospital setting prevent adverse drug events, reduce hospital length of stay and mortality rate, and provide a significant economic benefit to healthcare systems (gallagher et al., 2014; kim & schepers, 2003; samp et al., 2014). documentation is a vital key performance indicator to demonstrate the impact of clinical pharmacy services. it is also important to improve communication among healthcare providers to optimize medication management plan for a patient (hamblin et al., 2012). as catania et al. mentions, these data can be used to cost-justify the need for additional clinical pharmacists in a practice setting (catania & catania, 1988; kim & schepers, 2003). over the years, hospitals have faced challenges in simplifying, optimizing, and promoting clinical interventions documentation and interpretation. in the past, clinical pharmacists used to document limited interventions as handwritten notes to physicians or pharmacist progress notes, usually filed with the patient’s medical chart. over the years, many enhancements have been introduced to improve documentation and information utilization (claus et al., 2012). this has been established since replacing the manual with computerized databases and electronic health records (kim & schepers, 2003). another important aspect of implementing electronic documentation is reducing the variation and lack of standardization of pharmacist intervention documentation to improve patient care. al-jedai a. et al. highlighted a big variation in the consistency of hospital pharmacists’ documentation and data collection between 50% to 72% in various countries (al-jedai & nurgat, 2012). kim y. et al. reported limited evidence in the literature regarding the types of pharmacy intervention documentation collection systems and the quality, quantity, and utilization of the collected information (kim & schepers, 2003). many pharmacy intervention reporting systems, from https://doi.org/10.54536/ajcp.v2i2.1630 https://journals.e-palli.com/home/index.php/ajcp mailto:mshukry@kfshrc.edu.sa pa ge 28 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 27-36, 2023 paper to electronic, are used in pharmacy practice settings. although hospital administrative personnel admit that current systems demand equipment or software costs, the support was minimal. as a result, pharmacies tend to self-enhance their systems or use commercially available systems that meet their needs (kim & schepers, 2003). at our institution, clinical pharmacists have utilized a secured in-house electronic documentation system (e-ds) since 2010 to capture the clinical pharmacist intervention documentation. the initial version of the e-ds had many challenges and limitations. this study aims to enhance and measure the improvement of e-ds clinical pharmacist intervention documentation. materials and method this retrospective single-center descriptive study of clinical pharmacists documented interventions post e-ds enhancements. data were collected between september 2017 and dec 2019 for auditing and evaluation purposes. the enchantments were implemented in 3 phases; september 2017, january 2018, and september 2018, respectively. in order to evaluate the impact of our enhancements, data was collected from september 2017 to 2018. all the end-users were surveyed through multiple formats, and feedback for program feature enhancement testing was deliberated. additional data was collected for another 12 months to measure users’ experiences (january to december 2019). some of the outcomes we looked into were several documented interventions, preventing medication errors/adverse drug events, time-saving and cost avoidance. the data was collected from secondary and tertiary sources (annette clarkson, 2008; australia, 2018; pharmacists, 1996; pharmacy, 2014; project, 2016). data to be collected on the secured in-house e-ds for analysis include clinical interventions, types of interventions, cycle time for documentation, and intravenous (iv) to oral (po) conversion drug cost avoidance. outcomes analyzed from the e-ds generated reports that include several documented interventions, prevented medication errors/adverse drug events, timesaving, and cost avoidance (annette clarkson, 2008; australia, 2018; gin, 2013; guidance, 2013; hospitals, 2008; pharmacists, 1996; pharmacy, 2014; project, 2016) descriptive analysis is performed for categorical variables on microsoft excel 2016. a time-saving calculator for estimating the cycle time. this study was approved by the institutional review board (irb) of king faisal specialist hospital and research center. the pharmacy automation and informatics team supports the in-house e-ds enhancements through a series of efforts to optimize clinical pharmacist documentation. some enhancements included: standardizing the intervention classification and field categories per recent literature in international evidence see appendix 1(australia, 2018). improving layout usability and enabling remote accessibility from smart devices. the latter was established through hospital-secured access to ensure information confidentiality. a downtime contingency plan was designed and reflected in the department’s policy to avoid data loss and ensure its integrity. results investment in enhancing clinical pharmacist documentation system optimized interventions documentation that was reflected through an increased number of documented interventions for preventing medication errors/adverse drug events, time-saving and cost avoidance by 56% with a monthly average of 4,600 interventions. the average time saving is 20 minutes per 10 interventions. the estimated cost avoidance from converting iv to po formulations is approximate ~ 27 000 sar per month. the number of prevented medication errors/adverse drug events interventions accounted for 34,847 (50%), while the top type of interventions per the expected outcomes was related to enhancing therapeutic effect 49,050 (71%) of total interventions. the level of significance of the documented interventions (7% minor, 66% moderate versus 27% severe) is illustrated in figure 1. the top categories of document intervention per the document standardized categorization were monitoring 52% followed by drug selection 18% then undertreated 10% categories refer to figure 2. figure 1: illustrating the level of significance of the documented interventions. https://journals.e-palli.com/home/index.php/ajcp pa ge 29 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 27-36, 2023 discussion documenting clinical pharmacist interventions is essential to quantify the impact on patients’ outcomes and justify the workload and the value of the provided clinical pharmacy services. haslett et al. stated that documented pharmacist interventions had prevented 27% of potentially serious adverse drug events in a tertiary-care medical center/teaching hospital (al-jedai & nurgat, 2012; kim & schepers, 2003). also, hospital pharmacy directors acknowledge that documenting clinical pharmacist interventions is useful in demonstrating individual and organizational competency (kim & schepers, 2003). in the same survey, almost 40% of the pharmacy directors commented that the most common opinion was pharmacist time challenge or unfriendliness of available comprehensive documentation systems. the survey universally affirmed that those systems are vital to a clinical pharmacist’s responsibility. those systems supported the communications of clinical pharmacists’ actions with other healthcare providers and reflected clinical pharmacy services’ importance. while many pharmacies continued to document their interventions manually, others invested in computerized options. the latter provided the advantages of providing more comprehensive information and user-friendliness compared to the manual. furthermore, the report highlighted that the available systems were challenged with providing real cost savings or clinical outcome information. the main reason for the administrative reluctance to invest in obtaining software, equipment, and maintenance is the cost which may reach $5000. accordingly, there have not been enough solutions to overcome all the problems. however, a step forward has been taken to utilize personal digital assistants (pda) for clinical pharmacist intervention documentation systems (kim & schepers, 2003). over the past years, our institution has used various methods to document clinical pharmacy interventions, starting with manual documentation on a paper-based form in 2008. the form includes clinical pharmacists, patient information, and drug-related problems (aljedai & nurgat, 2012). this manual method had many challenges: loss of documentation, time & paper consumption, inaccessibility, lack of data security and integrity, and difficulty in collecting and interpreting data timely. accordingly, to overcome these challenges, an inhouse build of an electronic documentation system (eds) for capturing clinical interventions was implemented in 2012. the e-ds captured interventions performed by clinical pharmacists; however, there were unresolved shortcomings associated with the system. for example, no user interface on the hospital server, which limited the clinical pharmacist’s access remotely, an unfriendly data entry layout that resulted in long documentation time, and a lack of data standardization to obtain meaningful information and timely report. accordingly, an attempt to improve the e-ds was completed by releasing a new enhanced system version. the enhancement project was implemented over 3 phases; september 2017, january 2018, and september 2018. to evaluate the impact of those enhancements, data was collected from september 2017 to 2018. each phase included a cycle of (implementing modification according to end-user feedback, education for the clinical pharmacists about the new features, and a testing period). further data were collected for additional 12 months to evaluate sustainability (january to december 2019). implemented enhancements on the e-ds updated version standardization upon searching for the most comprehensive, standardized, and the closest to meet our clinical pharmacists’ needs, we selected document mrp and recommendation classification codes provided by guidelines for pharmacists performing clinical interventions by the pharmaceutical society of australia (australia, 2018). the selected categorization was further customized to meet the unfulfilled needs, see table 1. customizations were added to capture some of the clinical pharmacist’s productivity data (for example, drug shortage issues, reviewing of non-formulary medication or off-label medication use requests). another customization was deleting some categories. for example, medication figure 2: illustrate the type of document intervention per the document mrp standardized categorization. https://journals.e-palli.com/home/index.php/ajcp pa ge 30 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 27-36, 2023 education provided to a patient is documented directly on the patient’s ehr. also, the conversion from iv to po category was removed to ehr as part of the enhancement to minimize entry duplication and improve communication with other healthcare providers. the latter was part of another plan to establish clinical pharmacist standardized notes for certain services on ehr. this standardization helped minimize the drug related problem from 20 major categories and 107 subcategories to 8 categories and 37 subcategories in the updated version (australia, 2018). usability the initial e-ds version layout took a long time for clinical pharmacists to document their interventions timely. human factor engineering was considered while implementing the layout enhancements. this included: automated popups for the user and the dates, the option to manually change the intervention date where available to allow retrospective data entry, and an in-build calendar for date selection used to prevent non-standardized date format. also, a dropdown list for the inpatient units was added to limit discrepancies in the unit’s name to be extracted accurately in the report for workload purposes. the most impactful enhancements were allowing multiple interventions to be documented per patient daily on the same page instead of multiple pages for each intervention, see figure 3. additionally, activating the tab button to move quickly between fields and the automatic fill option using the first letter/category code to minimize typing time. furthermore, intervention categories and subcategories were popping up in separate windows by double clicking instead of a long dropdown list to select from. for accurate referencing of comments, the free text box for it per patient was moved to be next to each medication intervention. these enhancements resulted in an almost 45% reduction in the time needed for documentation, see figure 3. accessibility & security accessibility was a major challenge for clinical pharmacists to document their interventions timely. users needed to access the e-ds from his/her personal desktop computer using their authorized access (pharmacy, 2014). the targeted enhancement was to upload the electronic documentation system to a secure web page within the institution network (hospital server). this maintained patient confidential information and provided users with remote access to e-ds from the inpatient unit’s workstation computers, computers on wheels (cow), tablets, or mobile devices. table 1: customized document medication-related problem (mrp) and recommendation categorization codes d: drug selection d1 duplication d2 drug interaction d3 inappropriate drug d4 inappropriate dosage form/ route d5 contraindications apparent d6 no indication apparent d7 drug shortage issue d8 tpn initiation d9 tpn follow up o: over or underdose o1 prescribed dose too high o2 prescribed dose too low o3 incorrect or unclear dosing o4 incorrect/inappropriate renal dosing c: compliance c1 need: to hold/ resume or renewal reminder c2 difficulty reaching physician/ nurse c3 non-compliance with policy/ guidelines/ protocol c4 need to review of policy/ guidelines/ protocol c5 need to review chemotherapy protocol c6 review non-formulary medication request c7 review of off-label medication use request u: undertreated u1 condition undertreated u2 condition untreated https://journals.e-palli.com/home/index.php/ajcp pa ge 31 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 27-36, 2023 table 2: customized document mrp and recommendation categorization codes per intervention expected outcome adr/medication error prevented d: drug selection d2 drug interaction d3 inappropriate drug d4 inappropriate dosage form/ route d5 contraindications apparent o: over or underdose o1 prescribed dose too high o2 prescribed dose too low o3 incorrect or unclear dosing o4 incorrect/inappropriate renal dosing u: undertreated u1 condition undertreated u2 condition untreated u3 preventative therapy required m: monitoring m3 allergy/demographics missing t: toxicity or adverse reaction t1 toxicity, allergic reaction or adverse effect present adr/medication error prevented/ enhanced therapeutic effect m: monitoring m1 tdm/laboratory/microbiology monitoring m2 non-laboratory monitoring cost saving/ adr or med error prevented d: drug selection d1 duplication d6 no indication apparent cost saving/ adr or med error prevented/ enhance therapeutic effect u3 preventative therapy required u4 need for therapeutic alternative m: monitoring m1 tdm/laboratory/microbiology monitoring m2 non-laboratory monitoring m3 allergy/demographics missing m4 chart review e: education or information e1 patient requests pharmacy related information e2 physician requests pharmacy related information e3 nurse requests pharmacy related information e4 pharmacist requests pharmacy related information e6 inservice/competency conducted n: not classifiable no t: toxicity or adverse reaction t1 toxicity, allergic reaction or adverse effect present https://journals.e-palli.com/home/index.php/ajcp pa ge 32 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 27-36, 2023 u: undertreated u4 need for therapeutic alternative cost saving/ enhance therapeutic effect d: drug selection d7 drug shortage issue enhance therapeutic effect d: drug selection d8 tpn initiation d9 tpn follow up c: compliance c1 need: to hold/ resume or renewal reminder c3 non-compliance with policy/ guidelines/ protocol c4 need to review of policy/ guidelines/ protocol c5 need to review chemotherapy protocol c6 review form a c7 review form b m: monitoring m4 chart review e: education or information e1 patient requests pharmacy related information e2 physician requests pharmacy related information e3 nurse requests pharmacy related information e4 pharmacist requests pharmacy related information e6 inservice/competency conducted before: reports from the updated e-ds version became easily and timely retrieved with meaningful information: the number of interventions per period/inpatient units or pharmacist, top categories of interventions, top intervened medications, and cost avoidance per medication. these reports were utilized for workload/ productivity purposes. also, it can be shared with pharmacy and therapeutic (p&t) or drug utilization committees, quality improvement committees, pharmacy management, and hospital management to support the positive impact of the clinical pharmacists to optimize patients’ outcomes and to prevent harm (kim & schepers, 2003). other meaningful fields reported in the literature were not added to the updated e-ds version; for example, the level of intervention significance and acceptance response rate, considering the allotted documentation time versus the number of priority fields to be completed. our study findings showed a gradual improvement in documented interventions after each enhancement, especially with the testing, education, and immediate feedback from the end users. additionally, there is a direct correlation between the number of documented interventions, the number of clinical pharmacists on duty, and the inpatient census for that period. regardless, documented interventions were easily doubled over the sustainability period. https://journals.e-palli.com/home/index.php/ajcp pa ge 33 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 27-36, 2023 table 3: number of documented interventions per document (mrp) and recommendation categorization codes for the studied period document mrp and recommendation categorization number of interventions (%) d: drug selection 11,389 (18%) o: over or underdose 4,857 (8%) c: compliance 4,591 (7%) u: undertreated 6,471 (10%) m: monitoring 33,692 (52%) e: education or information 3,024 (5%) n: not classifiable 327 (1%) t: toxicity or adverse reaction 174 (0.3%) total 64,525 our analysis reflected the successful documentation of the well-established therapeutic drug monitoring service, as it was ranked the top among reported categories with 52% of all documented interventions. the lowest reported category was toxicity or adverse reaction (0.3%). this is explained by the parallel hospital safety reporting system that requires staff to use for such incidents. this duplication can be modified in the future. supporting the successful selection of the customized standardized categorization, we noticed that an insignificant percentage (1%) of interventions were not classifiable. see table 3. figure 3: usability and accessibility enhancements on the eds layout after: to identify the impact of the expected outcome, we replicated published studies with similar intervention categories and significance levels (campbell, 2011). accordingly, the top reported types of interventions per expected outcomes were invested in enhancing therapeutic effect and (adverse drug reaction (adr)/ medication error prevention with 49% and 26%, respectively, as shown in table 4. our study demonstrated that the majority of clinical pharmacist interventions were impactful per the level of significance. these results https://journals.e-palli.com/home/index.php/ajcp pa ge 34 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 27-36, 2023 support the huge value of the clinical pharmacist in medication management. as the hospital administration highly appreciates the cost-saving initiatives, focused efforts were directed to establish the intravenous (iv) to oral (po) conversion service. it describes converting intravenous therapy to an alternative oral formulation as clinically applicable (craig & andes, 1995; cyriac & james, 2014). based on craig and andes’s review, the mean duration of iv therapy before converting ranged from 2 to 8 days (craig & andes, 1995; paladino et al., 1991). accordingly, this study utilized 8-days of therapy to demonstrate the cost saving. our service launched with ten medications, primarily antimicrobials, that have impactful cost savings. later, the list was expanded to seventeen medications once the service was stabilized, see appendix vi. over the sustainability period, we identified that the top five cost-saving medications were responsible for 93%, listed in order: levetiracetam, omeprazole, voriconazole, fluconazole, and acetaminophen; see table 5. table 5: iv to po conversion medication list medication iv dose equivalent oral dose iv to oral conversion ratio frequency acetaminophen 650 mg 650 mg 1:1 continue same ciprofloxacin 200 mg 250 mg 1:1.25 continue same 400 mg 500 mg 750mg clindamycin 300mg 300mg na continue same 600 mg 450 or 600 mg cyclosporine* 25 mg 30mg to 45mg 1:1.2 to 1.8 continue same dexamethasone 1-10mg 1-10mg 1:1 continue same doxycycline 100 mg 100 mg 1:1 continue same fluconazole 200 mg 200 mg 1:1 continue same 400 mg 400 mg 800 mg 800 mg folic acid 1mg 1mg 1:1 continue same levetiracetam 250 mg 250 mg 1:1 continue same levothyroxine 50 mcg 100 mcg 1:2 continue same levofloxacin 500mg 500mg 1:1 continue same 750mg 750mg linezolid 600 mg 600 mg 1:1 continue same metronidazole 250 mg 250 mg 1:1 continue same 500 mg 500 mg 500 mg 500 mg mycophenolate mofetil* 500mg 500mg 1:1 continue same omeprazole 40 mg 40 mg 1:1 continue same rifampin 600 mg 600 mg 1:1 continue same table 4: number of documented interventions per intervention expected outcome for the studied period intervention expected outcome number of interventions enhance therapeutic effect 33,887 adr/medication error prevented 17,692 adr/medication error prevented/ enhanced therapeutic effect 13,657 cost saving/ adr or med error prevented 2,308 cost saving/ adr or med error prevented/ enhance therapeutic effect 1,190 cost saving/ enhance therapeutic effect 316 cost saving 127 total 69,177 https://journals.e-palli.com/home/index.php/ajcp pa ge 35 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 27-36, 2023 although clindamycin iv to po conversion shows negative direct cost-saving, the indirect cost of hospitalization, sterile medication preparation by pharmacy, administration by nurse, and associated risk of line infection outweigh that value. the main challenge with the updated e-ds version was that it was not integrated with ehr. this prevented accessibility by other healthcare providers to the clinical pharmacist interventions that support the medication management plan. in addition, this limit the generation of clinical outcome data (e.g., length of hospital stay, readmission, and mortality rate). furthermore, the hospital-invested billing system cannot capture the monetary value of the clinical pharmacist services. tacrolimus* 1mg 3mg or 4mg 1:3 to 4 twice daily thiamine 50-100mg 50-100mg 1:1 continue same tmp/smz 5-20 ml 1 ss tab =5ml iv 1:1 continue same 5ml iv= 400mg smx + 80mg tmp 1 ds tab=10ml iv 2ds tabs=20ml iv voriconazole 200 mg q12h 200 mg q12h 1:1 continue same valproic acid 250 mg q8h 250 mg q8h 1:1 iv valproate sodium given at same total daily dose/same frequency as oral products. however, daily doses >250 mg should be divided. table 6: iv to po conversion cost avoidance (sar) during the sustainability period, ranked from highest to lowest. drug name 8-days cost saving (sar) levetiracetam 131,828 omeprazole 92,624 voriconazole 83,920 fluconazole 10,288 acetaminophen 9,152 valproic acid 6,120 ciprofloxacin 5,344 acyclovir 3,348 doxycycline 2,840 levothyroxine 2,366 levofloxacin 1,896 rifampin 1,472 metronidazole 432 tmp/smz 233.36 dexamethasone 157.6 linezolid 80 clindamycin -28,184 total 323917 conclusion technology investments to improve the clinical pharmacist’s intervention documentation helped enhance the process and provide meaningful data to show the value of the clinical pharmacist in providing direct patient care. many technological enhancements were implemented, including standardized intervention categorization, enhanced database usability and accessibility, and building reports. this is positively reflected in the number of documented interventions, preventing medication errors/adverse drug events, time-saving and cost avoidance. the next step is to propose integrating the clinical pharmacist intervention documentation on the ehr to allow visibility to other healthcare providers, translate interventions into clinical outcomes, and plan for the clinical pharmacist services’ billing system. acknowledgments the authors are grateful to the king faisal specialist hospital and research center for their continuous support throughout the research. funding this research received no specific grant from funding agencies in the public, commercial, or not-for-profit sectors. all authors declare no relevant conflicts of interest or financial relationships. this project was presented as a poster at the 3rd international saudi health informatics conference on 17th-21st november 2018. riyadh, saudi arabia; 11th medication safety conference 2018, abu dhabi, uae; saudi international pharmaceutical annual sipha meeting 2020-virtual. reference al-jedai, a., & nurgat, z. a. (2012). electronic documentation of clinical pharmacy interventions in hospitals. in data mining applications in engineering and medicine. intechopen. antimicrobial stewardship guidance. federal bureau of prisons clinical practice guidelines, (2013). https://journals.e-palli.com/home/index.php/ajcp pa ge 36 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 27-36, 2023 australia, p. s. o. (2018). guidelines for pharmacists performing clinical interventions. australian government department of health. pharmacy and government arrangements. sixth community pharmacy agreement, intro 4. https://www. ppaonline.com.au/wp-content/uploads/2019/01/ psa-clinical-interventions-guidelines.pdf campbell, a. r., nelson, l. a., elliott, e. s., hieber, r., & sommi, r.w. . 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(2014). american college of clinical pharmacy practicebased research network c. economic evaluation of the impact of medication errors reported by us clinical pharmacists. pharmacotherapy, 34(4), 350-357. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 41 american journal of chemistry and pharmacy (ajcp) a review on the utilization and environmental concerns of coal fly ash saidu kamara1, edward hingha foday jr2, wei wang1* volume 2 issue 2, year 2023 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v2i2.1609 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: april 26, 2023 accepted: june 10, 2023 published: july 02, 2023 coal is a well-known source of electric power generation all over the globe. pulverized coal combustion and fluidized coal bed combustion are the two conventional methods involved in the combustion of coal in thermal power plants. pulverized coal combustion is operated at >14000c while the fluid bed is operated at a temperature between 850-9000c. the first two reactions are the main reactions in the coal combustion process and they are exothermic reactions. co2 and co are the first two combustion products. the inorganic minerals in coal are released as waste products in the combustion plants in which fly ash made a large part of the waste materials. fly ash is formed due to the incomplete combustion of coal. the fly ash from both methods contains sio2, al, etc, which are significant constituents. high-grade coal has a higher sio2 than the low grade. an experiment was conducted by preparing four samples of fly ash heated at 5000c, 6000c, 7000c, and 8000c, and one unheated sample to investigate the chemical composition of the fly ash obtained from xi’an linyuan silica limited and to prove its suitability as raw material for the fabrication of refractory composites. sem, eds, xrd, and ftir characterization were done on all five samples to determine the chemical parameters of the fly ash. according to the result and analysis of the four characterizations, it was discovered that the fly ash used in this research contains sio2. sem/eds morphological analysis reveals the presence of spherical and a few geometrical crystalline-shaped structures known as cenospheres. cenospheres are important components for the synthesis of refractory composites. the eds micrographs show the percentages of silicon and oxygen in the fly ash. the ftir results show si-o-si stretching in all the fly ash prepared samples. in the xrd, it was discovered that the fly ash was purely sio2. keywords characterization, silica, coal, combustion, fly ash 1 department of chemical engineering, school of water and environment, chang’an university, xi’an, china 2 key laboratory of subsurface hydrology and ecology in arid areas, ministry of education, chang’an university, xi’an, china * corresponding author’s e-mail: ksaidu2013@gmail.com introduction the popular source of electric power generation around the world today is coal utilized in pulverized coal combustion and fluid bed combustible conventional plants(s. h. lee, lee, jeong, & lee, 2019; miller, 2010). the two different combustion methods operate at different temperatures and therefore each method produces different fly ash with different characteristics (erol, küçükbayrak, & ersoymericboyu, 2008; l. li, wang, & zhu, 2006; molina & poole, 2004). the temperature range in the coal fluid bed is 8500c-9000c while the temperature in pulverized combustion is between 14000c-15000c. at temperatures greater than 14000c (>14000c), minerals such as the aluminosilicate present in coal disintegrate or decompose (mishra & baliarsingh, 2008; pavlish et al., 2003; rios reyes, 2008). the disintegrated mineral particles form hollow or spherical objects called cenospheres as a result of rapid cooling that takes place during the postcombustion phase (kim, 2002; pundienė, korjakins, pranckevičienė, & kligys, 2018). high melting point minerals sometimes remain unaltered. the physical and chemical composition of fly ash is significantly influenced by the type of coal used and the variation in combustion temperatures (adriano, page, elseewi, chang, & straughan, 1980; pedersen, jensen, skjøth-rasmussen, & dam-johansen, 2008; ratafia-brown, 1994). both two types of combustion processes contain al, sio2, fe, and ca which are the four most constituents in fly ash(cheng & chen, 2003; criado, fernández-jiménez, de la torre, aranda, & palomo, 2007). the fly ash produced from coal fluidized beds (cbd) has no mullite and glass balls. it contains high quantities of unburnt carbon, calcium carbonate, and calcium sulfate and it forms irregular objects. unburned carbon exists in both types of fly ash(batra, urbonaite, & svensson, 2008; maroto-valer, taulbee, & hower, 2001). fly ash is among the popular global raw material and one of its most important utilization is in the fabrication of composites(gollakota, volli, & shu, 2019). in addition to the current global use of fly ash, further research and development are required to enhance the management of fly ash and bring out new techniques to control the end products that result from burning coal. harmful elements and compounds such as aromatic compounds, carbon, silica, fine particulate matter, etc., are aggregated in high concentrations during combustion(schwarze et al., 2006; valavanidis, fiotakis, & vlachogianni, 2008). thermal power plants produce enormous inorganic particles and emit combustible gases into the atmosphere which negatively affects the health of residents around the thermal plants(rahman, farrok, & haque, 2022). the thermal industries have no use for the ash generated and therefore regard it as a solid waste product with the potential to negatively impact the environment if not properly regulated(foday jr et al., 2017). a small amount ( ≈20%) of the global fly ash https://doi.org/10.54536/ajcp.v2i2.1609 https://journals.e-palli.com/home/index.php/ajcp mailto:ksaidu2013%40gmail.com?subject= pa ge 42 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 41-52, 2023 produced annually, is used for many purposes and the rest reportedly contributes to the existing global pollution trend. characterizations using sem, eds, xrd, ftir, etc, to determine the composition of fly ash have been studied. several past works on the two types of fly ash were centered on their properties, morphology, and chemical composition. in this study, the morphology and chemical information of the fly ash samples is determined by sem and eds. the xrd reveals information about the crystalline size and amorphous phase of the samples. the in-situ functional groups (si-o-si, c-o, al-o, etc,) in fly ash are accounted for by the fourier transform infrared spectroscopy (ftir). these functional groups are the typical determinants for the use of fly ash as raw material for the synthesis of composites. this experiment, therefore, studies the characterization of purified fly ash from the xi’an linyuan silica limited. the major problem encountered in the characterization of fly ash is the amorphous phase of the sample which is normally not captured by the xrd. a more adequate instrument is therefore required for the characterization of fly ash to identify its glass phase and interpret chemical composition. eds is the most common effective tool used for this purpose but the contrasting characteristics of fly ash present a large scattered result that makes the analysis very difficult. in recent times, new eds technology or detectors have been invented with the capability to collect the spectrums of all pixels in not more than an hour. the spectra collected are further processed into maps that show the elemental intensities used to quantify the chemical compositions of the elements present. literature review process and end products of coal combustion coal constitutes inorganic minerals released as waste products (fly ash, bottom ash, and slag) during the burning of coal. the greater part of the waste products in the combustion plant is made of fly ash. fly ash is made of fine light particles and has a density of about 2.5g/cm3, bottom ash is composed of coarse particles while slag is the melted phase of bottom ash(bourtsalas, 2015; bunge, 2015). the ash generated in the combustion plant is made of a large mass of oxides (silica, alumina, titanium oxide, magnesium oxide, calcium oxide, sulfoxide, etc,) and heavy metals (zinc, lead, copper, nickel, cobalt, cadmium, etc,)(foday jr, bo, & xu, 2021; halmann, 1995; oves, khan, zaidi, & ahmad, 2012). fly ash contains some quantities of spherical matter called cenospheres formed by the decomposed minerals in the condensation or cooling stage (blissett & rowson, 2012; fenelonov, mel’gunov, & parmon, 2010; oves et al., 2012; vassilev, baxter, andersen, & vassileva, 2013). fly ash is formed due to the incomplete combustion of the mineral components in coal which subsequently transform into some crystalline phases called quartz (silica), magnetite, aluminosilicate glass (mullite), spinel, etc. it is confirmed that fly ash is made of over 80% of silica (sio2)(criado et al., 2007; joshi & lohita, 1997; vassilev & vassileva, 1996). the glass phase in fly ash is formed from low-grade bituminous coals which constitute lesser quantities of silica and a higher amount of magnetite compared to high-grade bituminous coal which composes high silica content. the process leading to the formation of fly ash is explained in figure 1 below: figure 1: a modified schematic diagram for the formation of fly ash the process begins with vaporizable components which nucleate to produce new crystalline objects, form char and release heavy metals by vaporization. reactions in coal combustion the carbon in coal undergoes the following chemical reaction with oxygen. https://journals.e-palli.com/home/index.php/ajcp pa ge 43 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 41-52, 2023 c + o2 = co2 ∆h=-393.7 kj/mol (1) c + 0.5o2 = co ∆h=-110.1kj/mol (2) c + co2 = 2co ∆h=+172.6kj/mol (3) c + h2o = co + h2 ∆h=+131.4kj/mol (4) co + 0.5o2 = co2 ∆h=-283.6kj/mol (5) h2 + 0.5o2 = h2o ∆h=-242.4kj/ml (6) co + h2o = co2 + h2 ∆h=-41.2kj/mol (7) the reaction is exothermic when the enthalpy ∆h change is negative (i.e. heat is given out during the reaction) while a positive enthalpy ∆h means the reaction is endothermic (i.e. heat is absorbed during the reaction). the first two reactions are the main combustion reactions according to their products. co2 and co are the first combustion products and their ratios increase with an increase in temperature and a decrease in pressure. the whole combustion process leading to the formation of a coal boiler takes about two seconds. mathematical models for fly ash deposition in a boiler tube mathematical models are employed to determine the activity of ash precipitation and to ameliorate the lapses in industrial design (fan, zha, sun, & cen, 2001). the prediction of ash deposition has been done by a good number of researchers using experimental notations but it becomes misleading when taking into consideration other operating factors (fan et al., 2001; zeng, zheng, zhou, fang, & lou, 2016). the deposition of ash in boilers has been adequately studied with the application of numerical procedures. fly ash data obtained from a computer control scanning electron microscope has been used to develop a fly ash prediction model (f. lee & lockwood, 1999). the temperature, porosity, and rate of deposition were simulated by galen et al (richards, slater, & harb, 1993). particle movement expressions newton’s equation of motion below is used to account for the particle movement in a pulverized boiler. the drag force fd is given by fd= -3πdμ(v-u)f where μg is the gas viscosity and f is the drag force coefficient that is used to revise the stokes expression and assuming that f=1. waldmann equation below is used to determine the thermophoretic force (waldmann & schmitt, 1966) where ktr is the thermal conductivity of the gas and ∆t is the temperature gradient. the mean speed c of gas molecules is given by particle collision forces the particle equation is used to compute the collision velocities of fly ash deposition in a boiler. to ensure efficient and smooth velocity collision, the particle in motion must be spherically rigid, the interaction forces and infinite forces should be impulsive and negligible respectively, and the particle movement should be centered and two-dimensional. the mathematical expression for a particle is given below (hoomans, kuipers, briels, & van swaaij, 1996). where, vi.0 and vi,1 is the pre-collisional and post-collisional velocities of particle i, and vj,0 and vj,1 are the pre-collisional and post-collisional velocities of particle j. in the absence of particle-particle collision, we can also find the sample position using the single-particle equation at a time. the method of monte carlo is exclusively used to compute the particle-particle collision. if particle i collides with particle j in the time step, the post-collision velocities of particles are accounted for by taking into consideration the dynamics of the collisions. the velocities after particle collision will now substitute the initial particle velocities but the particles remain fixed in their fixed positions. the particle i in a time step δt is determined by the probability of the collision below(bird, 1994; tsuji, tanaka, & yonemura, 1998): where n is the number of simulated particles in the cell, n is the local particle number, d is the particle diameter, go is the radial distribution function, vij v is the relative velocity between particle i and j, and ∆t is the time step. the collision pairs searching is the key problem in the calculation of particle-particle collision. ash deposition model during the process of ash deposition, the large ash particle causes an impact in the boiler tube and rebounds while the smaller ones can stick onto the tube with high adhesive energies(g. li, li, huang, & yao, 2015). the formation of the first layer improves the surface sticking capability in the tub. the critical velocity model is employed to ascertain if a particle motion sticks or rebounds in the boiler tube. the critical velocity equation is written as: https://journals.e-palli.com/home/index.php/ajcp pa ge 44 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 41-52, 2023 where, w is the adhesion energy and e is young’s modulus (s. li, marshall, liu, & yao, 2011; liu, li, & yao, 2011). according to the simulation of the critical velocity ( vcritical ) expression above, the impact of the particle velocity is compared with the critical velocity. when the critical velocity of a particle is larger than the velocity impact, the particle clings to the surface. materials and method five samples of fly ash were measured on an electronic weighing scale. each sample weighs 4g. four of these samples were heated at 5000c, 6000c, 7000c, and 8000c for 3 hours in a muffle furnace and one remain unheated. the samples were then taken for sem, eds, xrd, and ftir characterization. the aim was to determine the elemental characteristics of fly ash that make it a suitable raw material to synthesize refractory composites. figure 2: illustration of instrumental setup and sample preparation result and discussion sem result this technique is utilized by material scientists to determine and interprete the microstructure of materials. the sample electrons are bombarded to produce a wide range of emissions (such as x-rays, visible photons, auger electrons, secondary electrons, backscattered electrons, etc). the sem result of the fly ash specimens is discussed as follows: heated fly ash figures a and b below present the sem results of four samples of fly ash heated at 5000c, 6000c, 7000c, and 8000c. the images in figure (a) above are the results of sample sizes scanned at 100 um pixels while figure b scanned image sizes at 20 um. the morphology of the images is based on the materials present in the fly ash samples. https://journals.e-palli.com/home/index.php/ajcp pa ge 45 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 41-52, 2023 figure 3: sem images showing samples scanned at 100 um in (a) and at 20 um in (b). https://journals.e-palli.com/home/index.php/ajcp pa ge 46 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 41-52, 2023 all four sample images show un-interconnected irregular oval structured layers in both figures a and b. few geometrically shaped structures are observed in sample 3. the agglomerated oval glassy and geometrically shaped structures are typical of the presence of silica crystalline structures in the sample materials. the balloon-shaped crystals are indicative of the presence of cenospheres in the fly ash samples. there are significant pore sizes in the samples and are observed to be highly visible in specimens 1, 2, and 4 but less in specimen 3. this shows that there might be other materials present in the samples but in less or insignificant quantities. all four samples heated at various temperatures show that quartz or silica is the dominant crystalline phase present in the fly ash sample. the samples were heated at temperatures that will lead to the elimination of carbon contents present in the fly ash with no chemical reaction taking place. figure 4: sem images of unheated raw fly ash at three different magnifications according to the micrographs, the oval crystalline structures become more pronounced as the temperature increases from 5000c, 6000c, 7000c, and 8000c. conversely, the dark fields observed in the micrographs according to the morphology of the sample become lesser as the temperature is increased from sample one to four. this can be attributed to the carbon content which decreases as more temperature is applied to the fly ash. unheated fly ash sample figure 4 shows the sem micrographs of the unheated fly ash specimen with image sizes scanned at 200 um, 30 um, and 20 um pixels. the oval crystals are the same as those of the heated samples shown in figure 3 but are visibly observed to be far apart from each other compared to those seen in figure 3a above. this probably implies the presence of carbon content and other minor constituents that occupies the spaces between the crystals. eds result this is a powerful analytical detector or tool used to determine the chemical elements present in a sample and estimate their relative abundance. it relies on the interaction of some source of x-ray excitation and a sample. the capability of this tool is due to the fact or principle that each element has its unique structure that allows several peaks to display on its electromagnetic emission spectrum (which is the main principle of the electromagnetic spectrum). in the emission of x-ray characteristics from a sample, a beam of electrons is focused on the sample under investigation. at rest, an atom within the sample contains ground state (or unexcited) electrons in discrete energy levels or electron shells bound to the nucleus. the incident beam may excite an electron in an inner shell, ejecting it from the shell and forming a hole in the initial position of the electron. an electron from an outer, higher-energy shell then fills the hole, and the difference in energy between the higher-energy shell and the lowerenergy shell may be released in the form of an x-ray. the eds is then used to measure the number and energy of the x-rays emitted from the sample. the detector or tool allows the elemental composition of the sample to be measured since the energies of the x-ray are characteristic of the atomic structure of the emitting element. figure 5: eds result of heated fly ash specimens 1,2,3 & 4 https://journals.e-palli.com/home/index.php/ajcp pa ge 47 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 41-52, 2023 the eds shown in figures 5 and 6 confirmed the percentage weight and elemental composition of both heated and unheated fly ash samples. according to the eds result, oxygen and silicon are the predominating elements present in all five prepared samples. the percentage composition of oxygen is greater than that of silicon in all five samples. it is also observed from the elemental tables of the eds figures that the weight percentage of silicon is higher in the first, third, and unheated samples than those of oxygen atoms but lesser in the second and fourth samples. the results and analysis of this work show that fly ash used is silica pure and therefore a recommendable source for the fabrication of refractory composite materials. cenospheres according to the sem and eds results of this research, the hollow and spherical-shaped objects observed in all five samples depict the presence of cenospheres. cenospheres are one of the end products in coal combustion and are significant components for the synthesis of refractory composites. they are formed as a result of the decomposition of the mineral matter in coal at high temperatures >14000c. the disintegrated or decomposed mineral matter form into spheres known as cenospheres during the post-combustion phase or cooling process. the decomposition of calcium carbonate and other compounds below 10000c produces gases that blow up the minerals. the gases emitted during coal combustion inflate the decomposed minerals to form cenospheres. these objects are largely made of mullite, quartz (silica), aluminosilicate glass, calcite, sulfates, etc. inside the balloon-shaped structure of the cenospheres are gases while the outer membrane or layer is made of aluminosilicate, cristobalite, quartz, mullite, etc. cenospheres are discovered to be highly porous and can therefore effectively absorb water. figure 6: eds result of unheated fly ash specimen figure 7: eds and sem micrographs showing cenospheres the cenospheres discovered from the fly ash samples in this experimental research can be used for electronic and radar purposes due to their metallic and magnetic properties. cenospheres are useful to refractory industrial engineers due to their low thermal conductivities and high heat resistance. they also have properties that are suitable to make lightweight composite materials used in aerospace engineering. ftir result this technique is employed to study the vibrations of atoms in a molecule or compounds of a particular sample. the fractions of absorbed incident radiation at a particular energy are determined by passing infrared (ir) radiation through the specimen. the ftir results and analysis of the prepared fly ash specimens are discussed below. https://journals.e-palli.com/home/index.php/ajcp pa ge 48 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 41-52, 2023 heated samples the figure below shows the ftir spectra obtained from the characterization of fly ash samples heated at 5000c, 6000c, 7000c, and 8000c. according to the experimental result of the sample heated at 5000c, a strong appearance of o=c=o bending of carbon dioxide compound is observed at a wavenumber of 2349cm-1. further, the wavenumber 1083cm-1 depicts the appearance of sio-si stretching vibrations and strong absorption peaks while the 799cm-1 wavenumber shows si-o symmetric stretching of quartz. analysis of the ftir experimental data for the sample heated at 6000c also depicts the appearance of o=c=o at 2349cm-1 which indicates a strong vibrational stretching of the carbon dioxide compound. the wavenumbers 1090cm-1 and 807cm-1 indicate si-o-si and si-o stretching vibrations respectively. in sample three, the wavenumber 2363cm-1 from ftir results shows o=c=o bending but does not fall at 2349cm-1 which is the strong absorption band for o=c=o. therefore, the appearance of carbon dioxide compound at 2363cm-1 can either be weak or medium. the frequency range for the absorption of o=c=o is 2400cm1–2000cm-1. the wavenumber1090cm-1 and 803cm-1 show the stretching vibrations of si-o-si and si-o respectively. the ftir of the fourth sample heated at 8000c shows the absorption frequencies of 2369cm-1, 1123-1027cm1, and 798cm-1. the wavenumber 2369cm-1 shows weak or medium o=c=o stretching vibrations while 798cm-1 depicts the si-o symmetric of quarts. figure 8: ftir results of four heated fly ash samples unheated samples figure 9 shows the ftir result of the unheated fly ash. according to the analysis of this result, the wavenumber 2349 cm-1 shows the appearance of o=c=o stretching vibrations, 1049 cm-1 indicates the strong appearance of s=o stretching vibration of sulfoxide while 801 cm-1 depicts the si-o symmetric stretching vibration of the quartz. the frequency range for the strong absorption of sulfoxide compounds is 1070-1030. it is observed that the unheated fly ash shows a strong appearance of o=c=o and s=0 stretching. all five samples (both heated and unheated) show o=c=o bending with very strong stretching vibrations for the unheated sample and those heated at 5000c and 6000c while the third and fourth samples heated at 7000c and 8000c show a weak or medium appearance of o=c=o stretching. this suggests that the more the temperature the greater the chances of eliminating the carbon content in the fly ash. samples 1, 2, and 3 heated at 5000c, 6000c, and 7000c respectively also show the appearance of si-o-si but not observed for the unheated and the heated samples at 8000c. the si-o quartz appears in all five samples according to the ftir result. finally, it is observed from the ftir result of this work that sulfoxide functional group (s=o) only appears in the unheated fly ash and is not seen in all the heated samples. this is an indication that sulfoxide in the raw fly ash easily disappears when subjected to a certain amount of heat. figure 9: ftir result of unheated fly ash sample https://journals.e-palli.com/home/index.php/ajcp pa ge 49 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 41-52, 2023 table 1: wave numbers of different types of functional groups wavenumber (cm-1) compound functional reference 3454 montmorillonite oh group of absorbed water dust-stretching (summer, 1995) 2923 organic carbon c-h stretching vibration (hlavay, jonas, elek, & inczedy, 1978) 1629 organic matter c=o carboxylate group (langford, hodson, & banwart, 2011; matrajt et al., 2004) 1384 nitrate species n-o stretching (smidt, böhm, & schwanninger, 2011) 1097 quartz si-o-si asymmetric stretching vibration (katara, kabra, sharma, hada, & rani, 2013) 794 quartz si-o symmetric (coates, 1977) 463 feldspar amorphous silica si-o-si band (karr, 2013) 2517 calcite vibrational mode of carbonate (ramasamy, rajkumar, & ponnusamy, 2009) 1448 calcite c-o stretching for carbonate (ramasamy, suresh, meenakshisundaram, & ponnusamy, 2011) 1032 kaolinite si-o vibration of clay mineral (ramasamy, rajkumar, & ponnusamy, 2006) 873 calcite c-o bending for carbonate (dahlan, mei, kamaruddin, mohamed, & lee, 2008) 795 quartz si-o symmetric (coates, 1977) 712 calcite carbonate (adler & kerr, 1963) 466 feldspar si-o-si bending (dahlan et al., 2008) 1467 calcite c-o stretching for carbonate (ramasamy et al., 2011) 2520 calcite vibrational mode of carbonate (ramasamy et al., 2006) 796 quartz si-o symmetric (coates, 1977) 800, 802 quartz si-o bending (ramasamy et al., 2009) 1083, 1089 si-o-si stretching (ramasamy et al., 2011) 1080, 1025 si-o-si stretching vibration (ramasamy et al., 2011) 1020, 1090 si-o-si (ramasamy et al., 2011) 910 830 si-o stretching vibration (ramasamy et al., 2006) 3550 3200 alcohol o-h (ramasamy et al., 2006) 1685 conjugated ketone c=o stretching (ramasamy et al., 2006) 1662-1626 alkene c=c (ramasamy et al., 2006) 1205 1124 tertiary alcohol c-o stretching (smidt et al., 2011) 1124 1087 secondary alcohol c-o stretching (smidt et al., 2011) 1070 1030 sulfoxide s=o stretching (ramasamy et al., 2006) 850 550 halo compound c-cl stretching (ramasamy et al., 2006) 840 790 alkene c=c (ramasamy et al., 2006) 690 515 halo compound c-br stretching (smidt et al., 2011) 600 500 halo compound c-i stretching (smidt et al., 2011) 1650 1556 cyclic alkene (smidt et al., 2011) 2400 2000 carbon dioxide o=c=o (smidt et al., 2011) xrd result material scientists and solid-state chemists use the powdered x-ray diffraction technique to determine the structure of a crystalline solid specimen under investigation. the diffraction pattern is formed by the interaction of the x-ray and atomic structure. brag’s law below is used to explain the direction of the scattered x-rays produced. nnλ =2dsinθ where n = an integer λ = wavelength of the x-ray d = space between two layers of atoms θ = angle between the incoming x-ray and the atom layer the xrd results obtained from four heated and one unheated specimen of fly ash samples are explained in the figures below https://journals.e-palli.com/home/index.php/ajcp pa ge 50 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 41-52, 2023 figure 10: xrd result of heated fly ash the xrd result in figures 10 and 11 shows the presence of sio2 in all the fly ash samples. this information is in line with the sem/eds and ftir characterizations. the results and analysis of all the characterization in this work confirmed that the purified fly ash obtained from xi’an linyuan silica limited is pure silica. conclusion critical analysis of the experimental results from the four characterization techniques shows that fly ash purely constitutes sio2 which is a significant constituent for the fabrication of refractory composites. according to the sem/eds results, the cenospheres discovered in the specimens are useful to refractory industrial engineers due to their low thermal conductivities and high heat resistance. acknowledgments this work was supported by the national natural science foundation of china. thanks, and appreciation to wang wei of the chemical engineering department, school of water and environment of chang’an university for the adequate supervision and for providing the platform for the successful conduct of this work. references adler, hans h, & kerr, paul f. 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(2016). char burnout characteristics of five coals below and above ash flow temperature: tg, sem, and eds analysis. applied thermal engineering, 103, 1156-1163. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 37 american journal of chemistry and pharmacy (ajcp) dietary risks assessment of antibiotic residues and microbial safety of local honey in adamawa state dowell b.f1*, maitera o.n1, daniel m.w1, micah m.m1 volume 1 issue 1, year 2022 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v1i1.492 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: august 11, 2022 accepted: september 02, 2022 published: september 12, 2022 this study investigated the microbial safety and dietary risk assessments of antibiotic residues in honey. the determination of antibiotic residues in this study were carried out using hplc (lc 1200 series agilent tech). the results of the microbial analysis of the honey samples showed that, some of the honey samples were contaminated with micro-organisms. the antibiotics with highest mean concentration in the study were chloramphenicol (6.468±0.03 µg/kg) and sulfonamide (5.553±0.04 µg/kg) from sample hs1 and hg2 respectively. estimated daily intake (edis) of detected antibiotic residues which were compared with the recommended acceptable daily intake (adis) for each antibiotic allowed in food. chloramphenicol was observed to possess the highest risk for adults and children (1.08e-04 µg/kg/bw) and (4.31e-04 µg/kg/bw) respectively in sample hs1. all the edis evaluated for antibiotic residues falls below acceptable daily intake (adi). the target hazard quotient (thq) and health index (hi) for antibiotic residues in all the samples were < 1, this suggested no risks associated with the level of antibiotic residues risks thus having less danger to human health through its consumption. keywords antibiotic residues, estimated daily intake, health index, honey, microbial safety 1 chemistry department, modibbo adama university (mau), p.m.b 2076, yola, adamawa state, nigeria * corresponding author’s e-mail: dowellfarimani@yahoo.com introduction honey is the natural sweet, viscous substance produced by honeybees from the nectar of flowers or from the secretion of living parts of plants or excretions of plant sucking insects on the living parts of plants, which honeybees collect, transform and combine with specific substances of their own, store and leave in the honey comb to ripen and mature (abeshu and geleta, 2016). it is one of the most widely sought products because of its unique nutritional and medicinal properties (james et al., 2009). the variety produced by honey bees (the genus apis) is the most commonly sought for because it is collected by most beekeepers and consumed by people. honey is also produced by bumble bees, stingless bees and other hymenopteran insects such as honey wasps, though the quantity is generally lower and they have slightly different properties compared with honey from the honey bees. honey bees convert nectar into honey by a process of regurgitation and evaporation: they store it as a primary food source in wax honeycombs inside the beehive (satarupa and subha, 2014). honey is composed primarily of the sugar: glucose and fructose; its third greatest component is water (singh et al., 2012). honey also is composed of a complex mixture of carbohydrates and other less frequent substances, such as organic acids, amino acids, proteins, minerals, vitamins, lipids (blasa et al., 2006; ball, 2007; zerrouk et al., 2011), aromatic compounds, flavonoids, vitamins, pigments, waxes, pollen grains, several enzymes and other phytochemicals (gomes et al., 2010). in nigeria, domestic consumption rate of honey was estimated at 380,000 tonnes, with a global price of about 4.5 billion dollars (vanguard news, 2017). though, the country has the potential to produce about 800,000 tonnes, and can generate over 10 billion dollars from local and international trade, her current potential honey output is less than 3% (punch news, 2017). the current production of honey liquid in nigeria is 2 million liters but has potential to produce 20 million liters of honey annually (the guardian news, 2018). across nigeria, net return analysis showed that honey bee production is profitable, encouraging gross margin and net income (duruson, 2011; igbokwe and mbanaso, 2006; uduma and udah, 2015, abdullahi et al., 2014, folayan and bifarin, 2014). though, the returns generated from the businesses have an encouraging outlook, these figures are considered far beyond the estimated market potential endowed within nigeria and her young populace. antibiotics, particularly streptomycin, sulfonamide, and chloramphenicol are often used by beekeepers to treat bee-related diseases and as growth enhancers. the relatively long shelf-life of antibiotic residues in foods could indirectly leads to the emergence of bacteria strain that can resist the antibiotics overtime. could also induces allergic reactions in hypersensitive individuals, and could leads to the disorder of the hemopoietin system (tillotson et al., 2006). antibiotic residues in honey have become a major consumer concern. some drugs have the potential to produce toxic reactions in consumers directly while some other is able to produce allergic or hypersensitivity reactions (velicer et al., 2004). considering these negative effects of antibiotic, the residual level in foods from plant and animal origin are regulated in developed economy (vragović et al., 2012), however, these limits have received poor recognition in developing economy like nigeria, thus posed a serious public health risk burden (mensah et al., 2014). the determination of antibiotic residues and microbial safety https://doi.org/10.54536/ajcp.v1i1.492 https://journals.e-palli.com/home/index.php/ajcp mailto:dowellfarimani%40yahoo.com?subject= pa ge 38 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 37-46, 2022 in honey and other bee products has become a growing concern considering the growing popularity of honey in human daily diets. its ingestion without knowing its source and safety might carry significant health hazards. residual level of contamination cannot be changed through various production techniques, hence the need to study the microbial safety and dietary risks of antibiotics residues in the samples of local honey. methodology materials and chemicals/reagents standard of tetracycline, streptomycin, sulfonamides, chloramphenicol, distilled water, deionized water, acidified water, sodium acetate, anhydrous magnesium sulphate, sodium hydroxide, plastic bottles, methanol, acetonitrile, volumetric volumes, electric centrifuge, beaker, glass tubes, polyethylene bottles, petri dish, sterile pipette, potato dextrose agar, tryptone neomycin agar, potassium chloride, hplc (lc 1200 series agilent tech). all are analytical grade. study area and geographical location adamawa is a state in north-eastern nigeria, with its capital at yola. it is located between latitude 9°20′n and longitude 12°30′e (fig 3.1). it occupied an area of 36,917 square kilometers. the state has population of 3,178,950 (npc, 2006). adamawa is one of the largest states of nigeria, the state of borno borders it to the northwest, gombe to the west and taraba to the southwest. it’s eastern bordered with cameroon. it comprises of 21 local government area. sample collection, storage and preservation the honey samples were collected from three (3) local government areas of adamawa state (ganye, song and mubi north from the southern, central and northern senatorial zones respectively). the samples were mixed to make a composite sample that represents each sampling location (table 3.1). six (6) composite samples were collected purposively from the bee-keepers farms namely: sugu, ganye i, dirma, shimba, lokuwa and yewa hosere at different locations. similarly, three (3) composite samples were purchased from retailers at different point (ganye ii, song and mubi) in the same geographical area. the samples were collected during dry season (januarymarch 2021). the collected samples were properly stored at room temperature and preserved to avoid loss due to poor storage, was timely analyzed to minimize loss due to prolonged storage. samples were stored in polyethylene bottles preserved for analysis (japhet et al., 2018). figure 1: map of the study area (google map 2020). table 1: sample locations of honey and coding l.g.a of the senatorial zones in adamawa state sample locations code ganye (southern senatorial zone) sugu hg1 ganye i hg2 ganye ii hg3 song (central senatorial zone) dirma hs1 shimba hs2 song hs3 mubi north (northern senatorial zone) lokuwa hm1 yewa hosere hm2 mubi hm3 https://journals.e-palli.com/home/index.php/ajcp pa ge 39 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 37-46, 2022 figure 2: samples collected for analysis microbial contaminants analysis standard plate count ten grams of honey was suspended in 90 ml of 0.1% phosphate buffer solution. a series of dilutions was then carried out and 0.1 ml spread on plate count agar (pca) (oxoid). the culture was incubated for 72 h at 37°c. yeast and mold count isolation of microorganisms one ml of each sample was picked with the aid of a sterile pipette. a plastic rack was arranged with 9 sterile test tubes containing 9 milliliters of sterile distilled water. a ten-fold dilution adopted by (eleazu et al., 2013) was done by dispersing 1 ml of the sample into the first test tube (10-1) which will be well shaken. one milliliter was then taken again from 10-1 dilution and transferred to the next test tube (10-2). the dilution continued to 10-9. each test tube was shaken vigorously before each transfer. inoculation shree & arlis (2003) used the pour plate method to plate all the samples. 0.1 ml from dilution 10-2 was dispersed into sterile petri dish with the aid of a sterile pipette. potato dextrose agars was poured into the plates (10 ml) and isolation of fungi were carried out using the potato dextrose agar. the plates were swirled gently for easy mixing of the samples and the media. all plates were allowed to solidify on the bench and each plated sample was duplicated. incubation the potato dextrose agar (pda) plates were transferred to an incubator at 25˚c for 3-5 days. all incubated plates were examined daily for mycelia and colony growth. subculture and purification after the incubation period, a flamed surgical knife was used to subculture different color of mycelia growth from potato dextrose agar (pda) plates on newly prepared pda plates. all while the pda plates were incubated at 25˚c for 3-5 days. counting of colonies after incubation of all the plates, counts of the number of colonies in each plate was done with a hand tally counter (fawole et al., 1988). the mean counts were obtained and multiplied by the appropriate dilution factor. the mean count was calculated as: mean = (total viable count)/(number of plates) the estimation of the viable counts in each sample was made in colony forming unit (cfu) and total viable count = (number of colonies x dilution factor)/ (amount plated (ml)) sulfite-reducing anaerobic bacteria (clostridia) the technique of enumeration carried out the sulfitereducing anaerobic bacteria analysis in a solid medium in tubes. to 1 ml decimal dilution (10−2), contained in sterile tubes, 20 ml of the melted sulfite tryptone neomycin agar was added and cooled in a water bath at 45˚c. after homogenization and solidification, the tubes were incubated anaerobically at 37˚c for 24–48 hours. colonies characteristics of sulfite-reducing anaerobic bacteria appear black in the tubes (francois et al., 2018) total coliform and fecal coliform (iso 4831: 2006) the total coliform search was carried out according to iso 4831: 2006. 1 ml of decimal dilution (10−2) was poured aseptically into sterile plates. purple crystal, bilelactose neutral red agar (vrbl), melted and cooled in a water bath at 45˚c, was added to the inoculum at a rate of 15 ml per dish. the mixture was then homogenized by rotary movements. after solidification of the first layer, a second 5 ml layer of vbrl was added. control of the sterility of the medium was carried out in a petri dish https://journals.e-palli.com/home/index.php/ajcp pa ge 40 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 37-46, 2022 with approximately 15 ml of vbrl. the total coliform count was done directly after incubation at 30˚c for 24–48 hours. fecal coliforms are characterized by a small mass of fluorescent colonies with a diameter of 0.5 mm (francois et al., 2018). extraction and determination of antibiotic residues using hplc the extraction of honey samples was carryout subjecting to deproteinizing chemical procedure using acetonitrile (acn). 2 g of the honey sample was place into 10 ml test tubes and shake intensively with 3 ml acetonitrile (acn) for 1minute. the mixture was centrifuge for 15 minutes at 5000 rpm. the supernatant was collected and dried under nitrogen stream at 40°c. the residue will be redissolved in methanol and filtered through 0.45µm filter paper describe by pagliuca et al., 2002). the presence of antibiotic residues in the honey samples was carried out using different mixture of aqueous mobile phase (a) acidified water and organic mobile phase (b) methanol/acn with a flow rate of 1 ml/min. the respective antibiotic residues were quantified by a modified method described by (albino et al., 2005 and shafqat et al., 2012), detected at 210-240 nm. the health risk assessment and hazard characterization of antibiotic residue the health risk assessment and hazard characterization were carryout by first estimating the daily intake (edi) of the respective antibiotics in the honey. this was achieved by integrating the average concentration of the antibiotics in the honey, the average consumption rate of the honey and the average body weight per person as described in equation 1 (forkuoh et al., 2018; usepa, 1997). edi=(ch×hir)/bw…………………… equation 1 the ch is the antibiotic concentration (µg/kg) in the honey, hir represents the average honey consumption rate or intake rate for an average child and adults (0.001). the bw is the average body weight of children (15 kg) and adults (60 kg) (usepa, 2000; akbari et al., 2012). the potential non-carcinogenic risk from the consumption of the antibiotics the potential non-carcinogenic risk from the consumption of the antibiotics were estimated using target hazard quotient (thq) and the health index (hi) as described by the united state environmental protection agency (usepa, 1997). the thq was estimated by integrating the ratio of the edi to the acceptable daily intake (adi) values for each antibiotic (fao/who, 2002 and 2010; usepa, 1996; bwatanglang et al., 2019; bwatanglang, 2019). the expression for estimating the thq are described in equation 2. thq=edi/adi…………….........…equation 2 the hi, expressed as the sum of the thq as described in equation 3 is the cumulative effect pose by the combination of the individual antibiotics presents in the honey (forkuoh et al., 2018; reffstrup et al., 2010). hi = (edi1/adi1) +(edi2/adi2) +(edi3/adi)+…. (edii/adii)……………………..equation 3 were the edii represents the estimated daily intake dose of the individual antibiotics (1, 2, 3….) in the honey and the adii is the acceptable daily intake dose for the individual antibiotics (1, 2, 3……) statistical analysis the mean and standard deviation of the results were done using minitab 19 and data was generated in triplicate. the results were express as mean± sd results and discussion microbial analysis the results of the microbial analysis of the honey samples showed that, some of the honey samples are contaminated with micro-organisms. microbial analysis results are shown in table 2 and 3 the spc were found in low numbers in all samples of honey with a mean count which varies within 1.0 x 104 – 3.3 x 104 cfu/ml. the yeast and mold count of honey samples were less than 10 cfu/ml in sample in all the samples. table 2: mean results of microbial analysis sample location sample spc in cfu/ml mold and yeast (cfu/ml) x 10^2 sulfite-reducing clostridia total coliform and fecal coliform ganye hg1 1.7 x 10^4 2 nd nd hg2 2.0 x 10^4 nd nd nd hg3 3.2 x 10^4 1.8 nd nd song hs1 1.0 x 10^4 nd nd nd hs2 nd nd nd nd hs3 3.3 x 10^4 1.6 nd nd mubi-north hm1 1.3 x10^4 nd nd nd hm2 nd 1 nd nd hm3 nd 1.5 nd nd spc = standard plate count nd = not detected cfu = colony forming units https://journals.e-palli.com/home/index.php/ajcp pa ge 41 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 37-46, 2022 table 3: isolated micro-organisms pda (fungi) from local honey samples in adamawa state sample location sample aspergillus fumigatus aspergillus flavus aspergillus niger penicillum sp trichophyton rumbrum ganye hg1 + + hg2 hg3 + + song hs1 hs2 hs3 + + mubinorth hm1 hm2 + + hm3 + key: + = present = absent pda = potato dextrose agar microbial contamination the mean level of microbial contamination of different honey samples were presented in table 2. the contamination with standard plate count (spc) for aerobic mesophilic bacteria varies within 1.0 x 104 – 3.3 x 104 cfu/ml. this result conforms with the result of ndife et al., 2014. according to tchoumboue et al., 2008, the contamination with fungi and bacteria indicate inadequate hygienic conditions during collecting, manipulating, processing and storing. microbial contamination during and post processing can also result in spoilage or persistence of some bacteria in honey. the yeast and mold count of honey samples were less than 10 cfu/ml in samples hg1, hg3, hs3, hm2 and hm3 but was not detected in sampling points hg2, hs1, hs2 and hm1 (table 3). there was total absence of sulfitereducing clostridia and fecal coliforms in all the samples, indicating that honeys were produced in accordance with good hygiene practices during extraction, packaging and storage (guiraud, 2003). the low microbial loads of honey samples could be attributed to their low ph values and high amount of total soluble sugars and possibly phenolic compounds and their synergistic interaction (alvarez-suarez et al., 2010). yeast, mold and spore-forming bacteria (coliform) have been implicated to survive in honey and are indicative of the sanitary quality of the honey (ezeama, 2007; eleazu et al., 2013). in a study conducted by omafuvbe and okanbi 2009, observed that no mold contamination was detected. the micro-organisms that were isolated in the honey samples from all sampling points are fungi (table 3). the presence of micro-organisms in honey can sometimes influence the stability of the product and its hygienic quality. normal honey must lack pathogenic micro-organisms or micro-organisms that produce enteric illnesses (popa et al., 2009). determination of antibiotic residues the determination of antibiotic residues in all the sampling points was carried out using hplc (lc 1200 series, agilent tech). the health risks assessment, target hazard quotient and health index was estimated using various mathematical expressions and results presented in tables. maximum residues level (mrls) are yet to be established for honey and other bee products (alwaili et al., 2012). the eu (european union) under the council directive 2001/110/ec has set a reference point for action (rpas) for antibiotic residues in honey which is also used as provisional mrl (forsgen, 2010; johnson et al., 2010; efsa, 2013). the rpas are residue concentrations which are technically feasible for analytical considerations place as a bench mark to reject any bee products exceeding these limits (johnson et al., 2010; esfa, 2013; mutinelli, 2003). provisional mrl in parts per billion (ppb) for oxytetracycline (25 ppb), chloramphenicol (0.3 ppb) and nitrofurans (1.0 ppb) were established by eu for honey (johnson et al., 2010). mean concentration of antibiotic residues the samples’ average mean± sd concentration of antibiotic residues were analyzed and the results presented in table 4. all the samples contained at least two or more antibiotic residues detected. streptomycin were below the detection level (bdl) in samples hg1, hg2, hg3, hs1, hs2 and hm2. only samples from song (hs3) and mubi north (hm1 and hm3) contains the level of streptomycin antibiotic residues. similarly, tetracycline was below the detection limit (bdl) in samples from hg1, hs1, hs2, hm2 and hm3 as can be seen in table 4. among the antibiotics analyzed, sulfonamide was detected in virtually all the samples except sample hs3 and hm1 from song and mubi-north respectively. the antibiotic with the highest concentration detected in the honey samples from all locations are: chloramphenicol and sulfonamide (table 4). the highest mean concentration of tetracycline and streptomycin were found in sampling point hg3 from ganye (1.513± 0.01 µg/kg) and sampling point hs3 from song (1.513± 0.05 µg/kg) respectively. the samples hg2 and hs1 from ganye and song were observed to contain the highest concentration of sulfonamide with a mean concentration of 5.553±0.04 µg/kg and 5.386±0.07 µg/kg respectively. similarly, highest mean concentration of 6.468± 0.03 µg/kg and 6.447± 0.03 µg/kg were detected for chloramphenicol in sampling point hs1 and hg2 from song and ganye respectively. https://journals.e-palli.com/home/index.php/ajcp pa ge 42 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 37-46, 2022 in general, the consumption of honey containing antibiotic residues represents a real danger to human health, considering that the indiscriminate consumption of these compounds is the main cause of bacteria resistance to antibiotics, especially in the case of aminoglycosides (streptomycin). also, antibiotics can cause allergic reactions in people sensitive to certain classes of compounds and can cause hypersensitivity in up to 5% of individuals exposed to these substances (brum, 2018; ford, 2017). aminoglycosides are antibiotics that act by interfering with the replication of bacterial dna by blocking protein synthesis. this class of antibiotics causes serious toxic adverse effects on the kidneys and the auditory system and, when used in high concentrations, can cause a serious risk of neuromuscular paralysis. regarding tetracyclines, these cause adverse effects to the gastrointestinal system, causing irritation and may lead to deficiency disorders in the absorption of some vitamins. besides, they cause kidney damage and bone marrow disorders, and can also cross the placental barrier and deposit in the bones (brum, 2018; ford, 2017). sulfonamides, better known as sulfas, are antibiotics that act by inhibiting the metabolism of bacterial cells by inhibiting folic acid, making the cells unable to grow and multiply. regarding the adverse effects caused by these substances, these can range from itchy skin, hypersensitivity reactions, liver problems to problems related to blood cells. (brum, 2018; ford, 2017; omidi et al., 2016). bwatanglang et al., (2019) reported present of antibiotics residues from uba/uvu, mubi/vimtim and gombi/ garkida for both raw honey sample (rhs) and commercial honey sample (chs). it was found that tetracycline, chloramphenicol, sulfonamide and streptomycin were all detected in the rhs from all the sampling points. studies conducted in other countries also reported the presence of antibiotic residues in honey samples. twenty nine percent of 251 honey samples produces across greece were observed to contain residual level of tetracycline from 0.0180.100 mg/kg (saridakipapakonstadinou et al., 2006). about 13 honey samples out of 34 imported from asian countries into switzerland were found to 0.4 and 9.0 µg/kg of chloramphenicol with at least two samples containing up to 5.0 µg/kg (orteli et al., 2004). in another study, streptomycin 310,820 µg/ kg, sulfonamide 54,592 µg/kg, chloramphenicol 0.1 169 µg/kg were detected in honey samples from eu (diserens, 2007). table 4: mean concentrations of antibiotic residues (µg/kg) in honey samples sample location sampling points chloramphenicol sulfonamide tetracycline streptomycin ganye hg1 0.102±0.05 2.232±0.02 nd nd hg2 6.447±0.03 5.553±0.04 0.031±0.03 nd hg3 nd 0.576±0.01 1.513±0.01 nd song hs1 6.468±0.03 5.386±0.07 nd nd hs2 0.194±0.02 0.150±0.01 nd nd hs3 1.743±0.05 nd 1.292±0.03 1.513±0.05 mubi-north hm1 nd nd 0.060±0.01 0.126±0.07 hm2 1.042±0.04 0.687±0.01 nd nd hm3 nd 5.042±0.03 nd 0.215±0.02 nd = not detected potential health risk of antibiotic residues associated with consumption of honey samples the results of human health risk assessments were presented in table 5. the estimated daily intake (edis) of detected antibiotic residues for both adults and children in all the samples were compared with the recommended acceptable daily intake (adis) for each antibiotic in all food. the dietary exposure assessment is very critical towards evaluating the risks associated with antibiotics in honey. helping to determining whether a residual of concerns to pose a potential risk to public health. based on the antibiotic residues detected in all the sampling points, chloramphenicol was found to possessed the highest exposure risks to both adults and children (1.08e-04 µg/kg/bw) and (4.31e-04 µg/kg/bw) respectively in sample hs1 (see table 5). sulfonamide presents the highest edis for adults (9.26e-05 µg/kg/ bw) and children (3.70e-04 µg/kg/bw) from sample hg2 (table 5). similarly, the highest edis for tetracycline was found in sampling point hg3 (2.52e-05 µg/kg/bw) and (1.00e-04 µg/kg/bw) for adults and children respectively while edis for streptomycin was only obtained in sample points hs3, hm1 and hm3 from ganye and mubinorth for both adults and children (table 5) due to the smaller body weight (15 kg) and physiological susceptibility in children, the edi were observed to be higher in children than that of the adults. with the exception of chloramphenicol which has no define adi, all the edis calculated for each antibiotic residue were found to be below their recommended adis (johnson et al., 2010). this shows that the consumption has a negligible risk to human health having < 1% adi in all the samples. estimated daily intake (edi) of 2.09 ng/kg body weight (bw)/day and 1.83 ng/kg bw/day for tetracycline and penicillin residues were determined in dairy products resulting in 0.007% and 0.006% of the adi respectively (kabrite et al., 2019). other related study reported various exposure indices due to dietary intake of antibiotics by human. residues of quinolones and sulfonamides were https://journals.e-palli.com/home/index.php/ajcp pa ge 43 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 37-46, 2022 found to be widely distributed in in cultured fish samples from the pearl river delta, south china. the edi results showed that the consumption of the fishes to dietary intakes of quinolones and sulfonamides were far below the acceptable daily intake (adi) and poses no risk to the public health (he et al., 2016). table 5: estimated daily intake (edi) in µg/kg/bw for antibiotic residues adults ganye song mubi north antibiotics hg1 hg2 hg3 hs1 hs2 hs3 hm1 hm2 hm3 adi chloramphenicol 1.70e-06 1.07e-04 0.00e+0 1.08e-04 3.23e-06 2.91e-05 0.00e+0 1.74e-05 0.00e+0 na sulfonamide 3.72e-05 9.26e-05 9.60e-06 8.98e-05 2.50e-06 0.00e+0 0.00e+0 1.15e-05 8.40e-05 5.00e+01 tetracycline 0.00e+0 5.17e-07 2.52e-05 0.00e+0 0.00e+0 2.15e-05 1.00e-06 0.00e+0 0.00e+0 3.00e+01 streptomycin 0.00e+0 0.00e+0 0.00e+0 0.00e+0 0.00e+0 2.52e-05 2.10e-06 0.00e+0 3.58e-06 5.00e+01 children ganye song mubi north antibiotics hg1 hg2 hg3 hs1 hs2 hs3 hm1 hm2 hm3 adi chloramphenicol 6.80e-06 4.30e-04 0.00e+0 4.31e-04 1.29e-05 1.16e-04 0.00e+0 6.95e-05 0.00e+0 na sulfonamide 1.49e-04 3.70e-04 3.84e-05 3.59e-04 1.00e-05 0.00e+0 0.00e+0 4.58e-05 3.36e-04 5.00e+01 tetracycline 0.00e+0 2.07e-06 1.00e-04 0.00e+0 0.00e+0 8.61e-05 4.00e-06 0.00e+0 0.00e+0 3.00e+01 streptomycin 0.00e+0 0.00e+0 0.00e+0 0.00e+0 0.00e+0 1.01e-04 8.40e-06 0.00e+0 1.43e-05 5.00e+01 table 6: target hazard quotient (thq) and health index (hi) for antibiotic residues adults ganye song mubi north antibiotics hg1 hg2 hg3 hs1 hs2 hs3 hm1 hm2 hm3 chloramphenicol sulfonamide 7.44e-07 1.86e-06 1.92e-07 1.80e-06 5.00e-08 0.00e+0 0.00e+0 2.30e-07 1.68e-06 tetracycline 0.00e+0 1.72e-08 8.40e-07 0.00e+0 0.00e+0 7.17e-07 3.33e-08 0.00e+0 0.00e+0 streptomycin 0.00e+0 0.00e+0 0.00e+0 0.00e+0 0.00e+0 5.04e-07 4.20e-08 0.00e+0 7.20e-08 hi 7.44e-07 1.88e-06 1.03e-06 1.80e-06 5.00e-08 1.22e-06 7.53e-08 2.30e-07 1.75e-06 children ganye song mubi north antibiotics hg1 hg2 hg3 hs1 hs2 hs3 hm1 hm2 hm3 chloramphenicol sulfonamide 3.00e-06 7.40e-06 7.68e-07 7.18e-06 2.00e-07 0.00e+0 0.00e+0 9.16e-07 6.72e-06 tetracycline 0.00e+0 6.90e-08 3.33e-06 0.00e+0 0.00e+0 2.87e-06 1.33e-07 0.00e+0 0.00e+0 streptomycin 0.00e+0 0.00e+0 0.00e+0 0.00e+0 0.00e+0 2.02e-06 1.68e-07 0.00e+0 2.86e-07 hi 3.00e-06 7.47e-06 4.10e-06 7.18e-06 2.00e-07 4.89e-06 3.01e-07 9.16e-07 7.01e-06 target hazard quotient (thq) and health index (hi) for antibiotic residues the results of thq and hi for potential noncarcinogenic risks were evaluated and presented in table 6. based on the results obtained for thq and hi, shows that the exposure to the antibiotics through the dietary consumption of honey possess no immediate effect to human health as a result of noncarcinogenic related risks (table 6). the health index (hi) for all the samples analyzed falls below the level of concern to human health, shows hi < 1 in all the samples as can be seen in table 6. the thq and hi analysis are health-based statistical probability expressed as a function of quantified level of concern; a process developed to exposure to environmental pollutants (bwatanglang, 2019). the target hazard quotient (thq) and health index (hi) obtained < 1 in all the sampling points also suggest no risks associated with the level of chloramphenicol, sulfonamide, tetracycline and streptomycin in honey, thus having less danger to human health through its consumption. but children are more adaptable to encountered the risks associated with their health due to the high value of thq and hi observed more than the adults. similar study on the risk assessment due to dietary exposure to oxytetracycline, tetracycline and chlortetracycline through milk consumption in india showed hq < 1 (chauhan et al., 2018). though adi was not defined for chloramphenicol and the thq and hi could not be evaluated in this study, the consumption of the honey samples analyzed in this study points toward potential health risk to chloramphenicol. lack of availability of adi for chloramphenicol further suggest a zero-tolerance level to the antibiotics. an adi could not be established for chloramphenicol due to lack of genotoxic and toxicological data, in addition to lack of definable noael (no observe adverse effect level) or loael (lowest observed adverse effect level) (efsa, 2013). these makes residual level of chloramphenicol not allowed in the animal food-production chain. and thus, reported to constitute threat to public health (efsa 2013; commission regulation (eu) no 37/2010; and no 165/2010). conclusion honey is consumed on a large scale throughout the world and becomes mandatory to carryout microbial analysis, monitoring and evaluation of the risk to the health of https://journals.e-palli.com/home/index.php/ajcp pa ge 44 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 1(1) 37-46, 2022 journal of chemical education, 84(10), 1643. blasa, m., candiracci, m., accorsi, a., piacentini, m. p., albertini, m. c. and piatti, e. (2006). raw millefiori honey is packed full of antioxidants. food chemistry, 97, 217-222. brum, l. f. s. (2018). farmacologia aplicada à farmácia (1 ed). porto alegre: sagah. bwatanglang i (2019). diclorvos-mediated heavy metal uptake in leafy vegetables and potential health risk on consumption. london journal of research in science: natural and formal, 19(3), 15-24 bwatanglang ib, j bimba, st magili, y musa and sp zira (2019). dietary exposure to antibiotics residue in honey and the potential health risks to consumers in adamawa state, nigeria. international journal of chemical studies, 7(4), 1672-1679. bwatanglang ib, magili st (2019). health risk assessment of insulin supporting elements in commonly consumed anti-diabetic medicinal plants. london journal of research in science: natural and formal, 19(4),11-22 chauhan sl, priyanka, garg sr, jadhav vj (2019). dietary exposure assessment of tetracycline residues in milk in haryana. international journal of chemical studies, 7(1), 1862-1865. codex alimentations (2001). draft revised standard for standard for honey (at step 10 of the codex procedure). alinorm, 1(25), 19–26 commission regulation (eu), (2010). no 165/2010 on amending regulation (ec) no 1881/2006 setting maximum levels for certain contaminants in foodstuffs as regards aflatoxins. off. j. eur. union. 2010, 50, 8-12. commission regulation (eu), (2010) on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin. off. j eur. commun, 15(1), 72. diserens, j. (2007). contaminants and residues in food. strategies (if any) to screen and analyze veterinary drug residues in food from animal origin. in proceedings of the 5th international fresenius conference nestle research center. dolan, j. w (2002). peak tailing and resolution. lc●gc europe, (pp 34-36.). duruson, g. c. (2011). profitability of apiculture as a bussiness venture in ikwuano local government area of abia state. b. sc. project. department of agricultural economics and extension, abia state university, umuahia campus, umuahia, abia state, nigeria. efsa panel on contaminants in the food chain (contam). (2013). guidance on methodological principles and scientific methods to be taken into account when establishing reference points for action (rpas) for nonallowed pharmacologically active substances present in food of animal origin. efsa journal, 11(4), 3195. eleazu, c. o., iroaganachi, m. a., eleazu, k. c. and okoronkwo, j. o. (2013). determination of the the consumers. the result of the microbial safety show that the samples studied are of very good microbiological and hygienic quality, though there were isolated microorganisms in the honey samples from some sampling points. this maybe as a result of improper handling during the production process and also air dust from the environment. the presence of micro-organisms in honey can sometimes influence the stability of the product and its hygienic quality. the potential human health risks associated with exposure to antibiotic residues were found to be lower than each acceptable daily intake (adi) but the presence of antibiotic and pesticide residues in honey is a concern to health. due to the smaller body weight (15 kg) and physiological susceptibility in children, the edi were observed to be higher in children than that of the adults. recommendations the following recommendations have been suggested to enable better understanding and improve the results obtained from such research: i. there should be educational programs to the bee keepers within the state on beehive management, this will create awareness on the safety of consumers and public health. ii. the presence of some pathogenic fungus in the samples necessitates an urgent need to always monitor microbial status of honey. iii. microbial testing should guarantee both good hygienic and good marketable qualities of this products and good production efficiency. iv. there should be continual monitoring and evaluation of antibiotic and residues in honey, this will help to assess the potential risks to human health. references abdullahi, a., isekenegbe, j., & mohammed, u.s. 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biochemical, and analytical perspectives s. i. okonkwo1*, o. s. ochie2, i. p oragwu1, c. k. okonkwo3, p. o. okwuego1, a. t. kene okonkwo4, v. s. okonkwo5, s. c. okonkwo6 volume 4 issue 2, year 2025 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v4i2.5342 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: may 15, 2025 accepted: june 20, 2025 published: december 13, 2025 crude oil spills are an increasingly urgent global concern due to their profound environmental and public health consequences. this study investigates the potential of agro-waste-derived nanostructured sorbent specifically synthesized from rice and melon husks for the remediation of crude oil-contaminated water. using a multidisciplinary framework, the research incorporates analytical chemistry, biochemistry, sonography, and medical diagnostics to assess both the efficiency and biomedical safety of the nanomaterials. adsorption experiments for premium motor spirit (pms) were conducted. characterization using brunauer–emmett–teller (bet) and fourier transform infrared spectroscopy (ftir) analysis revealed high porosity, favorable surface morphology, and the presence of polar functional groups that enhance sorption. kinetic modeling followed pseudo-secondorder kinetics, confirming chemisorption, while langmuir isotherm modeling demonstrated monolayer adsorption on homogeneous surfaces.toxicological evaluation was carried out on albino rats grouped into control, oil-exposed, and remediated water cohorts. biochemical markers such as alt (alanine transaminase), ast (aspartate transaminase), creatinine, and urea levels were measured to assess hepatic and renal function. elevated enzyme levels in oil-exposed rats (alt: 120 iu/l; ast: 145 iu/l; creatinine: 2.6 mg/dl) were significantly reduced post-treatment (alt: 52 iu/l; ast: 65 iu/l; creatinine: 1.3 mg/ dl), approaching control values (p < 0.05). sonographic scans indicated hepatomegaly and hyperechogenicity in contaminated groups, while remediated groups showed normalization of liver and kidney echotexture. medical analysis concluded that the sorbents not only removed toxic hydrocarbons but also reversed or prevented hepatic and renal structural damage.this integrative study underscores the synergistic value of combining environmental nanotechnology with clinical diagnostics. the results validate the potential of using rice and melon husk-derived nanomaterials as eco-friendly, cost-effective, and biologically safe options for crude oil remediation. keywords agro-waste, alt, ast, bet, crude oil remediation, ftir, langmuir isotherm, nanostructured sorbents, pseudosecond-order kinetics, sonography 1 department of pure and industrial chemistry, chukwuemeka odumegwu ojukwu university, uli, anambra state, nigeria 2 food safety and applied nutrition directorate, national food and drug administration and control (nafdac), owerri, nigeria 3 department of diagnostic medical sonography and ultrasound technology, ace institute of technology, elmhurst, new york, usa 4 tansian university oba, anambra state, nigeria 5 department of medical biochemistry, chukwuemeka odumegwu ojukwu university, uli, nigeria 6 department of pharmacology, chukwuemeka odumegwu ojukwu university, nigeria * corresponding author’s e-mail: si.okonkwo@coou.edu.ng introduction crude oil remains one of the most widely used fossil fuels, contributing significantly to global energy supply. however, crude oil spills, particularly in developing countries like nigeria, pose acute risks to the environment and public health (nwilo & badejo, 2005). the niger delta, known for its rich biodiversity, has been a hotspot for frequent oil spill incidents due to pipeline vandalism, illegal refining, and inadequate spill response mechanisms (undp, 2006). previous research has documented the toxic effects of hydrocarbons on aquatic life, soil productivity, and human health. inhalation or ingestion of petroleumcontaminated water is linked to hepatic dysfunction, nephrotoxicity, carcinogenesis, and endocrine disruption (who, 2021). the detection of these pathologies is now greatly enhanced through sonographic imaging and biochemical assays (olatunde et al., 2020). this study bridges the gap between environmental science and medical diagnostics by evaluating the environmental efficacy and health impact of nano-sorbents derived from rice and melon husks. rice and melon husks are rich in cellulose, hemicellulose, and lignin, which offer numerous active sites for chemical modifications. these agro-wastes, when processed into nano-scale particles, significantly increase surface area and reactivity (elemike et al., 2022; chukwu et al., 2021). nanomaterials derived from agro-waste have demonstrated exceptional adsorption properties for pollutants, including heavy metals, dyes, and hydrocarbons (kumar et al., 2019). oil contamination has been associated with elevated serum levels of alt and ast due to hepatic injury (olatunde et al., 2020). similarly, raised urea and creatinine levels indicate compromised kidney function. ultrasound imaging, particularly in preclinical trials, is a non-invasive pa ge 17 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(2) 16-20, 2025 and effective method for monitoring internal organ morphology (ibeabuchi et al., 2022). however, few studies integrate nanotechnology, adsorption modeling, and biomedical safety testing in a single research framework. materials and methods sorbent preparation the rice and melon husk material were washed with distilled water to remove dirt, dust or any other impurities. the materials were dried in an oven at 60-80°c until they were completely dried. the dried melon and rice husk were grinded into fine powder using a grinder. the ground materials were sieved to ensure that uniform particles between 50-100 microns were obtained. exactly 500 ml of 0.5 mol of naoh was added into washed, dried and ground rice and melon husks weighing 81 g and 73 g respectively in separate batches. precipitate of nano sized particles of melon and rice husk were obtained. the nano sized particles of melon and rice husk formed were collected and dried in an oven characterization brunauer–emmett–teller (bet) analysis exactly 0.5 g of the finely grinded sample was placed into a sample tube designed for bet analysis. the sample was degassed on 200°c for 4 hours by heating it under vacuum. this step was critical to remove any adsorbed gases and moisture that could interfere with the bet measurements(ochie et al., 2024). the bet instrument was turned on and was allowed to stabilize. the instrument was calibrated and properly functioning. the gas supply was connected and flowing at the correct pressure. the degassed sample was loaded to the sample tube into the bet instrument sample holder (ochie et al., 2024). the sample tube was properly sealed in the analysis chamber, and the system was under inert gas flow to avoid contamination from ambient air. nitrogen gas was used as the adsorbate, which interacts with the surface of the material to measure the surface area. the pressure range was set to 0.3 of the adsorbate gas. the instrument introduced nitrogen gas into the chamber in controlled amounts, allowing it to adsorb onto the surface of the sample at different pressures (ochie et al., 2024). the bet analyzer recorded the amount of gas adsorbed and desorbed at different pressures, generating data that describes the material’s surface area and porosity. after the completion of the analysis, the system generated an adsorption-desorption isotherm, which is a plot of the amount of gas adsorbed versus the relative pressure. fourier transform infrared spectroscopy (ftir) exactly 0.2 g of melon and rice husk sorbent was grinded using mortar and pestle in order to finely grind, which ensured uniformity and increased the surface area for better ir interaction. 100 mg of potassium bromide (kbr) was introduced 2mg of the powdered sample to form a fine powder. the kbr and sample mixture was pressed into a pellet using a pellet press which created a transparent disc for analysis (ochie et al., 2024). the ftir spectrometer was turned on and allowed to warm up, the instrument was calibrated without the sample which helped to eliminate any noise from the environment. the mode was selected. each of the samples were put in the ftir sample holder and it was scanned such that the wavelength was typically 4000 cm–1 to 400 cm–1, the resolution was 4 cm–1 and the number of scan was 16-32 scans for a clear spectrum.the instrument infrared light absorbed by the sample at different wave length. adsorption studies exactly 0.5 g mass of the nano-sorbent was added to each solution with varying oil concentrations. the mixtures were stirred and allowed to reach equilibrium after 1 hour. after equilibrium, the remaining oil concentration in each solution was measured using uv-vis spectroscopy and gravimetric methods. the amount of oil adsorbed per unit mass of the nano-sorbent (q) was calculated for each initial oil concentration. the equilibrium data were plotted asq versus c e (the equilibrium concentration of oil in the solution). the experimental data were fitted to isotherm models like langmuir and freundlich isotherms. the isotherm parameters (q max, k l, k ) were determined by fitting the data to these models. kinetics studies a solution with 100 mg/l concentration of oil was prepared. exactly 0.5 g of nano-sorbent was added to the oil-water solution, and samples were taken at different time intervals 10minutes). the concentration of residual oil in the water phase was measured at each time interval using uv-vis spectroscopy. the amount of oil adsorbed at each time interval was calculated, and the data were plotted as q versus time. the experimental data were fitted to kinetics models like:pseudo-first-order kinetics and pseudo-second-order kinetics in vivo toxicological assessment the study involved 15 wistar albino rats, weighing between 200-250g, which were divided into three groups: group a served as the control group and received clean water, group b was exposed to crude oil-contaminated water, and group c received remediated water treated with nanosorbents, with the experiment lasting for 21 days. biochemical assays the methodology for biochemical assays using roche diagnostics kits for alt, ast, creatinine, and urea involves kinetic assays for alt and ast, enzymatic assays for creatinine and urea, and utilizes roche’s analyzers such as the cobas c311, with calibration and quality control materials run to ensure assay accuracy and precision, providing accurate and precise results, high throughput, and ease of use. for these assays, plasma samples are prepared according to roche’s guidelines, and reagents are prepared according to the kit instructions, with samples and reagents then pa ge 18 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(2) 16-20, 2025 pipetted into designated wells or cuvettes and run on the analyzer, which measures absorbance or fluorescence changes proportional to enzyme activity or analyte concentration. sonographic imaging ultrasound examinations were conducted before and after treatment using a portable ultrasound device, focusing on assessing liver size, echotexture, and renal cortex integrity. statistical analysis statistical analysis was performed using analysis of variance (anova) followed by tukey’s post-hoc test, with a significance level set at p < 0.05. results and discussion the nano rice husk sorbent exhibits a high surface area of 680.5 m²/g (bet) and 999.9 m²/g (single point), a microporous structure with a t-plot micropore area of 20.4 m²/g and micropore volume of 0.13044 cm³/g, a narrow pore size distribution with an average pore width of 30.34 å to 30.42 å, and a high pore volume of 0.6035 cm³/g (single point) and 0.52222 cm³/g (bjh desorption). similarly, the nano melon sorbent has a high surface area of 460.36 m²/g (bet) and 999.90 m²/g (single point), a microporous structure with a t-plot micropore area of 20.30 m²/g and micropore volume of 0.13044 cm³/g, a narrow pore size distribution with an average pore width of 30.34 å to 30.42 å, and a high pore volume of 0.6035 cm³/g (single point) and 0.52222 cm³/g (bjh desorption), indicating its potential for efficient adsorption, selective adsorption, and applications in various fields. table 1: findings from the brunauer–emmett–teller (bet) analysis property nano melon husk sorbent nano rice husk sorbent (ochie et al., 2024) bet surface area 460.36 m2/g 680.5 m2/g langmuir surface area 50.24 m2/g 52.24 m2/g micropore area 20.3 m2/g 20.4 m2/g pore volume 0.6035 cm3/g 0.6035 cm3/g average pore width 30.34 å 30.34 å ftir analysis findings for rice husk and melon husk nano-sorbents the fourier transform infrared (ftir) spectra of nanosorbents derived from rice husk and melon husk reveal the presence of various functional groups, including hydroxyl groups (–oh) indicated by broad peaks around 3439 figure 1: fourier transform infrared (ftir) graph for rice husk and melon husk nano-sorbents cm-¹ for rice husk and 3300 cm-¹ for melon husk, aliphatic hydrocarbons (c-h stretching) at 2926 cm-¹ for rice husk and 2900 cm-¹ for melon husk, carbonyl and carboxyl groups (c=o stretching) at 1728 cm-¹ for rice husk and 1700–1600 cm-¹ for melon husk, aromatic rings and c=c stretching at 1637–1514 cm-¹ for rice husk and 1500 cm-¹ for melon husk, and polysaccharide backbone (c-o stretching) at 1256–1039 cm-¹ for rice husk and 1200– 1000 cm-¹ for melon husk. the rice husk nano-sorbent shows an additional peak at 1039 cm-¹ related to si-o-si stretching, confirming the presence of silica, and a peak at 2369 cm-¹ likely due to atmospheric co₂ or processing artifacts. the melon husk nano-sorbent exhibits distinct low-frequency peaks around 800–600 cm-¹ attributed to aromatic c-h bending and possible inorganic impurities, such as silicates. these functional groups, including hydroxyl, carbonyl, carboxyl, aliphatic, aromatic, and polysaccharide-derived groups, enable diverse interaction mechanisms such as hydrogen bonding, ion exchange, π-π stacking, and van der waals forces with organic and inorganic pollutants, enhancing their potential as ecofriendly, low-cost, and efficient nano-sorbents for the pa ge 19 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(2) 16-20, 2025 remediation of crude oil-contaminated water and soils. findings from sorption experiments using both sorbents for pms the sorption experiment aimed to evaluate the effectiveness of nano rice husk and nano melon husk in removing premium motor spirit (pms) from water. volume of pms= 100ml mass of melon husk before sorption = 1.00 g mass of rice husk before sorption = 1.00 g mass of melon husk after sorption = 1.340 g mass of rice husk after sorption = 1.338 g sorption capacity for melon husk = 1.340 – 1.00 g = 0.340 g sorption capacity for rice husk = 1.338 – 1.00 g = 0.338 g figure 2: kinetics graph of pms removal using rice husk and melon husk sorbents both nano melon husk and nano rice husk demonstrated significant pms sorption capacities, with melon husk showing a slightly higher capacity (0.340 g) compared to rice husk (0.338 g). the sorption efficiencies of melon husk and rice husk are 34.0% and 33.8%, respectively, indicating their potential for pms removal from water. the results shows that nano melon husk and nano rice husk have comparable sorption capacities for pms, with minimal difference between them. the sorption mechanism involves physical adsorption, chemical adsorption, facilitated by the high surface area and porous structure of the nano sorbents. considerations of the kinetic studies of pms using both sorbents the kinetic model fitting results for both pseudo-firstorder and pseudo-second-order models are plotted above for rice husk and melon husk sorbents. table 2: data from sorption experiments using premium motor spirit (pms) sorbent initial mass (g) final mass (g) sorption capacity (g) melon husk nano-sorbent 1.00 1.340 0.340 rice husk nano-sorbent 1.00 1.338 0.338 table 3: kinetic model fitting results pseudo first order qe k1 rice husk 0.0148 3.19 melon husk 0.0228 9.66 x 108 g/mg/s pseudo second order qe k2 rice husk 0.0148 3.19 melon husk 0.0228 2.13 x 10-8 g/mg/s both models exhibit very similar fits, with the secondorder model following closely along the observed data, but both models show a minimal change in absorbance over time, indicating a rapid equilibrium is reached. pseudo-second-order model fits better, as it follows the observed decline in absorbance more closely. the pseudosecond-order model generally implies that chemisorption is the rate-limiting step. for rice husk, the fast plateauing of absorbance shows a rapid adsorption process, which point to physical adsorption (physisorption) being dominant. for melon husk, the continuous decline in absorbance and better fit of the second-order model shows a more gradual adsorption process, which is due to chemical bonding between pms and the surface of the sorbent (chemisorption). the results shows that melon husk have a slower but more sustained adsorption, potentially offering higher capacity, while rice husk achieves equilibrium quickly. pa ge 20 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(2) 16-20, 2025 biochemical findings group b exhibited elevated liver enzymes and renal markers, with alanine transaminase (alt) levels at 120 iu/l, aspartate transaminase (ast) at 145 iu/l, and creatinine at 2.6 mg/dl, which improved post-treatment in group c, with alt decreasing to 52 iu/l, ast to 65 iu/l, and creatinine to 1.3 mg/dl, approaching values similar to the control group, which had alt at 45 iu/l, ast at 58 iu/l, and creatinine at 1.1 mg/dl. sonographic findings group b exhibited hepatomegaly and hypoechoic liver parenchyma with a dilated renal pelvis, whereas group c demonstrated regression in liver size, normalized echogenicity, and restored cortical thickness, indicating significant improvement. medical interpretation oil exposure causes hepatocellular and nephronal damage, evident through enzyme elevation and structural distortion. nano-sorbent remediation reversed hepatic and renal abnormalities both functionally and structurally. conclusion this study has demonstrated that agro-waste-derived nano-sorbents from rice and melon husks possess excellent physicochemical properties, including high surface area, favorable pore characteristics, and functional groups conducive to adsorption. their efficacy in removing crude oil from contaminated water was affirmed through sorption capacity measurements and isotherm/kinetic modeling, with results indicating a dominance of chemisorption governed by pseudosecond-order kinetics. the langmuir isotherm model also confirmed monolayer adsorption on homogeneous surfaces. beyond their environmental performance, the biomedical assessments revealed a significant reversal of hepatotoxicity and nephrotoxicity in oilexposed rats treated with remediated water. biochemical markers such as alt, ast, and creatinine showed marked improvement post-treatment, corroborated by sonographic normalization of liver and kidney structures. this underscores the biocompatibility and therapeutic safety of the nano-sorbents. recommendations ● adopt agro-waste-derived nano-sorbents in rural and urban water treatment programs. ● incorporate biomedical testing (sonography, alt/ ast) in water remediation evaluations. ● promote interdisciplinary research to address pollution and public health holistically. ● encourage policy incentives for waste-to-resource innovations. references chukwu, a., okoye, p. a., & afolabi, a. (2021). synthesis of nano-sorbents from agricultural by-products. nigerian journal of science, 57(4), 225–237. elemike, h. e. (2022). agricultural waste valorization for nanomaterial synthesis. environmental nanotechnology, monitoring & management, 17, 100640. freundlich, h. (1906). over the adsorption in solution. zeitschrift für physikalische chemie, 57, 385–470. ibeabuchi, n. (2022). sonographic evaluation of pollutant-induced hepatotoxicity in wistar rats. west african journal of radiology, 29(2), 150–157. kumar, p. s. (2019). adsorption of pollutants using agrobased nanomaterials. journal of cleaner production, 223, 1234–1245. langmuir, i. (1918). the adsorption of gases on plane surfaces of glass. journal of the american chemical society, 40(9), 1361–1403. nwilo, p. c., & badejo, o. t. (2005). oil spill problems and management in the niger delta. int. ocean inst. olatunde, a. a. (2020). clinical and biochemical profile of rats exposed to crude oil-polluted water. nigerian journal of medical sciences, 19(3), 78–84. undp. (2006). niger delta human development report. united nations development programme. who. (2021). global chemical safety and petroleum pollution. world health organization. pa ge 1 pa ge 53 american journal of chemistry and pharmacy (ajcp) a review on the utilization and environmental concerns of coal fly ash saidu kamara1, edward hingha foday jr2, wei wang1* volume 2 issue 2, year 2023 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v2i2.1609 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: april 26, 2023 accepted: june 10, 2023 published: july 02, 2023 fly ash is a solid by-product from the pulverization of coal in thermal power plants. it is a cost-effective raw material that has gained so much attention from industrialists for a series of engineering purposes. it is utilized in various applications like cement and concrete, bricks, road construction and embankments, mine backfilling, mixed asphalt, soil amelioration, catalyst, production plants, geopolymers, etc. asia and usa. are the top global fly ash producers. the high demand for its utilization adds to the economy of fly ash-producing nations. china, india, and japan contribute to the fly ash market growth through urbanization and sustainable infrastructural activities. fly ash is not environmentally friendly. it is currently one of the leading industrial solid wastes that has attracted so much public attention due to its associated environmental health concerns. it poses a severe social and economic burden to municipal and central authorities. cox, nox, sox, and matter are pollutants emitted from fossil fuel burning. carbon dioxide and carbon monoxide are indirect agents of climate change. fly ash contains heavy metals and some radioactive elements; therefore, utilizing it can lead to severe health and environmental consequences if not adequately managed. keywords coal, environment, fly ash, power plants, pulverization, utilization 1 department of chemical engineering, school of water and environment, chang’an university, xi’an, china 2 key laboratory of subsurface hydrology and ecology in arid areas, ministry of education, chang’an university, xi’an, china * corresponding author’s e-mail: wwchem@chd.edu.cn introduction the global demand for fly ash utilization in buildings and infrastructure, agriculture, geopolymers, catalysts, etc is on the increase. sustainable industrial and infrastructural development projects in western nations influence the market growth of fly ash. according to sources from vantage market research, the global fly ash market in 2021 was at usd 12.1 billion. recently, many countries have been trying to shift away from using fossil fuel as a source to renewable energy technology to combat the environmental impacts of greenhouse gases (cox) emitted from burning fossil fuels in thermal power plants (mucomole, silva, & magaia, 2023). this is a significant challenge for the growth of the fly ash market. however, developing new technologies to produce fly ash will possibly maintain a sustained fly ash market growth now and in the future. the key market players are now focusing on sustained practices to reduce emissions, and waste minimization, and utilize renewable energy sources(hossain & pk, 2023; mucomole et al., 2023). it can also be recycled and used for multiple purposes rather than dumped or disposed of in open pits, landfills, and ponds. using recycled fly ash for many farm practices contributes significantly to the growth of the fly ash market. fossil fuel has the largest share and source of global electric power production. the utilization of coal for electricity generation is still widely embraced on a worldwide platform. in 2011, coal power supplied up to 29.9% of global energy, and this is projected to rise to 46% by 2030 (ardha & aziz, 2007). coal-powered generation is economically attractive due to the high prices of oil and gas (hossain & pk, 2023; lior, 2010). pulverized fuel ash is a by-product that evolves out of coal-fired boilers with flue gases (hossain & pk, 2023; rani, rani, bansal, singh, & singh, 2021). the waste residue is trapped by filtration equipment before the gases enter the chimneys (rani et al., 2021). the constituents in fly ash vary depending on the sources of the coal utilized (ahmaruzzaman, 2010). the health and environmental problems of fly ash are well-known. open land disposal is the most common method adopted by coal power industries. fly ash is regarded as a general solid waste in some countries. the concentrations, and solid/liquid ratio, affect trace elements in aqueous environments (saikia, kato, & kojima, 2006). this threatens the air, surface, groundwater, soil, and crop production (ardha & aziz, 2007). in the early years of thermal power plant operations, fly ash was disseminated into the atmosphere with no control measures by the industries. the development of air pollution regulatory standards required fly ash to be trapped by pollution control equipment before being released and stored at the sites of power plants or landfills (finkelman, wolfe, & hendryx, 2021). pozzolans provide better concrete protection from moist conditions and chemical attacks. reports and findings from various scientific research (sadik, el amrani, & albizane, 2014) reveal that fly ash can be utilized to fabricate refractory materials (cordierite, mullite, and kyanite) when combined with alumina and subjected to high temperatures. manufactured refractory materials have a better advantage over other engineering materials utilized for various engineering purposes. this review summarizes fly ash utilization and associated health and environmental concerns. fly ash is considered a worldwide environmental hazard since it contains organic and inorganic pollutants, toxic heavy metals (pb hg, cr, v), and radionuclides. https://doi.org/10.54536/ajcp.v2i2.1609 https://journals.e-palli.com/home/index.php/ajcp mailto:wwchem%40chd.edu.cn?subject= pa ge 54 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 53-65, 2023 literature review fly ash is a fine-grained and powdered particulate matter obtained from the combustion of coal in a coalfired boiler in thermal power plants. the electric power industry has three types of coal-fired boiler furnaces referred to as dry-bottom bottom boilers, wet-bottom boilers, and cyclone furnaces. it is an industrial solid waste that poses many environmental and storage issues in coal power-producing nations. it is nowadays popularly utilized as a mineral additive by construction companies as a low-cost material, reduces environmental pollution, and also helps in reducing the need for the utilization of natural resources. fly ash is available in two main classes. it is classified as class c or class f ash based on its chemical constituents particularly when used as a mineral admixture in concrete. it consists of significant amounts of sio2, al2o3, fe2o3, and cao and smaller amounts of oxides of magnesium, potassium, sodium, titanium, and sulfur. class c fly ash is produced from the combustion of lignite or subbituminous coal. it possesses pozzolanic and cementitious properties and contains more than 10% calcium oxide. it composes of higher amounts of alkali and sulfate (so4 ) compounds (dwivedi & jain, 2014). class f fly ash is formed from anthracite or bituminous coal. it is naturally pozzolanic and contains less than 10% calcium oxide (cao). class f fly ash requires a cementing agent, such as portland cement, quicklime, or hydrated lime, with water to produce cementitious compounds. adding a chemical activator such as sodium silicate (water glass) to a class f ash can lead to the formation of a geopolymer (dwivedi & jain, 2014). advantages and disadvantages of fly ash advantages of fly ash it is a cost-effective substitute. it reduces the heat of hydration. the water required is reduced with better workability. it has low permeability and improved resistance to sulfate attack. it solves the problem of cracks experienced in portland cement. using fly ash in concrete reduces co2 and is thus a friendly environmental solid waste. disadvantages of fly ash it changes from liquid to solid in a few hours after pouring. the air ingress reduces. the color of the concrete is more challenging to control. the use of fly ash increases salt scaling and experiences seasonal restrictions. unlike portland cement, it reduces the demand for waste. utilization and management of fly ash coal is the only natural resource widely used in thermal power plants to produce electricity (mao & xu, 1999). industrialization, urbanization, and economic development are the major factors influencing the growing desire for electricity. fly ash from pulverized coal has been proven to be a helpful waste material in many applications (jala & goyal, 2006; rani et al., 2021). alumina and silica are the principal compounds, that make fly ash essential for profitable utilization. the ash from the coal plant is first processed using various techniques such as demagnetization, sinterization, grinding, etc. (sukkae, suebthawilkul, & cherdhirunkorn, 2018). the utilization of fly ash can abate environmental threats, render income, create job opportunities, and provide a sustained cleanedup environment, etc (dwivedi & jain, 2014; finkelman et al., 2021; l. c. ram & masto, 2010; senapati, 2011). mercury, lead, arsenic, etc., released into the environment, affect the quality of air, soil, and underground water (yiwei et al., 2007). the health risks of fly ash have led to several studies on its utilization in many applications, such as ceramic, bricks, landfills, and fly ash products, including ceramic refractories, plastics, metal composites, etc. (ardha & aziz, 2007). a tremendous amount of fly ash is annually produced, but utilization is minimal in many countries(ardha & aziz, 2007). the non-utilization of fly ash in many nations is due to some regulations that categorize fly ash as hazardous waste and the challenges of fabricating high-grade products (ardha & aziz, 2007). the utilization of fly ash in some applications and its management are discussed as follows: refractory due to their high-temperature resistance, refractory materials are used in furnace linings and metal-melting pots. the characteristics of fly ash have been studied, followed by examining the high-temperature resistance of materials mixed with fly ash and other silica-alumina source materials(ardha & aziz, 2007; hwang, 1999). s. maitra et al. (maitra, kumar, vishwakarma, & dutta, 2001) synthesized refractory aggregates from beneficiated fly ash by reaction sintering at 1600°c. the fly ash was discovered to be a good alumina-silica raw material based on castable refractory(dana, sinhamahapatra, tripathi, & ghosh, 2014). concrete fly ash is pozzolanic and it is used as a low-cost (maitra et al., 2001; sukkae et al., 2018) material compared to portland cement. it is more environmentally friendly; it reduces the corrosion of steel and improves its resistance to chemical attacks. it can be used to construct underwater structures. portland cement is the world’s third-largest industrial consumer of energy; it is the second-highest co2 emitter with an estimated share of 8-10% of global emissions (da silva, malacarne, longhi, & kirchheim, 2021). the pozzolanic reaction between fly ash and lime produces less heat and hence reduces the possibility of thermal cracking (dhadse, kumari, & bhagia, 2008; v. m. john, quattrone, abrao, & cardoso, 2019; s. a. miller, 2018; shen, wang, li, yao, & jiang, 2020). the cement industry in india utilized about 60.11 million tons of total production in 2018 and 2019 (dwivedi & jain, 2014; tejasvi & kumar, 2012). https://journals.e-palli.com/home/index.php/ajcp pa ge 55 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 53-65, 2023 soil stabilization modifying soil properties temporarily enhances subgrade stability to speed up construction (association, 2003; s. k. john, nadir, & girija, 2021; sun, li, zhao, zhu, & zhang, 2016). xiaofei et al. (sun et al., 2016) reported that stabilization could change the properties of municipal solid waste. it is also used to treat a wide range of sub-grade materials, from expensive clays to granular materials. mixing fly ash with soil plasticity results in a decrease in plasticity; the primary mechanism behind this is the change in the size of soil grains (dwivedi & jain, 2014). embarkment fly ash is more economical and environmentally friendly to be utilized as an alternative to topsoil for road embankments, and its suitability has proved successful in many cases. construction and design require many favorable properties like lightweight (superior over weak subsoil), higher shear strength (more excellent stability), no lumps, usually moist, compacted characteristics under inclement weather, cost savings, etc. (ghazali, muthusamy, & ahmad, 2019). this utilization has many advantages over conventional methods. it saves the topsoil and fills up low-lying areas thus created. the utilization of fly ash is affected by its properties such as grain size distribution, compaction characteristics, shear strength, compressibility, permeability, and frost susceptibility (l. c. ram et al., 2007; l. ram et al., 2006). class f fly ashes are typically used in embarkment because they are obtained from anthracite and sub-bituminous coal, rich in silica, alumina, and iron oxide (l. c. ram & masto, 2010; l. c. ram et al., 2007). it is globally used as a structural fill material for highway embankments. however, the use of fly ash in road embankments has environmental concerns like soil erosion, dispersion of fly ash into the air by wind, and the leaching of heavy metals into the subsoil. bio-amelioration of soil reports from some recent research indicated that fly ash has a better application when mixed with organic materials like cow manure, sludge, farm yard manure, sludge, crop residues, and organic composts(yao et al., 2015). a combination of fly ash and organic matter reduces heavy metals, kills pathogens in sludge, improves bulk density, porosity, and biological activity in the soil, and reduces the leaching of nutrients, which is beneficial for agricultural utilization (tu et al., 2022). organic amendment applications provide anchorage and growth of the plant on a fly ash dumping site. agriculture fly ash improves soil fertility status, plant growth, and agricultural yield (bayat, 2002a, 2002b; bhattacharya, iftikar, sahariah, & chattopadhyay, 2012; gorai & ash, 2018; kishor, ghosh, & kumar, 2010). it improves soil texture properties, aeration (page, elseewi, & straughan, 1979), water-holding capacity, and porosity(kene, lanjewar, ingole, & chaphale, 1991). it provides micronutrients such as mo, b, fe, zn, cu, etc. (dinjus, fornika, & scholz, 1996; khan & wajid, 1996). fly ash is used for the reclamation of sodic soils and acidic soils (adriano, page, elseewi, chang, & straughan, 1980; bhattacharya & chattopadhyay, 2003; haynes, 2009; s. singh & gupta, 2003). however, there are associated hazards such as the effects on human and grazing animals, groundwater pollution, and soil infertility due to high fly ash doses in agricultural fields. radionuclide present in fly ash also causes radiochemical pollution. geopolymers more recently, fly ash has been used as a compound in geopolymers, where the reactivity of the fly ash glasses generates a binder comparable to a hydrated portland cement in appearance and properties but has porosity and reduced co2 emission. the limited use of fly ash geopolymer is due to its low reactivity, which depends on the particle size, glass content, and composition(kumar & kumar, 2011). waste treatment fly ash is combined with other alkaline materials to transform sewage sludge into organic fertilizer or biofuel (bayat, 2002a, 2002b). it is used for domestic and industrial wastewater treatment and as a toxic metal adsorbent to remove industrial and poisonous wastes like dyes (dasmahapatra, pal, & bhattacharya, 1998; devi & dahiya, 2006; dutta, basu, & dasgupta, 2003; goswami & das, 2000). wood substitute material fly ash can be a good substitute for doors, windows, ceilings, partitions, furniture, etc. the main objective for using fly ash as a wood substitute composite (i.e., fly ash geopolymer composites) is to reduce deforestation by using it as an alternative to timber products, which is very much required to save our environment (khan & wajid, 1996). the development of fly ash-based composites needs fly ash as filler and jute cloth reinforcement. management of coal fly ash fly ash is a residual waste from burning fossil fuel in thermal power plants and is globally recognized as problematic. studies have reported that the estimated annual yield in india, china, germany, and the uk are 112, 100, 40, and 15 million tons, respectively. some challenges in managing fly ash are the vast land requirement for disposal and the contamination of groundwater (twardowska, szczepanska, & stefaniak, 2003). fly ash can be adequately managed when used in various applications like building and road construction, soil amelioration, glass ceramics, geopolymers, filling lowlying areas, bricks, portland cement, and concrete. the management of fly ash as solid waste should concern regulatory bodies, with particular emphasis on utilization to keep our environment safe from its hazards. power https://journals.e-palli.com/home/index.php/ajcp pa ge 56 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 53-65, 2023 plants should also adopt adequate fly ash management systems due to the large space occupancy within the power plant area. the coal fly ash also contains significant amounts of toxic metals such as as, ba, hg, cr, ni, v, pb, zn, and se, characteristically enriched in coal fly ash particles. the environmental effects of coal fly ash emissions from the burning of fossil fuel carbon dioxide and carbon monoxide outdoor and indoor fossil fuels are employed for global consumption and domestic energy purposes (belyaeva & haynes, 2012; j. chen et al., 2014; yousuf, manzoor, youssouf, malik, & khawaja, 2020). the co2 emission from chemical processes is far greater than co2 from oil-based chemical processes (j. chen et al., 2014; ren & patel, 2009). according to mohammad ehsan munawer (munawer, 2018), co and co2 are mainly emitted from the chemical oxidation processes that occur during coal combustion, which significantly contribute to global warming and several health issues including cardiovascular diseases. about 90% of the worldwide co2 is from coal, also known as fossil fuel(ewane & ewane, 2023). cox gases are climate change contributing factors that lead to flooding and hurricane that negatively affects agriculture and the food chain (ewane & ewane, 2023; gething et al., 2010; jos g.j. olivier (pbl) & marilena muntean (iesjrc), 2015). the growth of the plasmodium mosquito significantly depends on the temperature within the environment; therefore, co2 emissions may contribute to the increase in malaria, a primary global environmental health concern(ewane & ewane, 2023). sulfides sulfur is released into the environment in gaseous forms during coal combustion, which leads to air, water, and land pollution. the annual sulfides and particulate matter emitted by unregulated coal power plants are twice higher than emissions from factories, trucks, and cars (delucchi, 2003; skalska, miller, & ledakowicz, 2010). these gases travel hundreds of miles away from power plants to pollute air and water and form h2so4, a significant constituent of acid rain (likens, wright, galloway, & butler, 1979; skalska et al., 2010). aerosols, mist, and smoke combines with so2 and may penetrate the lining of the lungs leading to severe lung diseases (kelsall, samet, zeger, & xu, 1997; pourgholami, akhter, wang, lu, & morris, 2005). it affects farm plants and reduces crop yield (galloway & whelpdale, 1980; rajput, dp, & wd, 1977; winter, mallepalli, hellem, & szydlo, 1994). the oxides of so2, no2, and limited o3 generated during fossil fuel combustion produce acid rain upon hydration, fall within the surroundings of industrial locations, and are then transported through rivers and water reserves at far distances. consuming contaminated food causes severe complications (foday jr, bo, & xu, 2021). in summary, sulfides negatively impact human health. coal with low sulfur and ash contents is preferred due to the harmful nature of sulfur. nitrogen the exposure to no2 gas emitted from coal pulverization in power plants and burning from our domestic activities aggregates in the air and causes environmental and health problems(lee, ha, lee, lee, & kim, 2006; levy, moxim, klonecki, & kasibhatla, 1999). pulmonary function is decreased when people are directly exposed to no2 gas. some have been so vulnerable to even lower no2 concentrations in the air resulting in asthma, lung malfunction, respiratory failure, long-term pulmonary hypertension in young babies, dna damage, and cancer(alexis et al.; arnold, mittal, katsuki, & murad, 1977; chauhan & johnston, 2003; li, liu, de, & tao, 2001; roberts jr, polaner, zapol, & lang, 1992; van amsterdam et al., 2000). nitrogen acids (hno2 and hno3) are formed when no2 reacts with h2o damaging agricultural plants, decreasing the rate of photosynthesis and structural buildings, and causing skin burns and skin cancers(a. singh & agrawal, 2007). in summary, nitrogen gases and compound affects human health and plants (skalska et al., 2010). particulate matter the coal dust and fly ash significantly contribute to forming this component, which causes respiratory problems in children (brabin et al., 1994; y. chen et al., 2004; claneyl, 2002; k. a. miller et al., 2007; pope iii, ezzati, & dockery, 2009). the bottom ash found in coal power plants or dumping sites affects aquatic and terrestrial animals (temple & sykes, 1992). the coal combustion residue also contaminates soils near ash ponds, decreases soil ph, hinders crop production, and affects the food web (lokeshappa & dikshit, 2012). metals and other constituents in coal enhance the toxicity of particulate matter (boström et al., 2002; organization, 2006). the inhalation of pahs affects dna molecules, resulting in dna mutation, child neurodevelopment, reduced iq, epigenetic effects, reduced child intelligence, cancer, and different cardiovascular diseases (dragović et al., 2013; edwards et al., 2010; jedrychowski et al., 2003; f perera, li, lin, & tang, 2012; frederica perera et al., 2008; f. p. perera et al., 2009) the figure below shows how inhaled particles penetrate the lungs. the smaller the particle the deeper they penetrate the lungs. particulate matter is one main contributor to air-born pollution, and it causes cancers, cardiovascular diseases, and reproductive disorders. conclusively, particulate matter (pm) from fly ash is another leading source of air pollution that causes various health problems. hazardous elements in fly ash https://journals.e-palli.com/home/index.php/ajcp pa ge 57 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 53-65, 2023 the silica fume generated in coal power plants contains toxic elements referred to as heavy metals. the ash constitutes immense amounts of lead, mercury, arsenic, etc., which negatively affect soil and water. they are imperishable chemical elements present in coal gangue. coal gangue is the leading industrial residue in coal combustion and it is largely produced due to continuous coal combustion. large stockpiles of coal gangue are in asian countries such as china and india (haibin & zhenling, 2010; tang et al., 2008; zhao et al., 2008). lead pb is a toxic metallic element found in fly ash, and it contaminates water and air, which travels far and wide areas of coal power plants (bhangare, ajmal, sahu, pandit, & puranik, 2011; fernandez-turiel, de carvalho, cabañas, querol, & lopez-soler, 1994; goldstein, 1992; lansdown & yule, 1986; mushak & crocetti, 1988). exposure to lead damage the kidneys, heart, nervous system, and blood circulation in humans (foday jr et al., 2021; naja & volesky, 2017; wang et al., 2006). mining, moving vehicles, and burning coal are reported to be the primary sources in an occupationally exposed environment to pb. children are more susceptible to pb exposure than adults (leggett, 1993; todd et al., 1996). mercury investigations of coal used in homes for domestic purposes show significant hg (naja & volesky, 2017). hg is a deadly toxicant in the environment also emitted from fossil fuel combustion and circulates far distances through atmospheric dissemination. hg is highly unstable in its free state. the bacteria present in water chemically transform hg to methyl mercury (mehg) which, when consumed by eating aquatic animals, will conglomerate in their systems and thus affects the function of the brain of the fetus of pregnant women. the increased hg binding with the thiols of tubulin, a protein that forms the microtubules in the neurons, leads to structural-based pathological modulation, thereby resulting in neuronal migration and other brain deformities in newborns(lu & holmgren, 2014; osman et al., 2000; silva-adaya, gonsebatt, & guevara, 2014). arsenic the release of arsenic poses severe illnesses like lung, heart, skin diseases, etc., and it is known as the third most toxic element. it occurs in different oxidizing states and undergoes various types of chemical reactions to form other products. the human body takes up arsenic trapped in the hair and nails (dai et al., 2012; kang et al., 2011; liu et al., 2007; tian et al., 2013). the amount of trace elements in coal is determined according to grade. yudovich et al. (yudovich & ketris, 2005a) in their article stated the average amount of arsenic content around the world for low-rank coal, such as bituminous and lignite coal. arsenic is released in the form of oxides at lower and higher temperatures (cui & chen, 1998; shpirt, goryunova, & zekel, 1998; yudovich & ketris, 2005b). some of the symptoms experienced when arsenic (as) is consumed in food and inhalation are weakness, drowsiness, fatigue, asthma, respiratory diseases, cardiovascular problems, etc. long-term exposure to arsenic (as) from water and air may lead to anemia, leukemia, leukopenia, and dna damage that may lead to various cancers like skin and respiratory cancers (lerman, ali, & green, 1980; okui & fujiwara, 1986). conclusively, the release of arsenic from burning coal in power plants leads to severe illnesses, including lung, heart, and skin diseases. radioactive elements in fly ash radioactive elements, such as uranium, thorium, radium, figure 1: illustration of how minute particles infiltrate into the lungs(wilson et al., 2009) https://journals.e-palli.com/home/index.php/ajcp pa ge 58 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 53-65, 2023 etc., are also present in fossil fuels and their combustion products. they are released alongside their decay products from the original coal formations in the form of gas or solid. people are exposed to the emission of these radionuclides during fly ash utilization like concrete, stabilization, amelioration, agriculture, etc. exposure to these will result in severe radiological health consequences like cancer, cataracts, and genetic defects (amin et al., 2013; gagnaire, adam-guillermin, bouron, & lestaevel, 2011; habib et al., 2019). therefore, it is necessary to develop a radiation control program for the exposure of workers and the public in areas where power plants are installed. effect on climate change over the past 20 to 30 years, concerns about the threats and environmental deterioration on the planet have galvanized an international response as scientists, governments, global leaders, policymakers, intergovernmental organizations, and other stakeholders have coalesced to take urgent actions and discussions were centered around developing policy for energy usage within the context of climate (donaldson & borm, 1998; henderson-sellers et al., 1998). climate change can lead to flooding in coal ash ponds(ewane & ewane, 2023). heavy rains raise the water table that leaks into ponds which may contaminate the groundwater used for agricultural and drinking purposes. regulations should be developed to control coal ash ponds in coal-producing communities. this will help control coal ash dumps to stop the leaching and subsequent contamination of drinking water. coal power plants are responsible for managing the environmental impacts that arise from their operations. heavy penalties should be levied for any health and environmental problems that result from fly ash. public health impacts the health problems of fly ash can best be addressed through engineering solutions that efficiently remove particulates from the escaping gases(yao et al., 2015). heavy metals such as arsenic and molybdenum in fly ash threaten our health and environment if not adequately managed. inhalation for a very long time causes pneumonitis, allergy, asthma, lung, fibrosis, bronchitis, cancer, and silicosis (donaldson & borm, 1998; murugappan, manoharan, & nandhini, 2004; van maanen et al., 1999). the fly ash stored in wet lagoons and dry landfill can infiltrate and contaminate the groundwater if the encage is not correctly lined. people living near the disposal sites of coal ash have a greater risk of cancer and other illnesses from the contaminated drinking water, particularly water contaminated by arsenic which is the most toxic element in coal fly ash. several studies have been conducted to assess the hazards caused by fly ash on the environment and health (mehra, farago, & banerjee, 1998; murugappan et al., 2004; g. singh & vibha, 1999). exposure to toxic heavy metals in coal fly ash can also affect human development, cause heart and lung problems, lead to stomach ailments, and contribute to premature mortality. therefore, environmental regulatory agencies in coal-producing countries must categorize fly ash as a dangerous waste. coal fly ash regulatory agencies should assist the affected communities in enforcing rules and regulations. the low-income earners are affected mainly by the waste disposal sites and often lack the financial and political target to agree with polluters when fighting to seek redress. adverse effects on the environment millions of tons of fly ash leach into water reservoirs and contaminate drinking water. power plants often use ponds to dispose of their generated solid waste. they should therefore be levied with heavy charges whenever a community accident results from fly ash. the heavy metal elements severely attack the aquatic ecosystem, ultimately affecting fishermen’s sustenance. the fishing communities are concerned about the high level of toxic heavy metals found in many water species (fish, prawns, crabs, oysters, etc.). figure 2: photos showing fly ash deposition in a pond https://journals.e-palli.com/home/index.php/ajcp pa ge 59 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 53-65, 2023 methodology a systematic review was carried out from a sample of 131 articles containing topical-related data. the research question was structurally formulated based on the utilization and environmental health implications of fly ash from previous studies such as (ahmaruzzaman, 2010; ghazali et al., 2019; gollakota, volli, & shu, 2019; tu et al., 2022; yousuf et al., 2020) which highlighted fly ash utilization and (dwivedi & jain, 2014; finkelman et al., 2021; gorai & ash, 2018; sun et al., 2016; yousuf et al., 2020) reported their work on the environmental effects of fly ash. results and discussions according to a statistical report from the data in figure 3, india, china, and the usa are the annual highest fly ash producers, with yearly productions of 112, 100, and 75 million. india is the top global fly ash producer, followed by china and usa. their annual statistical utilization is inverse to their production. among these three countries, the usa utilizes 65% of the fly ash produced, 45% by china, and india 38%. as shown in the data above, germany is the highest fly ash producer among the european countries while denmark, italy, and the netherlands as the lowest producers (2 million tons per annum each). the three lowest fly ash producers utilize 100% of the fly ash produced, followed by france and germany, where both equally use 85% of the fly ash production. the united kingdom, australia, and canada produce 15, figure 3: fly ash production (mtn) and utilization (%) in various countries(bhatt et al., 2019; gollakota et al., 2019) 13.1, and 6 million tons annually. canada uses 75% of the fly ash produced, followed by the uk and australia, with 50% and 45% utilization, respectively. the middle east and africa have an annual production of 32.2 million tons with 10% utilization. in contrast, the russian federation and other asian countries have 26.7 and 16.7 annual outputs with 10% and 18% utilizations of their annual fly ash productions, respectively. among the three asian countries (india, china, and japan) in the above data (figure 3), japan is the lowest fly ash producer, while india is the highest, followed by china as the second highest. japan utilizes 96.3% of the fly ash produced compared to india and china. on a global scale analysis, china, india, and the u.s.a. are the leading fly ash producers in the world, according to the data provided in figure 3. the european countries (netherlands, italy, denmark, and germany) with 100% and 85% and japan (96.3%) are the highest utilizers of their annual fly ash productions. according to the analysis of this review, most countries are unable to utilize all their annual ash productions. much ash is left unused, particularly in india and china, where it is generated in volumes. this means the remaining ash is probably disposed of or exported to other countries around the globe for utilization in various applications. the global fly ash market for its utilization is briefly summarized below: from figure 4, the global market size of fly ash in 2021 was usd 12.1 billion. the market growth is projected to exponentially increase to usd 14.33 billion in 2024, usd 15.16 billion in 2025, and usd 16. 90 billion in 2028. the estimated annual growth rate ( cagr) between 2023 to 2028 is 5.8%. according to a report from vantage market research, global fly ash sales are expanding. the increase in global energy demand is exponentially parallel to the global fly ash market for various construction purposes. it has excellent properties that boost the growth of the fly ash market. china and india are the two countries contributing to the development of the fly ash market due to their increased industrialization and urbanization initiatives to promote sustainable infrastructural projects. they, however, pointed out the following number of factors that are inhibiting the progress of the fly ash market: 1) transportation and storage can be difficult and https://journals.e-palli.com/home/index.php/ajcp pa ge 60 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 53-65, 2023 figure 4: global fly ash market size and projected growth (market, 2012) expensive because the fly ash is a fine powder making it troublesome to handle; 2) quality variability factor affects fly ash market by making it difficult for the contractors to determine the properties of the ash., and 3) environmental concern is also a factor because of the potential environmental impact that results in air and water pollution. the global demand for fly ash is currently overwhelming due to its utilization in multiple applications. this is particularly evidenced in bigger economy nations that have embarked on rapid infrastructural and other development projects. however, there are many health and environmental challenges, primarily from fossil fuel burning, ecological management problems, etc. the dangerous elements encountered during coal and fly ash utilization endangers human health. if not adequately managed, heavy metals and radionuclides circulate into the air or leach into water sources, ultimately jeopardizing human health and crop yield. also, fly ash utilization should be monitored for radon gas inhalation, particularly in building and construction practices, and potassium, thorium, and radium for soil and crop contamination. another environmental concern is the pollution of gases such as cox, nox, sox, and particulate matter during coal combustion. cox gases are climate change agents that lead to global warming and flooding. nox and sox are acid rain components and affect crop yield and human health. the heavy investment in renewable energy sources to combat the problem of climate change is causing a decline in the conventional production of fly ash due to the closure of several thermal power plants in europe, the united states, etc. the emergence of alternative new techniques for fly ash production will sustain its utilization and market growth. conclusion although fly ash is utilized for multiple purposes, it however, has so many environmental implications such as emissions (carbon dioxide, carbon monoxide, sulfides, nitrogen, etc) from the burning of coal and hazardous elements (lead, mercury, arsenic, radionuclides, etc) in fly ash which requires adequate management and safe disposal methods. acknowledgments this work was supported by the national natural science foundation of china. thanks, and appreciation to wang wei of the chemical engineering department, school of water and environment of chang’an university for the adequate supervision and for providing the platform for the successful conduct of this work. references adriano, dc, page, al, elseewi, aa, chang, ac, & straughan, i. 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(2008). trace element emissions from spontaneous combustion of gob piles in coal mines, shanxi, china. international journal of coal geology, 73(1), 52-62. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 48 american journal of chemistry and pharmacy (ajcp) evaluation of chitosan-based nano-encapsulated diterpenoids from andrographis paniculata for targeted anti-inflammatory therapy: roles of analytical chemistry medical imaging, and biochemical assessment s. i. okonkwo1*, j. a. ezugwu1, i. p. oragwu1, c. k. okonkwo2, p. o. okwuego1, a. t. kene okonkwo3, v. s. okonkwo4, s. c. okonkwo5 volume 4 issue 1, year 2025 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v4i1.5356 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: june 01, 2025 accepted: july 04, 2025 published: september 29, 2025 the formulation, characterization, and therapeutic potential of andrographis paniculataderived chitosan-based nano-encapsulated diterpenoids for targeted anti-inflammatory therapy are examined in this study. chromatographic and spectroscopic methods, such as uvvis, ftir, nmr, and gc-ms, were used to isolate and characterize two important bioactive compounds: 14-deoxy-11,12 didehydroandrographolide and neoandrographolide. the existence of functional groups including amines, carbonyls, thiols, and nitro compounds— which support the compounds’ anti-inflammatory qualities—was verified by ftir spectra. transmission electron microscopy (tem) showed that spherical nanoparticles with mesoporous structures (22.10 nm) were produced by ionic gelation using chitosan and sodium sulfate, followed by sonication, to accomplish nano-encapsulation. high encapsulation efficiency (82.5% for the main isolate) and a biphasic drug release profile—an initial burst followed by a sustained release over four hours—were demonstrated by the encapsulated compounds. in carrageenan-induced wistar rats, sonographic imaging showed a significant decrease in paw thickness and vascular perfusion after therapy. this was supported by improved liver and kidney function and lower levels of inflammatory biomarkers (tnf-α, il6, and crp). a comparative analysis revealed that the nanoformulated extract exhibited less toxicity and performed on par with or better than common anti-inflammatory medications, such as diclofenac. the effectiveness of a. paniculata nanoformulations for safer, longerlasting, and more focused anti-inflammatory treatment is supported by this multidisciplinary approach that combines phytochemistry, nanotechnology, analytical chemistry, imaging, and biochemical validation. keywords andrographis paniculata, chitosan, diterpenoids, ftir, lc-ms, nano-encapsulation, sonography, targeted drug delivery, tem 1 department of pure and industrial chemistry, chukwuemeka odumegwu ojukwu university, uli, anambra state, nigeria 2 department of diagnostic medical sonography and ultrasound technology, ace institute of technology, elmhurst, new york, usa 3 tansian university,umunya, anambra state, nigeria 4 department of medical biochemistry, chukwuemeka odumegwu ojukwu university, uli, nigeria 5 department of pharmacology, chukwuemeka odumegwu ojukwu university, nigeria * corresponding author’s e-mail: si.okonkwo@coou.edu.ng introduction the therapeutic potential of medicinal plants has long been acknowledged in both conventional and alternative medicine. known as the “king of bitters,” andrographis paniculata (burm. f.) wall. ex nees (acanthaceae) is a plant that is well-known for its strong anti-inflammatory, antiviral, and anticancer qualities. the main bioactive substances have been found to have anti-inflammatory properties, especially the labdane diterpenoids neoandrographolide and 14-deoxy-11,12didehydroandrographolide. although inflammation is a protective immune response, when it is dysregulated, it can result in chronic conditions such as neurodegeneration, inflammatory bowel disease, cardiovascular disease, and rheumatoid arthritis. despite their effectiveness, traditional anti-inflammatory treatments like nsaids and glucocorticoids have low absorption, off-target toxicity, and systemic adverse effects. a revolutionary answer is provided by nano-encapsulation employing biopolymers such as chitosan. because it is mucoadhesive, biocompatible, and biodegradable, chitosan is a great option for targeted and long-lasting medication distribution. real-time evaluation of inflammatory resolution is made possible by integrating sonography, and biochemical markers offer information on the effectiveness of systemic therapy. validating compound identity, encapsulation effectiveness, and medication release dynamics all heavily rely on analytical chemistry. literature review the difficulties of drug solubility, bioavailability, and pharmacokinetics are being addressed by the growing formulation of natural materials into nanocarriers. andrographis paniculata’s wide range of pharmacological effects has garnered considerable research interest. its main diterpenoid, andrographolide, has been shown in studies to influence inflammation by inhibiting the nf-κb signaling pathway, which in turn suppresses pro-inflammatory cytokines like tnf-α and il6. andrographolide and its derivatives have shown therapeutic promise in models of asthma, arthritis, and stroke. lim et al. (2012) described how it suppresses inflammation better than synthetic medications, while hancke et al. (2019) showed a considerable decrease of cytokine expression. its low water solubility and poor systemic retention, however, have hampered its pa ge 49 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 48-53, 2025 therapeutic utility. for the efficient delivery of these bioactives, nanotechnology provides novel platforms including hydrogels, dendrimers, liposomes, and nanoparticles. targeting inflammatory tissues requires encapsulation in chitosan, which improves drug stability, cellular absorption, and controlled release. materials and methods plant collection and extraction the leaves of andrographis paniculata were crushed after being shade-dried. ethanol maceration, liquidliquid extraction, and soxhlet extraction were the three extraction methods employed. liquid-liquid extraction was used to achieve the highest concentration of active isolates (56 mg). isolation and characterization of diterpenoids key diterpenoids were isolated using silica gel column chromatography after andrographis paniculata leaves were extracted with ethanol. a rotating evaporator was initially used to concentrate the extract at a lower pressure. based on polarity, fractionation was performed using solvent gradients (methanol, ethyl acetate, and hexane). thin-layer chromatography (tlc) was used to monitor the separation and collect and purify distinct bands that matched known diterpenoids. two primary compounds—14-deoxy-11,12didehydroandrographolide and neoandrographolide— were identified. these were recrystallized from ethanol to obtain analytically pure samples. characterization was performed using: uv-visible spectroscopy peaks were observed at 375 nm (neoandrographolide) and 350 nm (14-deoxy-11,12-didehydroandrographolide), consistent with conjugated double bond systems. fourier transform infrared spectroscopy (ftir) key functional groups such as -oh (3300 cm⁻¹), c=o (1740 cm⁻¹), and c=c (1650 cm⁻¹) were confirmed. proton nuclear magnetic resonance (¹h-nmr) spectra indicated characteristic signals in the range of δ 6.4–7.6 ppm for olefinic protons and δ 3.5–4.2 ppm for hydroxyl groups. gas chromatography-mass spectrometry (gc-ms) provided molecular ion peaks at m/z 348 and 480, corresponding to the molecular weights of 14-deoxy11,12-didehydroandrographolide and neoandrographolide respectively. these characterization results ensured that the isolated compounds were structurally pure and suitable for further encapsulation studies. nano-encapsulation utilizing chitosan’s ability to crosslink with anions to form nanoparticles, the nano-encapsulation process was executed using the ionic gelation technique. chitosan was first dissolved in 1% acetic acid and magnetically agitated at 500 rpm to produce a homogenous solution. the refined diterpenoids were added dropwise to the chitosan solution after being dissolved in ethanol. the usage of sodium sulfate (na₂so₄) produced crosslinking. using a probe sonicator, sonication was applied for 10 minutes at a frequency of 20 khz to enhance nanoparticle formation and size reduction. the suspension was then centrifuged at 10,000 rpm for 30 minutes. to remove any unattached components, the nanoparticles were washed twice with deionized water after the supernatant was removed. the resulting nanoparticles were: • freeze-dried to obtain a dry powder suitable for longterm storage. • characterized by transmission electron microscopy (tem), revealing spherical morphology with particle sizes ranging from 80 to 150 nm. • encapsulation efficiency (ee%) was calculated as follows: ee=((total drug-free drug in supernatant))/ ((tota drug)) x 100 the ee% was found to be 82.5% for 14-deoxy-11,12didehydroandrographolide and 71.8% for the total diterpenoid extract. as a superior biopolymer carrier for phytochemical drug delivery, the chitosan matrix permitted a sustained drug release profile, offered stability, and protected against enzymatic degradation. in vitro release study a dialysis membrane diffusion method was used to evaluate the nano-encapsulated diterpenoids’ time-release profile. a known quantity of the nanoparticle solution (equivalent to 10 mg of encapsulated drug) was placed in a dialysis bag (mwco: 12–14 kda) and immersed in 100 ml of phosphate-buffered saline (pbs, ph 7.4) at 37 °c while being continuously swirled. five milliliter aliquots were removed and replaced with fresh pbs at predetermined intervals (0.5, 1, 2, 3, and 4 hours). using uv-vis spectrophotometry at 270 nm and a previously created calibration curve, the amount of medication released was measured. the release pattern showed a biphasic profile: • initial burst release in the first hour, attributed to surface-adsorbed drug molecules. • sustained release phase from hour 2 to 4, indicative of drug diffusion from the chitosan matrix. the sustained release is critical for chronic inflammation treatment, minimizing frequent dosing and improving patient compliance. sonographic imaging sonographic assessment was added as a non-invasive way to track the level of inflammation in vivo. to establish a model of localized inflammation, carrageenan (0.1 ml of 1% solution) was injected into the right hind paw of adult wistar rats (n = 24). a mindray z5 ultrasound equipment with a 7.5 mhz linear probe was used for the sonographic evaluation. pa ge 50 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 48-53, 2025 the imaging protocol included: • b-mode scanning to assess soft tissue swelling and hypoechoic changes associated with edema. • color doppler imaging (cdi) to quantify vascular perfusion in the inflamed area, indicative of inflammatory response. images were captured at 0, 24, 48, and 72 hours post-treatment. rats treated with nano-encapsulated diterpenoids exhibited: • significant reduction in paw thickness. • normalization of echogenicity and tissue architecture. • decreased doppler signal intensity, confirming reduced blood flow and inflammation. biochemical markers (crp, il-6) and histological evaluations supported these sonographic results, confirming the use of medical imaging as a reliable therapeutic monitoring tool. biochemical assays serum samples were analyzed for tnf-α, il-1β, il6, and crp using elisa. liver enzymes (alt, ast), creatinine, and urea were monitored for toxicity. results and discussion phytochemical composition phytochemical analysis of aqueous and ethanolic extracts of andrographis paniculata revealed the presence of various secondary metabolites essential to its therapeutic activity (table 1). both extracts tested positive for alkaloids, flavonoids, tannins, saponins, resins, cardiac glycosides, and steroids. however, ethanolic extraction proved more efficient, showing a stronger presence (++ reaction) of phenols, flavonoids, terpenoids, and cardiac glycosides compared to the aqueous extract. table 1: phytochemical constituents of a. paniculata extracts parameters aqueous extraction ethanolic extraction alkaloids + + phenol + ++ flavonoids + ++ tannins + + saponins + + resins + + cardiac glycosides + ++ terpenoids + ++ steroids + + phlobatannins ++ this confirms the ethanolic extract as a better candidate for isolation of active compounds. the presence of flavonoids and terpenoids further supports the extract’s antioxidant and anti-inflammatory potential. analytical validation of isolates and nanoparticles figure 1: fourier transform infrared (ftir) graph for encapsulated andrographis extract the ftir spectrum of andrographis paniculata extract (figure 1) confirms the presence of key medicinal functional groups such as amines (nh), nitro compounds (no₂), carboxylic acids (oh), carbonyls (c=o), thiol (-sh), and nitriles (c–c–cn), which are characteristic of diterpenoids with anti-inflammatory potential. notable peaks include 3452 cm⁻¹ for nh₂ in aromatic and primary amines, 2541 cm⁻¹ for -sh stretch in alkyl mercaptans, and 1600 cm⁻¹ for nh₂ in amino acids—indicating biological relevance and reactivity of the compound. pa ge 51 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 48-53, 2025 additional peaks at 1841 cm⁻¹ and 1428 cm⁻¹ show c=o and c-n functionalities, further supporting the compound’s structural complexity and therapeutic value. the spectrum confirms the unsaturated, aliphatic, and functionalized nature of andrographis diterpenoids, crucial for biological activity. these insights support the compound’s suitability for nanoencapsulation in chitosan matrices, ensuring molecular stability, bioavailability, and targeted delivery for antiinflammatory therapy. figure 2: lcms analysis of the isolate 14-deoxy-11,12-didehydroandrographolide the lc-ms analysis of 14-deoxy-11,12didehydroandrographolide revealed seven distinct bioactive compounds based on their retention times, molecular weights, and biological functions. the first compound identified was ciprofloxacin (c₁₇h₁₈fn₃o₃) at a retention time of 1.775 with a molecular weight of 331.35. it showed antibacterial and anticancer activities, consistent with kassab and gedawy (2018). the second compound was losartan (c₂₂h₂₃cln₆o), eluting at 2.178 with a molecular weight of 422.91, known for its angiotensin receptor antagonist properties, as also reported by zhang et al. (2012). tryptophan (c₁₁h₁₂n₂o₂), found at 2.757, exhibited angiotensin antagonistic effects, aligning with the findings of chen et al. (2016). menthone (c₁₀h₁₈o) appeared at 3.316 and showed antifungal, anti-inflammatory, antibacterial, and antiviral properties, similar to zhao et al. (2022). telmisartan (c₃₃h₃₀n₄o₂) was identified at 3.521 and displayed antihypertensive activity, as supported by brittain (2020). safrole (c₁₀h₁₀o₂), with antiangiogenic activity, was prominent at 3.717, consistent with the work of zhao et al. (2005). finally, linoleic acid (c₁₈h₃₂o₂), found at a retention time of 6.060, exhibited anticancer, immuneboosting, weight-reducing, and antiatherogenic effects, similar to the findings of aydin (2005). figure 3: tem micrograph for encapsulated chitosan isolate 1 (14-deoxy-11,12-didehydroandrographolide) at 100 nm transmission electron microscopy (tem) images (figures 2-3) of encapsulated 14-deoxy-11,12didehydroandrographolide and neoandrographolide within a chitosan-cholesterol matrix revealed welldispersed, predominantly spherical nanoparticles with pore sizes around 22.10 nm. these fall within the mesoporous range (2–50 nm), as reported by jiaxun liu et al. (2024), indicating suitability for drug delivery applications. the spherical morphology observed, especially in figure 2, enhances uniform drug release and is ideal for cellular uptake, biodistribution, and improved bioavailability—key advantages for anti-inflammatory drug delivery (jindal, 2017; kulkarni & feng, 2013; yameen et al., 2014). chitosan provides biocompatibility, biodegradability, and protection of the active compound, while cholesterol improves nanoparticle stability and mimics cell membranes, enhancing cellular interaction (de oliveira andrade, 2016) pa ge 52 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 48-53, 2025 overall, the tem results confirm effective encapsulation and optimal nanoparticle characteristics, making the formulation promising for targeted inflammatory treatment. encapsulation efficiency encapsulation efficiency (ee%) was calculated based on the amount of drug entrapped in chitosan nanoparticles. uv-vis analysis at 270 nm showed high efficiency values: • 14-deoxy-11,12-didehydroandrographolide: 82.5% • total extract encapsulation: 71.8% high ee values reflect the effectiveness of the ionic gelation method and compatibility of chitosan with diterpenoid structures. the strong electrostatic interactions between protonated amine groups of chitosan and negatively charged sulfate groups ensured optimal entrapment. this encapsulation is critical for achieving sustained release and targeted drug delivery, reducing the frequency of administration and minimizing systemic side effects. in vitro release profile the drug release profile, measured using a dialysis membrane method in pbs (ph 7.4), was plotted against time. uv-vis spectrophotometry revealed a biphasic release: • initial burst phase (0–1 hr): attributed to surfacebound drug molecules. • sustained release phase (1–4 hr): due to diffusion from the chitosan matrix. compared to non-encapsulated extract, which showed a rapid release within 30 minutes, the encapsulated drug extended release up to 4 hours. this is advantageous for anti-inflammatory therapy requiring prolonged drug presence at the inflammation site. the kinetics followed a quasi-fickian diffusion model, suggesting diffusion as the primary release mechanism. sonographic evidence of inflammation resolution sonographic imaging played a vital role in monitoring the anti-inflammatory effect of the formulation. in the carrageenan-induced rat paw edema model: • b-mode ultrasound showed reduced paw thickness and improved tissue echotexture after 24–48 hours of treatment. • color doppler imaging revealed decreased vascular perfusion in the inflamed area, indicating reduced inflammation. compared to diclofenac-treated groups, the chitosanencapsulated diterpenoids showed equivalent or superior reduction in vascular signal intensity and paw volume. these results were quantified using standardized imaging scales and validated against histopathological findings. thus, sonography offered a real-time, non-invasive assessment of drug efficacy and biodistribution, demonstrating the formulation’s clinical applicability. biochemical and clinical observations the anti-inflammatory effect was further confirmed by measuring serum levels of inflammatory biomarkers: • tnf-α, il-6, and c-reactive protein (crp) levels significantly decreased in the treatment group (p < 0.01). • liver and kidney function parameters (alt, ast, creatinine, urea) remained within physiological limits, confirming safety. these results corroborate the findings from sonographic and histological assessments. additionally, white blood cell (wbc) count and neutrophil/lymphocyte ratios normalized after treatment, supporting systemic antiinflammatory action. this multidisciplinary validation confirms both the safety and therapeutic efficacy of the nanoformulated drug. comparative assessment with standard drugs comparative studies were conducted with standard antiinflammatory drugs—chymoral and diclofenac sodium. results indicate: • the encapsulated a. paniculata isolates produced comparable or superior edema reduction. • lower hepatotoxicity and longer duration of effect were observed. • sonographic profiles were more stable over time, and doppler indices showed consistent vascular improvement. histological examination of paw tissues confirmed reduced neutrophil infiltration and minimal fibrosis, aligning with biochemical and imaging data. thus, this formulation holds promise as a plant-based alternative to synthetic anti-inflammatories, particularly suitable for chronic conditions requiring long-term treatment. conclusion this work effectively illustrates how the transport, stability, and anti-inflammatory effectiveness of the bioactive chemicals are much improved when andrographis paniculata diterpenoids are nanoencapsulated utilizing a chitosan matrix. the sustained drug release and high encapsulation efficiency of the chitosan-based nanoformulation decreased the frequency of administration and enhanced treatment compliance. analytical validation using ftir, sem/tem, xrd, and uv-vis verified the nanoparticles’ morphological and structural integrity. the in vivo anti-inflammatory impact was demonstrated by better histological architecture, normalized organ function measures, and decreased blood levels of inflammatory biomarkers (tnf-α, il-6, and crp). sonographic imaging supported the incorporation of radiological assessment in preclinical investigations by providing a new and real-time way to evaluate inflammation. the nanoformulation not only matches but may surpass traditional treatments in terms of efficacy and safety, according to comparative data with pa ge 53 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 48-53, 2025 standard medications. the importance of integrating imaging, clinical science, nanotechnology, and phytochemistry in medication development is highlighted by this interdisciplinary work. clinical trials, large-scale formulation, and prolonged pharmacokinetics in human models should all be investigated in future research. references akbar, s. (2011). andrographis paniculata: a review of pharmacological activities and clinical effects. alternative medicine review, 16(1), 66–77. https:// pubmed.ncbi.nlm.nih.gov/21438646 beg, s., javed, s., kohli, k., ahmad, f. j., & aqil, m. (2010). nanotechnology-based approaches for the delivery of bioactive compounds in cardiovascular disorders. current vascular pharmacology, 8(2), 168–179. https://doi.org/10.2174/157016110790226645 choudhury, h., pandey, m., hua, c. k., mun, c. s., jing, j. k., kong, l., … & sisinthy, s. p. (2021). an update on natural compounds in the remedy of diabetes mellitus: a systematic review. journal of traditional and complementary medicine, 11(2), 204–229. https://doi. org/10.1016/j.jtcme.2020.10.004 hancke, j. l., burgos, r. a., & ahumada, f. (1995). activity of andrographis paniculata in the treatment of upper respiratory tract infections. phytomedicine, 2(4), 293–298. https://doi.org/10.1016/s09447113(11)80052-1 jin, l., quan, c., hou, x., & fan, s. (2016). potential pharmacological resources: natural bioactive compounds from marine-derived fungi. marine drugs, 14(4), 76. https://doi.org/10.3390/md14040076 kayser, o., lemke, a., & hernandez-trejo, n. (2005). the impact of nanobiotechnology on the development of new drug delivery systems. current pharmaceutical biotechnology, 6(1), 3–5. https://doi. org/10.2174/1389201053167148 kwon, h. c., kauffman, c. a., jensen, p. r., & fenical, w. (2006). marinopyrrole a: a cytotoxic and antibiotic compound from a marine streptomycete. journal of the american chemical society, 128(5), 1622–1632. https:// doi.org/10.1021/ja057153z lim, j. c., chan, t. k., ng, d. s., sagineedu, s. r., stanslas, j., & wong, w. s. (2012). andrographolide and its analogues: versatile bioactive molecules for combating inflammation and cancer. clinical and experimental pharmacology and physiology, 39(3), 300–310. https://doi.org/10.1111/j.1440-1681.2011.05619.x palanisamy, s., liu, x., & wang, y. (2017). marine-derived alkaloids and their anti-inflammatory activities. marine drugs, 15(9), 261. https://doi.org/10.3390/ md15090261 tabas, i., & glass, c. k. (2013). anti-inflammatory therapy in chronic disease: challenges and opportunities. science, 339(6116), 166–172. https:// doi.org/10.1126/science.1230720 pa ge 1 pa ge 66 american journal of chemistry and pharmacy (ajcp) unveiling the sars-cov-2 spike protein: a comparative analysis of vaccine development approaches and glycosylation implications leonel c. mendoza1* volume 2 issue 2, year 2023 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v2i2.1634 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: may 05, 2023 accepted: june 25, 2023 published: july 03, 2023 in december 2019, a mysterious pneumonia-causing sickness frightened the world. sarscov-2 caused the acute respiratory illness. since march 11, 2020, 220,563,227 covid-19 cases and 4,565,483 deaths have been reported worldwide as of october 2021. sarscov-2, like all coronavirus, appears to have crowns due to its s proteins and enters host cells using highly glycosylated spike (s) proteins. s1 and s2 are sars-cov-2 spike protein subunits. s2 controls transmembrane fusion, while s1 controls receptor binding. antibodymediated neutralization targets sars-cov-2 spike (s) proteins, which are essential for viral entry and fusion. this paper summarized how s protein was used in newly created and distributed sars-cov-2 vaccines and the implications for future advancements given the emergence of more lethal sars-cov-2 variants in this paper. it also discussed the role of s protein glycosylation in the viral entry and binding mechanism of sars-cov-2 and the implications for developing adaptive immunity and vaccines. the review was carried out through a deductive search strategy with keywords: covid-19 vaccines, ncov-2019 vaccines, coronavirus, covid-19 vaccine development, s protein, and protein glycosylation using google scholar. the emergence of more transmissible and potentially more lethal sars-cov-2 variants, such as the delta variant, highlights the need for continued research on vaccine development. future research should focus on understanding the mechanism of the spike protein and how vaccines can effectively target the mutated regions. continued monitoring and adaptation of vaccination strategies are essential to control the ongoing covid-19 pandemic. keywords coronavirus, covid-19 vaccines, covid-19 vaccine development, ncov2019 vaccines, protein glycosylation, s protein 1 college of teacher education, mindoro state university-calapan city campus, philippines * corresponding author’s e-mail: leonel.mendoza@minsu.edu.ph introduction coronaviruses are single-stranded rna viruses classified into three genera (alpha, beta, and gamma coronaviruses), which correspond to groups 1, 2, and 3 of the coronavirinae subfamily, coronaviridae family, and nidovirales order or superfamily (weiss & leibowitz, 2011). coronaviruses infect a wide range of animals, both domestic and wild, and humans (pedersen & ho, 2020). six strains, namely, hcov‐229e, hcov‐oc43, hcov‐nl63, hku1, sars-cov, and mers-cov, are known to cause respiratory diseases (hasöksüz et al., 2020). sars-cov and mers-cov variants infect the lower respiratory tract and are therefore more harmful (mcintosh & peiris, 2009); the others, on the other hand, infect the upper respiratory tract but only in moderate cases (fehr et al., 2015). in december 2019, the globe was alarmed by reports of an emergent illness of unknown origin, characterized by pneumonia, and epidemiologically related to a seafood market in the chinese city of wuhan (zhu et al., 2019). a novel coronavirus (sars-cov-2) has been identified as the source of the acute respiratory infection by employing a “pneumonia of unknown etiology” surveillance system built in the aftermath of the 2003 sars outbreak to allow early detection of new infections (li et al., 2020a; zhu et al., 2019). covid-19 was declared a “public health emergency of international concern” (li et al., 2020b) by the world health organization (who) on the 30th of january 2020, and then a pandemic on the 11th of march 2020. (olson et al., 2020). on september 7, 2021, it was reported that there were 220,563,227 confirmed covid-19 cases and 4,565,483 confirmed deaths globally (who, 2021). sars-cov-2 is a member of the betacoronavirus genus (perlman & netland, 2009; schoeman & fielding, 2019; vlasova et al., 2007), which is one of the genera of the orthocoronavirinae subfamily (hasöksüz et al., 2020). it is quite similar to the sars-cov virus, which caused a worldwide epidemic in 2003. sars-cov-2 share the identical tropism and mode of entry as sars-cov– about 80 percent (zhou et al., 2020)– because they use the same cellular receptor, angiotensin-converting enzyme 2 (ace2) (yan et a., 2020). like all coronaviruses, sarscov-2 appears to have crowns because of the s proteins that protrude on its surface (zhao et al., 2021). sarscov-2 uses these highly glycosylated spike (s) proteins (wrapp et al., 2020). the purpose of this research is to describe how the sars-cov-2 s protein was employed in the development of vaccines given its critical function in receptor binding and membrane fusion in the sars-cov-2 virus and discuss the glycosylation of its proteins. specifically, this paper compared each approach on which s protein is given focus in vaccine development. by doing so, this paper can yield implications for future studies. literature review structure the s protein is a class i fusion protein that helps the virus connect to the angiotensin-converting enzyme 2 https://doi.org/10.54536/ajcp.v2i2.1634 https://journals.e-palli.com/home/index.php/ajcp mailto:leonel.mendoza%40minsu.edu.ph?subject= pa ge 67 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 66-73, 2023 (ace2) receptor on the host cell surface, causing the virus and cell membrane to fuse (pardi et al., 2015; hoffman et al., 2020; rauch et al., 2018). the overall length of s protein in sars-cov-2 is 1,273 amino acids, including a signal peptide at the n-terminus (zhao et al., 2021). two subunits, namely, s1 and s2, composed the s protein of sars-cov-2. these subunits are cleaved from a furin site in the virus’ s protein, containing multiple essential amino acids. the s1 subunit is apical v-shaped and harbors one ace2-recognition motif per monomer, the receptorbinding domain (rbd) (sterberg & naujokat, 2020). sars-cov-2 uses a spike (s) protein that is highly glycosylated to gain entry to host cells, as previously stated (wrapp et al., 2020). the s protein is glycosylated through the secretory pathway and is glycosylated by the host cellular glycosylation mechanism (zhao et al., 2021). with this process comes two benefits for the virus itself. to begin with, the mannose residues within these glycans are crucial components in interacting with cell surface attachment proteins (li et al., 2017; tortorici et al., 2019; robson, 2020) before attaching to ace2. second, it helps protect the virus from host antibodies by hiding the underlying polypeptide epitopes (doores, 2015; bagdonaite & wandall, 2018). function as mentioned, the s protein in sars-cov-2 is responsible for receptor identification and membrane fusion (gallagher & buchmeier, 2001), which are carried out by functional domains near the s1 and s2 termini (hasöksüz et al., 2020). the s2 subunit is in charge of transmembrane fusion, whereas the s1 subunit controls receptor binding (chen & guo, 2020; hasoksuz et al., 2002). the trimeric s protein is cleaved into s1 and s2 subunits after viral infection and s1 subunits are released during the transition to the postfusion conformation (song et al., 2018; simmons et al., 2013; belouzard et al., 2009; simmons et al., 2004). because it contains the receptor-binding domain (rbd), which directly connects with ace2, a peptidase domain, the s1 subunit can bind to receptors (li et al., 2005). to engage a host cell receptor, s1’s receptor-binding domain (rbd) undergoes hinge-like conformational movements, briefly hiding or exposing the receptor binding determinants (wrapp et al., 2020). when s1 connects to the ace2 receptor on the host, another cleavage site on s2 is revealed and cleaved by host proteases, a step that is essential for viral infection (belouzard et al., 2009; millet & whittaker, 2015; simmons et al., 2005). the polybasic cleavage site of s may contribute to sars-cov-2’s high virulence, as furin and furin-like proteases, which are necessary for s’s proteolytic activation, are widely expressed in humans, allowing sars-cov-2 to infect a broader range of tissues. furthermore, multiple investigations have suggested that the s protein of sars-cov-2 may employ ace2 to infect the host (zhou et al., 2020; hoffman et al., 2020; kuba et al., 2005; li et al., 2003). ace2 is important because it aids in the maturation of angiotensin, a hormone that regulates blood pressure and vasoconstriction (yan et al., 2020). cardiovascular illnesses are very likely when ace2 expression in the lungs, heart, kidneys, and intestines is reduced (crackower et al., 2002; zisman et al., 2002; raizada et al., 2007). the s protein is a target for antibodymediated neutralization because of its essential function. furthermore, studying the prefusion s structure can help with vaccine development. materials and methods the review was carried out through a deductive search strategy with keywords: covid-19 vaccines, ncov-2019 vaccines, coronavirus, covid-19 vaccine development, s protein, and protein glycosylation using google scholar. articles were included if they were published in english, appeared in peer-reviewed publications, and were related to the issue of the sars-cov-2 virus and vaccine development. statistics from the world health organization were also included. the flowchart outlining the search technique is presented in the figure below. figure 1: search strategy employed results and discussion glycosylation of sars-cov-2 s protein viral protein glycosylation is a successful virus strategy for modifying its proteins using the host-cell machinery. as a result, glycans play critical roles in viral infection and immune response (zhao et al., 2021). glycans on viral surface proteins play a role in viral entry, fusion, epitope shielding, viral protein folding, stability, and protection. for instance, by shielding surface antigens with glycan envelopes, glycosylation on the surface proteins of viruses can prevent antibodies from binding, which plays a significant role in viral infection (shajahan et al., 2021). as mentioned, the s protein of sars-cov-2 is highly glycosylated. sars-cov-2 has 22 n-glycosylation sites and several o-glycosylation sites on each protomer of the transmembrane homotrimeric protein (watanabe et al., 2020). sars-cov-2 infection is influenced by the glycosylation process of both the virus and the target cells on several levels, according to recent reports (reis https://journals.e-palli.com/home/index.php/ajcp pa ge 68 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 66-73, 2023 et al., 2020). given this, it’s worth examining how this process works and how it affects sars-cov-2 because it could lead to vaccine development breakthroughs. for replication and protein glycosylation, sars-cov-2 uses the machinery of the host cell. as a result, the viral surface glycans are composed of host glycans that the immune system recognizes as self, suppressing the anticarbohydrate immune response (crispin et al., 2015). moreover, t cell activation and cytokine production are affected by differential hemagglutinin n-glycosylation, which poses a challenge for vaccine development (hütter et al., 2013). furthermore, as shown in the case of influenza viruses, mutations in protein sequences can alter glycosylation by generating new or removing existing glycosylation sites (altman et al., 2019; zost et al., 2017). this could lead to the emergence of new virus strains (altman et al., 2019). this paper summarizes how protein glycosylation plays a role in sars-cov-2 virus replication, binding and entry, immune response, therapy, and vaccine development in the table below. table 1: biologic and pathologic effects of glycosylation of sars-cov-2 s protein and other insights from literature aspect role of glycosylation in biology and pathogenesis of sars-cov-2 s protein and additional insights reference/s viral entry glycosaminoglycan heparan sulfate of the cellular glycocalyx is required for sars-cov-2 infection of these target cells. heparan sulfate was found to interact with the receptor-binding domain of the sars-cov-2 spike glycoprotein, which is located next to ace-2, causing the spike structure to open up and allow ace-2 to bind to it. clausen et al. (2020) the conformation of the s protein’s receptor-binding domain has been shown to be modulated by glycans at n165 and n234 of the protein. casalino et al. (2020) a sialic acid-binding pocket was discovered at the n-terminus of the s protein from sars-cov-2, suggesting that it may play a role in viral binding. verma (2020) human lectins galectin-3, 7 and 8, siglec-10, macrophage galactose lectin (mgl), and dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (dc-sign) bind to two n-glycans of the rbd of the s protein at n331 and n343, which may provide insight into sars-cov-2 tropism and binding. lenza et al. (2020) adaptive immunity when compared to other blood groups, the proportion of seropositives in group o individuals was significantly lower, implying that group o individuals have a lower risk of infection. gallian et al. (2020) sulfate glycans on the sars-cov-2 s protein could act as selectin ligands and play a role in immune regulation. lowe (2002) variations in virus and host glycosylation are likely to influence tissue tropism and individual infection susceptibility; differences in virus infectivity and patient susceptibility may be caused by sars-cov-2 and ace 2 mutations that alter n-glycosylation sites. reis et al. (2021) vaccine development sars-cov-2 glycosylation can affect each of the current vaccine strategies in different ways (i.e., vector-based vaccine, mrna-based vaccines). reis et al. (2021) suitable glycosylation of the recombinant protein will at least partly determine how immunogenic polypeptide epitopes of vaccine glycoproteins fold. reis et al. (2020) the presence of glycans on protein antigens influences cellular uptake, proteolytic processing, mhc presentation, and subsequent t-cell priming, providing insight into how proper folding of recombinant vaccine glycoprotein can affect the development of an adaptive immune response wolfert & boons (2013) the presence of non-human glycans on recombinant therapeutic glycoproteins may induce antibodies directed against non-human glycan epitopes. zhou & qiu (2019) vaccines based on the sars-cov-2 s protein because the s protein is involved in receptor binding and membrane fusion, vaccinations may induce antibodies that prevent virus binding and fusion or neutralize virus infection. s protein is the major antigenic component of sars-cov. it is essential to generate host immunological responses, neutralizing antibodies, and protective immunity against virus infection among all structural proteins (du et al., 2009). this paper reviewed and compared each approach on which s protein is given focus in vaccine development. some of these vaccines are already being used and distributed worldwide. table 1 summarizes the findings. https://journals.e-palli.com/home/index.php/ajcp pa ge 69 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 66-73, 2023 mrna-based three mrna-based vaccines, biontech/pfizer (bnt162b2 mrna), moderna (mrna-1273), and curevac (cvncov), have been reported (sadarangani et al., 2021; kowalzik et al., 2021). this type of vaccine is said to be different from the traditional kind of vaccine. in an mrna vaccine, a transcript encoding one or more immunogens is delivered into the host cell’s cytoplasm, which is translated into immunogenic proteins (kowalzik et al., 2021). mrna vaccine manufacture does not entail infectious elements or the risk of stable integration into the host cell genome; instead, the vaccine rna strand is destroyed (sadarangani et al., 2021). because of this, this type of vaccine is safer. in the case of biontech/pfizer, a prefusion stabilized, membrane-anchored sarscov-2 full-length spike protein is encoded (polack et al., 2020; walsh et al., 2020). two proline mutations modify this encoding s protein to lock protein in the pre-fusion conformation (pardi et al., 2015; kariko et al., 2005; wrapp et al., 2020). mrnabased vaccines, particularly the biontech/pfizer vaccine, elicit high sars-cov-2 neutralizing antibody titers and robust antigen-specific th1-type cd4+ and cd8+ t-cell responses, similar to dna-based vaccines developed for sars-cov, which induce neutralizing antibody and t-cell responses and protective immunity (wang et al., 2005; huang et al., 2007; liu et (sadarangani et al., 2021). the s1-binding antibody is also present after the first of table 2: summary of key findings vaccine developer nature of the vaccine s protein utilization approach reference/s biontech/ pfizer mrna-based prefusion stabilized, membrane-anchored sarscov-2 full-length spike protein is encoded polack et al., 2020; walsh et al., 2020 moderna mrna-based encodes the sars-cov-2 stabilized prefusion spike glycoprotein trimer baden et al., 2021 university of oxford/ astra-zeneca viral vector-based formulated with simian adenovirus vector encoding the full-length s protein with a tpa leader sequence that is recombinant and replication-deficient folegatti et al., 2020 gamaleya research institute viral vector-based made up of full-length s protein-expressing recombinant and replication-deficient human adenovirus 26 (dose 1) and human adenovirus 5 (dose 2) logunov et al., 2020 janssen viral vector-based formulation was characterized by a recombinant, replication-deficient human adenovirus 26 producing full-length s protein with two amino acid alterations in the s1/s2 junction that eliminate the furin cleavage site and two proline substitutions in the hinge region that keep the protein in the pre-fusion conformation bos et al., 2020 novavax protein subunit uses a recombinant nanoparticle of full-length s protein with protease resistance mutations at the s1/s2 cleavage sites and two proline substitutions to stabilize protein in a pre-fusion conformation, together with a saponinbased adjuvant (matrixm1) keech et al., 2020 sinovac biotech whole cell inactivated virus the attenuated or inactivated whole sars-cov-2 virus is administered to individuals to elicit the immune responses, which target the sars-cov-2 s protein. li et al., 2021 two doses (walsh et al., 2020). like biontech/pfizer’s the moderna vaccine encodes the sars-cov-2 stabilized prefusion spike glycoprotein trimer, which is necessary for host cell attachment and viral entry (baden et al., 2021). s-binding antibody was also detected 14 days after the first dose, with levels increasing marginally by 28 days and significantly after the second dose (jackson et al., 2020). after the second treatment, cd4+ t cells secreting th1 cytokines (tnf>il-2>ifnγ) increased significantly (jackson et al., 2020). viral vector-based incorporating immunogenic full-length or truncated viral surface proteins into viral expression has been a common strategy for vaccine development against infectious agents, primarily viruses (lundstrom, 2020). various viral vector expression techniques have been used to target viral surface proteins. adenoviruses (ads), alphaviruses, flaviviruses, measles viruses (mvs), rhabdoviruses, retroviruses (rvs), lentiviruses (lvs), and poxviruses are all examples of expression vectors (sm wold & toth, 2013; lundstrom, 2019). four viral vector-based vaccines, namely, university of oxford/astra-zeneca (chadox1 ncov-19), gamaleya research institute (gam-covidvac), janssen (ad26.cov2.s), and cansino biologics (ad5ncov), have been reported (sadarangani et al., 2021; knoll & wonodi, 2021). https://journals.e-palli.com/home/index.php/ajcp pa ge 70 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 66-73, 2023 the university of oxford/astra-zeneca vaccine is formulated with a simian adenovirus vector encoding the full-length s protein with a tpa leader sequence that is recombinant and replication-deficient (folegatti et al., 2020). when mice and rhesus macaques were immunized with the chimpanzee ad vector chadox1 ncov-19, which was designed to produce the sars-cov-2 s protein, it induced robust humoral and cellular immune responses and prevented pneumonia in macaques (van doremalen et al., 2020; folegatti et al., 2020). in humans, s-binding antibody was detected 14 days after the first dosage, and levels increased by 28 days; there was a significant rise after the second dose, with a peak at 14 days. also, t cell responses peaked 14 days following the first dosage but were slightly greater 28 days later (folegatti et al., 2020). meanwhile, the gamaleya research institute vaccine is made up of full-length s protein-expressing recombinant and replication-deficient human adenovirus 26 (dose 1) and human adenovirus 5 (dose 2) (logunov et al., 2020). s-binding antibody was found in 85–89% of people 14 days after the initial dose (logunov et al., 2020). fourteen days after the second dose, s antibody levels are boosted (logunov et al., 2021). based on proliferation assays and antigen-specific ifn production, cd4+ and cd8+ t cell responses were seen 14 days after the initial dosage (lugonov et al., 2021). for the janssen vaccine, the formulation was characterized by a recombinant, replication-deficient human adenovirus 26 producing full-length s protein with two amino acid alterations in the s1/s2 junction that eliminate the furin cleavage site and two proline substitutions in the hinge region that keep the protein in the prefusion conformation (bos et al., 2020). after stabilizing substitutions were inserted, the ratio of neutralizing vs. non-neutralizing antibody binding increased, indicating that the s protein was in a prefusion conformation, as observed in its experimental stages. s-binding and neutralizing antibodies were detectable in 99 percent of people 28 days after immunization, and antibody levels were maintained for at least 84 days (janssen biotech, 2021; sadoff et al., 2021). protein subunit to elicit an immune response, protein subunit-based vaccinations combine a protein (or part of a protein) from the targeted virus with an immune-boosting chemical termed an adjuvant (wadman, 2020). adjuvants are frequently required for this type of vaccine to increase the immune response and improve vaccine efficacy (brito et al., 2013). when a protein-based vaccination is taken up and processed into several epitopes by cells, it provides a more focused response to a particular antigen (li et al., 2021). the antigen in most of these vaccinations is either the s protein or its receptor-binding domain (rbd). the novavax vaccine, one of the vaccines that utilize protein subunit, uses a recombinant nanoparticle of fulllength s protein with protease resistance mutations at the s1/s2 cleavage sites and two proline substitutions to stabilize protein in a pre-fusion conformation, together with a saponin-based adjuvant (matrixm1) (keech et al., 2020). antibody to s-binding was identified 21 days after the first treatment, and significantly increased after the second dose. meanwhile, based on ifn, il-2, and tnf production in response to s protein stimulation, cd4+ t cell responses were detectable seven days after the second dosage, with a substantial bias towards a th1 cell phenotype (keech et a., 2020). whole cell inactivated virus sinovac biotech (coronavac), sinoparm (bbibpcorv; wibpcorv), and bharat biotech are three sarscov-2 vaccines that use the whole-cell inactivated virus method (bbv152) (sadarangani et al., 2021). inactivated vaccines, also known as killed vaccines, are made by growing the virus in a culture medium and then treating it with chemicals, heat, or radiation to inactivate it (li et al., 2021). the majority of these vaccines contain aluminum hydroxide (flanagan et al., 2020; who, 2021). the sinovac biotech vaccine, for example, is made from sars-cov-2 produced in vero cells, inactivated using β-propiolactone, and adsorbed onto aluminum hydroxide. in the case of vaccines developed against covid-19, the attenuated or inactivated whole sars-cov-2 virus is administered to individuals to elicit immune responses (li et al., 2021). these immune responses target the sars-cov-2 s protein. inactivated vaccines have a long history of use and can benefit many people, even individuals with advanced immunological senescence (iversen & bavari, 2021). conclusion sars-cov-2 mutations have been on the rise recently. the delta variant, for example, has been linked to a surge in cases in india and has now been found worldwide, including a significant increase in cases in the united kingdom (bernal et al., 2021). the spike protein mutations t19r, 157-158, l452r, t478k, d614g, p681r, and d950n distinguish the delta variant (ecdpc, 2021). several of these mutations may affect immune responses directed at the receptor-binding protein’s key antigenic regions (452 and 478) and the deletion of a portion of the n-terminal domain (li et a., 2020). p681r is located at the s1–s2 cleavage site, and it appears that strains with mutations there has greater replication, resulting in higher viral loads and transmission (johnson et al., 2020). this advent of more lethal sars-cov-2 variations raised questions about the efficiency of existing vaccines, which were meant to combat the initial sars-cov-2 strain. as a result, future vaccine development research must focus on how vaccines can effectively battle these developing strains, focusing on the mechanism of the sars-cov-2 s proteins, which is the site of mutations in the delta variant. references altman, m. o., angel, m., košík, i., trovão, n. s., zost, s. j., gibbs, j. s., ... & yewdell, j. w. 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(2017). contemporary h3n2 influenza viruses have a glycosylation site that alters binding of antibodies elicited by egg-adapted vaccine strains. proceedings of the national academy of sciences, 114(47), 12578-12583. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 27 american journal of chemistry and pharmacy (ajcp) influence of nano copper oxide addition on the thermal, rheological and tribological properties of locally sourced avocado oil based nanofluids idris iliyasu kirim1*, samuel ishaya2, amos danlami agbu3, tani yavala1, paul notani4, mohammed gambo abdullahi1 volume 3 issue 1, year 2024 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v3i1.3869 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: october 05, 2024 accepted: november 11, 2024 published: december 30, 2024 the goal of this study is to turn waste into value by producing lubricant from abandoned avocado fruits, as the globe looks for more environmentally friendly and sustainable lubricants than those derived from fossil fuels. in response to the growing need for highperformance, environmentally friendly products, we created a lubricant enriched with copper oxide nanoparticles (ncuo) using avocado oil as the foundation. the nano copper oxide was elucidated using xrd. to find out how these nanoparticles impact the oil’s viscosity, thermal stability, and other important characteristics, we introduced trace amounts of ncuo at concentrations of 0.4 wt%, 0.8 wt%, and 1.2 wt% in our studies. the nanofluids underwent 3.5 hours of ultrasonication at 75°c. the formulation with 0.8 weight percent ncuo proved to be the most effective, with a higher flash point of 220°c and viscosities of 7.5 cst at 40°c and 2.7 cst at 100°c, all of which satisfied the astm d445 standards. furthermore, as compared to pure avocado oil, the ideal formulation demonstrated a 13.6% decrease in the coefficient of friction (cof) and a 16.9% decrease in wear rate. good lowtemperature performance was indicated by the pour point of -1°c and the cloud point of 15°c, respectively. according to the thermal conductivity measurements, the oil’s capacity for heat transport was improved by the addition of ncuo. these results imply that avocado oil-based nanofluids, especially those containing 0.8 weight percent ncuo, have great potential as high-performing, environmentally friendly substitutes for lubricating light-duty engines and other industrial uses. keywords avocado oil, nanofluids, nano copper oxide, tribological properties 1 department of mechanical engineering, taraba state polytechnic, suntai, nigeria 2 works department, taraba state polytechnic, suntai, nigeria 3 department of electrical electronic engineering, taraba state polytechnic, suntai, nigeria 4 department of science lab. technology, taraba state polytechnic, suntai, nigeria * corresponding author’s e-mail: kirimsid@gmail.com introduction rising awareness of environmental sustainability and dwindling fossil fuel reserves have driven a worldwide movement towards eco-friendly alternatives across various industries. the lubricant sector, in particular, faces mounting pressure to lessen its dependence on petroleum-based products due to their non-biodegradable nature and harmful impact on the environment. conventional lubricants made from petroleum not only damage ecosystems but also accelerate the depletion of finite resources. to address these issues, bio-based lubricants especially those sourced from vegetable oils are increasingly recognized as promising substitutes. avocado oil, known for its high viscosity index, outstanding thermal stability, and abundant fatty acid content, has gained attention as potential base oil for biolubricants. its natural attributes make it ideal for uses that demand consistent lubrication at diverse temperatures. nevertheless, similar to other vegetable oils, avocado oil struggles to meet the demanding performance standards of industrial applications especially in wear resistance, friction reduction and oxidative stability which have impeded the broader adoption of biolubricants in sectors with rigorous performance criteria. nanotechnology presents a promising solution to these challenges. integrating nanoparticles like nano copper oxide (cuo) into bio-lubricants has been proven to significantly improve their tribological properties. with its high surface area and distinct physical characteristics, nano cuo enhances anti-wear and anti-friction qualities while offering excellent thermal conductivity. these improvements position nano cuo as an excellent additive for enhancing the performance of lubricants derived from avocado oil, potentially closing the gap between environmental sustainability and industrial efficiency. although the outlook is promising, there is limited comprehensive research on avocado oil-based lubricants formulated with nano cuo additives. current studies mostly address either the base properties of avocado oil or the overall impact of nanoparticles on lubricant performance, which leaves a significant gap in understanding this unique combination fully. this study aims to fill that gap by developing an eco-friendly and high-performance lubricant using avocado oil as its base combined with nano cuo additives. by employing systematic formulation, thorough characterization, and extensive performance testing methods, our research seeks to advance sustainable lubricant technologies while providing a viable alternative to conventional options that align with both environmental and industrial standards. traditional petroleum-based lubricants, though effective, present considerable environmental challenges due to their non-biodegradability and the emission of harmful byproducts. with nigeria being a signatory to the paris accord and international environmental regulations becoming stricter alongside increasing demand for pa ge 28 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 27-34, 2024 sustainable industrial practices globally, reducing reliance on fossil fuel-based products has become crucial. this shift necessitates finding eco-friendly alternatives. however, bio-based lubricants made from vegetable oils often fail to meet performance requirements in heavyduty or high-temperature applications, which hinder their widespread use. large quantities of avocado fruit are being transported from gembu in the sardauna local government area of taraba state to other regions, especially jalingo, the state capital. however, due to long travel distances and a lack of specialized transportation methods, many avocados spoil before reaching their destination markets. this results in waste that needs disposal and underscores an urgent need to transform this wasted produce into valuable products. avocado oil is known for its high thermal stability and viscosity index which make it promising base oil for bio-lubricants. nevertheless, its natural properties do not yet match up with the superior performance offered by synthetic lubricants alone. advances in nanotechnology have shown that adding nano copper oxide (cuo) additives could significantly boost anti-wear qualities while reducing friction and enhancing thermal stability when used within avocado-based oils as lubricants. despite these possibilities, there remains insufficient research on formulating such improved lubricant solutions featuring enhanced characteristics through nano scale cuo integration marking out crucial gaps needing bridging toward establishing sustainable high-efficiency lubrication standards systematically vetted via appropriate testing methodologies under various conditions. in their study, shafi et al. (2018), investigated the tribological performance of copper (cu) nanoparticles mixed with avocado oil as a lubricant. they used a pinon-disc tribometer to evaluate both the lubricating capabilities of pure avocado oil and the enhanced effectiveness when cu nanoparticles were added. stribeck curves were plotted for both base oil and oils containing cu nanoparticles at concentrations of 0.5 wt. % and 1 wt%. the worn surfaces on aluminum alloy 6061 pins were examined using scanning electron microscopy (sem). their findings revealed that incorporating cu nanoparticles reduced friction and wear noticeably. a minimum coefficient of friction was observed at a concentration level of 1 wt%, while specific wear rate reached its lowest point at the concentration of 0.5 wt%. this enhancement in overall tribological properties was attributed to the film-forming ability offered by adding these particles to bootstrap those advantages over ordinary base oils. in another study by guzman et al. (2018), they explored the tribological properties of copper nanoparticles. they incorporated these particles at concentrations of 0.3 wt% and 3.0 wt% into both mineral oil and synthetic ester base oils. the team utilized a pin-on-disk tribometer to measure the coefficients of friction (cof). from their investigations into friction and wear, they arrived at several conclusions: (1) they discovered that copper nanoparticles are unsuitable for use in synthetic ester base oil. the addition of these particles increased wear by 7.5 times, and there was no change observed in the friction behavior. (2) incorporating copper nanoparticles into mineral base oil significantly decreased friction and wear. at a concentration of 0.3 wt% of copper nanoparticles, the reduction in wear was observed to be as much as 64%. ultimately, they concluded that while copper nanoparticles are effective in mineral base oils, they do not perform well in synthetic base oils. akl et al. (2018) had examined the tribological properties of engine lubricants enhanced with nano-copper oxide as an additive. they incorporated copper oxide nanoparticles into mobil 1 sae15w-40sf engine oil at a concentration of 0.1% by weight. two new engines were utilized for testing over a period of 1000 hours; one was supplemented with the nanolubricant while the other used standard lubricant. periodic sampling resulted in twelve samples from each engine to assess wear particles using astm-d6595 spectrometry standards. further analysis on selected oil samples employed laser net fines analyzer tests, revealing improved friction characteristics through decreased wear rates when nano-additives were applied. specifically, reductions in aluminum, iron, and chromium wear particles were observed at rates of 48%, 11.5%, and 42%, respectively alongside average declines in specific types: cutting wear reduced by around 39%, severe sliding wear by approximately 36%, and fatigue wear by about 60%. additionally, a notable decrease in engine temperature was achieved. in their study, rajaganapathy et al. (2021) investigated the tribological properties of vegetable oil enhanced with nanoparticles using a pin-on-disc tribometer. they utilized palm and brassica oils in their experiments, incorporating copper oxide (cuo) and titanium oxide (tio2) nanoparticles as additives at weight percentages of 0.1% and 0.5%. the findings revealed that palm oil containing a lubricant sample with 0.5% cuo exhibited a significant reduction in both coefficient of friction and wear compared to other lubricant samples tested. additionally, the viscosity and thermal conductivity properties increased as nanoparticle concentrations were raised within these vegetable oils. materials and method materials the materials used to carry out this research safely are listed below: • waste avocado fruit. • copper acetate • sodium hydroxide • oleic acid • ethanol • phosphoric acid • water • potassium hydroxide • sodium hydroxide pa ge 29 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 27-34, 2024 method preparation of base oil waste avocado fruits were collected from gembu and jalingo. they fruits were cleaned, with the stones separated from the skins. the pulp was placed in a blender and bitten until it becomes thick paste. the paste was then thin spread onto a tray to ensure a wider surface area. the tray was lodge into an oven initially set at the temperature of 450c and held for 5-6hrs until it turns dark green. the dark green paste was transferred in small batch onto thin cheesecloth, while applying pressure, the oil is extracted slowly into a cleaned container. this procedure was repeated until 1000cm3 of avocado oil was obtained. finally, the extracted oil is transferred into plastic bottle and labeled avooil for storage. this process is painstaking and the oil yield is small as compared to the industrial process. purification of base oil determination of the acid value a digital balance was used to weigh 1.11g of base oil, while another measurement recorded 0.56g of koh, which was then dissolved in 100ml of water to create a 0.1 molar solution. this koh solution was titrated against the oil until the endpoint indicated by the oil turning pink was reached at a volume of 2.7 ml, determining what is known as either the acid value or neutralization number. determination of % total fatty acid the %tfa was obtained from equation 1 below, %tfa= (total fatty acid)/2 (1) preparation of copper oxide nanoparticles an aqueous solution containing 0.02 mol of copper acetate was prepared in a round-bottom flask. to this, 1 ml of glacial acetic acid was added and the mixture heated to 100°c with constant stirring using magnetic stirrer. subsequently, 0.4 g naoh was gradually introduced into the warmed solution until its ph reaches neutral (ph = 7), resulting in a copious formation of black precipitate. this precipitate was then subjected to centrifugation and washed four times using deionized water before being air-dried for up to one day. to enhance surface functionality, cuo nanoparticles were modified by adsorbing oleic acid as a carboxylate onto their surfaces; this chemical modification facilitates superior integration within blends acting as solid-liquid suspensions ultimately fostering strong intermolecular forces at interfacial zones according van der waals theory by minimizing molecular agglomerations. this was achieved by dispersing 5g of nanoparticle into a preheated 100ml of ethanol at 600c. the solution was then stirred for 15 minutes to attain even dispersion, and then 0.25 ml oleic acid was added and stirred for 2h. the solution was centrifuged to separate the ethanol-oleic acid mixture. the resulting nanoparticles were oven dried at 80 0c to remove the excess surfactant as adopted from (oparanti et al., 2021). bio nanolubricant preparation (bnnl) the nano-lubricant was formulated into one sample by dispersing the treated cuo nanoparticles (np) into the purified avocado fruit oil in weight percentages from 0.4 wt. % and stirred. the dispersion of nanoparticles in the base oil was done by ultra-sonication for 3.5h, at fixed interval of 30 minutes throughout the process. the sonication was conducted at 59 hz frequency, 100% power, and temperature of 75 0c. the precipitates at the bottom of the beaker were checked to make sure that the dispersion process is going in accordance with the protocols by (lawal et al., 2023). after the sonication, the blend was vacuum dried for 24hrs to exclude the effect of gas bubbles and moisture generated during the process. the biolubricant produced was stored into a bottle and labeled avoncuo4. figure 1: graphical abstract pa ge 30 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 27-34, 2024 evaluation of chemical properties xrd of nano copper oxide the phase composition of the nano copper oxide powders were investigated through data that were collected on x-ray powder diffraction (xrd) patterns machine (rigaku miniflex diffractometer operating a copper tube at λ=1.5418a, which was generated at a voltage of 40 kv and a current of 30 ma. viscosity measurement kinematic viscosity at 40°c and 100°c was measured using a brookfield dv-ii+ pro viscometer following astm d445. viscosity index was calculated based on the measured values. cloud point and pour point measurement the cloud point and pour point were determined using astm d2500 and astm d97, respectively. cloud point visual observation of wax formation while cooling the nanofluid. pour point the lowest temperature at which the nanofluid remained flowable. flash point measurement flash point was measured using the pensky-martens closed-cup apparatus according to astm d93. tribological test the pin-on-disk anton paar tribometer, which measures friction and wear with extreme precision under controlled loading circumstances, was chosen to evaluate the tribological characteristics of the nanofluids. the test was devised to assess the impact on wear behavior and friction of varying quantities of copper oxide (cuo) nanoparticles distributed in avocado oil. friction and wear measurements during the tribological test, the following properties were measured: coefficient of friction (cof) the cof was continuously recorded using the anton paar tribometer’s high-resolution friction force sensor (0.06 mn resolution), which accurately captured small variations in frictional forces throughout the test. wear scar diameter after each test, the wear scar on the stationary balls was measured using optical microscopy. this allowed for an estimation of the wear resistance of the nanofluids. results and discussion xrd pattern of the ncuo particles from figure 2 below, the crystalline planes of nano copper oxide, or nano cuo, are generally represented by discrete peaks in the x-ray diffraction (xrd) pattern. the peaks at 2θ values of 36° and 39° in a nano cuo xrd figure 2: xrd patterns of ncuo particles pattern are particularly interesting to examine because they can reveal information about the sample’s phase composition and crystal structure. prominent peaks at 36° and 39° usually suggest that monoclinic cuo makes up the majority of the sample’s composition. the lack of further prominent peaks may indicate phase purity, which would indicate the presence of none or very little other copper oxides (such as cu2o or metallic copper). the size of the cuo nanoparticles can be inferred from the peak broadening. larger crystallites are indicated by sharper peaks, and smaller nanocrystallites are indicated by broader peaks. the degree of crystallinity can be inferred from the relative intensities of these peaks in relation to other peaks in the xrd pattern. relative to the background noise, high-intensity peaks indicate wellordered crystal formations. pa ge 31 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 27-34, 2024 kinematic viscosity the kinematic viscosity of the nanofluids was measured at both 40°c and 100°c for three different concentrations of ncuo: 0.4 wt%, 0.8 wt%, and 1.2 wt%. the results are summarized in table 1 and illustrated in figure 3. table 1: viscosity of ncuo nanofluid at different concentrations conc. (wt%) viscosity @40oc viscosity @100oc 0.4 6.2 2.1 0.8 7.5 2.7 1.2 8.9 3.5 figure 3: viscosities of ncuo particles at different conc figure 3 illustrates a steady rise in viscosity with increasing ncuo concentration. at 40°c, the viscosity rose to 8.9 cst at 1.2 wt% from 6.2 cst at 0.4 wt%. likewise, across the same concentration range, the viscosity at 100°c increased from 2.1 cst to 3.5 cst. these findings support the notion that the presence of nanoparticles, particularly at greater concentrations, increases fluid resistance to flow. these viscosity values are within permissible bounds for lubricants, per astm d445, particularly for industrial applications where temperature control is crucial. ali et al. (2015) showed a similar trend in which the addition of metal oxide nanoparticles raised the viscosity. cloud and pour point the cloud and pour points of the nanofluids were measured to assess the low-temperature performance of the fluids. table 2 and figure 4 summarize the results. table 2: cloud and pour points of ncuo nanofluids conc. (wt %) cloud point in oc pour point in oc 0.4 15 -1 0.8 13 -3 1.2 11 -4 figure 4: cloud and pour points of ncuo nanofluids pa ge 32 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 27-34, 2024 figure 4 illustrates how cloud and pour points gradually decrease as ncuo concentration rises. the cloud point was 15°c at 0.4 weight percent and 11°c at 1.2 weight percent. the pour point also dropped, going from -1°c at 0.4 wt% to -4°c at 1.2 wt%. these decreases imply that raising the ncuo concentration improves the fluid’s low-temperature performance and increases its suitability for cold situations. the observed pour points satisfy astm d97 specifications for hydraulic fluids and industrial lubricants that must continue to function at lower temperatures. the findings of sedighi et al. (2014), who found that metal oxide nanoparticles decreased the pour point of oil-based fluids, are likewise consistent with this trend. flash point the flash point is an important safety measure for fluids that operate at elevated temperatures. table 3 and figure 5 summarize the results of the flash point measurements for the nanofluids. tribological properties table 3: flash points of ncuo nanofluids conc. (wt %) flash point (oc) 0.4 217 0.8 224 1.2 231 table 4: tribological properties of nanofluid sample cof wear scar diameter (mm) pure avocado oil 0.096 0.45 0.4wt% ncuo nanofluid 0.081 0.38 0.8wt% ncuo nanofluid 0.073 0.32 1.2wt% ncuo nanofluid 0.078 0.35 figure 5: flash points of ncuo nanofluids the flash point rose as the concentration of ncuo increased, as figure 5 above illustrates. from 217°c at 0.4 wt% to 231°c at 1.2 wt%, the flash point increased. this increase in flash point suggests that as nanoparticle concentration rises, so does the nanofluid’s thermal stability. for industrial applications, like engine oils or heat transfer fluids, that need stability at higher operating temperatures, these values are acceptable, under astm d93, which describes the closed-cup test procedure for flash point determination. vajjha and das (2009) observed a similar trend, with metal oxide nanoparticles improving base oils’ thermal stability. the friction coefficient is marginally lower at 0.4 weight percent ncuo than base oil, indicating a minimal degree of lubrication enhancement. more nanoparticles suspended in the base fluid at 0.8 weight percent cause the friction coefficient to further decrease, improving boundary lubrication. because of an even thicker coating of protective nanoparticles that lessens direct metal-tometal contact, the friction coefficient for the 1.2 weight percent sample is at its lowest. this decrease is in line with earlier research (li et al., 2015), which demonstrates that friction in nanofluids is much reduced at increasing nanoparticle concentrations. it is clear from the data that tribological performance improves with increasing ncuo concentration. the wear scar diameter, wear rate, and lowered frictions are all well-balanced in the 0.8 weight percent sample. the 1.2 weight percent sample, on the other hand, had the lowest wear rate, friction coefficient, and wear scar diameter, offering the best overall performance. when taking into account both tribological and economic factors, 0.8 weight percent may be the best option because it offers significant improvement without the higher costs and possible handling complications associated with a higher concentration of nanoparticles, even though 1.2 weight percent offers superior tribological performance. conclusion the potential of avocado oil-based nanofluids augmented with nano copper oxide (ncuo) as a workable substitute for traditional lubricants is well demonstrated by this study. the viscosity, tribological performance, and thermal stability of the lubricant were all markedly enhanced by the addition of ncuo at an ideal concentration of 0.8 weight percent. according to the findings, the improved formulation significantly reduced the coefficient of friction and wear rate when compared to pure avocado oil, in addition to showing a higher flash point and excellent viscosity characteristics. these results demonstrate how well nano additives work to improve the performance of bio-lubricants, which qualifies them for use in high-performance lubrication applications. the combination of the advanced qualities of ncuo with avocado oil, a renewable and biodegradable resource, is in line with the growing need in a variety of industrial sectors for environmentally friendly lubricants. pa ge 33 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 27-34, 2024 additionally, this work opens the door for future studies into the stability and long-term performance of avocado oil-based nanofluids and advances our knowledge of the synergistic impacts of nanotechnology on bio-lubricants. in summary, this research highlights the significance of creative methods for improving environmentally friendly lubrication technology, encouraging a change in the lubricant sector toward more environmentally friendly options. recommendations i. it is recommended to explore additional concentrations of ncuo beyond 0.8 wt% to identify any potential enhancements in lubrication performance and thermal stability. this may help determine the optimal concentration for specific applications. ii. conduct long-term stability and aging studies of avocado oil-based nanofluids to evaluate their performance over extended periods. this will help ascertain their viability for real-world applications and identify any degradation issues that may arise. iii. implement field tests in various mechanical systems, such as light-duty engines and industrial machinery, to assess the performance of the optimized nanofluids under operational conditions. this will provide insights into their practical effectiveness and durability. iv. investigate the environmental impact of using avocado oil-based nanofluids in comparison to conventional lubricants. this assessment should include studies on biodegradability and toxicity to ensure that these formulations align with sustainability goals. v. consider investigating other bio-based oils as potential base fluids for nanofluid formulations. this could expand the range of sustainable lubricant options available and enhance the understanding of how different oils interact with nano additives. vi. encourage collaboration with industry stakeholders to facilitate the adoption of advanced bio-lubricants. this partnership can foster innovation and support the development of new technologies that enhance the performance and sustainability of lubricants. vii. educational outreach: promote awareness and education on the benefits of using bio-lubricants, particularly those enhanced with nanotechnology, within both the industrial sector and academic circles. this will help drive interest and investment in sustainable lubricant solutions. funding this work was sponsored under the tetfund’s institution based research (ibr) funds, (2024). reference akl, s. y., abdel-rehim, a. a., & khafagy, e. a. 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(2019). thermal and rheological properties of vegetable oil-based nanofluids. energy conversion and management, 198, 111755. https://doi.org/10.1016/j. enconman.2019.111755 pa ge 1 pa ge 12 american journal of chemistry and pharmacy (ajcp) the formulation of aloe vera gel herbal soap and characterized its physicochemical and antibacterial activities with market available herbal soap, butwal, nepal arjun bhandari1*, bipana sapkota1, gautam prasad chaudhary2 volume 3 issue 1, year 2024 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v3i1.3410 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: june 30, 2024 accepted: august 24, 2024 published: november 21, 2024 the incredible properties of formulated and commercial aloe vera herbal soaps have been thoroughly investigated revealing their potent phytochemical and antibacterial effects. the samples were collected from rupandehi, lumbini province. aloe vera soap was carefully formulated using a cold process. through disc diffusion, the soap antibacterial effect was tested against harmful gram-positive and gram-negative bacteria, including escherichia coli and staphylococcus aureus. the impressive results showed a maximum zone of inhibition (zoi) of 8 mm and 7 mm for formulated soap, respectively. this was due to the presence of important bioactive compounds such as tannins, saponins, and reducing sugar, which are abundant in aloe vera extract. interestingly, the zoi was less noticeable at lower concentrations of 100 mg/ml, 50 mg/ml, and 25 mg/ml. the formulated soap still exhibited similar properties to the commercial soap, with a ph of 9.7, moisture content, and foam stability. additionally, the formulated soap had a mixture content of 4.6%, 0.52% free caustic alkali, and a foam stability test measurement of 6.5 cm. these remarkable results highlight the antibacterial potential of aloe vera-based soap, proving its effectiveness against harmful microorganisms. keywords antimicrobial test, antiseptic soap, escherichia coli, phytochemical screening, qualitative analysis, staphylococcus aureus 1 department of chemistry, butwal multiple campus, tribhuvan university, butwal, nepal 2 department of pharmacy, crimson college of technology, pokhara university, butwal, nepal * corresponding author’s e-mail: bhandariarjun112@gmail.com introduction as a cleaning agent, soap is produced as granules, bars, flakes, or liquid and is made by the reaction of potassium or sodium salt with different naturally occurring fatty acids. any water-soluble salt of fatty acids with eight or more carbon atoms is also known as soap. soaps are made for many uses, such as cleaning, bathing, taking medication, etc. the negative ions on the hydrocarbon chain that is connected to the carboxylic group of the fatty acids are what give soap its cleaning properties. because soap’s carboxylic group is more soluble in water than it is in oil or grease, soap is mostly utilized in conjugation with water for cleaning reasons (edah et al., 2017). triglycerides are saponified to generate soap; this process reacts with a strong alkali, like koh or naoh to form glycerol and fatty acid salts. the long hydrocarbon chain of a cleaning soap molecule contains an ionic interaction with a metal ion, typically potassium or sodium, at one end of the chain, where a carboxylic acid group is located. the ionic end is soluble in water, but the hydrocarbon end is non-polar and extremely soluble in non-polar substances. because of their capacity to combine water-insoluble materials and retain them within the water suspension, soaps have the power to clean (shehu et al., 2020). the structure of the soap molecule is represented below: (structure of soap, 2024). ch3 (ch2)16 coona+ non-polar hydrocarbon chain ionic end (soluble in non-polar substances) (soluble in water) the fatty acids required to produce soap, such as stearic acid, myristic acid, palmitic acid, lauric acid, and oleic acid, are what give it its lathering and washing qualities. even in seawater, soaps produced with fatty acids with 12 or more carbons are highly soluble and readily lather. however, because they irritate the skin and have offensive scents, fatty acids with 10 or fewer carbons are seldom employed in soaps. because soaps can emulsify or scatter things that are insoluble in water and hold them in a suspension of water, they have cleaning properties. this capability is demonstrated by the molecular makeup of soaps. the molecules of soap envelop the oil droplets when they are introduced to water containing oil or other substances figure 1: a general scheme of soap preparation (saponification) process (besty et al., 2013) pa ge 13 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 12-19, 2024 that are insoluble in water. the ionic end of the oil permits it to dissolve in water, but the alkyl groups of the soap molecules dissolve it. consequently, the oil droplets should scatter throughout the water and be removed by washing (habib et al., 2016). the general saponification reaction used in soap preparation is shown figure 1. producing natural and handmade soap is a completely creative process that calls for a range of abilities, materials, creativity, and careful thought in order to produce soap of the highest caliber. fatty acids and alkali salts derived from plants or vegetables, as well as organic materials or natural smells, are characteristics of herbal soaps. it takes the presence of fatty acids and bases like potassium and sodium hydroxides for the hot-and-cold process that makes soap (sindhu et al., 2019). natural, organic ingredients like herbs and skin-beneficial ingredients are used to make herbal soaps. there is very little chance that these soaps will cause any harm because they are entirely chemical-free. rather, they contain essential oils like saffron, sandalwood, strawberry, and rose water, along with natural oils like castor and almond oil, all of which are great for smoothing, brightening, and whitening skin (hrushikesh & hingane, 2022). herbal soap preparation is a pharmaceutical or medicinal that is generally used to treat illnesses and injuries and to boost general health using plant components such as leaves, stems, roots, and fruits. it has antifungal and antibacterial qualities as well. this product can be administered topically and possesses antibacterial effects. it is available in many different forms, such as ointment, soap, gel, lotion, cream, and solvent extract. the qualities of creams and soaps have been used to cure a variety of skin issues (dwiyanti et al., 2021). the evergreen green herb aloe vera, as aloe barbadensis, is a plant that has fruits that are loaded with seeds, yellow tubular blooms, and thick, triangular leaves with sharp edges. each leave is composed of three layers: inside is a translucent gel that contains 99% water and additional ingredients such as vitamins, lipids, sterols, amino acids, and glucomannans. aloe vera leaves are used to create a mucilaginous, colourless gel that is frequently used in cosmetic and therapeutic applications (sanchez et al., 2020). in the past, this medicinal herb was used to cure skin ailments like inflammation, burns, and sores. aloe vera has also been shown to have antioxidant, anticancer, antidiabetic, and anti-hyperlipidemic properties. aloe vera contains over 75 different compounds, including vitamins a, c, e, and b12, enzymes like amylase, catalase, and peroxidase, minerals like zinc, copper, selenium, and calcium, sugars like monosaccharides like mannose6-phosphate and polysaccharides like glucomannans, anthraquinones like aloin and emodin, fatty acids like lipol and campesterol, hormones like auxins and gibberellins, and other substances like salicyclic acid, lignin, and saponins (sopan et al., 2023). this study was conducted to manufacture the herbal soap in assess its biological activity against both grampositive and gram-negative bacteria in addition to its physicochemical features. materials and methods the study was conducted in sainamaina municipality, rupandehi, lumbini province, nepal. according to rupandehi (wikipedia, 2024), the local government area is located between 27º 43’ 12’’ north latitude and 83º 18’ 36’’ east longitude. the aloe vera leaves collected from home garden. to keep the leaves out of the sun, they were kept in the refrigerator for further use. figure 2: aloe vera plant source: fieldwork 2023 data analysis and interpretation the primary data was collected after the experiments and observation in the laboratory work. the data was arranged in tables and were evaluated by using appropriate statistical methods such as bar diagrams, graphs, and lines and other tools. table 1: analysis and methods parameters methods employed ph ph measurement pa ge 14 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 12-19, 2024 moisture content change in initial and final weight free caustic alkali titration foam stability height of the foam saponification value titration chemicals and standards every chemical utilized was of the best grade available on the market. laminar air flow and antimicrobial activity incubator and autoclave were provided by indosati scientific lab equipment, delhi, and s.m. scientific instrument (p) ltd. extraction of aloe vera gel initially, aloe vera leaves that were just cut were cleaned with water to rid of any dirt or yellow material known as aloe latex that leaked out of the leaves. 400 g of aloe vera leaves were precisely weighed using an automated weighing balance following washing. to gently cut away the clear gel in the center, the tops of the leaves and the spines at the edges of each leaf were removed. to achieve the pure gel, the residual gel was spoonfully scooped into a mixture and then ground and filtered off. after that, the gel was placed in a beaker and kept at 4ºc in the refrigerator for further study. formulation of aloe vera soap the formulation of aloe vera soap was carried out using (upadhyay et al., 2021). after weighing 185 g of aloe vera gel using an electronic scale, the gel was transferred into an ice cube tray to freeze. subsequently, the beaker was filled with the frozen gel of aloe vera. then, gradually add 65.65 g of naoh. the temperature of the aloe vera gel increased when naoh was added. i had chilled the aloe vera gel to stop it from burning for that reason. the flakes of naoh were dissolved. an r.b. flask was filled with 450 g of mixed oils (350 g olive oil and 100 g of coconut oil). subsequently, the r.b. flask was filled with the aloe vera gel and naoh combination. after that, three items were thoroughly combined with a shaker machine. using the glass rod, the mixture and the aloe vera gel were rapidly mixed. aloe vera gel, a vegetable oil, and naoh solution had an exothermic reaction. there would be a longer saponification procedure. thus, the mixture was stirred for two to three minutes, and then it was allowed to thicken for ten minutes. the mixture was thickened after one hour of this operation. to make the mixture thick, it was put into the mold and allowed to rest for a few days. a thick bar started to form in the mold after one to two days. after the prepared soap had been soaked in the filter paper, it was ready to use. figure 3: a-formulated aloe vera and b-commercial herbal soap physiochemical analysis of soap prepared the properties of formulated and commercial soap can be characterized in terms of ph, moisture content, free caustic alkali, and foam stability. determination of ph in soap to determine the ph of the produced soap, the beaker holding 20 ml of distilled water was filled was 2 g of the soaps that had been manufactured. using the glass rod, it was aggressively swirled until the soap was completely dissolved. using a ph meter, the soap solution’s ph was determined after 12 hours (kareru et al., 2010). determination of moisture content the empty clean crucible dishes were dried in the oven at a temperature of 105ºc for 30 minutes and cooled for 10 minutes in a desiccator. 5 g of the prepared soap sample were taken in a dried, and tarred dishes. the sample was then dried for two hours at 101ºc to determine the moisture content (benjamin et al., 2022) with some modification. a watch glass filled with soap was removed from the oven after two hours, and the total weight was determined. the following formula was used to calculate the moisture content; % moisture content= weight loss/sample weight * 100 (1) pa ge 15 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 12-19, 2024 determination of free caustic alkali (fca) dissolved 5 g of aloe vera soap into 30 ml of ethanol. 10 ml of 20% bacl2 were added along with a few drops of phenolphthalein indicator. then, titrate the solution against 0.05 m h2so4, by changing the color of the indicator endpoint is recorded (habib et al., 2016) with some modification. the free caustic alkalinity was calculated by using the following formulae: fca = 0.31/w × va (2) (betsy, 2013) where, va = volume of titrated acid w = weight of aloe vera soap fca = free caustic alkali foam stability in a 100 ml measuring cylinder, 1 g of the aloe vera soap was weighed and dissolved in 20 ml of distilled water. after giving the mixture a good shake for two to 2 minutes, it was left to stand for an additional 2 minutes. after that, the height of the foam was measured and noted (kareru et al., 2010). determination of phytochemicals tannins, flavonoids, saponins and reducing sugars were determined using qualitative methods described by imohiosen, (2023) with some modifications. test for tennins in a beaker, 20 ml of distilled water and 0.5 g of aloe vera extract were brought to a boil before being filtered. to the filtrate, a few drops of 0.1% ferric chloride were applied. a blue-black or brownish-green coloration suggested the presence of tannin. test for flavonoids sulphuric acid (h2so4) was added to a portion of the plant extract’s aqueous filtrate after 5 ml of diluted ammonia solution was added. the presence of flavonoids was shown by the appearance of a yellow solution that disappears with standing. test for saponins 2.0 g of plant extract was boiled in a water bath with 20 ml of distilled water, then filtered. for a stable, persistent fourth, 10 ml of the filtrate and 5 ml of distilled water were combined and forcefully shaken suggest the saponins. test for reducing sugars 1 ml of fehling’s solutions a and b was added to 1ml of each sample’s aqueous filtrate, which was then heated in water. the presence of non-reducing sugar was indicated by the red precipitate. antimicrobial activity test for the research, gram-positive (s. aureus) and gramnegative (e. coli) bacteria were obtained from the crimson college of technology’s department of b.sc. mlt in butwal, rupandehi, nepal. the gram-positive and gram-negative commercial standards gentamycin and ciprofloxacin were utilized. preparation and dilution of soap samples extract (chaudhari, 2016) in a sterile container 1 g of each soap sample was weighed and dissolved in 5 ml of distilled water. the resulting concentration of formulated and commercial soap of 200 mg/ml, 100 mg/ml, 50 mg/ml and 25 mg/ml were then made serially and utilized for the disc preparation. preparation of culture medium (rahama & sani, 2020) muller hinton agar (mha) media preparation activation of culture plates the previously prepared, frozen at 5ºc media plates were sufficiently dried during incubation. in a sterile airflow hood, the plates were thereafter allowed to cool. preparation of filter paper disc whatman’s no. 1 filter paper was used to create filter paper discs. 5 mm discs were produced in petri plates and autoclaved for 15 minutes at 121ºc to sterilize them. a formulated and commercial soap solution of 200 mg/ ml, 100 mg/ml, 50 mg/ml and 25 mg/ml was added one by one to each sterile disk. paper discs soaked in the soap solution were allowed to stand for a period of one hour to ensure full saturation of the soap preparations. after being aseptically withdrawn from the soap solution, the discs were left to dry in an oven at 25ºc. assay of antimicrobial activity overnight cultures were kept ready for anti-microbial activity. assay of the antimicrobial activity of soap were done by disc diffusion method. disc diffusion assay to find antibacterial assay, the agar disc diffusion method (chaudhari, 2016; rahama & sani, 2020) was employed. the surface of sterile muller-hinton agar plates was inoculated with a standardized 0.1 ml saline suspension of test organisms. using sterile forceps, aseptically transfer sterile filter paper discs made from varying concentration i.e. 200 mg/ml, 100 mg/ml, 50 mg/ml and 25 mg/ml respectively of the individual formulated and commercial soap samples directly onto the plate surface and incubation at 37ºc for 24-48 hours. measurement of zoi the zone of inhibition (zoi) was determined by using a ruler or vernier caliper in millimetre (mm.) caused by the aloe vera formulated soap and commercial soap and compared with standard antibiotics following a 24 hours’ incubation period. pa ge 16 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 12-19, 2024 results and discussion the physicochemical parameters of the soap were categorized based on multiple factors, including ph, moisture content (mc), free caustic alkali (fca), and foam stability of both prepared and commercial aloe vera soap. table 2: the physiochemical results obtained from prepared soap and commercial soap characteristics prepared soap commercial soap ph 10.75 9.18 free caustic alkali (%) 0.527 0.806 moisture content (%) 4.6 8.8 foam stability (cm) 6.5 10 figure 4: the bar graph of physicochemical analysis of prepared and commercial herbal soap physicochemical properties the ph values of the herbal soap that was produced and sold were found to be within the permitted range of 9 to 11, which was in line with earlier research conducted by rama and sani (2020). based on the measured ph, the soap should be alkaline, less corrosive, safe for the skin, and cause fewer skin reactions when used. other characteristics that were indicative of the produced and commercial herbal soap were measured, such as the moisture content, formability, and percentage of free alkali content. the prepared aloe vera soap had a moisture level of 4.6%, according to the analysis’s results, while commercial soap had a high moisture content while produced soap had a low moisture content. low moisture indicates that the material won’t hydrolyze if stored for an extended amount of time (chitkara et al., 2020). the aloe vera soap that was prepared had a free caustic alkali of 0.57%, whereas commercial soap had a free caustic alkali of 8.8%. the prepared soap’s low free caustic alkali rating indicates that it is suitable for sensitive skin types. the amount of alkaline-free ingredients in the soap that, in excess, might irritate the skin is known as the free caustic alkali value. it is a sign that the soap won’t be too rough on the skin or fabric. according to the trial approach. homemade aloe vera soap had the second-highest foam height at 6.5 cm, after commercial soap with a height of 10 cm. for foam height, liquid viscosity is the most influential factor. the type of oil used to make the soap, particularly palm kernel oil, whose main fatty acid is lauric acid and is well-known for its high formability, is responsible for the amount of foam height (rahama & sani, 2020). the height of the foam aids in removing grime, oil, and bacteria from the skin. phytochemical screening table 3: results of phytochemical screening compounds results tennis + flavonoids + saponins + reducing sugar + (+) = present table 3 shows the phytochemical screening result of the aloe vera extract which indicated the presence of all phytochemicals tested. in the previous literature (kareru et al., 2010) almost all the phytochemical screening of the aloe vera leaves extract. antibacterial susceptibility test table 4: the antibacterial activity of produced and commercial soap of their zoi s zone of inhibition against fractions (mm) produced soap (mg/ml) commercial soap (mg/ml) 200 100 50 25 200 100 50 25 pa ge 17 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 12-19, 2024 e. coli 8 7 8 7 s. aureus 10 9 8 ciprofloxacin (e. coli) 34 33 s. aureus 20 17 gentamicin (e. coli) 24 25 s. aureus 15 13 control (-) = no zone of inhibition figure 5: the bar graph of antimicrobial activity of prepared and commercial herbal soap with e. coli and s. aureus bacteria figure 6: different concentrations of solution prepared by formulated and commercial herbal soap pa ge 18 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 12-19, 2024 a = zoi of formulated soap against s. aureus; b = zoi of commercial soap against s. aureus; c = zoi of formulated soap against e. coli; d = zoi of commercial soap against e. coli the prepared soap and the commercial soap exhibit strong inhibition at the highest concentration (200 mg/ ml), which is followed by a steady decline in inhibitory character down to the lowest dosage (25 mg/ml) in the given anti-bacterial screening results. according to sharma et al. (2022), the zoi of gram-positive (b. subtiles) and gram-negative (s. typhi) at 400 mg/ml were, respectively, 0.82 and 1.62 mm for manufactured herbal soap and marketed herbal soap. higher concentrations of zoi of produced soap a-500, 250, 125, 62.5, and 32.25 mg/ml were achieved for s. aureus 20, 16, 12, 9, and 7 as well as e. coli 15, 11, 9, 8, and 7 mm, respectively, showing anti-bacterial action (kareru et al., 2010). the finding shows that the commercial aloe vera soap had a higher concentration of anti-bacterial activity than the produced soap. this is because the commercial aloe vera soap had extra ingredients that enhanced its activity, while the produced aloe vera only contained olive and coconut oil and naoh. also by preventing bacterial development, this showed that both soaps have anti-bacterial activity. conclusion in addition to its moisturizing and anti-inflammatory qualities, aloe vera seems to enhance the skin’s ability to absorb some pharmaceutical molecule. since, external application of aloe vera on intact skin is generally considered harmless and does not appear to have any side effects, the use of this natural resource as a penetration enhancer is intriguing (sharma et al., 2015). this study shows that formulated herbal soap from aloe vera leaves can be successful. when compared to commercially available herbal soap, the formulated soap’s physicochemical qualities show a significant improvement. the results that are now available indicate that aloe vera extract has antibacterial properties and has been successfully employed in the formulated soap. recommendation to prevent adverse effects on the skin, make sure the right amount of necessary ingredients is used when making aloe vera soap. we can create a variety of herbal soaps with easy-to-use methods that don’t include dangerous chemicals for everyday usage. • before using soap on our bodies, it is important to inspect its quality. • determining the minimum inhibitory concentration (mic) of aloe vera soap. • determining the results of various soap’s minimum bactericidal concentration (mbc) may also be conducted. • determine the antioxidant character of herbal soaps. data availability the data used to support the findings of this study are included in the article. authors’ contributions all author prepared the materials, collected the data, and conducted the analysis in addition to contributing to the study’s conceptualization and design. gautam p c has specially supported the anti-bacterial analysis procedure. moreover, arjun b wrote the draft of the work, offered feedback on earlier iterations, and read and approved the completed version. acknowledgments the authors would like to thank butwal multiple campus, tribhuvan university and crimson college of technology, pokhara university for their invaluable assistance in testing the study samples and greatly enhancing the correctness of our results. references ajegi, j. o., benjamin, i., igbawase, s. d., kukwa, r. e., ngunoon, t. p., samoh, t. f., & iortile, t. m. 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(2021). formulation and evaluation of aloe vera containing herbal soap. journal of engineering technologies and innovative research, 8(5), d411-d416. pa ge 1 pa ge 2 american journal of chemistry and pharmacy (ajcp) a systematic review: methods of gingivectomy for esthetic marginal periodontal tissue conditioning maha munshi1* volume 2 issue 2, year 2023 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v2i2.1681 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: may 24, 2023 accepted: june 22, 2023 published: july 21, 2023 this review compared the prognosis of gingivectomy procedures carried out through a scalpel or alternative techniques. an extensive search was performed in the electronic database. articles were identified according to inclusion criteria. the extracted data from the selected studies were organized in tables and assessed for risk of bias. ten clinical trial studies were identified. nine studies used the laser method for gingivectomy, and only one trial was based on the electrosurgical method. seven studies were at low risk of bias, and three were at high risk. no significant differences were presented in pain and discomfort in the electrosurgical studies. one study reported non-significant differences between electrosurgery and scalpel regarding clinical healing. based on limited data in this systematic review, the diode laser showed superiority over the scalpel in pain and discomfort postoperatively. in contrast, for pi and gi, the data were insufficient to conclude the results. keywords gingivectomy, electrosurgery, scalpel, diode laser, esthetic marginal periodontal tissue, tissue conditioning, periodontal 1 malmo university, sweden * corresponding author’s e-mail: mahamunshi321@outlook.com introduction a healthy periodontium should be maintained, especially during tooth restoration, by keeping a safe zone of healthy gingival tissue called “biological width” between the edge of the sub-gingival filling and the bone margin or during the excision of excess gingival tissues for esthetic purposes. the term “biological width” was replaced by “supracrestal attached tissue” in the new classification of periodontal and peri-implant disease and conditions in 2017(jepsen et al., 2018). it is histologically composed of the junctional epithelium and supracrestal connective tissue attachment. the clinicians can create a healthy supracrestal attached tissue or reposition existing and maintain biological width by gingivectomy, including an apically repositioned flap. gingivectomy in crown lengthening procedures is one of the methods proven effective in maintaining the supracrestal tissue attachment intact. it includes removing a pre-determined deep periodontal pocket using a scalpel, electrosurgery, and laser surgery. this systematic review compares the conventional method for gingivectomy (i.e., surgical scalpel) with other methods described for gingivectomy in the literature based on precision cutting, cost-effectiveness, hemostatic function, patient’s perception of pain, discomfort during and after the surgery, and periodontal parameters, including pi, gi, and pocket depth (pd). methods of gingivectomy surgical scalpel the surgical scalpel has been used as a gold standard for many years. despite the ease of use, cost-effective, precision, and less harmful to the surrounding tissues, bleeding during the surgery obscures the surgical field, and the risk of scalpel injury has been reported. the risk of scalpel injury was about 18% of all sharp injuries, and about 31% of scalpel injuries occurred during direct scalpel use, according to the exposure prevention information network “epinet” report in 2003(perry et al., 2003). to obtain a clear surgical field, the clinician needs a surgical instrument that cuts efficiently like a scalpel, causing less bleeding during the surgery and minimal risk of injury. william cameron developed this surgical instrument in 1928. it was his first dental electrosurgical unit to assist in achieving hemostasis. electrosurgical unit harris defined an electrosurgical unit (es) as “the use of specially designed electronic equipment that produces a limited variety of high-frequency waveforms to cut or remove soft tissue”(hs., 1976). while oringer mj. defined it as “the application of electrically generated heat energy to living tissue to alter or destroy it for therapeutic purposes” (oringer, 1975). there are two types of es units, monopolar and bipolar. the bipolar type has a forceps-like electrode, whereby the electrical current passes from one tip to the other, with the targeted tissue placed between them. the monopolar type is the most common use in dentistry, as described by gnanasekhar jd. et al. (gnanasekhar & al-duwairi, 1998; yalamanchili et al., 2013). there are three different waveforms: fully rectified and filtered, fully rectified and unfiltered, and partially rectified. the fully rectified, filtered waveform produces excellent tissue separation with little hemostasis and the least lateral heat. this waveform can be used for soft tissue surgeries, such as a frenectomy, incision and drainage, and gingival trough procedures around the teeth. the fully rectified, unfiltered waveform produces good tissue separation and hemostatic effect but causes tissue shrinkage and https://doi.org/10.54536/ajcp.v2i2.1681 https://journals.e-palli.com/home/index.php/ajcp pa ge 74 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 74-85, 2023 generates additional lateral heat, avoiding bone contact. it is suitable for gingivectomy, gingivoplasty, and pulpotomy. the partially rectified waveform causes much more lateral heat than the fully rectified, unfiltered; therefore, it can be used only as a hemostatic effect in soft tissue and as a heat source for bleaching agents (gnanasekhar & al-duwairi, 1998; yalamanchili et al., 2013). moreover, electrosurgery has an alternating current that passes through the tissue. the tissue resists the electrical current, generating lateral heat, which causes the intracellular water to boil, increasing intracellular pressure and rupturing cell membranes. chemical method chemosurgery is another method to perform a gingivectomy. two chemicals were used to remove the gingiva, such as 5% paraformaldehyde and potassium hydroxide. schwarz first used formaldehyde n. et al. (n, 1917). paraformaldehyde is a white powder that is not readily soluble, so it needs to be mixed with zinc oxide powder and eugenol as a liquid. this mix is called a “medicated cement pack”(orban, 1943). special care should be taken in applying medicated cement packs in the gingival pockets because of possible abscess formation, bone necrosis, and allergy to formaldehyde(orban, 1943). potassium hydroxide was advocated by harndt e.(e, 1951). however, it is a strong alkali, and it has a necrotising effect on the gingiva and alveolar bone. laser system laser is another method for gingivectomy. “laser” is an abbreviation of light amplification by the stimulated emission of radiation. the laser was invented first by theodore maiman, usa, in 1960(maiman, 1960). four common types of lasers are used now in dental practice, including the carbon dioxide laser (co2), the diode laser (dl), the neodymium: aluminum-yttrium-garnet (nd: yag), and the erbium: aluminum-yttrium-garnet (er: yag). laser wavelengths in medicine and dentistry generally range from 193 nanometers (nm) to 10,600 nm. american, in 1989, produced the first dental laser for commercial use, using nd: yag as an active medium, but only for soft tissue surgery(keller & hibst, 1989). after a few years, the uk 1990 developed a machine laser using nd: yag as an active medium that can cut through the enamel, dentin, and bone. shortly followed by a similar er, cr: ysgg (erbium chromium: yttrium scandium gallium garnet) laser in 1997(parker, 2007a). the laser has a clear and precise cut, good visibility related to its ability to seal blood vessels and lymph vessels during the procedure, minimal damage to adjacent tissues, and fewer postoperative complications such as pain and swelling, which increase patient satisfaction, accelerate wound healing, and less surgical time. carbon dioxide laser kumar patel invented the co2 laser in 1964. the wavelength ranges from 9.4 to 10.6 micrometres. about 98% of the energy is converted to heat and absorbed at the tissue surface with little penetration. for decades, the co2 laser was the gold standard for intraoral soft tissue surgeries. the laser medium consists of water or air-cooled gas discharge (carbon dioxide, nitrogen, hydrogen, xenon, helium) that helps produce a beam of infrared light by activating the footswitch (viraparia et al., 2012). the depth of the laser incision is related to the power setting and the exposure’s duration. higher energy removes the tissue, while lower energy is used for hemostasis and photocoagulation. continuouswave mode forms char during the procedure. char accumulation during the surgery leads to increased tissue temperature up to >200°c, so it is recommended to clean the surgical site to avoid thermal damage(low & mott, 2014). in addition, this system has no tactile feedback. clinical applications of co2 laser (ishikawa et al., 2009; viraparia et al., 2012) a) oral and maxillofacial surgery; b) oral medicine; c) pre-prosthetic procedures; d) periodontal procedures; e) endodontics: pulpotomy, filling material removals such as gutta-percha or resin; f) restorative procedures. nd: yag the nd: yag laser was developed by geusic et al. in 1964. the medium is a crystal of yttrium-aluminium-garnet doped with neodymium. have a wavelength of 1064 nm. in 1973, data were calculated for the water absorption spectrum of different lasers, such as argon, diode, nd: yag, co2, er, cr: ysgg, and er: yag by hale and querry (hale & querry, 1973). they showed that the energy of nd: yag scattered rather than being absorbed by water like a diode laser, which led to greater penetration in the soft tissue. in addition, the wavelength is attracted to colours, so this system is ideal for ablating potentially hemorrhagic abnormal tissue and hemostasis of small venous vessels and capillaries. nd: yag laser device has contact or non-contact probes, giving tactile feedback. clinical applications of nd: yag laser(ishikawa et al., 2009) a) oral and maxillofacial; b) periodontal procedures: gingivectomy, gingivoplasty, operculum removal, bactericidal effect (cobb, 2006), suppressing or eradicating putative periodontal pathogens from periodontal pockets (parker, 2007b; verma et al., 2012), as an adjunctive or alternative treatment to conventional mechanical therapy in periodontitis (cobb, 2006). er: yag and cr: ysgg as for the co2 laser system, it is essential for er: yag and cr: ysgg that the tip be at least 1-2 mm away from the target tissue to have a good result. the erbium laser has two distinct wavelengths, 2790 2940 nm cr: ysgg lasers and er: yag lasers, respectively, and has a high affinity for hydroxyapatite and the highest absorption of water(ishikawa et al., 2009). consequently, it is the https://journals.e-palli.com/home/index.php/ajcp pa ge 75 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 74-85, 2023 laser for treating dental hard tissues (harashima et al., 2005). it can also be used for soft tissue ablation because it produces a little heat into the underlying tissues and elevates the pulpal temperature compared to a co2 laser. clinical applications of er: yag and cr: ysgg laser(ishikawa et al., 2009) (bader & krejci, 2006) a)endodontic; b) periodontal procedures: srp, ablation of bone tissue, bactericidal effect on implant surface; c) restorative procedures. diode laser (dl) the diode laser has energy absorbed by the pigmentation in the soft tissue and haemoglobin, making this system an excellent hemostatic agent (fornaini et al., 2016). it is a solid semiconductor operating at 810-980 nm wavelengths. the energy is either a continuous or a pulsed mode. using the pulsed mode during the soft tissue procedure and lowering the power setting is recommended. in contrast to other laser systems, it table 1: pico model population patients in need of marginal tissue conditioning intervention gingivectomy comparison use a scalpel as a standard method, and compare it to other methods (e.g., electrosurgery and different types of laser). outcome patient's reported outcomes and clinical periodontal parameters directly contacts the target tissue so the clinician can feel tactile feedback (kravitz & kusnoto, 2008). clinical applications of the diode laser (verma et al., 2012) (bm s, 2017) a) oral and maxillofacial; b) periodontal procedures; c) restorative procedures; d) orthodontic; d) endodontic. material and methods research question the research question was, “for marginal tissue conditioning, is gingivectomy with alternative methods superior to conventional gingivectomy with a scalpel regarding patient-reported outcomes and clinical periodontal parameters?” the protocol was constructed according to “prisma” (preferred items for systematic reviews and metaanalyses) criteria (liberati et al., 2009), and a question was formulated according to “pico.” model as mentioned in table 1. table 2: search strategy sr. no search strategy 1 gingivectomy [abstract & keywords] or electrosurgery [abstracts & keywords] or diode laser [abstract & keywords] or gingival enlargement [abstract & keywords] or gingival hypertrophy [abstract & keywords] or scalpel [abstract & keywords] or gingivoplasty [abstract & keywords] 2 gingival hypertrophy [tw] or gingivectomy [tw] or electrosurgery [tw] or diode laser [tw] or supra crestal attached tissue [tw] or gingival enlargement [tw] or gingivoplasty [tw] or crown lengthening [tw] or treatment [tw] or scalpel [tw] 3 gingivoplasty [abstract & keywords] or gingivectomy [abstract & keywords] or gingival enlargement [abstract & keywords] or supra crestal attached tissue [abstract & keywords] or scalpel [abstract & keywords] or diode laser [abstract & keywords] or electrosurgery [tw] or gingival hypertrophy [tw] 2): “gingival hyperplasia” or “gingival hypertrophy” or “gingival overgrowth” and gingivoplasty or gingivectomy or “crown lengthening”. search strategy up to august 2019, the following mesh keywords and free text phrases were searched in the electronic databases: medline (pubmed), scopus, and web of science (table assessment of the eligibility of studies and the data extraction method the author assessed the research based on the inclusion and exclusion criteria. inclusion criteria for studies were selected based on articles in the english language, available as full-text, consisting of human trials, controlled clinical trials (cct), or randomised controlled clinical trials (rcts) with a minimum of 10 patients per study. it also includes articles based on patients who underwent surgery for gingivectomy by using the scalpel compared with the alternative methods. in addition, exclusion criteria exclude articles based on languages other than english, animal or vitro studies, case reports, clinical notes/letters, and editorials. materials were then screened by reviewing the titles and excluding irrelevant articles for our investigation. the remaining studies’ abstracts were evaluated, and the complete texts of the published literature were received. if a full-text article could not be retrieved, the author was contacted and requested the complete version. a search was also conducted in the reference lists of the selected papers. lastly, a forward search of the listed studies was also conducted using the science citation index. https://journals.e-palli.com/home/index.php/ajcp pa ge 76 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 74-85, 2023 appraisal of methodological quality risk of bias assessment (rob) the methodological quality of the studies included in the systematic review was appraised as part of the data extraction process. this was performed using the cochrane collaboration’s risk of bias assessment tool (cochrane handbook for systematic reviews of interventions) (sterne et al., 2019). the following criteria were evaluated at “low,” “high,” or “unclear” risk of bias: randomisation process, deviations from intended interventions, missing outcome data, measurement of the outcome, selection of the reported result, and overall. like all surgical procedures, “gingivectomy,” performed using the scalpel, electrosurgical method, or other alternative procedures, requires anaesthetising the periodontal and surrounding structure. this could be done through anaesthetic gel or induction of local anaesthesia. all the included studies were evaluated individually: high risk if only one high risk were present, and unclear risk if only one of the criteria was unclear and no high-risk present. data synthesis and analysis the extracted data from the selected studies were arranged in four tables. the first one is about the general characteristics of the selected studies, including author, year of publication, study number in roman style, study design, randomisation procedure, participant number, age, gender, type of intervention, follow-up, co-morbidity, and drop-out. the second table presents the data of the patient’s related outcome measures (proms). the third table reports the data, including author, year of publication, type of interventions, and periodontal parameters like pi, gi, and bleeding rate (br). finally, the last table shows the data of the other parameters, like the clinical observation of healing and crevicular fluid measurements. results and discussion electronic database search the database searches led to the retrieval of 552 articles from the computerised database (figure 1), with 159 duplicated studies identified and eliminated. in addition, 10 full-text studies were examined after excluding 286 papers based on the title and 97 studies based on the abstract. a further search was conducted per the reference lists from full-text, and previous review and scientific citation index, which included three full-text articles. the ultimate number of studies considered for full-text evaluation was 13. following a comprehensive articles examination, 3 publications were excluded for differing reasons listed in table 3. 10 publications met the eligibility criteria and were included in the present systematic review. figure 1: flow diagram table 3: reasons for exclusion of 3 full-texts study (year) reasons for exclusion petersen et al. (1993) no control group ize-iyamu et al. (2013) less than 10 patients inchingolo et al. (2010) less than 10 patients risk of bias assessment the risk of bias was done for the final included studies, and all the studies were assessed as low risk, except three studies, iii, vi, and ix, which were high risk (figure 2). study characteristics ten studies were included in the systematic review, seven were rcts, and three were ccts; all were prospective studies. the rcts used different randomisation techniques (see table 4). most studies used simple ones like coin toss or computer generator random methods. only one study used the stratified random sample technique. the designs were variables between parallel in eight studies and splitmouth in two studies. the number of participants ranges between 11 and 58. the age of the samples ranged between 11 and 48 years old. the follow-up duration for https://journals.e-palli.com/home/index.php/ajcp pa ge 77 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 74-85, 2023 the samples was variable, between 2 hours and 6 months, but one study did not report the follow-up period. all the included studies reported that a full explanation of the clinical procedure was provided before participants and signed informed consent was obtained from the patients, except two studies (ii, ix) that did not report. six studies reported that the ethics committee approved them. unfortunately, heterogeneity in the data collected was found regarding study design, type of intervention, outcome measures, and observational periods. figure 2: risk of bias assessment table 4: general characteristics of included studies author(year) (ref. no.) study no. study design randomization procedure participant no. age(years) gender (f/m) typesof intervention follow up co-morbidity dropout koppolu et al. (2016) (29) i prospective rct parallel group computer general randomization 14 18-30 8/6 c: scalpel t: diode laser 1 month none none aremband et al. (1973)(30) ii prospective rct split mouth randomisation 27 18-35 14/13 c: scalpel t: electrosurgical unit 1, 2, 3 weeks none none durham et al. (2009) (31) iii prospective cct split mouth 15 12-17 10/5 c: scalpel t: diode laser 2 months none none prasad et al. (2018) (32) iv prospective rct parallel group coin toss method 20 16-40 9/11 c: scalpel t: diode laser 7 days none 2 patients did not follow up öncu et al. (2017)(33) v prospective rct parallel group coin toss method 20 35.2 12/8 c: scalpel t: diode laser nr none none akram et al. (2017) (34) vi prospective cct parallel group 50 25-45 nr c: scalpel t: diode laser 1 week none none sobouti et al. (2014) (35) vii prospective rct parallel group 30 17-29 18/12 c: scalpel t: diode laser 2 hours none none https://journals.e-palli.com/home/index.php/ajcp pa ge 78 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 74-85, 2023 lione et al. (2019)(59) viii prospective rct parallel group computer generated randomization 58 11.7-19.8 26/32 c:non-surgical t:scalpel t:diode laser 6 months none 2 patients mussa et al. (2017) (37) ix prospective cct parallel group 30 17-34 14/16 c:scalpel t:diode laser 1, 2 week none none kumar et al. (2015) (38) x prospective rct parallel group 10 20-40 nr c:scalpel t:diode lasr 24, 36h 1, 2, 3 week none none types of reported outcome variables proms most studies performed quantitative and qualitative assessments regarding proms and clinical periodontal outcomes, but the primarily quantitative assessment was performed. proms were reported in the form of a scale and as a subjective assessment. vas scales of pain and discomfort were reported together in one study (iv) and as separate scales; e.g., the vas scale of pain was presented similarly (10cm) in five studies (i, iii, v, vii, x). in contrast, the vas scale of comfort was presented only in one study. in addition, one study (iv) reported the frequency of pain at different time intervals. another study (x) presented patients’ satisfaction levels. a qualitative assessment regarding the pain and discomfort was performed in one study (vi). patients were investigated if they experienced the pain of equal severity on both sides or more severe on one side, if they needed to use any pain killer, and if there was any discomfort and difficulty during speech and eating. another qualitative assessment was done in one study (i) regarding the cosmetic outcome by evaluating the patient’s perception of cosmetic change and expected outcome. effect of diode laser: pain two studies performed the mean vas scores [rob: low (i, x)] on the first day and were significantly lower in the laser groups than in the scalpel groups. another study reported lower pain values in the laser than in the scalpel in the first two days [rob: high (vi)] with no statistical analysis. three studies [rob: low (i, iv, vii)] presented the vas scores on the third and seventh days of follow-up and were significantly low in the laser groups in comparison with the scalpel groups in two studies only (iv, vii). one study [rob: low (v)] presented that the mean vas values for the whole evaluation period and significantly lower in the test groups than in the control groups. only one study [rob: high (iii)] evaluated the vas of pain daily for seven days. it showed a lower significant value in the laser group than in the scalpel group on the third day. study (iv) evaluated the frequency of pain on the third, fifth, and seventh days, and it was significantly low in the laser group than in the scalpel group. studies (vi, vii) presented how many analgesics the patient consumed. it was significantly high in the control groups than in the test groups after 82 minutes postsurgical in the study (vii), while the other study (vi) table 5: comparison b/t diode laser & scalpel regarding proms. author (year) (ref. no.) study no. prom/proms koppolu et al. (2016) (29) i vas of pain patient’s perception of cosmetic patient evaluation for perception regarding expected outcome from the treatment after 4th week day 1.29* 3 day 0.25 7 day 0.0 1 day 3.1 3 day 1.98 7 day 0.25 1 w mild=1 moderate=3 sufficient=3 2 w mild=0 moderate=3 sufficient=4 4 w mild=0 moderate=2 sufficient=5 moderate=3 sufficient=2 2 w mild=2 moderate=3 sufficient=2 4 w mild=1 moderate=3 sufficient=2 unsatisfied=0 p. satisfied=2 t. satisfied=5 unsatisfied=2 p. satisfied=2 t. satisfied=3 https://journals.e-palli.com/home/index.php/ajcp pa ge 79 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 74-85, 2023 durham et al. (2009) (31) iii vas of pain 1 day (1.9) 2 day (0.5) 3 day (0.3)* 4 day (0.2) 5 day (0.1) 6 day (0.1) 7 day (0.1) 1 day (1.4) 2 day (0.9) 3 day (0.4) 4 day (0.4) 5 day (0.2) 6 day (0.1) 7 day (0.1) prasad et al. (2018) (32) iv vas of pain & discomfort frequency of pain at different time intervals (intermittent pain/no pain) 3 day 0.50* 5 day 0.10* 7 day 0.30* 3 day 2.0 5 day 1.10 7 day 1.00 3 day 50%/50%* 5 day 10%/90%* 7 day 30%/70%* 3 day 100%/0% 5 day 100%/0% 7 day 90%/10% öncu et al. (2017) (33) v vas of pain 2.4* 7.1 akram et al. (2017) (34) vi pain (n) discomfort analgesic consumption (n) experience some pain for the first two days (15) experience some pain for the first two days (21) experience some discomfort in speech & eating (0) experience some discomfort in speech & eating (25) (60%) (84%) kumar et al. (2015)(38) x vas of pain vas of comfort patient’s satisfaction during surgery (3) 24-36h (2)* during surgery (1) 24-36h (4) 1 week (19)* 1 week (16) 3 week (8) 3 week (6) sobouti et al. (2014) (35) vii vas of pain analgesic consumption (n) postsurgical 0.0* postsurgical 5.2 (0) (14)* after 82 minutes ref reference, no number, grey shade box= test group, vas visual analogue scale, *statistical significant between test & control groups, w week, p partial, t total, (n) number of patients, h hour. showed high analgesic consumption in the control group than in the test group but did not perform statistically analysis (see table 5). discomfort two studies [rob: low (iv); high (vi)] evaluated discomfort values, and one study [rob: low (x)] evaluated the vas of comfort. study (vi) reported that all patients experienced discomfort during speech and eating in the control groups. another study (x) showed a higher significant value of vas of comfort in the laser group than in the scalpel group after a week. the other study (iv) showed lower significant discomfort values in the laser groups than the scalpel groups on the third and seventh days (see table 5). effects of electrosurgery pain the pain was evaluated in one study [rob: low (ii)] by asking about the patient’s experience after one week and showed no significant difference between the electrosurgery and scalpel groups. however, the vas scale of discomfort was not performed (table 6). table 6: comparison b/t electrosurgery & scalpel regarding proms. author (year) (ref. no.) study no. prom/proms aremband et al. (1973)(30) ii patient’s experience 6.67% patient’s experience 10% periodontal parameters we found six studies that reported the periodontal parameters. the pi and gi were presented in four studies [rob: high (iii, vi, ix); low (viii)]. the bleeding parameter was presented as a scale in two studies [rob: low (v, vii)] and as subjective data in one study (ix). https://journals.e-palli.com/home/index.php/ajcp pa ge 80 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 74-85, 2023 one study presented the pd and the clinical crown length (viii). another study (ix) assessed the swelling and presented subjective data (see table 5). effect of diode laser pi and gi were performed in four studies [rob: low (viii); high (iii, vi, ix)]. three studies (iii, vi, ix) evaluated the pi after a week postoperatively. no significant difference was recorded between the laser and scalpel groups (iii, ix). study (vi) presented a higher significant postoperative value in the control group compared to the preoperative value. the pi was evaluated after two weeks postoperatively (ix), after a month in two studies (iii, viii), and after six months in another study (viii), and showed no significance between the laser and scalpel groups. in contrast, the gi was presented significantly higher in the control groups in comparison to the test groups after a week (vi, ix) and also after two weeks (ix), but study (iii) showed no significance. the evaluation of gi after a month (iii, viii) and after three and six months (viii) did not show any significant values. other periodontal parameters like pd and clinical crown height were evaluated in one study (viii) after one, three, and six-month observation periods and showed a significant reduction in the pd. a significant increase in the clinical crown height after one month versus baseline values in the two test groups was observed compared to the control groups, but no significant difference between the two test groups. after three months of follow-up, a relapse occurred in the test groups with a decrease in the clinical crown height and an increase in the pd, but not significant when compared with the control groups or between the test groups. when observing six months versus three months, no significant difference was found between the groups in pd and clinical crown height. the bleeding was assessed in three studies [rob: low (v, vii); high (ix)] in two different ways. study (ix) evaluated the bleeding by grading the following criteria: none, self-limiting, require pressure, coagulation, and require ligation or hemoclip. all the patients required pressure in the control group, while in the test group, only one patient required pressure, two patients were self-limiting bleeding, and eleven patients did not bleed. studies (v, vii) reported a significantly lower bleeding rate in the test group than in the control group. only one study (ix) evaluated the swelling according to the following: none, slight, moderate, and severe. no swelling was reported in the test group compared to the control group, while three patients showed slight swelling in the control group (table 7). table 7: comparison b/t diode laser & scalpel regarding periodontal parameters a ut ho r ( ye ar ) (r ef . n o. ) st ud y n o. periodontal parameters li on e et al. (2 01 9) (3 6) v ii i pi gi pd (mm) ccl (mm) d l sc al pe l n on e su rg er y d l sc al pe l n on e su rg er y d l sc al pe l n on e su rg er y d l sc al pe l n on e su rg er y b 0.4 1 m 0.6 3 m 0.7 6 m 0.6 b0.4 1 m 0.6 3 m 0.5 6 m 0.6 b 0.6 1 m 0.5 3 m 0.4 6 m 0.7 b 0.3 1 m 0.5 3 m 0.6 6 m 0.7 b 0.3 1 m 0.5 3 m 0.6 6 m 0.7 b 0.5 1 m 0.4 3 m 0.5 6 m 0.6 b 4.8 p.s 0.7 1 m 1.8* b 4.9 p.s 0.9 1 m 1.8* b 4.4 1 m 4.0 3 m 3.3 6 m 3.8 b 7.7 p.s 10.6 1 m 10.1* b 7.9 p.s 10.3 1 m 9.9* b 7.6 1 m 8.0 3 m 8.3 6 m 8.2 m us sa et a l. (2 01 7) (1 4) ix pi (n) gi (n) intraoperative bleeding (n) swelling (n) test control test control test control test control bl 0.51 1 week 0.38 2 week 0.28 bl 0.46 1 week 0.37 2 week 0.38 bl 0.72 1 week 0.55* 2 week 0.29* bl 0.85 1 week 1.45 2 week 0.68 none 11 self-limit 2 pressure 1 coagulation 0 ligation or hemoclip 0 none 0 self-limit 0 pressure 16 coagulation 0 ligation or hemoclip 0 none 14 slight 0 moderate 0 severe 0 none 13 slight 3 moderate 0 severe 0 https://journals.e-palli.com/home/index.php/ajcp pa ge 81 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 74-85, 2023 a kr am et a l. (2 01 7) (3 4) v i #scaling 1.22 #surgery 0.3 1 week 0.41 #scaling1.37 #surgery.0.39 1 week 1.31* #scaling 1.18 #surgery 0.44 1 week 0.53 #scaling 1.34 #surgery. 0.52 1 week 0.91* so bo ut i e t a l. (2 01 4) (3 5) v ii bleeding rate 0.36 out of 4* 1.15 out of 4 1.32 o nc u et al. (2 01 7) (3 3) v 1 week: 0 (26) 1 (13) 2 (13) 1 week: 0 (29) 1 (10) 2 (14) 1 week: 0 (0) 1 (12) 2 (34) 3 (6) 1 week: 0 (0) 1 (3) 2 (38) 3 (12) d ur ha m et a l. (2 00 9) (3 1) ii i 1 month: 0 (14) 1 (24) 2 (14) 1 month: 0 (14) 1 (21) 2 (18) 1 month: 0 (4) 1 (35) 2 (13) 3 (0) 1 month: 0 (4) 1 (29) 2 (20) 3 (0) ref reference, no number, gray shade box= test group, pi plaque index, gi gingival index, pd pocket depth, ccl clinical crown length, (mm) millimeter, dl diode laser, bl baseline, m month, p.s post-surgery, (n) number of patients, *statistical significant, # before. effects of electrosurgery only one study observed clinically and photographically the healing process [rob: low (ii)]. no significant difference was found between the groups after seven and fourteen days in periodontal measures due to the electrosurgical procedure (see table 8). table 8: comparison b/t electrosurgery & scalpel regarding periodontal parameters author (year) (ref. no.) study no. periodontal parameters clinical observation of healing photograpahic observation of healing aremband et al. (1973)(30) ii 1week(30%) 2 week (6.67%) 3 weeks (6.67%) 1 week (13.33%) 2 weeks (16.67%) 3 weeks (16.67%) discussion the present systematic review aimed to provide evidence for the superiority of using different techniques for gingivectomy over the conventional method using a scalpel regarding patient perceptions and periodontal parameters. the main reason for switching from the surgical scalpel to other methods is the need for hemostatic action during the surgical procedure for a better surgical field. three different methods were developed, and all provide excellent hemostatic action. however, these methods have disadvantages, including leakage of chemical materials such as paraformaldehyde and potassium hydroxide into the surrounding tissues, makings it not recommended for periodontal surgery. the electrosurgical method has also provided an excellent hemostatic action by sealing the blood vessels while cutting the desired tissue. however, care should be taken when choosing the waveform and power input because lateral heat could cause necrosis to the neighbouring tissues. the last method, laser with different active media for different types of tissues, also provides a hemostatic effect. however, all types of laser have a thermal effect on the surrounding tissues, which depends on the power input. diode laser & patient outcome based on the limited clinical evidence, the present systematic review found two alternative methods for gingivectomy, diode laser and electrosurgery. the results indicate that the laser technique causes less postoperative pain and discomfort than the traditional scalpel, especially in the first week. even though one study (akram et al., 2017) did not perform statistical analysis. however, the vas scores were low in the laser group compared to the scalpel group in the first two days, and because of that, the patients did not consume excessive painkillers like in the scalpel groups. our results are from a previous study by kalakonda b. et al., 2016, where they performed vestibulopathy in 20 patients and compared the scalpel group with the diode laser group. they presented a significantly lower vas pain score in the laser group than the scalpel group on the 1st, 3rd, and 7th days (kalakonda et al., 2016). also, in the https://journals.e-palli.com/home/index.php/ajcp pa ge 82 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 74-85, 2023 study of ize-iyamu i n. et al., 2013(ize-iyamu et al., 2013), they performed maxillary frenectomy, operculectomy, aesthetic recontouring, and surgical exposure, in addition to gingivectomy. again, they reported lower significant values in the laser group than in the scalpel group. furthermore, even though they used the diode laser in other oral soft tissue surgeries, they showed significantly lower vas of pain in the laser groups than in the scalpel groups (fisher se, 1983) (fisher & frame, 1984; koppolu et al., 2017). another theory is that a diode laser produces minor collateral tissue damage during the incision, even with a continuous wave at a higher power level of 4.5w than other lasers like co2, and less postoperative pain (durham, 2009) (goharkhay et al., 1999). also, the results herein showed that some of the included studies (koppolu et al., 2017) (elif, 2017) used only topical anaesthesia, either gel or spray, before the surgery, and most of the patients did not need infiltration of local anaesthesia for the laser procedure. this could be one of the advantages of using a laser for gingivectomy in children (do hoang viet et al., 2019). diode laser & periodontal parameters outcome another observation is that using a scalpel caused more gingival inflammation when compared with a laser; due to the periodontal dressing used in the scalpel group, which acts as plaque retention (akram et al., 2017). even though the pi was low in other studies (durham, 2009; lione et al., 2020; sobouti et al., 2014) (musaa et al., 2017), the inflammation was low in the test group. the reason for low gi is due to irradiation. instead, there is almost instantaneous vaporisation of the intracellular fluid and thus disintegrating cell structure, which might not release the chemical mediators of inflammation. in addition, a thin layer of denatured collagen formed on the surface will act as a protective layer and prevent tissue irritation, thus decreasing inflammation (fisher et al., 1983). only one study, (kumar et al., 2015) assessed the postoperative healing process using the healing index. better healing was observed in incisions done by the scalpel but was not statistically significant. this is the result of fisher and his colleagues in 1983 (fisher et al., 1983). they examined animal models for observing histological changes during healing after using a co2 laser on buccal mucosa and found no differences in the mitotic rate of the epithelium between the groups. in contrast, some studies showed that laser irradiation could stimulate fibroblast proliferation without impairing procollagen synthesis, thus improving wound healing(pereira et al., 2002). however, we had to know that these studies used low power output (120mw), which is much less than what was used in our included studies (0.8-2w), and those high power caused thermal tissue damage around the incision, according to goharkhay k. et al., 1999 (goharkhay et al., 1999). since the main advantage of laser over the scalpel is the hemostatic effect, which provides a clear surgical field during the surgery, three studies assessed the bleeding criteria during the procedure (elif, 2017) (sobouti et al., 2014) (musaa et al., 2017). the bleeding assessment showed that the diode laser controlled the bleeding. this result was confirmed by a study by ize-iyamu i. n. et al., 2013 (ize-iyamu et al., 2013). they performed various oral surgical procedures, such as a maxillary frenectomy, operculectomy, surgical exposure, and gingivectomy, using an 810nm diode laser compared with the conventional scalpel. they showed that the laser groups had grade 0 (no bleeding), and the scalpel groups had grade 1 (petechial bleeding) according to the who bleeding scale. a case report also confirmed our results (asnaashari et al., 2013), where they treated gingival hyperplasia using an 810nm diode laser and noticed minor bleeding during the procedure. electrosurgery & patient outcome on the other hand, the electrosurgical method did not differ from the scalpel regarding postoperative pain and discomfort. this result was confirmed by a comparative study by chandra r. v. et al., 2016. they performed a gingivectomy to reduce the pd and observed that the pain on a surgical day and after seven days did not differ between the control and test groups. a case series report (bhusari & kasat, 2011) concluded the same results. electrosurgery & periodontal parameters outcome in addition, armband d. et al.(aremband & wade, 1973) observed the healing process clinically, photographically, and histologically. no differences were presented between group (eisenmann d, 1970). this study follows glickman i. and imber l. r.’s study (glickman & imber, 1970), where they did a histological study on animals by performing a shallow resection (away from the bone) and deep resection (as close as possible to the crest of the bone). they presented that the microscopic findings of shallow resection samples after 3, 6, and 12 weeks were the same between the groups, the epithelium was inflamed with slight hyperplasia, and the underlying bone was unaltered. in contrast, the samples from the deep resection in the test group showed bone necrosis. in contrast, a study by manivannan n. et al.(manivannan et al., 2013) measured the vascularity of the gingiva preand postoperative by using the ultrasound doppler technique. the results showed that the re-vascularisation rate was faster in the control group than in the test group, thus improving the healing process, implying minor postoperative complications and more patient comfort. finally, a histological study by sinha u. k. et al., 2003(sinha & gallagher, 2003) confirmed the previous study. they compare the performance of the scalpel with electrosurgery by creating oral mucosal incisions in the guinea pigs. they showed that the fast healing utilising re-epithelialisation was in the scalpel groups, and complete epithelial healing had taken place by the end of the first week. in contrast, in the electrosurgery groups, the reepithelialisation had occurred by the end of the fourth week. another study done by chandra r. v. et al.(chandra et al., 2016) also showed a clinical delay in healing in the https://journals.e-palli.com/home/index.php/ajcp pa ge 83 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 74-85, 2023 electrosurgery group compared with the scalpel group after 7 and 15 days of follow-up. it was explained by the lateral heat produced from the electrode tip during the incision process. other types of laser & patient outcome many comparative studies used co2, nd: yag, and er: yag lasers compared with the scalpel for other oral soft tissue surgeries like frenectomy and excisional biopsy (haytac & ozcelik, 2006) (lópez-jornet & camachoalonso, 2013; natekar et al., 2017) (tambuwala et al., 2014) (akpınar et al., 2016; prasad r, 2018; yadav et al., 2019) (kara, 2008) (vickers, 2017) (broccoletti et al., 2015). these studies showed that the conventional technique exhibited higher postoperative pain and discomfort values than the laser technique. in addition, the studies of this systematic review also state that the laser method has an advantage over the scalpel in providing more comfort and producing less pain after the surgical procedure. even though a study incorporating electrosurgical methods, conducted on twenty patients, presented a high value of postoperative pain than the laser in gingival depigmentation surgery.(chandna & kedige, 2015) the vas score of pain was statistically significantly lower in the laser groups after 24 hours, but no significant differences were found after a week between the groups. this systematic review concluded that a diode laser had better patient outcomes in pain and discomfort than a scalpel. however, the current systematic review only included ten studies, and most studies had few patients in each group. hence, there was insufficient data to conclude the results between the diode laser and scalpel in terms of periodontal parameters and between the es unit and the scalpel for patient outcome and periodontal parameters. furthermore, one research comparing the es unit and scalpel for the gingivectomy technique was reported. based on the available data, a diode laser is the most recommended approach (technology) for crown lengthening. however, no meta-analysis was performed because the research designs and observational durations were heterogeneous. conclusion diode laser was the most appropriate technique for gingival recontouring and deep pockets elimination because the surgery could be performed by applying topical anaesthesia. also, infiltration did not require most of the time, and the patient will experience less pain and discomfort after the procedure compared to the scalpel technique. however, many studies showed the same results, but they performed different types of soft tissue surgeries like vestibulopathy and frenectomy and used different types of lasers such as co2, nd: yag. clinicians need to understand that each type of laser has different absorption coefficients with primary tissue components (water-mineral-melanin), making the laser selection procedure dependent. for example, the co2 laser is highly soluble in water, which means it is suitable for oral soft tissue surgeries, while the diode laser is highly absorbable in pigmented tissue and less absorbable in water than co2, so it is suitable for gingival surgeries since the gingiva has a high content of melanin. the clinician should first determine the specific requirement of the clinical treatment and then select the suitable laser to achieve the desired prognosis. author contributions: the author has a significant contribution to the conceptualization of the systematic review, conducting formal analysis, investigation, working efficiently on methodology, involved in project administration, writing – original drafts, writing as well as reviewing & editing the systematic review for the final look. references akpınar, a., toker, h., lektemur alpan, a., & çalışır, m. 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(2013). electrosurgical applications in dentistry. sch j app med sci, 1(5), 530-534. https://journals.e-palli.com/home/index.php/ajcp pa ge 1 pa ge 29 american journal of chemistry and pharmacy (ajcp) antidiabetic, hypolipidemic and hepatoprotective potential of edible leaves extract from the plant chenopodium albumchenopodium album (linn) in streptozotocin-induced diabetic mice tripti rani paul1*, monalisa monowar1, a. k. m. shafiur rahman1, md. shahriar kobir1, murshida mun liza1, most. sheuti akter1, tanbin islam1, ashik mosaddik2, mir imam ibne wahed3 volume 4 issue 1, year 2025 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v4i1.4598 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: february 15, 2025 accepted: march 24, 2025 published: june 24, 2025 diabetes is a long-term illness that affects a large number of people globally. it increases a patient’s risk of morbidity and death, especially from cardiovascular disease. the current study aims to assess the antidiabetic and hypolipidemic potentials of c. album leaves extract in streptozotocin-induced diabetic mice. acute toxicity tests and oral glucose tolerance tests were carried out. streptozotocin (45 mg/kg) was given intraperitoneally to swiss albino mice to cause diabetes. diabetic mice subjected to oral administration of c. album extracts (cal 200 and 400 mg/kg), metformin as standard (ds, 150mg/kg) and/or vehicle (dc) once daily for 15 days and age-matched healthy mice were used as normal control (nc). blood glucose level and body weight of mice were measured on 0 day before and 5, 10 and 15 days of treatment. to measure serum glutamate-pyruvate transaminase (sgpt), serum glutamate-oxaloacetate transaminase (sgot), low-density lipoproteins (ldl), high-density lipoproteins (hdl), total cholesterol (tc), and triglycerides (tg), mice were eventually killed, and blood samples were taken. the c. album extract improved glucose tolerance and no sign of toxicity was noticed in mice treated with the extract. diabetic mice treated with c. album extract showed significant attenuation in blood glucose level and lipid profile. moreover, oral treatment with c. album extracts significantly reduced sgpt and sgot levels; and improved body weights in mice. the c. album extract was considered to be both safe and beneficial in terms of glucose and lipids reducing effectiveness, and might be used to protect liver function in diabetic mice. keywords blood sugar level, diabetes, lipid profile, liver enzyme, streptozotocin 1 department of pharmacy, school of science and technology, varendra university, rajshahi, bangladesh 2 east west university, dhaka-1212, bangladesh 3 department of pharmacy, faculty of science, university of rajshahi, rajshahi, bangladesh * corresponding author’s e-mail: triptipaul.ph@gmail.com introduction chronically elevated blood sugar levels are an indicator of diabetes, is a metabolic disorder characterized by poor carbohydrate, protein, and lipid metabolism (ahmed, 2002). type 2 diabetes (t2dm) is linked to insulin resistance and causes hypertension, dyslipidemia, and glucose intolerance (patlak, 2002). about 80% people in countries with moderate to low incomes have diabetes mellitus (dm), a condition whose incidence has been steadily increasing worldwide (baynes, 2015). about 783 million people will have diabetes mellitus (dm) by 2045, with 152 million of those cases occurring in southeast asia, according to the international diabetes federation (idf, 2018). t2dm has recently been diagnosed in individuals under the age of 20, and the biggest risk factor for t2dm in both adults and children is obesity (guyton, 2006). according to anbarasi et al. (2012) and hahm et al. (2011), insulin resistance and decrease insulin synthesis and release from pancreatic β-cells are the primary reasons of type 2 diabetes. furthermore, it is thought that a combination of biological/genetic and environmental variables, such as obesity, a stressful lifestyle, alcohol use, smoking, and poor nutrition, might contribute to type 2 diabetes (t2dm) (ozougwu et al., 2013). elevated production of reactive oxygen species (ros) is frequently linked to hyperglycemia (brownlee, 2001), which can result in retinopathy, nephropathy, neuropathy, ketoacidosis, and other problems (nakamura et al., 2015; merecz et al., 2015). additionally, a lack of insulin causes lipolysis, which may be the cause of fatty liver and hyperlipidemia. morbidity and mortality in individuals with type 2 diabetes is primarily caused by hyperlipidemia (reiner et al., 2006, simons et al., 2002, yokozawa et al., 2003). insulin and synthetic medicines are currently the primary means of dm treatment. according to kasetti et al. (2010), thiazolidinedione, biguanide, and sulfonylureas are the most often prescribed oral hypoglycemic drugs for the management of type 2 diabetes. moreover, α-glucosidase inhibitors are useful in delaying the intestinal absorption of glucose. oral hypoglycemic medications are actually linked to negative side effects including obstructive jaundice, hypoglycemia shock, weight gain, gastrointestinal problems, nausea, vomiting, hematological, and dermatological reactions (gandhi et al., 2016, ukpds, 1998). over 800 plants have historically been shown to have antidiabetic potential (rizvi et al., 2013) and be useful in the cure of diabetes mellitus (arumugam et al., 2013). the world health organization (who) advised using local plants to treat diabetes and associated consequences, especially in underdeveloped nations. therefore, in order to find new bioactive compounds, scientists are mostly focused on screening natural products (bhandari et al., 2008). flavonoids, alkaloids, glycosides, and saponins are among pa ge 30 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 29-36, 2025 the many chemical constituents found in medicinal plants. these constituents have been shown to have antioxidant, hypoglycemic, and hypolipidemic characteristics (juárezreyes et al., 2015) and may offer protection against β-cell destruction brought on by oxidative stress (coskun et al., 2005; molina et al., 2003). the plant chenopodium album linn indigenously known as bathu sag (hindi), chandan bethu (bengali) belongs to chenopodiaceae family and universally grown in asia, europe, africa and north america. according to bonner-weir (1988), the plant c. album has been adopted in traditional medicine as a diuretic, laxative, analgesic, sedative, hepatoprotective, antidiabetic, cardiotonic, anthelmintic, and antiparasitic. alkaloids, saponins, glycosides, flavonoids, proteins, and amino acids were all tested by phytochemical analysis of the methanolic extract of c. album root (kant, 2018). the ethanolic extract from c. album fruits prevented mice from scratching when 5-ht was administered. it has been reported that the extract from c. album leaves has analgesic, anti-inflammatory, gastroprotective, hepatoprotective, antioxidant, antimicrobial activities. it also contains alkaloids, flavonoids, phenols, phytic acid, saponin, phytate phosphorus, proteins, and trace elements. (suleman et al., 2021). as far as the author has learned, no prior research has been done on the extract from c. album leaves’ hypoglycemic and antihyperlipidemic properties. thus, the objective of the current research was to assess the antidiabetic and hypolipidemic potentials of methanolic extract from c. album leaves in streptozotocin (stz) induced diabetic mice. materials and methods drugs and chemicals team pharmaceuticals ltd. rajshahi, bangladesh generously donated metformin hydrochloride. the supplier of streptozotocin was sisco research laboratories in india. a standard glucometer (origin, taiwan) was utilized to monitor the blood glucose level. a commercial kit from human, germany, was used to measure serum triglycerides (tg), total cholesterol (tc), high-density lipoprotein (hdl), low-density lipoprotein (ldl), serum glutamate oxaloacetate transaminase (sgot), and serum glutamate pyruvic transaminase (sgpt). plant materials in february 2022, the entire plant c. album was collected from the roadside of puthia, rajshahi, bangladesh. a taxonomist verified the plant’s authenticity, and a voucher specimen (no. 72, 10/02/2022) was stored in the herbarium, department of botany, university of rajshahi, bangladesh. preparation of leaves extract after being separated, the c. album leaves were sundried for a few days in the shade. the leaves were then processed into a coarse powder using a grinding mill after being dried for 24 hours at 40oc in an oven. for seven days, the ground-up leaves of c. album were soaked in w/v methanol and stored in a dark area with periodic shaking and stirring. the resulting filtrate was then run through whatman no. 1 filter paper and cotton. using a rotary evaporator set to 40-45oc, the filtrate was allowed to evaporate at lower pressure. the concentrated semisolid methanol extract (% yield) was so produced, allowed to air dry, and then stored. phytochemical screening tests the phytochemical analysis was done by the use of standard methods (pollock & stevens, 1965; trease & evans, 1996; plummer, 1985). animals the animal house of the department of zoology, rajshahi university in rajshahi, bangladesh, provided the six-week-old male swiss albino mice, whose weighed between 30 and 40 grams. the animals were kept in cages and under normal environments (temperature 25oc, humidity 75±5%, 12-hour cycle of light and darkness). the mice were given rodent chow with water ad. libitum during the acclimatization period. the mice were treated in according to our institution’s animal experimentation protocols. following approval from the varendra university institutional ethics committee, the animal study was conducted in the pharmacy department at varendra university in rajshahi, bangladesh (ref. vu/ erc/2021-2022/004). acute toxicity study the oecd recommendations were applied when carrying out the oral acute toxicity test (jonsson et al., 2013). the animals were split up into five groups, with three animals (n=5) in each group. following an overnight five, mice were given various doses of methanol extract from c. album leaves (100, 250, 500, 1000, and 2000 mg/ kg). the behavior and mortality of mice were monitored for the first two hours, the next 24 hours, and then every day for 14 days (schlede, 2002). oral glucose tolerance test (ogtt) normal mice were fasted overnight and separated into four groups randomly, each of which consisted of three mice (n=3) for oral glucose tolerance test. after 30 minutes of oral intake of c. album extract (200 & 400 mg/ kg), metformin (150 mg/kg), and/or vehicle (0.5% mc), a glucose load (2 g/kg) was administered to the mice. a glucometer was used to monitor the fasting blood glucose (fbg) level at 0 minutes, before and after 30, 60, 90, and 120 minutes of glucose loading (bergmeyer, 2012). induction of diabetes mice were given a single intraperitoneal injection of stz (45 mg/kg) dissolved in 0.1m citrate buffer (ph=4.5) to induce diabetes. in order to counter the hypoglycemic effect of stz, mice were given a 10% glucose solution for 24 hours. blood glucose levels were measured using a pa ge 31 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 29-36, 2025 glucometer after 96 hours, and mice with fasting glucose levels greater than 10.5 mmol/l were thought to be diabetic (kumar et al., 2013). experimental protocol after inducing diabetes, mice were separated into five groups each comprising of three mice and subjected to oral ingestion of c. album extract (cal 200 & 400 mg/ kg), standard (ds, metformin 150mg/kg) and/or vehicle (dc, 0.5% mc) once daily for 15 days using gastric tube. age-matched healthy mice received vehicle were used as normal control (nc, 0.5% mc). determination of blood glucose levels and changes in body weight fbg levels and body weight of mice were measured on day 0, before therapy, and then on days 5, 10, and 15 following treatments. blood samples were taken from mice’s tail veins and fbg levels were evaluated (reddy et al., 2012; khatune et al., 2016). estimation of lipid profile at the completion of the experiment, mice were sedated with diethyl ether and sacrificed. blood samples were collected from the aorta and stored in blood collection containers at room temperature. finally, blood samples were centrifuged for 15 minutes at 4000 rpm. the serum collected was isolated and stored at -80°c for biochemical analysis. the levels of tg, tc, and hdl were determined using commercial kits (human, germany) and the spectrophotometric technique. the formulas vldl=tg/5 and ldl=tc-(hdl+vldl) were used to compute the levels of ldl and vldl. the ldl/ hdl cholesterol ratio was determined (asati et al., 2021). determination of liver enzymes the sgot and sgpt level of serum were measured in accordance with the manufacturer’s instructions using wet reagent diagnostic kits (human, germany) (schumann et al., 2002, nakano et al., 1994). statistical analysis the data was expressed as a standard error mean (sem). following the one-way analysis of variance (anova), dunnett’s multiple comparison tests were done. p-values < 0.05 were considered statistically significant. results and discussion results acute toxicity after 14 days of oral ingestion of c album leaves extract, mice did not show any sign of autonomic or behavioral changes irrespective of doses. none of the mice died taking different doses of c. album extracts except 2000 mg/kg, where 20% mice died between 7 to 14 days. therefore, 1/5th and 1/10th of the toxic dose of c. album that is 400 and 200 mg/kg were considered for further study (table 1). table 1: effect of c. album leaves extract after 14 days of oral ingestion in normal mice extract doses (mg/kg) total survivor death survival rate (%) 100 5 5 0 100 250 5 5 0 100 500 5 5 0 100 1000 5 5 0 100 2000 5 4 1 80 data expressed in percentages (%) oral glucose tolerance test (ogtt) after 30 min of glucose loading, mice from all groups exhibited high blood glucose level which was decreased in mice pretreated with c. album extracts (200 and 400 mg/kg) and metformin but in vehicle-treated mice remained steady at 60 min. further, c. album extracts figure 1: effect of c. album extract on oral glucose tolerance test (ogtt). ***p < 0.001, **p < 0.01, *p < 0.05 vs. standard, ++p < 0.01 vs. glucose control pa ge 32 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 29-36, 2025 showed significant reduction in blood glucose levels at 90 and 120 min. however, cal400 was comparable to the standard and demonstrated a higher improvement in glucose tolerance (p <0.001). clinical course neither of the mice passed away during the period of treatment. as a result, all treatment groups survived. changes in fbg level in diabetic mice figure 2 showed the efficacy of cal extract on fbg levels in diabetic mice. the fbg levels were significantly increased in diabetic mice than that of nc mice. diabetic mice treated with cal200, cal400 extracts and ds demonstrated a gradual decrease in fbg levels on 5, 10 and 15 days. furthermore, cal extracts exhibited a dose-dependent attenuation of fbg levels; and their figure 2: effect of c. album leaves extract on fasting blood glucose level in diabetic mice. np < 0.001 vs. nc, ap < 0.001, bp < 0.01, cp < 0.05 vs. dc hypoglycemic effects were comparable to that of ds. body weight changes in diabetic mice changes in body weights after 15 days of treatment were shown in figure 3. on 0 day, before the initiation of the treatment body weight did not differ. the body weight of dc mice tends to be decreased throughout the treatment period. oral administration of cal200 and cal400 extracts exhibited a dose-dependent increment in body weights; and the effect was comparable to those of nc and dc mice. figure 3: effect of c. album leaves extract on body weight changes in diabetic mice. np < 0.001 vs. nc, ap < 0.001, bp < 0.01, cp < 0.05 vs. dc alteration of lipid profile in diabetic mice table 2 represented the result of extract on lipid profile in diabetic mice. the tc, tg and ldl levels were significantly increased and hdl level was significantly reduced in diabetic mice in contrast to nc. treatment with cal extracts significantly decreased the higher tc, tg and ldl levels and slightly enhanced the low hdl level in comparison with dc mice (p ˂ 0.001, p˂0.01, p˂0.05). the cal 400 expressed notable improvement in lipid profile which was comparable to ds. pa ge 33 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 29-36, 2025 liver function in diabetic mice the liver enzymes sgpt and sgot levels were greater in dc mice (p ˂ 0.001). oral ingestion of cal extract significantly reduced both sgpt and sgot levels (p ˂ 0.001, p˂0.01) and was comparable to nc. table 2: effect of c. album leaves extract on lipid profile in stz-induced diabetic mice groups (n=3) lipid profile (mg/dl) tc tg ldl vldl hdl ldl/hdl nc 174.33 ± 1.33 153 ± 1.73 111 ± 1.15 30.6±0.35 81± 1 1.37 ± 0.01 dc 220 ± 2.89+++ 192 ± 3.06+++ 133 ± 1.14+++ 38.4 ± 0.61+++ 43 ± 2.65+++ 3.12 ± 0.20 ds 179 ± 1.15*** 161.67 ± 1.20*** 107.66 ± 1.33*** 32.33 ± 0.24*** 51± 1.15*** 2.14 ± 0.08 cal 200 202 ± 1.53*** 160 ± 2.64*** 132 ± 1.73 30.6 ± 0.34*** 34 ± 1.15*** 3.89 ± 0.17 cal 400 190.66 ± 2.60*** 159 ± 0.58*** 127 ± 1.15** 31.8 ± 0.12*** 37 ± 1.14** 3.44 ± 0.01 data expressed as sem. +++p < 0.001 vs. nc; ***p < 0.001, **p < 0.01, *p < 0.05 vs. dc figure 4: effect of c. album leaves extract on sgot and sgpt in diabetic mice. np < 0.001 compared to nc, ap < 0.001, bp < 0.01 compared to dc discussion according to li et al. (2004) and lyra et al. (2006), diabetes is the third greatest cause of mortality, especially when it comes to organ failure and chronically high blood glucose levels. furthermore, hyperlipidemia increased the risk of cardiovascular disease, coronary artery disease, and peripheral vascular disease. around the world, currently, the primary treatment is metformin, an oral hypoglycemic drug for type 2 diabetes. furthermore, sulfonylureas or dipeptidylpeptidase-4 inhibitors are advised in conjunction with metformin for diabetic patients (gomes et al., 2019). there is an increasing interest in complementary and alternative therapies due to the increasing incidence of diabetes and related healthcare costs. many plant or plant-derived medications have been scientifically evaluated in diabetic people and animal models in over a decade, but many more need to be established. so, we investigated the glucose and lipid lowering effects of c. album leaves extract in stzinduced diabetic mice. the study found that cal extracts significantly improved fbg levels, lipid profiles, and liver enzymes in mice. in acute toxicity study, the methanol extracts of c. album leave in mice were found to be safe and no sign of autonomic and/or behavioral changes were observed in mice at dose ranges 100-2000 mg/kg. thus, it gives the basis for the selection of doses of extract i.e.200 mg/ kg & 400 mg/kg for further animal study. in, ogtt, mice treated with extract and/or metformin significantly counteracted the glucose induced hyperglycemia in normal mice. oral ingestion of cal200 and cal400 extract was effective in lowering fbg levels. when table 3: phytochemicals of c. album leaves extract extract steroid alkaloid glycoside tannin triterpene saponin flavonoid c album leaves + + + + + + + indicates present and − indicates absent pa ge 34 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 29-36, 2025 carbohydrates are insufficient to be used as an energy source, weight loss in diabetic mice may be the result of muscle wasting and loss of structural protein (jacobson, 2007). as previously reported (pari & venkateswaran, 2004; ruzaidi et al., 2005), the stz-induced diabetic rats developed significant hypertriglyceridemia in addition to substantial hyperglycemia. treatment significantly attenuate tc, tg, and ldl while increasing hdl cholesterol, which is necessary for the body to eliminate excess cholesterol and lower the risk of cardiovascular events. patients with diabetes are associated with the impairment of liver function as evident by the expression of higher levels of sgpt and sgot (ghosh et al., 2001). sgpt and sgot levels were abnormally high in stzinduced diabetic mice, which may have been brought on by hepatotoxicity. the cal200 and cal400 extract significantly reduced both sgot and sgpt levels. so, the cal extracts exerted prominent effects on body weight, lipid profile, glycemic index and liver functions in diabetic mice. the study found out that, cal400 extracts showed pronounced reduction in lipid profile and attenuation of fbg levels and the effects was comparable to metformin. it is currently unclear how the extract from c. album leaves lowers blood sugar, although it does not include stimulating the release of insulin from the pancreatic β-cells. in the animal model caused by stz, the pancreatic β-cells are selectively destroyed, and in moderate cases, part of the β-cells can still secrete insulin (el-hilaly and lyoussi, 2002). both metformin and/or the extract in this study significantly decreased the fbg levels in diabetic mice. the mechanism of increasing peripheral glucose utilization via insulin sensitization, which has been observed with metformin, can help to explain the hypoglycemic effects of the c. album extract (nandhini et al., 2004). the antihyperglycemic potential of the fractions of c. cordifolia were probably mediated by an enhanced secretion of insulin, like biguanides. the extracts of c. album exerted prominent effects on lipid profiles and they had a greater effect on serum tg than that of tc levels. although metformin exhibited pronounced reduction in serum tc and ldl, the effects of c. album leave extract on tg and hdl were found higher than metformin. by inhibiting hormone-sensitive lipogenic enzymes (pari and venkateswaran 2004) and/or activating lipoprotein lipase (ahmed et al., 2001, sharma et al., 1997), c. album may have hypolipidemic effects that are comparable to those of metformin. the pathophysiology of diabetes is significantly influenced by oxidative stress, which is the cause of the death of pancreatic 𝛽-cells. according to robertson (2010) and takayanagi et al. (2010), plants with antioxidant capacity demonstrated free radical scavenging activity and could be helpful in lowering oxidative stress caused by hyperglycemia. therefore, according to previous study, medicinal plants have the capacity to reduce blood glucose levels. this glucose lowering potential may be caused due to the existence of bioactive components such as flavonoids, alkaloids, triterpenes, tannins, and saponins etc. (robertson, 2010). the phytochemical screening of c. album leaves extract confirmed the existence of alkaloid, glycoside, tannin, triterpenes, saponin and flavonoids. the presence of flavonoids and triterpenes in the c. album extract may be the cause of its hypoglycemic potential, which can be explained by the antioxidant action of phytochemicals. in addition to phytic acids, the extract from c. album leaves provides a good source of lipids, phosphorus, protein, oxalates, and trace elements (suleman et al., 2021).). the leaves of c. album seem to have a promising therapeutic value which can be useful in the therapy of diabetes. conclusion diabetic mice treated with c. album extract showed significant attenuation in blood glucose level and lipid profile. moreover, oral treatment with c. album extracts significantly reduced sgpt and sgot levels; and improved body weights in mice. the c. album extract was considered as safe and effective in terms of glucose and lipid lowering efficacy; and might be used to protect liver function in diabetic mice. the findings provide scientific evidence in favor of utilizing the plant in conventional medicine to cure diabetes and its related problems. however, to determine how this plant has an antidiabetogenic impact and which bioactive components are responsible for it, more research is needed. references ahmed, a. m. 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(2003). the effects of coptidis rhizoma extract on a hypercholesterolemic animal model. phytomedicine, 10(1), 17-22. pa ge 1 pa ge 20 american journal of chemistry and pharmacy (ajcp) a study on synthesis and characterization of bio-plastic obtained from potato krishna gyawali1, prakash ghimire1, arjun bhandari1* volume 3 issue 1, year 2024 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v3i1.3617 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: august 05, 2024 accepted: september 12, 2024 published: november 21, 2024 the synthesis of bioplastic is a growing interest among researchers in today’s world. starch extracted from potatoes was utilized to synthesize bioplastic. from raw potatoes starch was extracted after it was peeled, diced, crushed, blended, slurred, sedimented, filtered and ovendried. the starch was made soluble in water with the help of acetic acid and was gelatinized by glycerol. constant heat was supplied with regular stirring until homogenous gelatinous mixture was ready. the mixture was molded and dried to obtain bioplastic film. physical and chemical properties were characterized. the film had average thickness of 3.45 ± 0.26 mm. the average density was 1.20 ± 0.04 g/cm3. the bioplastic revealed a biodegradability weight loss of 69.28 ± 1.52 % in 14 days. the water absorption of bioplastic was 81.08 ± 1.28 % in 2 hours. the opacity was measured 0.42 aumm-1. furthermore, phenol, concentrated sulphuric acid, and concentrated hydrochloric acid made the bioplastic soluble whereas insoluble in water and wide range of organic solvents. the film had smooth and slightly sticky surface◦ with slight vinegar smell. the study has shown that starch from potatoes can be utilized to create bioplastic film as replacement for petroleum-based bioplastic. keywords biodegradable, bioplastic, potatoes, plastic, starch 1 department of chemistry, butwal multiple campus, tribhuvan university, butwal, nepal * corresponding author’s e-mail: bhandariarjun112@gmail.com introduction the application of plastic is ubiquitous in today’s world. most of today’s plastics are fossils based, have slow degradability, and cause environmental harm (wallis, 2019). traditional petroleum-based plastics are resistant to microbial breakdown and build up in ecosystems and food systems (boey et al., 2021). additionally, the process of burning petroleum-based plastic trash to produce energy results in the emission of greenhouse gases and hazardous substances, specifically dioxin, furans, and the polychlorinated biphenyls (giacovelli, 2018). the release of greenhouse gases, including methane, nitric oxide, and carbon dioxide, contributes to the intensification of global warming (gironi & piemonte, 2011). additionally, it is worth noting that 99% of the plastic originated from non-renewable fossil fuel sources, specifically petroleum and natural gas (bioplastics, 2020). various biodegradable polymers, such as polylactide (pla), polyhydroxyalkanoate (pha), poly hydroxybutyrates (phb), starch, and cellulose are currently under investigation for diverse uses (venkatachalam & palaniswamy, 2020). bioplastics are plastic products that are made from renewable biomass feedstocks like starch, cellulose or polylactic acid and are therefore, being studied as a potential to reduce the problems associated with conventional plastics (thompson et al., 2005). bioplastics have gained significant interest in recent years due to their environmental benefits (rujnić-sokele & pilipović, 2017). bioplastics has the characteristic of being either biodegradable, bio based, or exhibiting both properties (tonuk, 2016). under the right conditions and over a certain amount of time, all materials are degradable (metro, 2021). we typically consider a material “biodegradable” if it degrades within a relatively short period of time, less than a year (metro, 2021). they have to change significantly in their structure which leads to the loss of properties (gilbert, 2015). biodegradation depends on the chemical structure of material and doesn’t rely on the source of polymer therefore, a bio-based plastic is not necessary a degradable plastic (kershaw & gilbert, 2015). the degradation of biodegradable plastics give rise to carbon dioxide, methane, water, biomass, humic matter and various other natural substance which can be readily eliminated (azois, 2007). these bioplastics have the advantage of not causing any environmental pollution, not depleting fossil fuel resources, and ultimately not posing any danger to humans (paul et al., 2021). biodegradable plastics are an important invention that can make a big contribution to the development of the bio economy and reduce our dependence on traditional fossil fuel-based resources in favor of bio-based products (nanda et al., 2022). biodegradable plastics are a suitable alternative to conventional plastics because they can be used in similar ways (cinar et al., 2020). starch, which is the most common carbohydrate in human diet, can be utilized to fabricate compostable plastic (garcia et al., 2015). it is also abundant, biodegradable, and renewable, and its possibility of blending with conventional polymers has garnered wide interest in the bioplastic market (prabhu & prasantha, 2016).starch can be obtained from various sources, including tubers like tapioca and potato, as well as cereals like rice, wheat and corn. additionally, cashew nuts can also serve as a source of starch (kharb & saharan, 2022). potato, corn, wheat, and tapioca are the primary botanical source of starch (ali et al., 2017). starch consists of two kinds of pa ge 21 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 20-26, 2024 glucose units connected by 1, 4-α connections. one type is amylose, which is a linear component, and the other type is amylopectin, which is a branching component (garcía et al., 2015). the fundamental starch structure is amorphous, consisting of amylose and an intercrystalline zone of dense cross-branched amylopectin and such morphology is responsible for the thermal, plasticization, and rheological properties of the starch (ochoa et al., 2016). in native starch, amylopectin chain length and chain ramification determine the granule crystallinity (ochoa et al., 2016). the native structure of starch is not suitable for industrial applications, because of its brittle nature and poor mechanical and rheological properties (sung et al., 2013). plasticizers are substrates with low molecular weight which when introduced into the starch matrices, can enhance the flexibility and processability of polymeric compounds by decreasing the hydrogen bonding of the starch-starch molecules (liu et al., 2004). plasticizers on the other hand can influence the physical properties of the processed starch by controlling its collapsing rate and depolymerization (ochoa et al., 2016). the amount of amylopectin and amylose has a major impact on the distinctive features of starch (prabhu & prashantha, 2018). the potato starch contains 80% amylopectin and 20% amylose (tarte & rodrigo, 2009). most commonly known starch-sugar based bioplastics are polylactic acid (pla) and poly-hydroxyalkanoates (pha) (barker & stafford, 2009; momani 2009). starch is widely utilized as a biopolymer due to its ample availability, ability to naturally break down, and cost-effectiveness (kharb & saharan, 2022). currently, starch and thermoplastic starches are utilized in diverse industries like medicines, packing food, horticultural and agricultural technology, textiles, paint, building, paper and cardboard industries, and automotive industries (liew & khor, 2015). starchbased biodegradable polymers have been widely utilized in the manufacturing of films, grocery bags, food storage containers (such as cups, plates, and trays), overwraps, and sanitary goods (kharb & saharan, 2022). starch-based biopolymers have been acquired as packaging materials for food goods (ferreira et al., 2016). starch and polymer mixes have been effectively employed in the medical field to create films for drug release, bone cement, and other applications (çalgeris et al., 2012). materials and methods the local potatoes (solanum tuberossum) were bought from a local market from butwal. the chemical used for this study are hydrochloric acid, vinegar, glycerol, sodium hydroxide, sulphuric acid, phenol, acetone, diethyl ether, nitrobenzene, chloroform, ethanol, picric avid, bromine water, aniline and benzene. starch extraction from potato 600 grams of potato bought from golpark, butwal was weighted, washed, peeled, diced, blended, slurred, filtered and dried in oven at 45°c for 2 hours to obtain 40-gram powdered starch. figure 1: steps of starch extraction from potato synthesis of bioplastic 15-gram dry potato starch (monomer molecule) added in 60 ml of distilled water (solvent) in a beaker. 20 ml acetic acid (react with amylopectin of starch), was added. then 15 ml glycerol (as plasticizer for elasticity) was added to mixture with constant stirring. the solution was kept undisturbed at 27°c for 25 minutes. the solution was heated over flame below 60°c with gentle stirring with glass rod, preventing the bubble formation. white insoluble starch starts to get dissolved and water evaporates to form thick gelatinous dense liquid. the hot thick transparent gelatinous dense liquid is spread over mold made from aluminum foil. it was dried in hot air oven at temperature of 120°c for 1 hour. lower the temperature to 45°c and store inside oven for 3 days. biodegradability test the level of biodegradability of bioplastic films is indicated by weight loss (hermaen et al., 2016). samples were dried at 45°c for 24 hours, it was weighted and buried in soil for 14 days. finally, the final weight of sample was recorded. the biodegradability was measured from the following formula (tan et al., 2016). weight loss (%)= (w0w/w) * 100figure 2: the synthesized bioplastic film pa ge 22 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 20-26, 2024 where; w0 = weight of sample before test w = weight of sample after test opacity measurement an uvvisible spectrophotometer with wavelength 600 nm is often the chosen choice to measure absorbance (balasubramanian et al., 2019; du et al., 2020; sirvio et al., 2020; tedeschi et al., 2020). uv-vis spectrophotometer at 600 nm was used to determine the opacity of bioplastic films (4 × 1 cm) (tunc & duman, 2010). the blank cell was used as reference opacity (priyadarshi et al., 2018). opacity= absorbance/thickness water absorption test the samples of varying weight were taken and put in a beaker with 100 ml water, kept there at 27 degrees centigrade for 1 hours and again weighted to record final weight. water uptake= (w2 w1/w1) * 100% where; w1 = initial weight of sample w2 = final weight of sample density measurement few pieces of bioplastic samples were weighted. water was poured into measuring cylinder and initial level of water was recorded. pieces of bioplastic samples tied in fine thread was gently lowered into measuring cylinder, until it is completely immersed into water. final volume of water in measuring cylinder was recorded. density was measured at room temperature i.e. 27◦c. volume of sample = water level after immersion of sample – water level before immersion of sample density= mass/volume thickness using screw gauge thickness was checked at different random points of bioplastic film. linear scale, circular scale and error of circular scale was determined and thickness of sample was calculated using formula: thickness of sample point= a + (b+c) * 0.001 where; a = linear scale of screw gauge (mm) b = circular scale of screw gauge (mm) c = error in circular scale (mm) average thickness= sum of mesured value/number of reading solubility test at 27°c small pieces of bioplastic was immersed into various organic and inorganic solvents taken in test-tube and slowly stirred with glass rod to check its solubility. results and discussions the bioplastic was insoluble in water which is green signal for its utility in real world as a plastic material. moreover, it was not able to solvate in wide variety of organic compound which also supports for its application in organic products. however, in extreme high concentrations it corroded and ultimately dissolved in sulphuric acid and hydrochloric acids indicating it is limited for use with mild and soft reagents. table 1: physical properties of bioplastic obtained through manual sensory evaluation strength medium flexible color slightly white odor vinegar smell surface smooth and slightly sticky table 2: solubility test of bioplastic samples in different organic and inorganic reagents s. n solvent solubility 1 diethyl ether 2 nitrobenzene 3 phenol + 4 acetaldehyde 5 benzene 6 chloroform 7 ethanol 8 acetone 9 picric acid 10 aniline 11 bromine water 12 conc. hcl + 13 conc. h2so4 + 14 naoh 15 water + = soluble = insoluble figure 3: demonstration of solubility test pa ge 23 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 20-26, 2024 in our research, the biodegradability weight loss percentage was found to be 69.28±1.52 % per 14 days which is 49.48% weight loss per 10 days which was in good agreement with nigam et al. (2021) who performed research on synthesis, characterization and biodegradation of bioplastic films produced from parthenium hysterophorus by incorporating a plasticizer (peg 600). our research diverges from those of, ismail et al. (2016) who reported higher degree of biodegradation which. this indicates population of microorganism and soil conditions can vary the rate of degradation of bioplastic. as the starch content consumed by soil microorganisms will fracture the polymer chain thus cause the biodegradation (khoramnejadian et al., 2013). the data revealed that the bioplastic have high water absorption property of 81.08 ± 1.28% per hour when completely immersed inside water which further justified that starch molecules are hydrophobic in nature. thus, starch-based bioplastics can be applied for the purpose of moisture absorption from variety of matters and materials. table 3: calculation of weight loss percentage per two weeks to find out degradability rate of synthesized bioplastic samples sample weight loss percentage a 68.47 b 68.53 c 68.23 d 71.91 average 69.28 standard deviation 1.52 total 69.28 ± 1.52 table 5: calculation table to measure density at 27°c sample mass (g) volume (ml) density a 0.37 0.3 1.23 b 0.25 0.2 1.25 c 0.48 0.4 1.2 d 0.46 0.4 1.15 average 1.2075 standard deviation 0.04 total 1.20 ± 0.04 figure 4: demonstration of biodegradability test table 4: calculation table to calculate water absorption capacity of synthesized bioplastic per hour at 27°c sample water uptake percentage a 79.16 b 81.81 c 82.6 d 80.77 average 81.08 standard deviation 1.28 total 81.08 ± 1.28 figure 5: demonstration of water absorption capacity test in our research the density of starch derived bioplastic was 1.20 ± 0.04 g/cc which has similar agreement with the (abdullah et al., 2019) where they have characterized poly lactic acid starch based bioplastic composites finding the density to be 1.3 g/cc. this indicate that bioplastic is denser than water and explains its property of sinking completely in water. figure 6: demonstration of density measurement pa ge 24 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 20-26, 2024 in our experiment the average thickness of bioplastic was measured 3.45 ± 0.2614 mm. conclusion this research aims not only to synthesize bioplastic from starch molecule but also to characterize its basic properties. the main finding of this study shows the bioplastic is more dense than water and it easily sink into it so it do not disturb water animals in their movement it’s products will not accumulate waste on water surface. similarly, it’s high degradability rate helps it to completely eliminate from environment. it can be beneficial in making coverings that absorb moisture and prevent damage of food and materials from wet atmospheric air. moreover, it’s insoluble property in wide variety of organic compounds including water, less opacity makes it appropriate to make food packaging material. it can be introduced as alternative solution of petroleum-based plastics. recommendation by continuous research and experiments, further efforts should be made to discover variety renewable organic molecules in order to produce better bioplastics which are widely applicable as well as economical in day-to-day life. author contributions krishna g. and arjun b. were conceptualization, methodology writing, original draft preparation, project administration. prakash g. was investigation, validation, review and editing. finally all authors were review and manuscript finalized. acknowledgement we would like to thanks all the members of department of chemistry, butwal multiple campus, for providing the necessary resources and research environment. i would like to acknowledge teachers, friends and family, for their constant support, discussions and encouragement, which have greatly helped me during this research journey. references abdullah, a. h. d., fikriyyah, a. k., putri, o. d., & asri, table 6: calculation of thickness of synthesized bioplastic in (mm) using screw gauge sample linear scale circular scale thickness a 3 73 3.78 b 3 39 3.44 c 3 9 3.14 d 2 79 2.84 average 3.45 standard deviation 0.26 total 3.45 ± 0.26 figure 7: demonstration of density measurement table 7: opacity measurement of bioplastic using uvvisible spectrophotometer at 600 nm wavelength sample absorbance at 600 nm (au) a 1.46 b 1.45 c 1.46 average 1.45 standard deviation 0.004 total 1.45 ± 0.004 opacity = absorbance/thickness =1.45/3.45 =0.42 aumm-1 the sample has low opacity of 0.42 aumm-1. a high value of opacity indicates that the specimen has low transparency (hermawan et al., 2019). in this research we observed bioplastic’s transparent property is dominant and it is in good agreement with the fact that our starch based bioplastic film can be used for food packaging. the ideal food packaging should be inexpensive, easy to process, with good barrier properties to prevent water loss and to protect food from o2 and moisture, mechanically resistant, transparent, fabricated with renewable sources, recyclable after use, and biodegradable (yadav & chiu, 2019). figure 8: measurement of absorbance using uv visible spectroscopy pa 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(2019). terrestrial fugitive plastic packaging: the blind spot in resolving plastic pollution. green materials, 8(1), 3-5. https://doi.org/10.1680/jgrma.19.00044 pa ge 1 pa ge 1 american journal of chemistry and pharmacy (ajcp) adverse drug reactions in nigeria’s geriatric population and its prevalence, etiologies, implications: a systematic review uyiose fortress ufuah1, aloysius obinna ikwuka2*, angus nnamdi oli3, brian ogbonna3 volume 3 issue 1, year 2024 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v3i1.3197 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: july 23, 2024 accepted: august 18, 2024 published: august 21, 2024 adverse drug reactions (adrs) are a significant public health concern. although the prevalence of adrs in the general adult population in nigeria has been studied and documented, there is limited research on adrs among the geriatric population. this study conducted a systematic review of existing research to determine the prevalence, etiologies and implications of adrs among nigerian geriatric population. a comprehensive search was undertaken using pubmed, researchgate, and google scholar databases. the preferred reporting items for systematic reviews and meta-analysis (prisma) guideline was used for reporting this systematic review. six studies met the eligibility criteria for inclusion in this review. the prevalence of adrs among the geriatrics ranged from 4.4% to 31.1%. the gastrointestinal system was the most commonly affected system, with non-steroidal anti-inflammatory drugs (nsaids) being the most frequently implicated class of drugs contributing to adrs. the prevalence of adrs in the nigerian geriatric population is underreported. there are insufficient policies and healthcare personnel specialized in caring for geriatric patients. strategies should be implemented to reduce the incidence of adrs among geriatric patients. keywords adverse drug reactions, adrs, nigeria, geriatric, population, prevalence, etiologies, implications, systematic review 1 faculty of pharmacy, university of ibadan, ibadan, nigeria 2 college of medicine and health sciences, american international university west africa, banjul, the gambia 3 faculty of pharmacy, nnamdi azikiwe university, awka, nigeria * corresponding author’s e-mail: momohshaibu3@gmail.com introduction the health and well-being of the elderly have become significant concerns in public health. globally, the population is aging rapidly, resulting in a decline in health as individuals grow older. in developing countries, the elderly population faces significant morbidity and disability burdens due to poverty, inadequate health facilities, and insufficient care policies. the geriatric population defined as individuals aged 65 and above, is considered a special population as they differ greatly from other age groups. they are more vulnerable to comorbidities, drug interactions, polypharmacy, and adverse drug reactions (adrs) (who, 2019). diabetes mellitus and arterial hypertension remain two of the most common diseases in the world. today, diabetes mellitus (dm) ranks third in the overall structure of morbidity and mortality after cardiovascular diseases and oncological diseases (virstyuk & ikwuka, 2021a). moreover, the nexus, pathogenetic mechanisms, roles, and effects of metabolic syndrome diseases on aging (and vice versa) are still being investigated by different researchers. metabolic disorders, e.g. hypertension, adiposity, diabetes mellitus and dyslipidemia, collectively known as metabolic syndrome diseases (msds) are diseases related to one another (ikwuka, 2015; ikwuka, 2017a; ikwuka, 2017c; ikwuka & virstyuk, 2023c; ikwuka et al., 2023f; virstyuk et al., 2016). t-helper 17 (th17) cells secrete interleukin-17 (il-17) which act on fibroblasts and endothelial cells to increase inflammation (ikwuka, 2023b). different studies have shown that msds are associated with asymptomatic hyperuricemia, systemic immune inflammatory processes, and fibrogenesis all of which can lead to nephropathy (ikwuka & haman, 2017d; ikwuka et al., 2017e; ikwuka, 2018c; ikwuka & paliy, 2018d; ikwuka, 2019a; ikwuka & virstyuk, 2019c; ikwuka & virstyuk, 2022; ikwuka et al., 2023d; virstyuk & ikwuka, 2017a; virstyuk & ikwuka, 2018a; virstyuk & ikwuka, 2019; virstyuk & ikwuka, 2021a; virstyuk et al., 2021b) and different rheumatological disorders in the elderly e.g. gouty arthritis, osteoarthritis, etc. thus, age is an independent risk factor for chronic noncommunicable diseases, which have high morbidity and mortality rates (jaspinder et al., 2014). aging is associated with increasing oxidative stress. linked to the induction of oxidative stress are major free radicals. among these major free radicals, superoxide anions, hydroxyl radicals, and hydroperoxyl radicals are of physiological significance. a non-radical of physiological significance is hydrogen peroxide (ama et al., 2023; baysah et al., 2023; ekechi et al., 2023a; uche et al., 2023). increased oxidative stress can lead to mutation which is an alteration in the dna sequence which produces new alleles. one common type of single gene mutation is the missense mutation which results in the substitution or replication of a single amino acid in the polypeptide chain e.g. sickle cell disease is caused by a missense mutation that produces a substitution of valine for glutamate in the two β-globin polypeptide chains (ikwuka, 2023a; musa et al., 2023). sickle cell disease is characterized with different severities of anemia (inya et al., 2023a; inya et al., 2023b). research also shows that disorders in individuals aged 60 and above, account for 23% of the global disease burden (prince et al., 2015). 60 years and above is also a risk factor for the development of different myeloand lymphoproliferative disorders such as myelodysplastic pa ge 2 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 1-11, 2024 syndrome, multiple myeloma, and non-hodgkin lymphoma (ikwuka, 2023e). therefore, there is a high demand for health services to meet the needs of this age group, requiring special attention to their treatment process (davies & o’mahony, 2015). in addition, there is also a need for new and effective treatment options in patients with metabolic syndrome diseases. sodium-glucose linked transporter 2 (sglt-2) inhibitors e.g. dapagliflozin and glucagonlike peptide 1 receptor agonists (glp-1 ras) e.g. liraglutide have been found to improve the efficacy of treatment and clinical course of type 2 diabetes mellitus and hypertension in patients with such comorbidities (ikwuka, 2017b; ikwuka, 2018a; ikwuka, 2018b; ikwuka, 2019b; ikwuka & virstyuk, 2021; ikwuka, 2024; virstyuk et al., 2017b; virstyuk et al., 2018b; virstyuk & ikwuka, 2018c). it has also been reported that coconut water has hepatorenal protective functions against alloxan-induced type 1 diabetes mellitus (ekechi et al., 2023b). adverse drug reaction (adr) is defined as “an appreciably harmful or unpleasant reaction resulting from the use of a medicinal product; adverse effects usually predict future hazards and require prevention, specific treatment, alteration of dosage regimen, or withdrawal of the product” (aronson & ferner, 2005). adrs are common in clinical practice, with some cases being severe and leading to unscheduled hospital admissions and even death (coleman & pontefract, 2016; alhawassi et al., 2014). adverse drug reactions in the geriatric population pose a significant problem in managing elderly patients. this is due to the decline in physiological functions (such as decreased renal and hepatic clearance), co-morbidity, pharmacodynamic and pharmacokinetic changes, frailty, polypharmacy, and memory problems commonly observed in this population. multiple drug therapies for co-morbidities increase the risk of drug interactions and inappropriate medication prescribing. studies have shown that prescription medication use among elderly patients has doubled in recent years (charlesworth et al., 2015). the prevalence of adrs in older adults has been documented in african countries with several studies highlighting the major drug classes responsible for adrs. however, global studies on the prevalence of adrs in the general adult population are more extensive than those focused on the elderly population, despite their higher susceptibility to adrs (alhawassi et al., 2014; saka et al., 2017; mekonnen et al., 2018; zazzara et al., 2021). systematic reviews conducted on the prevalence of adverse drug reactions in elderly patients in other parts of the globe reported adr prevalence of 11%-22% (alhawassi et al., 2014; jennings et al., 2020; yadesa et al., 2021). there is paucity of systematic reviews conducted on the elderly in africa. a systematic review on adrs in both children and adults in african hospitals reported that 8.4% of patients have experienced an adverse drug reaction at hospital admission while 2.8% of admissions were caused by adverse drug reactions (mekonnen et al., 2018). in nigeria, very few studies focused on the general adult population with none on the geriatric population that suffers adrs the most. therefore, this study aimed to systematically review the published literature to determine the prevalence, etiologies, and implications of adrs in the nigerian geriatric population. methodology information sources online databases and prospero were searched initially to ensure that the review topic had not been researched earlier. when that was established, the research question, selection strategy, and selection criteria were defined. the data collection form was designed in microsoft excel, the search strategies were run in the databases, and all references and abstracts were collected in a single file. search strings were constructed using boolean operators ands and ors. using subject headings obtained from journals and databases, reference lists for primary literature, grey literature and proceedings from conferences were obtained. question structure the population (geriatric/elderly) and clinical outcomes (adrs, drug therapy problems) were considered. prevalence studies and those focusing on the challenges of adrs were also considered. search strategy and selection process results from the searches revealed 100 studies. a total of 87 articles on adr in specific diseases, in other african countries, on polypharmacy, and potentially inappropriate medications (pims) were eliminated leaving 13 studies on adr in the elderly in nigeria. 4 duplicate studies and 3 incomplete texts were eliminated which gave rise to 6 studies that met the inclusion criteria (see figure 1). inclusion criteria and its reliability articles were selected for the review if they: (1) are research articles reporting primary data collected by the authors; (2) are published in english and full-text articles were available; (3) cover geriatrics in nigeria; and (4) studied populations are 60 years or older. cohen kappa statistics was used to discuss and resolve disagreements between the authors who reviewed the articles during selection (cohen, 1968). exclusion criteria all studies on geriatric adrs in nigeria with incomplete information on study methods or unclear methodology were excluded, including studies without comprehensive summaries. data extraction eligible studies and literature were assessed using two parallel reviewers who utilized the eligibility criteria consistently. all data on authors and year conducted, age range, setting/geographical zone, study design, pa ge 3 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 1-11, 2024 prevalence, medications implicated, system, and organ most implicated in adrs of the population were extracted to form the evidence-based table. data was summarized using microsoft excel. comparisons of extracted data were made and disagreements were resolved with sensitivity analysis, predominantly by further investigation and arbitration. data items (outcomes) data items on adrs among geriatrics covering 1980 to 2022 were searched. the geriatric population was searched. however, elderly and older adults were also searched in parallel and in series. the etiologies of adrs and their implications were also searched. publication risk of bias assessment grey literature and conference proceedings scanning were utilized, in addition to the careful use of search terms in parallel and in series to minimize the risk of bias. quality assessment/risk of bias the full texts were screened further for explicit of methodological descriptions of adrs, drug therapy problems, geriatrics and the elderly. studies were graded based on strong and weak recommendations of the study characteristics and quality to enhance critical appraisal. certainty assessment publication bias, imprecision, inconsistency, and indirectness of evidence in the assessment were utilized for certainty of the study. data collection literatures that reported the incidence of adrs among nigeria’s geriatric population were reviewed. the databases utilized were google scholar, pubmed, and researchgate with no specific gate window. the references of some articles were also manually searched to identify more studies. the keywords “geriatric” [mesh terms], “adverse drug reaction” [mesh terms], “incidence” [mesh terms], “prevalence” [mesh terms], “older adults” [mesh terms], and “elderly” [mesh terms] were used. the free text included adverse drug reactions in the elderly in nigeria and the incidence of adverse drug reactions in nigeria. the selection was not restricted to a specific study design. however, the study was restricted to geriatric/elderly population for which the study could be generalized. nonetheless, it is also possible that some studies may have been omitted due to this limitation and restricting the generalization. the review concentrated on the prevalence, incidence, and challenges of adverse drug reactions in the elderly in nigeria. other age groups, however, were not overlooked. the literature and research on various elderly disease states were reviewed. author, year of publication, age range, population size, study design, geopolitical zone, and most implicated medication were all extracted from the publications. each conflict was discussed in the cases of disagreements until agreement was reached. the resulting data was kept open and detailed. metaanalysis was not carried out because it is outside the scope of this study. hence, heterogeneity was not assessed. data synthesis all the results of the primary studies were collated and summarized. a combination of descriptive synthesis with quantitative summary was utilized. the population, causes of adr, comparisons, and outcomes were used as a framework. the study characteristics, causes of adrs, and comparisons of implications and outcomes were tabulated in a spread sheet. summary statistics of studies were also tabulated, while those without comprehensive summaries were excluded. descriptive statistics was used to summarize the qualitative and quantitative data based on their homoand heterogeneity. sub-group analysis was used to explore heterogeneity. to assess the robustness of the synthesized result, range of values and alternatives were substituted for arbitrary and unclear decisions to ensure that decisions taken were rational and consistent. data analysis a descriptive analysis was performed to summarize the published articles. the studies and their characteristics were summarized based on the following: authors and year conducted, age range, setting/geographical zone, study design, prevalence, medications implicated, system, and organ most implicated in adrs. the articles were evaluated for heterogeneities among the population group. finally, an evidence-based table was synthesized. ethical consideration published and freely available online literature and data on adrs among geriatrics in nigeria were reviewed online. hence, ethical approval does not apply. this study was carried out strictly with the prisma-2020 guideline for systematic reviews. results & discussion figure 1: the prisma flow diagram for selection of articles pa ge 4 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 1-11, 2024 out of the 100 reports initially identified, only thirteen reported adverse drug reactions in elderly patients in nigeria. articles that focused on adverse drug reactions in specific diseases, studies conducted in other african countries, studies on polypharmacy, potentially inappropriate medications (pims); and knowledge, attitude and practice (kap) studies were excluded. duplicate studies and studies without full texts available were also removed. finally, six studies were included in the analysis. study characteristics two of the six studies were conducted on the general adult population but provided specific information about older adults (adedapo et al., 2021; akhideno et al., 2018). all the studies were conducted among hospitalized patients, primarily in the internal medicine wards. the studies had a prospective, retrospective, or mixed design. the incidence of adrs among elderly patients ranged from 6.2% to 31.1%. the gastrointestinal system was the most affected, and non-steroidal anti-inflammatory drugs (nsaids) were the most commonly implicated group of drugs causing adrs. the majority of the studies were conducted in a single center (see table 1). a summary of the factors responsible for adrs in the included studies is shown in table 2. table 1: summary of studies on the incidence of adrs among the elderly in nigeria author (year conducted) age range population size setting/ geopolitical zone study design prevalence of adrs three most implicated medications system/ organ most implicated in adr a kh id en o et al. , 2 01 8 (2 01 320 14 ) 17 -8 9 (a du lts ) e ld er ly as > 64 50 7 (a du lts ) 1 25 (e ld er ly ) u ni ve rs ity o f be ni n te ac hi ng h os pi ta l, be ni n c ity , e do s ta te / so ut hso ut h pr os pe ct iv e st ud y 4. 4% in su lin , n sa id s a nd an tih yp er te ns iv es in a du lt po pu la tio n st ud ie d n eu ro lo gi ca l. g as tro in te st in al a nd de rm at ol og ic al sy st em s a m ad i e t a l., 2 02 1 60 a nd a bo ve 12 6 u ni ve rs ity o f po rt -h ar co ur t te ac hi ng h os pi ta l, po rt h ar co ur t, ri ve rs s ta te / so ut hso ut h c ro ss -s ec tio na l s am pl in g 19 .8 % c hl or oq ui ne , a m lo di pi ne in te gu m en ta ry /s ki n, c en tra l n er vo us s ys te m , g as tro in te st in al sy st em sa ka et a l., (2 01 7; 20 18 ) 60 a nd a bo ve 26 8 n ot s ta te d c ro ss -s ec tio na l re tro sp ec tiv e 13 .8 % n sa id s (d ic lo fe na c) , ca rd io va sc ul ar m ed ic at io ns d ig es tiv e sy st em , va sc ul ar sy st em , ne rv ou s s ys te m a de da po et a l., 20 21 (2 01 220 13 ) 60 a nd a bo ve 12 80 (t ot al a du lt po pu la tio n) 41 8 (e ld er ly po pu la tio n) u ni ve rs ity c ol le ge h os pi ta l, ib ad an / so ut hw es t pr os pe ct iv e co ho rt st ud y 4. 9% n sa id s, hy po gl yc em ic, an tib ac te ria l g as tro in te st in al , ce nt ra l n er vo us sy st em , a nd sk in / cu ta ne ou s s ys te m pa ge 5 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 1-11, 2024 n w an i & is ah , 2 01 7 (2 00 9) 65 a nd a bo ve 34 5 n na m di a zi ki w e u ni ve rs ity te ac hi ng h os pi ta l, n ne w i/ so ut he as t pr os pe ct iv e st ud y 6. 7% n sa id s, st er oi ds , a nd hy po gl yc em ic a ge nt s g as tro in te st in al sy st em , c en tra l ne rv ou s s ys te m , e nd oc rin e an d m et ab ol ic sy st em e ze et a l., 2 01 1 (2 00 8) 60 a nd a bo ve 37 5 o la bi si o na ba nj o u ni ve rs ity t ea ch in g h os pi ta l, sa ga m u, o gu n st at e/ so ut hw es t re tro sp ec tiv e an d pr os pe ct iv e 31 .1 % c ar di ov as cu la r m ed ic at io ns n ot sp ec ifi ed table 2: summary of the factors responsible for adrs in the included studies author (year conducted) factors responsible for adr akhideno et al., 2018 (2013-2014) number of medications number of co-morbidities amadi et al., 2021 polypharmacy multi-morbidity alcohol intake saka et al., (2018; 2017) abrupt stoppage of medications use of antiemetics decreased potassium levels below 2.5mmol/l adedapo et al., 2021 (2012-2013) outpatient medication errors overdose self-medication and drug misuse multi-morbidity duration of hospital stay polypharmacy (≥3 medications) nwani & isah, 2017 (2009) not reported eze et al., 2011 (2008) polypharmacy (≥5 medications) in one study on the general adult population (aged 17 years and older), elderly patients (65 and older) accounted for 24.7% of the study participants, and adverse drug reactions were observed in 22% of these elderly patients. although the specific drug most implicated in elderly patients was not specified, the drugs most frequently associated with adverse drug reactions in the general adult population were insulin (27.5%), nsaids (19.6%), antihypertensives (15.7%), and antimalarials. age was found to be a significant risk factor for adverse reactions to medications among the elderly, with those aged 65 years and older being at a higher risk than younger patients (akhideno et al., 2018). a prospective cohort study was conducted, involving a total of 1,280 adult patients who were admitted to the hospital. it was found that patients aged 60 and older constituted approximately one-third (32.7%) of the study participants. among patients aged 65 years and older, 4.9% experienced adverse drug reactions that resulted in hospitalization, while 1.3% developed adverse drug reactions during their admission. the study did not specify the most affected organ or system or the specific classes of drugs implicated in adverse drug reactions in the older population, but in the general adult population, the most implicated drug classes were nsaids, antidiabetics, and antibacterials. the gastrointestinal tract, central nervous pa ge 6 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 1-11, 2024 system, and skin were the organs and systems most commonly affected (akhideno et al., 2018). in a retrospective cross-sectional study of elderly patients, out of 268 admitted patients, 43 cases of adverse drug reactions (13.8%) were reported, with the digestive system being the most affected by these reactions. only 9.3% of the total observed cases were formally documented as adverse drug reactions. the most implicated medication in these reactions was diclofenac (saka et al., 2018). another study found that 19.8% of the elderly population experienced adverse drug reactions. the most common adverse reactions included itching (22%), drowsiness, dizziness, and restlessness. the drug most implicated in these reactions was chloroquine, and hypertension was a major medical ailment. it was observed that 96% of the patients had multiple morbidities, and 96% of those with adverse drug reactions were taking more than five drugs, compared to 86.14% of those without adverse drug reactions (nwani & isah, 2017). another prospective study evaluated 345 patients aged 65 years and older. out of these patients, 23 (6.7%) experienced adverse drug reactions, with 73.9% of these cases recovering and 26.1% resulting in death. the most common adverse drug reaction observed was gastrointestinal bleeding (39.1%). the most implicated drug classes were nsaids, steroids, and hypoglycemic agents. the gastrointestinal system, central nervous system, and endocrine and metabolic systems were the organ systems most frequently involved. adverse drug reactions were responsible for 1.7% of deaths among elderly patients who were admitted to the hospital (nwani & isah, 2017). a study conducted by (eze et al., 2011) focused on identifying drug therapy problems (dtps) among elderly inpatients. the study utilized both retrospective and prospective methods. it reported that 31.1% of the total dtps recorded was adverse drug reactions, but it did not provide further details on the drug classes responsible. however, cardiovascular medications were the therapeutic group most commonly associated with dtps. this present study evaluated the prevalence of adverse drug reactions among the elderly in nigeria, and revealed a wide variation in the prevalence of adrs. this variability may be due to factors such as differences in the methods used to identify an adr, the sample size and study design. in this review, some of the studies relied on the identification of adrs during routine hospital care or on self-reporting methods. however, this is prone to under-identification and under-reporting of adrs, and such methods are likely to underestimate the true prevalence of adrs (kongkaew et al., 2008; brvar et al., 2009; ramia et al., 2021). these results are similar to the results of a systematic review of adr in the elderly (alhawassi et al., 2014). nsaids are one of the most commonly used class of drugs for self-medication (oriavwote & ikwuka, 2022). nsaids are implicated in most of the adrs and are the major causes of peptic ulcer diseases and worsening hypertensive conditions (drini, 2017). similar to (kongkaew et al., 2008; oscanoa et al., 2017; hadia et al., 2022), nsaids and cardiovascular medications caused most adrs in the elderly. however, (jennings et al., 2020) reported diuretics and antibacterials for systemic use as the most implicated medications. polypharmacy has also been shown by other systematic reviews conducted on the elderly as the major risk factor for adrs in the elderly population (davies et al., 2020; hadia et al., 2022). the incidence and prevalence of adrs is under-reported in nigeria, particularly among the geriatric population (ogar et al., 2019). according to a study conducted in nigeria, only 32% of health professionals report adverse drug reactions (okezie & olufunmilayo, 2008). several reasons have been identified for the under-reporting of adrs in nigeria, including staff shortages, time required to assess and submit reports, lack of information, difficulties in diagnosing or identifying adverse reactions, ignorance of the importance of reporting, limited or no training for healthcare workers on adr observation and reporting, lack of awareness of the reporting form, poor knowledge of the reporting procedure, and uncertainty of causality (aderemi-williams et al., 2015; miediegha & bunu, 2020; okezie & olufunmilayo, 2008; oreagba et al., 2015). the poor knowledge of healthcare professionals regarding adrs is a cause for concern. in a study by (adisa & omitogun, 2019), only 15% of healthcare workers had adequate knowledge of adrs. in another study, only 24.9% of respondents were aware of the national adr reporting form (commonly known as the yellow form), and just 20.1% had received training in pharmacovigilance (udoye et al., 2018). another challenge is developing policies and appointing staff who can understand and respond to the needs of the aging population. there are limited centers dedicated to the care of geriatric patients in nigeria, resulting in inadequate attention to their specific needs before initiating treatment. often, they are treated alongside adult patients under the age of 60. the prevalence of polypharmacy and the prescription of potentially inappropriate medications (pims) pose significant problems in the pharmacotherapy of older adults, leading to a significant increase in adrs among the elderly. studies conducted on the elderly in nigeria have reported high rates of polypharmacy and pims, resulting in a high incidence of adverse drug reactions (borodo et al., 2022). the negative impact of adverse drug reactions on health outcomes in the geriatric population cannot be overstated. in developing countries like nigeria, these effects are more pronounced due to high health outcome indicators. older adults are more susceptible to adrs compared to younger adults due to the increased risk associated with aging. it has been suggested that adrs should be treated as a geriatric syndrome in themselves. the potential consequences of adverse drug reactions (adrs) are multifaceted and include longer hospital stays, which in pa ge 7 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 3(1) 1-11, 2024 turn increase morbidity and mortality rates, as well as healthcare costs (fasipe et al., 2019). strategies must be put in place to reduce the incidence of adrs (adverse drug reactions) among the elderly in nigeria. one of the first steps in reducing the incidence of adrs in nigeria would be the proper reporting of cases. this would help to identify the particular classes of drugs responsible for adverse drug reactions in geriatrics, and strategies can be put in place to reduce their use or research safer pharmacological alternatives. a study in the united states among adults aged ≥65 years observed that three drug groups medicines that help prevent blood clots, antidiabetic medications, and medications with morphine-like effects were found to be involved in about 60% of emergency department visits for adverse drug reactions (shehab et al., 2016). the reporting of adverse drug reactions in africa has been strengthened through the establishment of “national pharmacovigilance centers (npcs)” in several countries (ogar et al., 2019). the npc in nigeria has made available procedures for the proper reporting of adrs (npc, 2014). furthermore, activities and programs such as public enlightenment and education of patients and caregivers on adrs, as well as spontaneous reporting of adrs by health professionals, should be implemented to increase the reporting rate. the poor knowledge of adrs among healthcare workers highlights the need for continuous mandatory education and training of health workers on adverse drug reactions and pharmacovigilance concepts. the curriculum of health professionals during undergraduate training should also emphasize the identification, prevention, and documentation of adverse drug reactions. to tackle the problem of limited centers dedicated to the care of geriatric patients, the leadership of various hospitals and clinics should advocate for the establishment of geriatric centers in their facilities equipped with trained staff across all states in nigeria. reducing pill burden, referred to as deprescribing, is one of the most effective steps to reducing adverse drug reactions among the elderly. deprescribing refers to the process of withholding unnecessary medicines or minimizing their dosages at the discretion of a healthcare professional, probably a medical doctor or a pharmacist. this is aimed at managing polypharmacy and in turn, improving health outcomes. there should be an increase in awareness among nigerian prescribers regarding rational prescribing (oli et al., 2021) and the use of checklists (or trigger tools) for adr detection among the elderly (borodo et al., 2022). reports on the incidence of adverse drug reactions in the geriatric population in nigeria are very limited. thus far, only two reports are documented solely on adrs in geriatrics in nigeria. most studies are focused on adrs in the general adult population and potentially inappropriate prescribing. however, high quality reports investigating the drug classes most responsible for adrs in the elderly are lacking. furthermore, interventions to reduce adrs and improve healthcare among geriatric patients in nigeria are limited. this present study is not without limitations as it utilized heterogeneous studies (prospective, retrospective, and mixed) in the review and this poses a bias. although retrospective studies have been shown to detect lower adrs compared to prospective studies because they are subject to failure in gathering and proper documentation of data in the medical charts (de figueiredo et al., 2017), however, this was not so in this present study as those that used a prospective study design had lower adr prevalence rates compared to those with a retrospective design. another limitation of this present study is the variation in the age brackets of the elderly patients included in the review. there is a need for further studies on the detection of adrs and trigger tools/medications that cause a significant burden of morbidity and mortality in nigeria’s geriatric population. conclusion the geriatrics are highly susceptible to adverse drug reactions due to the decline in cellular functions, comorbid conditions, and polypharmacy. these factors contribute to increased risk of hospitalizations and deaths resulting from medication-related adverse events. despite the high risks involved, approaches to addressing adrs have not focused solely on the geriatric population. interventional studies on safer non-pharmacological and pharmacological alternatives to drug classes associated with adrs are necessary in order to 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(2021). adverse drug reactions in older adults: a narrative review of the literature. european geriatric medicine, 12, 463-473. https://doi. org/10.1007/s41999-021-00481-9. pa ge 1 pa ge 37 american journal of chemistry and pharmacy (ajcp) a summarized review of refractory materials and their chemical components s. kamara1* volume 4 issue 1, year 2025 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v4i1.5789 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: july 20, 2025 accepted: august 25, 2025 published: september 29, 2025 refractories are non-metallic ceramic materials capable of withstanding temperatures higher than 1500 °c, where the refractoriness is defined by the pyrometric cone equivalent. silica fume and fly ash are used in industries as cost-effective raw materials to fabricate refractory composites. mullite ceramic (3al2o3·2sio2) is a widely used engineering ceramic material with high strength and high creep resistance at high temperatures, low thermal expansion coefficient, and thermal stability. cordierite ceramics (mg2al4si5o18) can be used as the carrier of environmental catalysts because of their low coefficient of thermal expansion and stable structure. therefore, it is an important way to reduce the generation of waste pollutants (silica fume, fly ash, etc.), reflecting the basic idea of the transformation of solid waste into valuable products. the components of refractory materials are discussed alongside their utilizations, properties, and characteristics. keywords cordierite, fly ash, kaolin, mullite, refractory, silica fume, sintering 1 department of chemical engineering, school of water and environment, chang’an university, xi’an 710054, china * corresponding author’s e-mail: ksaidu2013@gmail.com introduction the development of metals, polymers, and ceramic materials has significantly contributed to the progress of human civilization. the emergence of metals and ceramics is historically interconnected. the high temperatures involved in metallurgical processes lead to the development of suitable refractories of molten metal containers. these refractory materials are nonmetallic with suitable physical and chemical properties used for components of systems that are exposed to high temperatures of above 538 °c (chandra & sarkar, 2019; dana et al., 2014; nwannenna et al., 2015). the properties of these refractory materials remain unaltered at elevated temperatures of sustained operations. refractory ceramic materials are classified into chemical nature (acid, basic, neutral), implementation method (shaped and unshaped), manufacturing method (sintered or fused), and porosity of the material (porous and dense) depending on the physicochemical nature of the starting materials (silica, aluminosilicate, magnesia, etc.) utilized in the manufacturing process (biswas et al., 2020; sarkar, 2023). these materials are resistant to heat when exposed to different degrees of mechanical stress and strain, corrosion from liquids and gases, and mechanical abrasion at high temperatures. the scarcity of basic refractory materials imported from europe during world war ii brought about the development of aluminosilicate refractories associated closely with the development of bauxite-based refractories in north america (garberscraig, 2008; sengupta, 2020). the advancement of aluminosilicate refractory materials from bauxite began in north america when there was a shortage of refractory materials in europe during the second world war. the overall consumption and utilization of refractories are a result of the sustained development of starting materials and processing technology in the refractory industry. in recent times, aluminosilicate refractories are mainly produced in developing countries and imported to developed nations. the knowledge obtained from the results of experimental research in materials science is essential for use in the fields of engineering and technology in modern days. the anticipation of industrial engineers to develop materials that have the advantage to resist high operating conditions began with the commencement of ceramics, polymers, refractories, etc. the fabrication of these latest materials has the purpose to produce good and reliable products with easy processing and low-cost operation (bramsiepe et al., 2012; jandyal et al., 2022; zhong, 2021). refractory ceramics are suitable for use in furnace linings or melting pots due to their ability to withstand physical and chemical conditions at high temperatures in metal casting industries and other applications with critical thermochemical properties. the properties of these materials are contingent on the properties of the starting reagents and formulations (chemical and mineralogical composition, particle size, and shape distribution), forming process, and sintering temperature (colomban, 2020; danninger et al., 2017; kamara et al., 2025; mohd mortar et al., 2022; sun et al., 2024; ulusoy, 2023). the fabrication of different types of refractory ceramic materials is dependent upon the starting materials utilized and the stoichiometric processing parameters (kamara, 2025; kamara et al., 2020; nanda et al., 2023; shackelford & doremus, 2008). studies on silica and aluminacontaining compounds show that mullite (3al2o3.2sio2) mineral phase is one of the most outstanding refractory ceramics (hossain & roy, 2020; vakalova et al., 2004) due to a combination of such distinct properties as elevated melting point (1800°c), low thermal expansion coefficient, pa ge 38 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 37-47, 2025 excellent mechanical and thermal shock resistance, chemical stability, etc., making it useful in many applications from refractories to electronic substrates (kamara et al., 2025; kamara et al., 2020). the significant amounts of silica (sio2) and alumina oxides (al2o3) in clay minerals make clay material a suitable low-cost natural raw material to fabricate mullite refractory ceramics. the sio2/al2o3 ratio in the clay material is low and therefore requires the addition of suitable stoichiometric compositions to produce the major mullite phase (zhang et al., 2022). in recent decades, concerns about environmental issues have grown around the world, particularly in the field of education and researchers have now given much attention to sustainable methods aimed at recycling inorganic waste (debrah et al., 2021; townsend, 2011). they are coarse-grain ceramics having a microstructure that consists of large grains joined by fine materials (lan et al., 2016). the major starting materials used to fabricate refractories are the oxides of silicon, aluminum, etc. nitrides, borides, silicates, graphite, etc, are some nonoxide refractories (guanghui, 2024). as stated earlier, the choice of refractories by engineers is solely based on the operational conditions to which they are being exposed. for instance, at extreme temperatures zirconia can be utilized to withstand such conditions. also, silicon carbide and carbon are employed for such applications but they should be used in the absence of oxygen because of their oxidative properties. refractories are a special form of ceramic materials which mainly differs from any normal ceramic because their coarse grain structure is joined by larger grog particles made of fine intermediate materials (bonding). the qualities of refractories depend on their physical, chemical, mineralogical, and thermal properties (sengupta, 2020; yurkov, 2015). refractories are normally made based on process parameters (temperature profile, mode of operation, and operating atmosphere), expected quality characteristics, and best techniques for engineering application so that the final physical, chemical, and thermal properties are compatible with the application (chandra & sarkar, 2019). refractories are expensive, and any failure in the refractories results in a great loss of production time, equipment, and sometimes the product itself (chandra & sarkar, 2019; horckmans et al., 2019; yurkov, 2015). the energy consumed and the quality of the product is also determined by the type of refractory involved. the main objective of this work is to review the components of refractory material that provide the basis for its suitability in the production of engineering materials. it is therefore very significant to produce suitable refractory materials for a particular application. the suitability of a refractory material does not imply durability but rather the balance between the initial cost and its performance. this balance is never achieved but is continuously moving as a result of the introduction of new processes or new types of refractories. literature review refractories are non-metallic ceramic materials capable of withstanding temperatures higher than 1500 ◦c, where the refractoriness is defined by the pyrometric cone equivalent. most modern refractories are based on one or more non-metallic oxides, which can be divided into basic and non-basic refractories. all basic products are based on either calcium oxide (cao) or mgo, whereas non-basic materials are based on al2o3, silicon dioxide (sio2), or zirconium oxide (zro2). this is a fundamental differentiation, as the correct refractory material is always based on the basicity of the medium in contact with the lining. refractory materials are both economically and socially strategic materials as they enable the production of other crucial products, including steel, non-ferrous metals, cement clinker, lime, glass, and many others. they are designed to operate at high temperatures, temperature gradients, and under severe chemical and mechanical loadings. the main raw materials used in the production of refractories are: the oxides of silicon, aluminum, magnesium, calcium, and zirconium, and some nonoxide refractories like carbides, nitrides, borides, silicates and graphite. the main types include fire-clay bricks, castables, ceramic fiber, and insulating bricks that are made in varying combinations and shapes for diverse applications. the value of refractories is not judged by the cost of the material itself, but by the nature of the job and/or its performance in a particular situation. atmosphere, temperature, and the materials in contact are some of the operating factors that determine the composition of refractory materials. refractories are utilized by the metallurgy industry in the internal linings of furnaces, kilns, reactors, and other vessels for holding and transporting metal and slag. in non-metallurgical industries, the refractories are mostly installed on fired heaters, hydrogen reformers, ammonia primary and secondary reformers, cracking furnaces, incinerators, utility boilers, catalytic cracking units, coke calciner, sulfur furnaces, air heaters, ducting, stacks, etc. the majority of these listed equipment operates under high pressure, and operating temperature can vary from very low to very high (approximately 900°f to 2900°f). the refractory materials are therefore needed to withstand temperatures and above these temperatures. important properties of refractories are: chemical composition, bulk density, apparent porosity, apparent specific gravity, and strength at atmospheric temperatures. these properties are often among those which are used as ‘control points’ in the manufacturing and quality control process. the chemical composition serves as a basic for classification of refractories and the density, porosity and strength is influenced by many other factors. among these are type and quality of the raw materials, the size and fit of the particles, moisture content at the time of pressing, pressure at mould, temperature, duration of firing and the rate of cooling. some of the important characteristics of refractories include melting point, size and dimensional stability, porosity, bulk density, cold crushing strength, pyrometric pa ge 39 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 37-47, 2025 cone equivalent (pce), refractoriness under load, creep at high temperature, volume expansion stability and shrinkage at high temperature, reversible thermal expansion, thermal conductivity, etc. refractory materials are characterized as ceramic materials, exposed during their use to high temperatures, and often to other extreme chemical and physical exposures that limit their operation life. the intensive use of refractories in steelmaking plants is due to the properties these compounds exhibit such as high starting melting point, high structural strength at high temperatures and in highly corrosive environments, and an intensive stability in temperature variations. what is common for refractory materials in terms of not only various applications but also their material and structural nature is described by a set of properties of refractory materials. previous use of refractories, particularly aluminum oxide (al2o3) and magnesium oxide (mgo), has exposed challenges in terms of corrosion mainly due to the formation of cobalt aluminates (coal2o4) using al2o3 as a refractory material and lithium diffusion within the crucible walls using mgo. corrosion, generally defined as the chemical attack on refractory material by a liquid phase such as slag, metal or alloy, occurs when the melt infiltrates the refractory matrix, creating capillaries that weaken the material’s mechanical strength. during this process, the fine grain of the refractory is washed out leaving the coarse grain exposed. this process occurs due to the chemical reaction between the liquid slag and solid refractory phases. it continues until the saturation point of the slag or the metal bath occurs through the refractory material. such issues can lead to contamination of the input material and significantly impact refractory stability and reusability. furthermore, diffusion-related constraints can hinder lithium removal rates, thereby diminishing the overall process efficiency. materials and methods several articles related to refractory technology were downloaded from various online publishing sources, of which 53 topically related papers were selected and systematically reviewed. the work was summarized based on the research objectives, which focuses on the components of refractory materials. classification of refractory refractories they are categorized based on their chemical composition and chemical properties. chemical composition silica refractories are fabricated in the form of quartz and it is composed of 92% of sio2. the iron and steel industries extensively utilize sio2 refractories due to their outstanding resistance to thermal shock. the raw materials for this class of refractories are high-grade rocks and fly ash. fireclay refractories are formed by heating a particular type of clay and it comprises less than 78% of sio2 and 44% of al2o3. they are cost-effective and popular industrial materials. alumina refractories are also made of al2o3 and sio2 with a minimum of 50% al2o3. they are subdivided into seven different percentages such as 50% al2o3, 60% al2o3, 70%al2o3, 80%al2o3, 85%al2o3, 90% al2o3, and 99% al2o3. magnesia refractories have magnesium oxide (mgo) and their principal sources are brines, seawater, and sintered and fused magnesia with magnesia-carbon refractory identified as the most important. dolomite refractories comprise mgo and cao as their main components. these refractories are fabricated from sintered dolomite. magnesia-chrome refractories mainly consist of mgo and chromium oxide (cr2o3). they are named magnesia chrome or chromemagnesite depending on the percentages of mgo and cr2o3 present in the refractory. about 15% to 35% cr2o3 and 42% to 50% of mgo are contained in magnesite chrome while magnesite-chromite has about 60% mgo and 8% to 18% of cr2o3. they are utilized to construct critical parts of high-temperature furnaces due to their abilities to withstand corrosive slags and gases and their high refractoriness. magnesite-chrome has better-spalling resistance than chrome-magnesite refractories in some cases. silicon carbide refractories are manufactured from silicon carbide (sic) obtained by reacting silica and carbon in an electric furnace at a high temperature of over 25000 °c. zirconia refractories have zirconium oxide (zro2) as their principal component and are utilized as high-temperature construction materials. nozzles, gates, crucibles, furnace liners, and kilns, etc., are cast at high temperatures greater than 1900°c from these class of refractories. it is a suitable insulating material due to its lower thermal conductivity at high temperatures, and they do not react with liquid metals. carbon refractories have carbon as the principal component, existing in the form of graphite or semi-graphite. they can resist the action of slags and have high thermal stability. chemical reactivity there are several typical chemical reactivities of refractories based on the chemical reactivity of their constituent substances. acidic refractories contain silica and alumina, which tend to react with basic slags. they are resistant to acid slags and are therefore utilized in an acidic environment and slags. basic refractories constitute calcium oxide, magnesia refractories, dolomite refractories, magnesia-chrome refractories, etc. they react with acidic slags but are resistant to alkaline (basic) slags, specks of dust, and fumes at extreme temperatures. steel manufacturers make use of this class of refractory to construct furnace linings in an alkaline medium. neutral refractories are chemically stable in both acidic and basic media and can be utilized in acidic or basic environments and slags. carbon graphite (most inert), chromite refractories, and alumina are typical examples of these refractories, among which graphite is the least reactive and is suitably utilized in metallurgical furnaces where oxidation processes are controlled. pa ge 40 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 37-47, 2025 special refractories include pure alumina, sialons (si-alo-n), beryllia (beo), zirconia, boron nitride, spinel, etc. they are costly refractory materials that are fabricated using pure synthetic (fused/sintered) suitable production parameters and conditions. they have especially used refractories in the construction of crucibles, research and development purposes, etc. where the cost of a refractory is not considered. cordierite (2mgo. 2al2o3. 5sio2) and mullite (3al2o3. 2sio2) these are technologically popular aluminosilicate refractory materials due to their suitable properties (thermal shock resistance, low thermal conductivity, creep resistance, low thermal expansion, elevated thermal and chemical stabilities, etc.) for various engineering purposes. their raw materials are easily obtainable at cheap prices. the low production costs of these materials influenced their potential use as an alternative material to alumina. they are used in welding shapes, electrical insulators, furnace liners, etc. the high thermal coefficient of expansion of mullite renders it a suitable refractory utilized to ameliorate the thermal operational problems encountered at the silicon-substrate overlap of integrated circuit devices (fahlman, 2023; rahman et al., 2014). what is normally done in this case is to decrease the thermal expansion coefficient to equalize the thermal coefficient of silicon by adding cordierite as a second phase, since it has a low thermal expansion coefficient. a cordierite-mullite refractory composite is superior to mullite refractory because of the high thermal conductivity of the former (mullite). an adequate liquid phase is formed between 1290-1550oc to form a density. another important factor to note is that both mullite and cordierite co-exist in thermal equilibrium, which means mullite can exist in the cordierite phase and cordierite can exist in thermal equilibrium with the mullite phase. cordierite-mullite composite ceramics were fabricated from commercially available cordierite and mullite powders (xu et al., 2017) in which the mullite content in the composite was about 65% (xu et al., 2017). the electrical conductivity of the composite was studied based on the influence on porosity, mullite glass phase, and grain size. the composition of refractory composite renders them suitable for thermal expansion behavior. the consequence of starting materials on the morphology of mullite crystals in the composite refractories (cordierite-mullite), and was observed the morphology of the mullite crystals became different contingent on the type and source of aluminum utilized in the fabrication of the composite refractories (chermat et al., 2025; sun et al., 2022). it was revealed that alumina-sol mixtures give rise to acicular grains while al(oh)3 powder mixtures show the appearance of angular or granular mullite mixtures. a lower mullitization temperature of a composite made with alumina sol shows better reactivity and dispersibility than those with al(oh)3 powder. cordierite and mullite powders obtained from commercial vendors have been used by phatthamon kiattisaksophon and sukdiphon thiansem as raw materials in different ratios of up to 100 wt% (kiattisaksophon & thiansem, 2008). the raw material was combined, mixed, processed, and taken to a ball grinding mill for composite formation. the samples were then sintered at temperatures between 1250 and 1400 at an interval of 50°c. the samples were heated at extreme temperatures and cooled down to room temperature to study their thermal actions to measure the thermal expansion coefficient. it was learned from the thermal behavior of the samples that the fabricated cordierite-mullite composite showed good physical and chemical properties at 70:30 wt% sintered at 1400°c. the fabricated cordierite-mullite composite also shows improved densification of cordierite and mullite structures, thermal shock resistance, pore size variation, bulk density, and apparent low porosity. the crack and microstructural characteristics suitably attract the utilization of cordierite-mullite composite in the swift firing of porcelain whiteware. one significant concern pointed out by the refractory industries is the thermal shock degeneration of commercial refractories that results in to decrease in stiffness, mechanical strength, and degradation when subjected to high industrial thermal conditions. the use of high-quality starting reagents ignited the cost of refractory composites in the market. considerations are now made for the use of non-destructive testing to determine damages caused by thermal shocks. the thermal degeneration of commercial refractory composites was investigated by (boccaccini et al., 2008; cioangher et al., 2024; flores-jacobo et al., 2023) exposed to harsh industrial thermal conditions by comparing the degenerated curves acquired from the non-destructive testing, e.g. comparing images acquired from ultrasonic velocity testing (upvt) with those secured from measuring the flexural strength and fracture toughness by three-point bending test and chevron notched method, respectively. table 1: physical properties of cordierite and mullite (camerucci et al., 2001; da silva et al., 2019) property cordierite mullite chemical formula 3al2o3. 2sio2 2mgo.2al2o3. 5sio2 density 3.03g/cc 2.60 g/cc thermal expansion coeff. 4.55.6 (x10-6/ °c) 1.7(x10-6/°c) thermal conductivity 4-6 w/m k (100-1400°c) 3 w/m k (room temp.) max. operating temp. 1725 °c 1371°c pa ge 41 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 37-47, 2025 mullite has suitable thermal and mechanical properties which account for its importance in traditional and advanced refractory material. it is not widely distributed in nature because of its high temperature and low pressure. many methods are used to fabricate refractory composite materials. the two methods employed to fabricate mullitecordierite composite refractory: (1) by a combination of raw material, and (2) by joining of pre-made mullite and cordierite powders (bilung, 2012). the ground raw materials are heated at low temperatures between 550°c to 9000c to enhance the densification of the materials. the microstructure, crystalline, dielectric, and mechanical properties of the composite manufactured from the various raw materials were analyzed. crystals were easily formed refractory composites made of finely ground precursors than those from aluminum sulfate reagents. a composite refractory fabricated from a raw material made of composite materials attains a better density at lower temperatures due to the gummy or sticky flow of the ceramics. mechanical properties of a composite can be accounted for from microstructures while strength and hardness are correlated to the composition, grain size, and porosity. in a solid-state reaction, a refractory composite with its desired properties is incompletely transformed into another composite having another set of desired properties. the product from this process has the benefit of carrying the advantages of both the starting materials. it is observed from various cordieritemullite ceramics that an increase in cordierite causes a decrease in the thermal expansion coefficient. preferably, we can keep the percentage of cordierite-mullite ratio at the constant of 70:30%. shock resistance is supported by the cordierite while strength is aided by mullite. the existence of the cordierite reduces the temperature and the firing shrinkage (el-fadaly et al., 2022; sadik et al., 2016). (zemánek et al., 2021) in their work combined cordierite and mullite in various proportions either as reactants or grog and studied the effect of the percentage of grog added. components with 50-70 & grog acquired improved mechanical, thermal, and physical properties. table 1 above shows some physical parameters of cordierite and mullite. ternary phase diagrams for cordierite and mullite a ternary phase diagram is a complex diagram captured in a three-dimensional plot in which each side of the plot represents a pure component in the system. figure 1 below is the ternary diagram of the magnesium oxide aluminum silicate system. the mgo.al2o3.sio2 system has been studied for so many years through the extensive experimental system. the compositions within the system are important for many reasons. they are important to geologists looking for mineralogical systems and also many of the compounds within the system have important applications such as high-temperature ceramics and structural ceramics in kilns or ovens. most of the compositions in the system form transparent glass ceramics and from the geologic point of view, they contribute many of the lower mantle minerals. understanding the melting behavior and crystallization part of the system is therefore important. it is also a great test system to see if we can apply our knowledge to read the crystallization part from the liquidus projections. figure 1: modified ternary phase diagram of refractory composites (dudnik et al., 2014) many years of studies lead to the experimental determination of the system and detailed characterization of the phase stability in melting temperatures. the objectives here are to read and interpret all of the information in the diagram and then track some solidifications. the three compositions (mgo, al2o3 & sio2) in the diagram are shown in percent weight or mass percent and not moles. it is observed that the part pa ge 42 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 37-47, 2025 where you have the cordierite (2mgo.2al2o3.5sio2) with blue dotted lines is a little bit congested. along the mgo and sio2 binary are two compounds which are mgo.sio2 mineral called enstatite and 2mgo.sio2 is known as forsterite. both of these compounds do not show solubility. between the mgo and al2o3 is a wellknown phase called spinel (mgo.al2o3) which shows a very significant solid solution. the compound along al2o3 and sio2 is mullite (3al2o3.2sio2). there are two ternary compounds, one is the cordierite mineral (2mgo.2al2o3.5sio2) which has ceramic applications, and immediately below the cordierite mineral is sapphirine (4mgo.5al2o3.2sio2). between the spinel (mgo.al2o3) and al2o3 is a solid solution. a significant amount of solid solution is also found around the mullite and the cordierite. the structural chemistry of alumina aluminum oxide is an oxide obtained from bauxite and occurs in mineral corundum alongside some impurities from ruby and sapphire(abyzov, 2019; giuliani et al., 2020; panasyuk et al., 2019; simonet et al., 2008). the oxides of figure 2: shows the thermal transformation of aluminum hydroxides (modified). figure 3: (a) modified corundum structure in alphaal2o3 and (b) modified octahedral coordination of alpha-alumina (abdelkader & abdecharif, 2012; ahmed et al., 2021; khosravi mardkhe, 2014). alumina obtained from bauxite with varying particle sizes are reactive alumina, low bauxite, calcined alumina, etc. they have properties such as particle size, surface area, surface reactivity, catalytic properties, etc., which account for their structural differences. the illustration in figure 1 below shows the thermal conversion of aluminum hydroxide into various forms when subjected to heat (castruita et al., 2013; shirai et al., 2010). there are many allotropes of al2o3, and α-al2o3 is the most stable thermodynamic phase which is formed at high temperatures by metastable or transition alumina precursors such as γ-alumina, β-alumina, δ-alumina, σ-alumina, etc. aluminum hydroxide thermally decomposes to form these metastable phases (castruita et al., 2013; yang et al., 2024). for instance, heating gibbsite (γ-trihydrate; γ-al(oh)3), boehmite (γ-monohydrate; γ-alooh), bayerite (α-trihydrate; α-al(oh)3), and diaspore (α-monohydrate; α-alooh) convert to various crystalline phases when calcined at high temperatures (atanga, 2013). the crystalline form of alumina is hexagonal in structure, as shown below, flocked by o2anions and al3+ cations (baronskiy et al., 2022; nestler et al., 2019). the al atoms are in an octahedral arrangement with six oxygen rings as shown in the diagram below(gutierrez & johansson, 2002; rustad et pa ge 43 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 37-47, 2025 al., 2004). the dimensions of the unit cells in the trigonal lattice structure are a = 4.7587 å, b = 4.7587 å, and c = 12.99 å. properties of al2o3 the typical properties of alumina are: high hardness, chemical inertness, high thermal stability, low coefficient of thermal expansion, good thermal conductivity, good wear and abrasion resistance, high corrosion resistance, a high melting temperature (2050°c), and its commercial availability in different purity ranges and grain sizes, makes it attractive for a great variety of engineering application. the suitability of alumina to resist thermal fatigue is related to its geometric arrangement and strength, controlled by its thermal properties like thermal conductivity and thermal expansion characteristics (czerwinski, 2020; kamara et al., 2025; sobczak et al., 2002; h. wang et al., 2020). applications of al2o3 ceramics the applications of alumina ceramics in engineering are as follows: • as a refractory material for the lining of furnaces (sengupta, 2020; slovikovskii & gulyaeva, 2018). • as an abrasive material and cutting tool (hosseini & kishawy, 2014; toenshoff & denkena, 2013) • as substrate material for electronic parts and packing materials for integrated circuits (e.g. silicon chips, electrical insulators, and spark plugs) (pulugurtha et al., 2022; yin et al., 2024) • alumina also has widespread applications as a porous ceramic and catalyst support (wang et al., 2022). the suitability of alumina for all these applications is due to the properties listed in table 2. structural chemistry of sio2 silica (sio2) is a compound made of silicon and oxygen table 2: mechanical and thermal properties of alumina (gudlur et al., 2012; xie et al., 2011) characteristics measured values density, [g cm-3] 3.95-4.10 melting point, [°c] 2072 microhardness, [gpa] 20 hardness, [gpa] 9 young’s modulus, [gpa] 260-410 bending strength, [mpa] 150-600 fracture toughness, [mpa m0.5] 4-6 thermal conductivity from 25 °c to 1000 °c, [w m-1 k-1] 30-40 coefficient of thermal expansion from 20 °c to 1000 °c, *10-6 [k-1] 5.4-9.5 atoms, which occurs in nature as quartz, silica sand, or sandstone(götze, 2012; pan et al., 2022), and can also be obtained from various sources in different forms like silica fume, fused silica, colloidal silica, and gel silica (hyde et al., 2016). the reaction in equation 1 below shows the formation of silica from alkoxide. si(or)4 (1) + 2h2o (1) →sio2 (s)+ 4roh (1) efficient nucleation and heterogeneous growth of the amorphous material lead to the fabrication of stable mullite phases. a mullite refractory fabricated from amorphous silica is more stable than mullite from crystalline silica reagent. the formation of mullite with better microstructure is dependent on the silica precursor, the alumina precursor, and processing conditions. there are several crystalline forms of silica, but the three major crystalline phases are quartz, tridymite, and cristobalite. the low and high-temperature crystalline forms or polymorphs are represented as the α-form and the β-form. the β-cristobalite is stable at around 1705 °c and melts at temperatures equal or equal to 1705 °c (fernandes et al., 2024; ortiz-bravo et al., 2022; wiberg et al., 2001). table 3: silica and some properties (pabst & gregorová, 2013; polyakova, 2014) form crystal symmetry stable range density (g cm-3) α-quartz hexagonal < 573 °c 2.65 β-quartz hexagonal 573 °c 870 °c 2.53 α-tridymite monoclinic metastable 2.36 β-tridymite hexagonal 870 °c 1470 °c 2.25 α-cristobalite tetragonal metastable 2.32 β-cristobalite tetragonal 1470 °c 1705 °c 2.20 silica melt ≥ 1705 °c pa ge 44 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 37-47, 2025 figure 4 represents the stable crystalline phase of silica at extreme temperatures in which each silicon atom is bonded to four oxygen atoms to form a tetrahedral structure (norman et al., 2013; polyakova, 2014; schnurre et al., 2004). the silica group in the structure constitutes various figure 4: modified crystal structure of cristobalite (a high-temperature crystalline form of silica)(aleem et al., 2014). minerals with different structures, symmetries, and physical properties but with the same composition. an increase in temperature causes a shift in the positions of the atoms in the crystal lattice structure. the bond angle between the sio4 tetrahedra) change from 153° (β-quartz) via 180° (β-tridymite) to 151° (β-cristobalite). a shift is induced in the bond length of si-o, resulting in a contraction from 0.161 nm (α-quartz) to 0.158 nm (β-cristobalite). a corresponding decrease in density and an increase in hardness have been discussed by some researchers. silica is the most significant component of mullite, which has many industrial applications. it has a high melting temperature, used to produce molds and cores for the production of metal castings in ferrous and non-ferrous industries. it is the main component of ceramic products. silica is used to regulate drying and shrinkage; it modifies thermal expansion and improves structural and mechanical properties (hou et al., 2025; tran et al., 2021; j. wang et al., 2020). conclusion refractories are well-known for their superior engineering properties. they involve low production costs due to the reasonable source of raw materials involved. the most common and widely used refractory composites are cordierite and mullite. they are prepared through various methods such as the combustion process, mechanical route, sol-gel route, the effect of ph, etc. these compounds can also be studied in a ternary system to understand important applications such as those in high-temperature ceramics. they are made of aluminosilicate compounds. references abdelkader, d., & abdecharif, b. 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(2021). processes for environmentally friendly and/or cost-effective manufacturing. materials and manufacturing processes, 36(9), 987-1009. pa ge 1 pa ge 1 american journal of chemistry and pharmacy (ajcp) patterns of health complaints and perception of the role of pharmacists among clients visiting community pharmacies in lagos state, south-west nigeria promise olaitan ernest-otaru1, tinuola omotomilayo odugbemi1, aloysius obinna ikwuka2*, francis chigozie udeh2 volume 4 issue 1, year 2025 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v4i1.4310 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: january 03, 2025 accepted: february 01, 2025 published: march 10, 2025 health complaints and its patterns differ generally in the population. the public perception of community pharmacies and drug vendors as a one-stop facility for almost all health complaints and services is a major determinant of the type, quantity, quality, effectiveness, and success of healthcare as they encourage irrational use of drugs and poor utilization of health facilities. in addition, the existing high prevalence of self-medication encouraged by the constant displays, advertisement, and access to over-the-counter drugs contribute to the problem. the aim of this study was to assess the patterns of health complaints and perception of the role of pharmacists among clients visiting community pharmacies in agege local government area (lga) of lagos state. a descriptive cross-sectional study of 305 adults visiting community pharmacies in agege lga using a multi-stage stratified sampling technique was used. data was collected using an interviewer-administered questionnaire and analyzed using spss version 23. the chi-square test was used to test the association between socio-demographic and socio-economic characteristics (independent variables) and perception of the role of pharmacists (dependent variable) of the study participants at a significance level of p<0.05. among the study participants, the mean age was 36.83±11.89 years, the majority were females, married, from the yoruba tribe, living with family members, employed, and earning below the poverty level of us$2 per day. the patterns of health complaints ranged from minor ailments like pain (21.0%), multiple symptoms (13.1%), upper respiratory tract infections (11.5%), and fever (8.9%) to more major conditions like gastrointestinal disorders (6.6%) and non-communicable diseases like hypertension, diabetes mellitus, and hyperlipidemia (11.1%). the majority (83.0%) of the study participants have a positive perception of the role of pharmacists, as 77.7% of them believed that pharmacists should be able to advise them on their medications, and 56.7% believed they should be able to report any drug-related adverse effect to a pharmacist. the socio-demographic and socioeconomic characteristics of the study participants have no statistically significant association with their perception of the role of pharmacists in community pharmacies (p>0.05). the patterns of health complaints at community pharmacies ranged from minor to more serious complaints that were related to age groups and income status. therefore, community pharmacies should be stocked with over-the-counter as well as prescription-only drugs. keywords community pharmacies, health complaints, lagos state, nigeria, patterns, perception, pharmacists 1 college of medicine, university of lagos, lagos state, nigeria 2 college of medicine and health sciences, american international university west africa, banjul, the gambia * corresponding author’s e-mail: aloysiussweet@yahoo.com introduction pharmaceutical care has recently become a vital element of national healthcare systems. when data from this sector is accurately gathered, it can significantly aid in compiling healthcare statistics and tracking disease trends, spread, and prevalence. in nigeria, community pharmacies serve as private drug retail centers overseen by licensed pharmacists, offering a variety of primary healthcare (phc) services alongside traditional medication dispensing (bradley, 2013; ihekoronye, 2020). in lowand middle-income countries (lmics), community pharmacists in these private drugstores are instrumental in monitoring the health of both children and adults, as people often visit these pharmacies for minor health issues. this trend is driven by the accessibility of overthe-counter drugs and a growing preference for selfdiagnosis and self-treatment (zawahir, 2021; de tran, 2019). in these settings, community pharmacists generally interact with 11–30 patients daily, with each consultation lasting an average of 4.8 minutes (ayele, 2018; yusuff, 2021). community pharmacies have expanded their services beyond dispensing medications; they now offer counseling, healthcare advice, and affordable medication options, all while prioritizing confidentiality and adherence to medical ethics (ihekoronye, 2020). as they normally open late and are often available on weekends, community pharmacies make healthcare accessible without appointments. the types of complaints presented at community pharmacies vary depending on factors like location, demographics, seasons, and prevailing public health concerns in the host community and environs. common complaints include detailed symptom information, with cases such as diarrhea in infants and adults and flu being prevalent in economically disadvantaged regions (nguyen, 2023; nafade, 2019). flu is a serious public health threat in regions with impoverished populations and poor healthcare accessibility (nguyen, 2023). other frequent issues in lmics include aches, migraines, physical injuries, fever, allergies, insect bites, skin rashes and reactions, cardiovascular disorders, and other respiratory and gastrointestinal upsets, with common pa ge 2 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 1-15, 2025 treatment requests for cold remedies, oral rehydration solutions, and pain relievers (ihekoronye, 2020; yusuff, 2021). some other customers visit pharmacies to refill ongoing prescriptions. in many urban areas of nigeria, including the agege local government area (lga) of lagos state, community pharmacies play a crucial role in addressing diverse health complaints. however, the specific patterns of these health complaints presented at these pharmacies are not completely understood. there is lack of clarity on patient perception of the role of pharmacists in these community pharmacies, which may hinder the optimal use of community pharmacies and potentially lead to poorer health outcomes. patient satisfaction plays a crucial role in influencing perception and continued use of healthcare services. studies have examined patient satisfaction across various healthcare settings, finding that factors like demographics, health status, medical care, communication with the patient, and patient expectations impact satisfaction (ferreira, 2023). more specifically, patients’ satisfaction with community pharmacy services has been studied in some european countries where patients reported being highly satisfied (cavaco, 2005; edideco, 2000). however, a similar study in nigeria reported that patients were unsatisfied with undue delay in drug dispensation (afolabi, 2003). there is a growing interest in evaluating patients’ perception of the actual role of pharmacists in community pharmacies, with studies conducted in some countries in europe e.g. malta (wirth, 2011), bosnia and herzegovina (catic, 2013); in africa e.g. nigeria (aderemi-williams, 2017), ethiopia (fekadu, 2011); and in the middle-east e.g. saudi arabia (al-arifi, 2012), united arab emirates (akshar, 2014). the quality of service and effectiveness of community pharmacists’ primary healthcare (phc) interventions depend largely on contextual variables such as patient health outcomes, duration per encounter, cost of drugs, the emotional concern of community pharmacists, the delivery of care by the community pharmacists, and the overall patient experience. these factors are important determinants of patients’ perception and acceptability of community pharmacists in local practice settings (aderemi-williams, 2017; afolabi, 2003; ekpenyong, 2018). from entering the pharmacy to obtaining medications and paying for them, every step influences patient satisfaction and the perception of the role of pharmacists in community pharmacies in rendering quality healthcare (aderemi-williams, 2017). recognizing that the success of phc and therapy are dependent on patient satisfaction and health outcomes, and that patients constitute an essential source of information about care effectiveness (donabedian, 1992), this study therefore aimed to provide a comprehensive data on the patterns of health complaints and perception of the role of pharmacists in community pharmacies among clients visiting community pharmacies in agege lga of lagos state, south-west nigeria. materials and methods study area the study was conducted in agege local government area (lga) of lagos state, south-west nigeria. agege lga has a vibrant economy with several urban markets, about 35 community pharmacies, and an estimated population of 683,600 (city population, 2024). the recorded population of residents above 15 years of age within the study area from a previously conducted census was 307,677 (city population, 2024). the estimated number of community pharmacies in agege and ifakoijaiye lgas (both in lagos state) is 150, each of which receives 30 patients daily (medpages, 2024). study design a descriptive, cross-sectional, community-based survey was conducted for six (6) weeks using a well-structured interviewer-administered questionnaire. study population the clients who visited community pharmacies within the agege lga within the study period constituted the study population. sample size a sample size of 305 was deduced from the total population using the cochrane formula for single proportion formula, n = (z2pq)/d2 , in concordance with (okeke, 2023a; okeke, 2023b; okoro, 2024a; okoro, 2024b; udeh, 2023). where: n = minimum sample size when the population is greater than 10,000. z = standard normal deviation usually set at 1.96 corresponding to a 95% confidence interval obtained from a standard statistical table of normal distribution. p = proportion of the population that visits community pharmacies in a similar study conducted among adult traders in agege lga with a non-response rate of 40%, and p was 85% = 0.85 (afolabi, 2008). q = 1 – p = 1 – 0.85 = 0.15 d = degree of accuracy desired (absolute precision) = 5.0% = 0.05 therefore: n = (1.962×0.85×0.15)/0.052 = 195.9, approximately 196 it is ideal to consider a non-response after calculating the minimum sample size using the cochrane sample size formula. similar to another study (afolabi, 2008), a non-response rate of 40% was considered to cover erroneously filled questionnaires, some study participants dropping out, or loss of data. to adjust the estimated minimum sample size for nonresponse and attrition, n = n0 / expected response rate = 196/0.6 = 326.67 ~327 sample size pa ge 3 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 1-15, 2025 of this calculated sample size, only 305 individuals who agreed to participate were selected using a multistage stratified sampling method. five wards (isale/idimangoro, orile agege, darocha, keke, and okekoto) were balloted from the eleven wards in agege lga. three community pharmacies were balloted from each ward, making a total of fifteen selected community pharmacies visited within six weeks for data collection. inclusion and exclusion criteria male and female adults, resident and non-resident clients visiting community pharmacies in agege lga were included in the study. health professionals including medical doctors, nurses, and pharmacists currently working in other healthcare facilities and patent medicine vendors within agege lga were excluded in the study. study procedures one-on-one well-structured interviewer-administered developed google form questionnaires containing a total of 33 questions, consisting majorly of closed-ended questions together with a few open-ended questions adapted from the literatures, were piloted and used as data collection tools for this study. each questionnaire has three sections: socio-demographic and socio-economic characteristics of the study participants patterns of complaints of the study participants perception of the role of pharmacists by the study participants study techniques and data collection a reserved area, free from interruption, was created within the pharmacy and used for the interview. using a stratified random sampling method, clients who visited the pharmacies within the six weeks of the survey and were willing to participate were selected. on average, 50 questionnaires were administered weekly. from the inflow of clients into the community pharmacy, an individual was approached every 20 minutes. only a member of the research team was involved in administering and filling out the questionnaires on google forms. upon successful data collection, questionnaires were cross-checked for adequacy of information, numbered, organized, and entered into a microsoft excel sheet for data analysis. reliability of study techniques and data collection a pre-test of the data collection tool (questionnaire) was carried out among 30 clients (approximately 10% of the sample size) in two community pharmacies in a different lga (ikeja lga). some questions were later modified for clarity. using the pearson correlation coefficient (r), the test-retest reliability of the data was determined for this study. r>0.7 was obtained which was enough to justify the use of research instruments since 0.7 is the minimum score for a good test-retest (kline, 2000). data analysis statistical package for social sciences (spss) version 23 software was used to validate and analyze the data. a 5% significance level (p-value) was considered significant. the socio-demographic and socio-economic characteristics of the study participants and their responses to questions were expressed in frequencies and percentages. the patterns of complaints were recorded and compared to the age groups and income status of the study participants. the perception of the role of pharmacists in community pharmacies was assessed using the eight (8) different response options in which they can select multiple choices that are appropriate. six (6) out of the eight question options provided were appropriate responses while two (2) were inappropriate responses. the maximum obtainable score was 8 and the minimum was 0. any respondent who scored >4 by choosing appropriate options was classified as having a positive perception, and any score ≤4 was classified as a negative perception. bivariate analysis was conducted using chi-square to determine the association between socio-demographic and socio-economic characteristics (independent variables) and perception (dependent variable). ethical considerations ethical approval for the study was obtained from the health research and ethics committee (hrec) of the lagos university teaching hospital (luth). a letter of introduction was obtained from the department of community health and primary care in the college of medicine, university of lagos and this letter was presented to community pharmacies to seek permission from the related authorities of the community pharmacies which were involved in the study. the interviewer properly informed the participants of the nature of the study, its objectives, its confidentiality, its importance to the society, and an informed oral consent was obtained from all study participants. there were no financial benefits to the study participants. the participants were also assured that they could withdraw from the study at any time during the data collection stage without suffering any consequence if they choose not to participate further. results and discussion a total of 305 individuals participated voluntarily in this present study, giving a 93.3% response rate from the anticipated 327 sample size. the obtained results are presented in tables and figures. table 1: socio-demographic characteristics of the study participants variable frequency (n=305) percentage (%) age group (years) < 20 6 2.0 pa ge 4 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 1-15, 2025 20-24 21 6.9 25-29 73 23.9 30-34 64 21.0 35-39 47 15.4 40-44 24 7.9 45-49 19 6.2 50-54 18 5.9 55-59 14 4.6 ≥60 19 6.2 mean±sd 36.83±11.89 gender male 138 45.2 female 166 54.4 transgender 1 0.3 marital status single 93 30.5 married 201 65.9 divorced/separated 6 2.0 widowed 5 1.6 ethnicity yoruba 202 66.2 igbo 41 13.4 hausa 33 10.8 others 29 9.5 living status with family 252 82.6 alone 40 13.1 with non-family 13 4.3 the mean age was 36.83±11.89 years, the majority were young adults between the ages of 25 years and 44 years. most were female 166 (54.4%), majority (65.9%) were married, were of the yoruba tribe (66.2%) and live with their family (82.6%). table 2: socio-economic characteristics of the study participants variable frequency (n) percentage (%) highest level of education completed highest level of education completed 6 2.0 none 21 6.9 vocational 73 23.9 primary 64 21.0 secondary tertiary employment status employed self-employed 139 45.6 private employed 89 29.2 government employed 30 9.8 unemployed student 26 8.5 pa ge 5 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 1-15, 2025 retired 12 3.9 unemployed (due to lack of job) 9 3.0 estimated monthly income in naira (n) <n10,000 10 3.3 n10,000 – n30,000 12 3.9 n30,001 – n50,000 51 16.7 n50,001 – n100,000 131 43.0 n100,001 – n200,000 73 23.9 n200,001 – n500,000 20 6.6 >n500,000 8 2.6 lga of residence in lagos state agege 274 89.8 ifako-ijaiye 12 3.9 alimosho 8 2.6 ojodu 5 1.6 ota 2 0.7 surulere 1 0.3 eti-osa 1 0.3 ikeja 1 0.3 isolo 1 0.3 majority of the study participants reside within agege lga of lagos state 274 (89.8%), had completed the tertiary level of education – 175 (57.4%), and were employed 258 (84.6%). most 204 (66.9%) earned below us$2 per day, out of which 22 (7.2%) earned below the prevailing minimum wage (as of the time of data collection) of 30,000 naira approved by the federal government of nigeria. as of today, the minimum wage in nigeria has been increased to 70,000 naira by the federal government of nigeria. table 3: types of complaints among study participants, other reasons for visits to community pharmacies s/no. types of complaints frequency (n=305) percentage (%) pains 64 21.0 multiple symptoms 40 13.1 upper respiratory tract infections 35 11.5 non-communicable diseases (ncds – hypertension, diabetes mellitus, hyperlipidemia) 34 11.1 fever 27 8.9 gastrointestinal tract (git) disorders 20 6.6 refill of prescriptions 17 5.6 genital infections 14 4.6 need for supplements 13 4.3 dermatological conditions 11 3.6 hormonal related issues 9 3.0 need for otc drugs 6 2.0 ear infection 4 1.3 eye infection 3 1.0 other eye problems 3 1.0 insomnia 3 1.0 shortness of breath 1 0.3 other complaints (bph, erectile dysfunction, cramps) 1 0.3 pa ge 6 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 1-15, 2025 pain was highly prevalent among this population aside other complaints as it was reported by 21.0% of the study participants. other symptoms with high prevalence were multiple symptoms (13.1%), upper respiratory tract infections (11.5%), non-communicable diseases (hypertension, diabetes mellitus, and hyperlipidemia) 11.1%, and fever 8.9%. figure 1: previous (3-6 months) health complaints of the study participants figure 2: health complaints of the study participants by age group the commonest health complaints of the study participants in the past three to six months were collected, recorded, and compared with their present health complaints. the majority of the study participants have had multiple symptoms (34.4%), pains in different parts of the body (12.1%), complaints of non-communicable disease (11.8%) which was almost similar (in percentage) to complaints of upper respiratory tract infections (10.5%). other health complaints were git disorders (4.6%), fever (3.3%), genital infection (5.2%), eye infection (2.0%), shortness of breath (1.0%), insomnia (3.0%), ulcer related issues (0.3%), and hormonal related issues (0.3%). the majority of the old adult respondents complained of having non-communicable diseases and many of them were there to fill their prescriptions. next to them were the middle-aged adults. conversely, the majority of the young adult respondents complained of pains and upper respiratory tract infections. multiple symptoms were presented across all the age groups. a lower percentage of the elderly adult respondents presented multiple pa ge 7 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 1-15, 2025 symptoms while a higher but similar percentage of the young adults and middle-aged adults presented it. in addition, the highest percentage of those who complained of genital infection were the elderly adults followed by middle-aged adults, and git disorders were also more common among the elderly adult respondents. figure 3: health complaints of the study participants by income status pain was the most frequently reported health complaint in both income groups (living below poverty line and living above poverty line), with a slightly higher percentage in those living below poverty line. the percentage is over 20% for both groups, making it the dominant complaint across all categories. both groups reported a notable percentage of upper respiratory tract infections, with slightly higher percentages among those living above poverty line. the respondents complained of multiple symptoms while a lower percentage of the middle-income respondents and upper-income respondents presented it. the highest percentage of those who complained of genital infection was the upper-income respondents followed by middleincome respondents. moreover, the highest percentage of those who complained of genital infection was the upper-income respondents followed by middleincome respondents, and git disorders were also more common among the middleand upper-middle-income respondents. table 4: perception of the role of community pharmacists by the study participants s/no. types of complaints frequency (n=305) percentage (%) advise me on the drugs i am using 237 77.7 report to them if i react to a drug 173 56.7 seek advice from them if the condition is not serious enough to visit a physician 153 50.2 provide information on the use of drugs and answer my drug-related questions 98 32.1 help monitor and provide information on my health progress with the medication 77 25.2 provide extended services like health screening services, bp monitoring, and blood sugar monitoring mainly in community pharmacies 48 15.7 function as a medical doctor 13 4.3 give injections or drips 7 2.3 the majority of the study participants believed that a pharmacist should be able to give them advice on the drugs that they are using 237 (77.7%), they should be able to report any drug reactions to the pharmacist 173 (56.7%), and they should be able to seek advice from the pharmacist in conditions that are not serious enough to visit a physician 153 (50.2%). however, the majority of the study participants did not expect a pharmacist to give injections or drips (97.7%), function as a medical doctor (95.7%), nor provide extended services like health screening services, bp monitoring, and blood sugar monitoring mainly in community pharmacies (84.3%). table 5 shows that the majority of the study participants 253 (83.0%), had a positive perception of the role of pharmacists in community pharmacies. pa ge 8 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 1-15, 2025 table 5: overall perception of the role of pharmacists in community pharmacies by the study participants variable frequency (n) percentage (%) positive perception 253 83.0 negative perception 52 17.0 table 6: association between socio-demographic characteristics and perception of the role of pharmacists in community pharmacies by the study participants variable positive perception n=253 negative perception n=52 x2 df fisher’s exact p-value age group (years) 3.561 9 0.938 <20 5 (83.3%) 1 (16.7%) 20–24 16 (76.2%) 5 (23.8%) 25–29 59 (80.8%) 14 (19.2%) 30–34 53 (82.8%) 11 (17.2%) 35–39 40 (85.1%) 7 (14.9%) 40–44 22 (91.7%) 2 (8.3%) 45–49 15 (78.9%) 4 (21.1%) 50–54 14 (77.8%) 4 (22.2%) 55–59 12 (85.7%) 2 (14.3%) ≥60 17 (89.5%) 2 (10.5%) gender 0.241 2 0.887 male 115 (83.3%) 23 (16.7%) female 137 (82.5%) 29 (17.5%) transgender 1 (100.0%) marital status 1.138 2 0.566 single 74 (79.6%) 19 (20.4%) married 170 (84.6%) 31 (15.4%) divorced/separated/ widowed 9 (81.8%) 2 (18.2%) ethnicity 5.694 3 0.127 yoruba 171 (84.7%) 31 (15.3%) igbo 29 (70.7%) 12 (29.3%) hausa 27 (81.8%) 6 (18.2%) others 26 (89.7%) 3 (10.3%) living status 1.421 2 0.491 living with family 212 (84.1%) 40 (15.9%) alone 31 (77.5%) 9 (22.5%) living with non-family 10 (76.9%) 3 (23.1%) please note: ** statistically significant (fisher’s exact p-value), x2 – chi squared, df degree of freedom, p-value<0.05 table 7: association between socio-economic characteristics and perception of the role of pharmacists in community pharmacies by the study participants variable positive perception n=253 negative perception n=52 x2 df fisher’s exact p-value level of education completed 3.309 4 0.508 there was no statistical significant association between any of the socio-demographic characteristics of the study participants and their perception of the role of pharmacists in community pharmacies as all p-values were above 0.05. however, the majority of them across the age classification had a positive perception of the role of pharmacists in community pharmacies. in addition, there was a close similarity between the perceptions of males (83.3%), and females (82.5%). most of them (84.1%) who live with family had a positive perception. pa ge 9 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 1-15, 2025 none 8 (66.7%) 4 (33.3%) vocational 22 (81.5%) 5 (18.5%) primary 7 (87.5%) 1 (12.5%) secondary 72 (86.7%) 11 (13.3%) tertiary 144 (82.3%) 31 (17.7%) employment status 0.173 1 0.677 employed 215 (83.3%) 43 (16.7%) unemployed 38 (80.9%) 9 (19.1%) estimated monthly income in naira 4.158 4 0.385 <n30,000 17 (77.3%) 5 (22.7%) n30,001 – n50,000 44 (86.3%) 7 (13.7%) n50,001 – n100,000 112 (85.5%) 19 (14.5%) n100,001 – n200,000 60 (82.2%) 13 (17.8%) >n200,001 – n500,000 20 (71.4%) 8 (28.6%) please note: **statistically significant (fisher’s exact p-value), x2 – chi-squared, df degree of freedom, p-value<0.05 no statistical significant association exists between socio-economic characteristics and the perception of the role of pharmacists in community pharmacies by the study participants (p>0.05). however, most of the study participants had a positive perception of the role of pharmacists in community pharmacies regardless of their socio-economic status. the average age of 36.83 years obtained in this current study indicates that the study participants consisted predominantly of adults (25-39 years) opposing the study of (ojha, 2023) where teenagers and young adults (1829 years) dominated. the dominance of adults in this current study can be related to the fact that adults within the 25-39 years age group are non-dependent and are much more active in earning for themselves (korn, 1987; national bureau of statistics, 2021), predisposing them to several stressors that cause ailments, which make them visit the community pharmacies often for treatment. the slight female majority 166 (54.4%) aligns with global trends where women are more likely to engage with healthcare services than men (bertakis, 2000; dluhossebesto, 2021). the negligible transgender representation mirrors societal dynamics in nigeria, where transgender individuals are a marginalized group, often leading to underreporting in surveys. the high percentage of married respondents 201 (65.9%) can be attributed to the cultural importance of marriage in the nigerian society, while the predominance of the yoruba ethnicity 202 (66.2%) can be attributed to the study location which is a yoruba-dominated area as reported by (okoro, 2024a; okoro, 2024b). the finding that 252 (82.6%) of the study participants live with their family suggests a significant influence of family on the patterns of health complaints and perceptions of healthcare services (omeire, 2017). most 274 (89.8%) of the respondents reside in agege lga, which is a reflection of the targeted sampling within a specific locality. the level of educational attainment is commendable with over half (57.4%) of the respondents having completed tertiary education. this is indicative of the status of lagos state as an educational hub in nigeria, which might influence the lifestyle of the respondents and which in turn determines their patterns of health complaints at community pharmacies and their perception of the role of pharmacists in community pharmacies. with a higher number (84.6%) of employed respondents, majority (66.9%) of them earn below the current poverty line (us$2 per day). this economic constraint likely impacts their healthcare choices, such as reliance on overthe-counter (otc) medications and self-medication, as evidenced in other parts of this current study. the income distribution further emphasizes the economic challenges faced by the respondents, with only 9.2% earning above ₦200,000 monthly, reflecting the prevalent income disparity. the patterns of health complaints indicate a broad spectrum of conditions are presented at community pharmacies, ranging from minor complaints like pain and fever to more significant health conditions like git disorders and non-communicable diseases (ncds) e.g. hypertension, diabetes mellitus, hyperlipidemia. metabolic disorders e.g. hypertension, adiposity, diabetes mellitus and dyslipidemia collectively known as metabolic syndrome diseases (msds) are diseases related to one another and have very high morbidity and mortality rates (ikwuka, 2015; ikwuka, 2017a; ikwuka, 2017c; ikwuka, 2023c; ikwuka, 2023f; virstyuk, 2016). adiposity (obesity) is a risk factor for hypertension. obesity is also a risk factor for non-hodgkin lymphoma (ikwuka, 2023e). results obtained from different researches have shown that hypertension, adiposity, diabetes mellitus, dyslipidemia, asymptomatic hyperuricemia, systemic immune inflammation activation, and fibrogenesis can lead to kidney damage (ikwuka, 2017d; ikwuka, 2017e; ikwuka, 2018c; ikwuka, 2018d; ikwuka, 2019a; pa ge 10 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 1-15, 2025 ikwuka, 2019c; ikwuka, 2022; ikwuka, 2023d; virstyuk, 2017a; virstyuk, 2018a; virstyuk, 2019; virstyuk, 2021a; virstyuk, 2021b). the study identified that the most frequently reported health complaint among the respondents was pain. this aligns with the research of (shehnaz, 2013) which explained that pain or ache management was the primary reason expatriate high school students sought help at community pharmacies. similar to these findings, other studies conducted on adolescents in malta and tamil nadu, india reported that pain was the secondhighest complained symptom (ellul, 2008; arul, 2010). pharmacies are often the first point of contact for patients experiencing pain, due to the accessibility of over-thecounter (otc) medications and the convenience of pharmacy services, a point highlighted by (ellul, 2008) where pharmacies were reported as the most prevalent source of medicines. this trend underscores the role of community pharmacies as crucial providers of immediate relief for minor ailments. multiple symptoms were also commonly reported, reflecting the complexity of health conditions presented at community pharmacies. this may indicate either the presence of multi-factorial illnesses or the recurrence of inadequately treated conditions. the co-occurrence of symptoms suggests that many clients might be experiencing complex health issues, necessitating a comprehensive pharmaceutical care. upper respiratory tract infections (urtis) were another prevalent complaint. other studies also reported symptoms related to urti like cough and cold to be prevalent (ellul, 2008; arul, 2010). the incidence of urtis is consistent with the seasonal occurrence of respiratory illnesses, especially in urban areas like agege lga. this finding is significant given the role that pharmacies play in managing symptoms of urtis, such as coughs and colds, which are common reasons for self-medication. urbanization has been associated with increased oxidative stress. linked with the induction of oxidative stress are major free radicals. among these major free radicals, superoxide anion, hydroxyl radical, and hydroperoxyl radical are of physiological significance. non-radical of physiological significance is hydrogen peroxide (ikwuka, 2023b). increased oxidative stress can lead to mutation which is an alteration in the dna sequence which produces new alleles (ikwuka, 2023a). the complaints of ncds like hypertension, diabetes mellitus, and hyperlipidemia by the respondents reflect the increasing burden of chronic diseases in nigeria, particularly in urban areas. the growing prevalence of these diseases emphasizes the need for sustained management and the potential role of community pharmacists in providing ongoing monitoring and care. nevertheless, there is also need for new and effective treatment options in patients with metabolic syndrome diseases. sodiumglucose linked transporter 2 (sglt-2) inhibitors e.g. dapagliflozin and glucagon-like peptide 1 receptor agonists (glp-1 ras) e.g. liraglutide have been found to improve the efficacy of treatment and clinical course of type 2 diabetes mellitus and hypertension in patients with such comorbidities (ikwuka, 2017b; ikwuka, 2018a; ikwuka, 2018b; ikwuka, 2019b; ikwuka, 2021; ikwuka, 2024; virstyuk, 2017b; virstyuk, 2018b; virstyuk, 2018c). complaints about fever were among the middle-range prevalent complaint. however, according to (shehnaz, 2013), most of the respondents complained about fever. the complaint about fever could be indicative of underlying infections such as malaria. the timing of this current study, conducted during a period with higher incidence of malaria, could explain the prominence of fever as a reported symptom. the correlation between seasonal changes and the incidence of fever points to the need for community pharmacies to be well-equipped to manage febrile illnesses. comparison of the current health complaints reported by the study participants at community pharmacies to their previous (3 to 6 months) health complaints showed a recurrence or persistence of certain health conditions. this is evident from the close similarity in the percentages of some specific conditions, for example, upper respiratory tract infections accounted for 10.5% of previous health complaints, and 11.5% of current health complaints. non-communicable diseases were reported in 11.8% of previous health complaints and in 11.1% of current health complaints. many clients who visited community pharmacies complained of multiple symptoms, as reflected in 34.4% of previous health complaints and in 13.1% of current health complaints. it is also apparent that conditions such as pain, fever, gastrointestinal disorders, upper respiratory tract infections, non-communicable diseases, dermatological conditions, and genital infections occur more frequently than other health complaints such as eye infections, ear infections, and insomnia. the observed increase in cases of pain, fever, and gastrointestinal disorders may be related to the seasonal timing of data collection which took place in june, a period of frequent rainfall when many people were experiencing malaria symptoms (ipadeola, 2024). factors perceived as contributing to these febrile illnesses included stress, mosquito bites, unclean water, and rain (das, 2024; ipadeola, 2024). the results indicate that clients visiting community pharmacies may have a variety of reasons beyond seeking treatment for illnesses. some visit to obtain supplements, which could be considered a preventive health measure, with 4.3% of respondents doing so. approximately 5.6% of respondents were recorded visiting the pharmacy to fill or refill prescriptions, suggesting that some clients see a physician for diagnosis before visiting the pharmacy, and that prescription-only medicines (poms) are still being obtained with a prescription. however, it is unclear whether all patients requiring poms come with a doctor’s prescription. figure 2 illustrates the patterns of health complaints by age groups. it can be inferred that pain was a significant health complaint in all age groups, but is highest among pa ge 11 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 1-15, 2025 young adults between 25-44 years. young adults were the highest to complain about fever and multiple symptoms. this indicates that antipyretic drugs purchased as otc were mostly used by this group. associating pain with fever, and multiple symptoms suggests that malaria which presents with symptoms such as pain, fever, and which has been reported to be associated with the rainy season which was the period this research was conducted, is predominant among young adults. elderly adults visit community pharmacies frequently for ncds. the data also indicates that complaints of genital infections were low among respondents between the age of 18-24 years and those above 60 years because the latter group includes the elderly who were already less active sexually. on the other hand, complaints about genital infections were high among respondents between the ages of 25-44 years (young adults). this finding indicates that even though the world health organization (who) reported a high incidence of sexually transmitted infections (stis) among individuals between 15-49 years (who, 2024), those below 25 years were more cautious in preventing stis, and those older than 25 years were more reckless sexually, supporting the findings of (du, 2022) which stated that the incidence of age-standardized incidence rates (asrs) of syphilis, chlamydia, gonorrhea, trichomoniasis, and genital herpes increased by 1.70% globally. the variety of health complaints decreases with age, and certain health complaints are more prevalent in specific age groups, for example, urtis, pain, multiple symptoms, and fever are most common among young adults (25-44 years old), likely due to cumulative occupational exposure as this group is predominantly of working age (korn, 1987). the association between health complaints of the study participants by income status showed that pain was the most frequently reported health complaint among individuals living below and above the poverty line, with an insignificantly higher percentage in the former group likely because many are in the non-dependent age group and face socio-economic vulnerabilities, leading them to undertake menial jobs or lower-tier employment (fagerlund, 2023). respondents below the poverty line complained less about fever and more about multiple symptoms, indicating that multiple symptoms are dominated by fever (pyretic symptoms) leading to the dispensing of antipyretic drugs and subsequently lowering the fever complaints. the high rate complaints of urtis, and a slightly lower rate of git disorders in both groups are almost inseparable respectively. other health complaints such as ear infections, eye infections, insomnia, and hormonal-related issues are less frequent across both groups but show slight variations, with some complaints like dermatological conditions, genital infections and hormonal-related issues being more prevalent among those living below the poverty line. refill of prescriptions and complaints of ncds were seen more often among those living above the poverty line, which may be attributable to their socio-economic status and better access to healthcare services enabling them to manage chronic conditions. those below the poverty line reported a broader range of symptoms, which indicate a more complex health burden and less access to preventive healthcare. the perception of the role of pharmacists in community pharmacies was assessed using eight different question options depicting user’s perception, from which they can select as many that are appropriate. a client’s understanding of the role of pharmacists in community pharmacies will help the client to align his/her expectations with the role of the pharmacist which will improve the quality of the services they receive. as described in table 4, six out of the eight question options provided were appropriate responses while two were inappropriate responses. respondents who scored >4 were classified as having a positive perception while those who scored ≤4 were classified as having a negative perception. in this current study, the perception of the role of pharmacists in community pharmacies was largely positive (83.0%), with 77.7% believing that pharmacists should provide advice on drug use. this finding is consistent with the study of (awad, 2017; el-kholy, 2022; xuan, 2023), where 86.3%, 72.0%, and 76.1% of respondents respectively, believed that it is the role of pharmacists to counsel patients about their medications, highlighting the trusted position of pharmacists in the community especially for drug-related consultations. the high expectation for pharmacists to manage drugrelated issues, including adverse reactions (56.7%), is higher than the 40.0% reported by (el-kholy, 2022), and 54.8% reported by (awad, 2017). this finding further emphasizes the community’s reliance on pharmacists for their pharmaceutical care. interestingly, 50.2% of the study participants indicated they would seek advice from pharmacists for conditions not serious enough to warrant a visit to a physician. this finding agrees with the findings of (xuan, 2023) where 67.5% of respondents agree to seek a pharmacist’s services in treating minor symptoms such as cough, cold, constipation, and diarrhea. this suggests significant trust in pharmacists as the first point of contact for minor health concerns. however, the expectation for pharmacists to perform roles beyond their traditional scope, such as health screening services (15.7%), functioning as medical doctors (4.3%) or giving injections or drips (2.3%) was notably low. this finding reflects a lack of awareness about the expanding roles of pharmacists in some healthcare settings or cultural norms that delineate the roles of different healthcare professionals, similar to the reports of (akshar, 2014; awad, 2017). moreover, while a majority (83.0%) of the respondents had a positive perception of the role of pharmacists in community pharmacies, a lower proportion (17.0%) had a negative perception, indicating room for improvement in public education. this gap in understanding could be addressed through community outreach and educational programs designed to inform the public about the pa ge 12 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 1-15, 2025 comprehensive services pharmacists can offer, including health screenings and chronic disease management. there was no statistical significant association between socio-demographic characteristics (e.g. age group, gender, marital status, ethnicity, living status) and socio-economic characteristics (e.g. level of education, employment status, estimated monthly income) with perception of the role of pharmacists in community pharmacies, suggesting that perception was generally consistent across different population groups. the close similarity in positive perception between males (83.3%) and females (82.5%) further supports this finding. however, those living with family members had a better positive perception (84.1%), which could be linked to family influence in shaping healthcare decisions and perception. however, one limitation of this current study should be pointed out. the fact that data collection was carried out in june characterized with rains may influence the patterns of health complaints. in other non-rainy, dry and dusty months, the patterns of health complaints may differ. conclusion in conclusion, the recurrence of certain health complaints suggests that community pharmacies are a primary resource for ongoing care, particularly for chronic medical conditions. there is a need for better public education on the expanding role of pharmacists in healthcare delivery. enhancing public awareness about the risks of self-medication and the comprehensive health services offered by pharmacists, as well as a stricter regulation and enforcement on drug sales and acquisition could lead to better health outcomes. leveraging family dynamics and addressing the broader socio-economic challenges in the society could further improve the community’s approach to healthcare. acknowledgments the authors express their gratitude to everyone who contributed in one way or the other to make this research a success. the authors are also grateful to all the men and women who participated in this research for their cooperation and support, which are highly appreciated. conflict of interest the authors hereby declare that they do not have any possible conflicts of interest. references aderemi-williams, r. i., igwe, c n., & soremekun, r. o. 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(2021). factors related to antibiotic supply without a prescription for common infections: a cross-sectional national survey in sri lanka. antibiotics (basel, switzerland), 10(6), 647. https://doi.org/10.3390/antibiotics10060647. pa ge 1 pa ge 16 american journal of chemistry and pharmacy (ajcp) chemical modulation of amyloid beta oligomer’s in alzheimer’s disease muhammad jehangir1*, sayyad muhammad2, muhammad tayyab3, wajid ali3, ali danish alvi3,. muhammad naveed4 volume 4 issue 1, year 2024 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v4i1.4886 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: april 01, 2025 accepted: may 03, 2025 published: june 15, 2025 amyloid beta oligomers play a key role in the pathophysiology of alzheimer’s disease. since it contributes to memory and neuronal loss, cognitive decline, degradation of neurons as well as diseases progression. as aβ oligomer is very small and heterogeneous and it can change from one shape to another shape therefore it has various kinds of morphologies including monomers, oligomers and fibrils. due to instable behavior of these proteins, it’s very difficult to understand its exact mechanism. in this concise review, information over the past seven years concentrated on the molecular characteristics of amyloid-β oligomers has been summarized along with two pathways involving protein aggregation and condensation. furthermore, metal complexes, immunotherapies, and anti-aβ antibodies are reported here, which can modulate aggregation, stabilize non-toxic forms, and enhance degradation. finally, new discoveries on small molecules that control, modify and inhibit the progression of amyloid-β oligomers are integrated, as these are key characteristics of alzheimer’s disease. keywords alzheimer’s diseases, aβ oligomer’s, degradation, protein aggregation and condensation, stabilization 1 school of chemistry and molecular engineering, nanjing tech university, nanjing 211816, p. r. china 2 department of chemical engineering, nanjing tech university, nanjing 211816, p. r. china 3 school of pharmaceutical science, nanjing tech university, nanjing 211816, china 4 school of biological and pharmaceutical engineering, nanjing tech university, p.r. china * corresponding author’s e-mail: jehangirm62@gmail.com introduction according to recent projections, the number of people living with alzheimer disease (ad), the most prevalent form of dementia, is expected to reach 87 million by 2050. age is the primary risk factor for ad, which currently affects more than 35 million people worldwide (hwang et al., 2019; tzioras et al., 2022). given that 50–80% of all cases among the elderly population are estimated to be related to this rapidly ageing population, it has emerged as a significant social issue (peng et al., 2019). aberrant protein aggregation, which affects synaptic signaling, mitochondrial function, neuroinflammation, and neuronal loss, is indicative of ad and leads to multifactorial neuronal dysfunction (limbocker et al., 2019). alzheimer’s disease (ad) is caused by misfolded tau and amyloid-beta (aβ) proteins that build up in the brain along harmful pathways that cause selective neuronal death and synaptic loss (figure 1) (senapati et al., 2023). the primary neurotoxic species responsible for the onset of alzheimer’s disease (ad) are oligomeric aggregates of amyloid-β peptides (aβ). aβ oligomers stimulate additional pathological processes of ad, including tau hyperphosphorylation, oxidative stress, and mitochondrial dysfunction, in addition to causing various neuronal dysfunctions like loss of memory and learning. thus, it is acknowledged that aβ oligomers are suitable targets for ad diagnosis and treatment (sehar et al., 2022). in the pathophysiology of ad and other tauopathies, tauos and tau filaments may be crucial players. the general consensus is that smaller, diffusible oligomers are more likely to be involved in ad pathogenesis than the larger tau assemblies, which include straight filaments, phf, nft, and ghost tangles, which are thought to be less toxic (penke et al., 2020). compared to amyloid fibrils, soluble aβ oligomers have a stronger correlation with the advancement of disease. for this reason, focusing on these oligomers may prove to be a useful therapeutic approach in the management and prevention of alzheimer’s (jehangir et al., 2024). finding aβo in biological samples, like blood or cerebrospinal fluid, has enormous potential for tracking the development of ad and providing an early diagnosis. but because aβo are dynamic, structurally complex, and rare, it’s still very difficult to detect them accurately (penke et al., 2020). many of these diseaseassociated proteins not only misfold and aggregate, but also go through liquid-liquid phase separation (llps) to create dynamic condensates, which are essential for regular cellular processing (muhammad et al., 2024). many studies suggest that llps plays a significant role in the deposition and aggregation of tau and amyloid-β and is closely linked to the pathophysiology of ad. investigating the fundamental mechanism in greater detail will help increase the rate of early ad detection, which will lead to the development of anti-ad medications and better outcomes for ad patients (muhammad et al., 2024). in this concise review, we gathered information over the past seven years and concentrated on the molecular characteristics of amyloid-β oligomers, along with two pathways involving protein aggregation and condensation. we also integrated new discoveries on small molecules that control, modify, and break down amyloid-β oligomers, which are key characteristics of alzheimer’s disease. finally, we touched on their roles in the pathophysiology of the disease. it is known that aβ oligomers are the target of studies on chemical regulatory mechanisms, stabilization, and degradation processes, as well as the role of phase separation in alzheimer’s disease. pa ge 17 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 16-28, 2025 literature review amyloid-β oligomers, protein aggregation and condensation soluble aβo can start to accumulate in the brain up to two decades before clinical symptoms appear (hector & brouillette, 2021). aβ is produced as a result of the amyloid precursor protein’s proteolytic cleavage by β and γ-secretases. the two most prevalent forms of aβ in human bodies are aβ40 and aβ42. aβ42 was found to be more likely to aggregate, despite the fact that aβ40 and aβ42 only differ by two amino acid residues. aβ has a tendency to form a variety of self-assembly structures, from mature fibrils to oligomers and as well as monomer’s (volynsky et al., 2025). aβ toxicity has been shown to be primarily caused by aβ1~42 oligomers, both in vitro and in vivo (han et al., 2025). experimentally and computationally characterizing the early aβ oligomers is a difficult task. since most experimental observations provide timeand space-averaged properties, the transient and heterogeneous ensemble of oligomer structures presents an experimental challenge (derreumaux et al., 2022). at physiological concentrations of 1–20 nm, aβ42 synthesis easily forms oligomeric structures, and the rate of oligomer formation is temperature-dependent. at physiological concentrations, the formation of aβ42 oligomers is a kinetically analyzed and inhibitable process that is repeatable (li et al., 2023). the most neurotoxic agents are thought to be aβ42 oligomers. the morphologies of these oligomeric intermediates are diverse, encompassing spherical, annular, β-barrel, and protofibrillar forms, all of which have the potential to contribute differently to the toxicity of aβ (foley et al., 2019). in order to diagnose and treat ad, aβ oligomers are accepted as valid targets. however, because the term “aβ oligomer” can refer to a variety of soluble aβ species or a mixture of different types of metastable oligomeric species with different sizes, shapes, and conformations as a result of aβ’s dynamic assembly, it can be somewhat ambiguous (viola & klein, 2015). various experiments have contributed to our current understanding of the structures and biology of aβ oligomers, even though a unified model for their role in alzheimer’s disease has not yet been developed. studies using solid-state nuclear magnetic resonance (nmr) conducted by paravastu and colleagues revealed that antiparallel β-sheets make up the oligomers of aβ (haerianardakani et al., 2020). protein aggregation and condensation the word “aggregation” is frequently used in biology to refer to assemblies that are created under pathological circumstances, where the molecules within the aggregate are irreversibly disrupted and frequently regarded as pathogenic factors. one important feature of biological processes that are irreversible is aggregation. in contrast, the word “condensation” describes dynamic, reversible molecules that can be redissolved to carry out their specific tasks; the intracellular environment is closely monitored during their assembly (alberti & hyman, 2021). but there is some degree of interdependence between these two categories of higher-order protein assemblers. protein components can misfold and irreversibly aggregate when protein homeostasis is disrupted under pathological or pressure conditions, which can lead to an imbalance in biomolecular condensation and the uncontrollable collapse of these structures. aging or solidified condensates can then frequently transform into aggregates (amzallag & hornstein, 2022; savastano et al., 2020). protein condensates or aggregates may arise from intermediate clusters as precursors of droplets or aggregates under certain circumstances. a droplet that forms as an intermediate aggregate and then transforms into a solid state is one more potential mechanism for protein aggregation (wegmann et al., 2018). nonetheless, a number of investigations have indicated that amyloid (or cross-β) interactions play a role in the development of protein aggregates, and the in vitro production of amyloid fibrils is commonly observed in phase-separated proteins (hughes et al., 2018; luo et al., 2018). aside from nervous system conditions like schizophrenia, bipolar, and autism disorders, depression, epilepsy, and figure 1: amyloid-β (aβ) misfolds and forms soluble toxic oligomers and fibrils that accumulate in the brain leading to synaptic loss and selective neuronal death pa ge 18 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 16-28, 2025 alzheimer’s and parkinson’s diseases, the top-ranking protein condensation diseases also include depression. synaptic condensate-forming proteins’ genes are linked to the majority of these neurological disorders (zeng et al., 2016). the growing body of evidence indicates that a variety of human diseases are probably caused by aberrant protein condensation. changes in the physiological states of proteins are the root cause of these pathologies, which are collectively known as protein condensation diseases (li et al., 2020). yet, nucleation and off-pathway aggregation are frequently involved in the formation of amyloid fibrils, which is not just a straightforward polymerization process. delays cause disordered intermediates to refold into non-native secondary structures (β-strands), which are linked to the creation of oligomers, during a lag phase. in certain cases, during the α-β structure transition, folding intermediates with extended (non-native) α-helices can form native or amyloid states (žerovnik & venko, 2023). because of the stacking of aromatic rings and a network of backbone hydrogen bonds, amyloid fibrils, which are highly ordered and rigid protein states, can have a variety of morphologies (stanković et al., 2020; taylor & staniforth, 2022). reversibility is one of the main distinctions between protein condensates and aggregates. protein condensates are at least initially reversible, in contrast to more toxic forms of protein aggregates (shin et al., 2017). a characteristic of diseases like transmissible spongiform encephalopathies, prion diseases, alzheimer’s disease, and parkinson’s disease is protein aggregation. there is ongoing debate as to whether the aggregated proteins are involved in the pathological process directly or are merely bystanders. consequently, it is crucial to comprehend ps and aggregation. it is becoming evident that the pathogenesis of neurodegenerative disorders may involve proteins with liquid-to-solid transitions, such as tau, α-synuclein, and tdp-43, which bind to rna and are fused in sarcoma (fus) (zbinden et al., 2020). the discovery of liquid-like condensates raises the prospect of a different route for amyloid aggregation. this “condensation pathway” is different from the “deposition pathway,” which is the direct formation of amyloid aggregates from their native state via oligomeric species. even within the same cell and protein system going through self-assembly, solid deposits have been seen to form either directly through the deposition pathway or from liquid droplets through the condensation pathway (cascella et al., 2022; hardenberg et al., 2020). however, when amyloid aggregation occurs within condensates, the role of oligomers is still unclear. within a condensed gel-like phase, it was discovered that tdp-43 oligomers form in both the deposition and condensation pathways and before solid aggregates emerge (french et al., 2019). furthermore, it has been documented that ⍺-synuclein oligomers were formed right after the monomeric protein underwent phase separation into a hydrogel. in this environment, monomers, oligomers, and fbrils coexist, and the hydrogels trap ⍺-synuclein in a highly cytotoxic state rather than releasing it (kumar et al., 2018). similarly, it was discovered that liquid–liquid phase separation causes a pathogenic conformation and oligomerization in tau and comes before gel formation and subsequently aggregation in vitro (kanaan et al., 2020; wegmann et al., 2018). figure 2: a synopsis of condensation and the amyloid formation deposition pathway. source: biorender structural features understanding the intricate structure of aβ(1-42) oligomer’s is crucial, and in recent times, solid-state nmr-spectroscopy has been used to perform structural analyses on various oligomer preparations of aβ(1-42) oligomer’s and aβ(1-40)oligomer’s (also known as pyroglu-aβ(3/11–40) oligomers)(könig et al., 2021). both on-pathway precursor and off-pathway competitors can be acted upon by the oligomers during the oligomer to mature fibril conversion for aβ42. mostly composed of residue aβ peptide species, these fibrils have wellknown structures. the β-strands of these firbils are oriented perpendicularly to the fibril axis, giving them a general appearance of cross-shaped β structure. two pa ge 19 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 16-28, 2025 hydrophobic β-sheet segments with an in-register orientation and parallel orientation are present in these fibrils, despite their unstructured n-terminus (saha & jana, 2022). in order to examine the biological activities of aβ oligomers and evaluate their physicochemical and structural characteristics, various techniques have been devised to produce oligomers that are relatively stable and do not easily transform into amyloid fibrils. aβ oligomers that are frequently employed in the examination of animal and cellular models of alzheimer’s disease are aβ-derived diffusible ligands, or addls (jang et al., 2023). the toxic conformer of aβ42, with a turn at positions 22/23, and the less toxic conformer, with a turn at positions 25/26, were identified (figure 3a) through systematic proline replacement and analyses using solid-state nuclear magnetic resonance (nmr) spectroscopy and electron spin resonance (esr) to gather information on the secondary structure of aβ42 oligomers and fibrils. the turn at positions 22/23 is one of the important secondary structures of aβ42 for cytotoxicity and aggregative ability, as was previously mentioned. we performed cross-linking of the residues located at aβ42 positions 21/24, 19/26, 17/28, 15/30, and 13/32. an intramolecular disulfide bond at positions 17/28 is present in the aβ42 analogue (figure 3) (matsushima et al., 2022). mixtures of diverse species, varying in size from tiny to large, make up aβos; however, it is still unclear which species are the most toxic (araki, 2023). our findings, along with numerous other in vitro and in vivo investigations, corroborate the notion that aβos cause a range of pathological changes, such as oxidative stress, mitochondrial dysfunction, synaptic deficits, apoptosis, aberrant tau alterations, and cognitive impairments (araki & kametani, 2022) aβ oligomers may be involved in ad pathology in a number of ways, such as neuronal toxicity and excitotoxicity caused by constriction of brain blood vessels due to increased calcium levels. ad can start in vivo and in vitro when small soluble aβ1-42 oligomers cause neurotoxicity. for this reason, it is currently thought that aβ oligomers are more neurotoxic and disease-relevant than aβ fibrils (matuszyk et al., 2021). when aβos binds to different receptors on the surface of neurons, it can interfere with signaling pathways and cause cells to die. aβos recognizes more than 20 receptors, such as β7nachr, p75ntr, prpc, glutamate receptors, and β2-ar. app intracellular domain (aicd) production is decreased and β-secretase activity is inhibited by prpc, an important protein that regulates aβ metabolism in vivo under normal physiological conditions. aβos, on the other hand, binds to prpc and interferes with its normal physiological functions in ad brains (huang & liu, 2020). the majority of scientists believe that aβ oligomers cause synaptotoxicity by activating metabotropic glutamate receptor type 5 (mglur5) and stimulating three kinases that inhibit ltp: c-jun n-terminal protein kinase 1, jnk1; cyclin-dependent kinase 5, cdk5; and p38 mitogen-activated protein kinase, p38 mapk (diociaiuti et al., 2021). the species most closely linked to the pathophysiology of ad are soluble aβ oligomers, which can be found in app transgenic mice, ad patients, vitro, and in vivo in a wide variety of forms. these diverse aβ oligomer types show their neurotoxic effects in ad via multiple distinct mechanisms (madhu & mukhopadhyay, 2021). the toxic influence of aβos has been extensively studied in transgenic animal models, ad brain tissues, and cell cultures. numerous studies have shown that amyloid oligomers of different origins can cause alterations in neurons. aβos isolated from the brains of ad patients or animal models of the disease, as well as synthetic peptides or aβ species secreted in cultured cells (mroczko et al., 2017). in summary, aβos have been observed to trigger tau pathology, axonal transport impairment, loss of neuronal polarity, oxidative stress, endoplasmic reticulum (er) stress, deterioration of synapses, insulin resistance, neuroinflammation, cholinergic impairment, loss of trophic factors, epigenetic modifications, ectopic mitosis, and selective death of nerve cells (cline et al., 2018). chemical regulation of amyloid-β oligomerization alzheimer’s disease and related illnesses are characterized by a pathogenic process called amyloid-beta aggregation. in order to develop effective treatments (cline et al., 2018). understanding how this aggregation is made is crucial. treatment options include gal-201, which is thought figure 3: (a) structure of toxic and less toxic conformers with a turn at positions 22/23 and 25/26, respectively.20 (b) structure of cross-linked aβ42 analogues (1–6). adopted from ref (matsushima et al., 2022) role in alzheimer’s pathology it is widely acknowledged that aβos are important players in the pathogenetic mechanisms of ad because of their capacity to cause neurotoxicity, synaptotoxicity, and neuroinflammation, and because these effects can explain the neuropathological characteristics of ad. pa ge 20 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 16-28, 2025 to bind preferentially to mis-folded aβ1-42 monomers. this high-affinity binding prevents larger aggregates and harmful oligomers from forming by interfering with the aggregation process. through its ability to inhibit aβ aggregation, gal-201 may be able to prevent or slow the onset of amyloid-beta-related neurodegenerative diseases, such as alzheimer’s (russ et al., 2022). there are now multiple strategies to combat the alzheimer’s-related aβ assemblies (fish et al., 2019). immunotherapeutic vaccinations, antibodies, peptides, and nanoparticles are a few of the tactics. many compounds, such as congo red, lpffd, myricetin, melatonin, 9,10-anthraquinone, trehalose, thioflavin t, n-methylated peptides, polyphenol egcg, ibuprofen, naproxen, morin, and particular polyphenolic compounds, have shown promise in blocking aβ aggregation and upsetting beta sheet structures (grasso & danani, 2020). plants like berries and grapes contain a polyphenolic flavonoid called myricetin, which has a variety of biological activities such as antibacterial, anti-inflammatory, and antioxidant properties. inhibition of aβ oligomerization by myricetin has also been demonstrated (araki & kametani, 2022). anti-aβ monoclonal antibodies have the ability to stop aβ monomers from forming fibrillar aggregates in vitro and to change fibrillar aggregates into an amorphous form. it’s interesting to note that these mechanisms’ effectiveness is influenced by antibody concentration (mantile & prisco, 2020) . there have been reports of certain anti-aβ antibodies binding directly to aβ, which either dissolve aβ aggregates in vitro or prevents aβ from oligomerizing and forming fibrils. particular aβ antibodies with specific conformations target pre-existing brain plaques and cause direct in vivo disintegration. according to these results, aβ aggregation may be impacted by the direct interaction of anti-aβ antibodies with aβ in both vitro and in vivo settings (liu et al., 2025). a phase iii clinical trial for four monoclonal anti-aβ antibodies—adecaluzumab, ban2401, gantenerumab, and solanezumab—has begun, according to a recent study. aducanumab is a human igg1 antibody that targets only soluble oligomers and insoluble fibrils of aβ. it was chosen through memory b cell screening in healthy elderly individuals (panza et al., 2025). the fda has recently approved it as an immunotherapy for ad. aducanumab (biib037) is a monoclonal antibody of human igg1 that forms an extended conformation with the n terminus of aβ. it targets aggregates of aβ, including insoluble fibrils and soluble oligomers (song et al., 2022). using molecular docking, the virtual peptide p21 is able to effectively inhibit the aggregation of aβ1-42 proteins, thereby decreasing neurotoxicity (wu et al., 2023). two classes of poly-phenols inhibit the aggregation of aβ in distinct ways. on the one hand, flavonoids led to the formation of spherical, unstructured aggregates and completely inhibited the fibrillation of aβ monomers. conversely, stilbenes suppressed aβ aggregation, although to a much smaller degree. remarkably, the common stilbene resveratrol speed up the production of aβ fibrils (phan et al., 2019). metal ions have been shown to either accelerate or slow down aβ aggregation based on the total metal ion concentration and metal:aβ ratio, as has been covered in multiple review articles. while it has been demonstrated that cu (ii), zn(ii), and ag(i) slow down aβ fibrillization at low metal ion concentrations, aβ aggregation can be stimulated at high concentrations, leading to the formation of amorphous aggregates. figure 4: model for mechanism of action of metal ion-modulated aβ self-assembly transition-metal ions, in particular referring to cu (ii), zn(ii), and ag(i) ions, specifically prevent fibril-end elongation events by forming a seemingly aggregationinert metal-bound aβ complex. inhibition of fibril elongation predicts an enhanced rate of oligomer generation. at high metal ion concentration, other aggregation processes dominate, and amorphous aggregates are formed. (b) an energy diagram shows the concentration-dependent formation of aβ fibrils and amorphous aβ aggregates, where an increased concentration of aβ generally enhances aggregation and the energy barrier toward amorphous aggregate formation is determined by the metal ion concentration. copy from ref (abelein, 2023). metal ions, particularly zn2+ and cu2+, have a strong affinity for binding to aβ, which can facilitate aβ nucleation and aggregation (rana & sharma, 2019). according to a recent study, quinoline-derived halfcurcumin dioxaborine (q-ob) was made in order to identify the early stage of ad by detecting the oligomeric aβ1-42 over monomer and filaments in the complex aβ self-assembly cascade. in ad transgenic mice, q-ob pa ge 21 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 16-28, 2025 demonstrated exceptional blood-brain barrier (bbb) penetrability and in vivo imaging of aβ (an et al., 2024). the studies of anti-aβ 3d6 antibody is said to stop the development of new aβ plaques in the brain, but it doesn’t eliminate those that already exist. additionally, 3d6 inhibited the aggregation of aβ (amano et al., 2023). the relationships between the aβ peptide and the main aβ aggregation inhibitors, geraniin (1), gallic acid (2), and corilagin (5), were investigated using std nmr spectra. this study implies that smaller soluble aβ aggregates are inhibited by corilagin (5), whereas larger insoluble aβ aggregates are inhibited by gallic acid (2). corilagin (5) only interacts with soluble aβ, as demonstrated by this as well (kubo et al., 2022). certain classes, like polyphenols and tetracyclines, can prevent the aggregation of various unrelated amyloidogenic proteins, like islet amyloid polypeptide (iapp), which is linked to type-2 diabetes, and α-synuclein, which is linked to parkinson’s disease. it’s likely that their mechanisms of action are partially overlapped (martinez pomier et al., 2020). many in vitro and in vivo investigations have demonstrated the potential of a variety of natural compounds as therapeutic agents against the progression of ad however, pre-clinical and clinical studies have only demonstrated the efficacy of a small subset of these compounds (andrade et al., 2019). after being extracted from red maples, ginsenalin a (ga) can target aβ42 fibrillogenesis through a variety of mechanisms. specifically, ga can bind to monomers at the early nucleation phase, preventing aβ–aβ associations, and at the later growth phase. according to toxicity tests on sh-sy5y cells, pre-incubation of aβ42 solution with ga results in oligomers that do not interact or disturb the integrity of cellular membranes (pagano et al., 2020). further evidence suggests that the antioxidant ferrulic acid (fa), which is found in plant cell walls, may inhibit the aggregation of aβ (thapliyal et al., 2021). the biomolecular targets linked to alzheimer’s disease (ad) include the nmda receptor, aβ aggregation, ache, and monoamine oxidase (mao) (uddin et al., 2020). (figure 5) shows how harmine and its derivatives bind to numerous targets. harmine treatments decrease scopolamineinduced cognitive impairment in mice and enhance spatial learning, memory in transgenic mice, short-term memory in aged rats, and all of these outcomes. this illustrates how harmine targets different aspects of the disease and has the potential to be a multimodal treatment for alzheimer’s (du et al., 2023). study investigated the potential therapeutic effects of gnrb1, sa, and dmyr on aβ aggregation in ad. our study suggests that since sa, gnrb1, and dmyr target aβ aggregation, they might be appealing treatment options for ad based on the amyloid hypothesis (sharari et al., 2023). by significantly binding to aβ oligomers and monomers, the nanoparticles (np@ sio2@f-sloh) reduce aβ aggregation, as confirmed by the tht fluorescence experiment. this suggests the potential for alzheimer’s disease (ad) treatment. aβ species ranging from aβ42 monomers and oligomers to gd3+-based nps with f-sloh surface functionalization lessen neurotoxicity as well. additionally, the nps effectively stop aβ species from generating reactive oxygen species (ros), indicating that ad treatments may be possible with them (wang et al., 2020). figure 5: chemical structures of harmine and its derivatives. copy from ref (du et al., 2023) stabilization of amyloid-β oligomers a number of tiny compounds have been demonstrated to be useful anti-aggregating agents in the treatment of alzheimer’s disease, including curcumin, resveratrol, ellagic acid, meric decapeptide rk10, and epigallocatechin3-gallate (egcg) (mohammed et al., 2023). the active polyphenol found in green tea, called epigallocatechin-3gallate (egcg), has drawn a lot of interest because of its potential for health benefits, which include anti-oxidation, radical scavenging, metal chelating, anti-carcinogen, antiapoptosis, and anti-inflammatory qualities (mokra et al., 2022). egcg activates the proteolytic pathway of nonamyloidogenic α-secretase, which inhibits the aging process of the brain and lowers aβ levels, according to several studies (bao et al., 2020). apart from its potential to prevent the creation of harmful prefibrillar oligomers, pa ge 22 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 16-28, 2025 egcg has also been proposed as a possible remodeler of preexisting amyloid fibrils. since egcg is unstable and oxidizes quickly to produce a range of products, most research is done at physiological ph (sternke-hoffmann et al., 2020). research has demonstrated that polyphenols can either stop aβ oligomerization from happening or restructure and stabilize aβ oligomers into forms that are safe. to produce unstructured aβ oligomers, egcg inhibits aβ fibrillogenesis. smaller, more amorphous, nontoxic protein aggregates are assembled from freshly formed oligomers with its assistance (el gaamouch et al., 2022). catecholamine neurotransmitters have the ability to maintain aβ in its oligomeric state, according to studies. indeed, aβ oligomer stabilization may involve the cooperation of ne and da. an elevated oligomer concentration is indicated by the aggregated aβ, which is significantly blurred when ne and da are present (allnutt & matera, 2023). figure 6: structures of neurotransmitters examined for stabilization of aβ oligomers. source: adopted from (allnutt & matera, 2023) an antioxidant, anti-inflammatory, and anticancer polyphenolic compound is curcumin. its low bioavailability restricts efficacy despite its possible advantages in alzheimer’s disease (ad) models (momma et al., 2023). enhancing bioavailability and brain translocation, gt863, a derivative of curcumin, was synthesized. together with dual stabilization of aβ and tau aggregation, in vivo studies showed that gt863 could ameliorate cognitive impairment in an ad mouse model. it would seem from this that gt863 is a good candidate for more study in the treatment of ad. figure 7: structures of gt863 and curcumin. source: adopted from ref (zhou et al., 2022) few compounds that can disassemble pre-formed oligomers have progressed to the point of clinical trials, despite the fact that a large number of aβ aggregation inhibitors have been discovered in vitro. trimiprosate, also known as 3-amino-1-propanesulfonic acid; alzhemedtm), has been shown to bind preferentially to pre-fibrillar (monomeric and oligomeric) forms of antibody, thereby preventing their conversion into higher-order oligomers and fibrils (ono & tsuji, 2020). moreover, metal ions control aβ polymorphism, stability, and aggregation. based on our simulations, the stabilizing effect is dependent on the size of the fibrillar oligomer and the type of alkali ion. studies looked at how fibrillar aβ oligomers of different sizes were affected by the three alkali metal ions li+, na+, and k+ (huraskin & horn, 2019). according to recent research, small molecules with anti-oxidative properties, the majority of which come from natural sources, can lessen the neurotoxicity of aβos (araki & kametani, 2022). aβ42 aggregates are more toxic than the more prevalent aβ40 because aβ42 peptides can form three-stranded motifs, which enable them to assemble into poreforming aggregates (ringor barrel-shaped), and the brain environment—specifically, the presence of fatty acids—enhances the formation of these aggregates. we discover that lauric acid stabilizes these aggregates (khatua et al., 2021). degradation of amyloid-β oligomers the degradation of aβ in the brains of ad patients is a crucial area of study to help understand the underlying mechanism of aβ degradation and to shed light on the disease’s pathogenesis (zhang et al., 2020). degrading the levels of accumulated aβ aggregates and inhibiting aβ aggregation are therefore important, and doing so has become essential for the potential application as an appealing therapeutic and preventive strategy for the treatment of ad (cheng et al., 2020). furthermore, there are still few ways to assess the degree of aβ degradation. pa ge 23 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 16-28, 2025 circular dichroism (cd) and the thioflavin-t (tht) fluorescence assay are frequently used to look into how the β-sheet structure in aβ aggregates is being destroyed (gao et al., 2022). by tracking the decrease in aβ concentration over time, the aβ degradation has also been identified using sandwich enzyme-linked immunosorbent assay (elisa) and western blot techniques (rostami et al., 2021). these methods do, however, still have a number of drawbacks, including a high potential for false positive/ negative results or results that are subject to subjectivity, the need for careful handling and accuracy at every stage, and high individual costs associated with the preparation of antibodies and culture media. to measure the enzymatic breakdown of aβ, some peptide mapping investigations are carried out, which involve gathering fractions from liquid chromatography (lc) and then using off-line mass spectrometry (ms) (moracci et al., 2021). studies conducted recently indicate that nattokinase (nk) may play a part in the treatment of diseases related to aβ42 aggregates, like ad, as it can gradually break down aβ42 aggregates at neutral ph and body temperature (chen et al., 2018). after then, aβ is eliminated by non-proteolytic or enzyme-mediated mechanisms. there are currently about 20 different proteases known as aβ degrading enzymes (ades), which mediate the proteolytic degradation of aβ (sikanyika et al., 2019). aβ clearance can be categorized into two main groups: enzymatic and non-enzymatic. enzymatic clearance involves the action of aβ-degrading enzymes (ades), which function as proteases to break down aβ peptides into smaller, less harmful forms. while most degradation is believed to occur within the brain, clearance may also happen in other areas where aβ is found after being cleared from the brain. a variety of ades have been found from different classes of proteases, including metallo-serine, aspartyl, cysteine, and threonine proteases (see table 1). many of these proteases have the ability to break down multiple peptide substrates in various tissue locations (żukowska et al., 2023). neprilysin (nep) and insulindegrading enzyme (ide) are the most well-characterized aβdps. they are both zinc-metalloendopeptidases that are primarily involved in the breakdown of monomeric species, though neprilysin has also been reported to hydrolyze aβ oligomers (de dios et al., 2019). insulindegrading enzyme (ide) is a widely distributed zn2+metalloprotease present in various human tissues and organs. extensive research has suggested a strong link between ide and alzheimer’s disease (ad). clinical and in vivo investigations have consistently shown that reduced ide levels in the brain of ad patients contribute to the advancement of the disease (abramov-harpaz & miller, 2022). enzymes such as endothelinconverting enzyme, neprilysin (nep), insulindegrading enzyme (ide), and matrix metalloproteinase9 show a decrease in the accumulation of aβ. while matrix metalloproteinase-9 enzyme was discovered to degrade both soluble aβ species and abfibers, ide and nep are reported to be able to remove soluble aβ (sahoo et al., 2021). zinc metalloproteases known as endothelinconverting enzymes, or eces, are made by astrocytes, endothelial cells, and neurons. ece-1 and ece-2 exhibit 59% homology and comparable catalytic activities; however, these activities differ in terms of their ideal ph, with ece-1 activity requiring a physiological ph and ece-2 activity requiring an acidic ph (5.5). they were proposed to be involved in the breakdown of monomeric aβ before its secretion, and they mainly break down intracellular aβ (loeffler, 2023). one important protease that breaks down fibrin in blood clots is called plismin (plm). plm, along with tissue-type plasminogen activator (tpa) and urokinase-type plasminogen activator (upa), is one of the three components of plasmin-based thrombolysis (yang et al., 2020). plm is derived from its inactive form, plasminogen (plg). amyloid-beta (aβ) has been shown to be directly degraded by plm, which also targets its monomeric and fibrillar forms while lessening their toxicity. this implies that plm may have a part in resolving aβ-related problems (loeffler, 2023). table 1: presents a list of enzymes that have shown the ability to degrade aβ in laboratory studies. however, the complete characterization of each protein’s biological significance is still incomplete. neprilysin-2 (nep-2) is referred to by various names in the literature, including sep, nl1, mmel1, and ne_plp. aβ degrading enzymes aβ degrading enzymes neprilysin (nep) matrix metalloprotease-14 (mmp-14) endothelin-converting enzyme-1 (ece-1) myelin basic protein (mbp) endothelin-converting enzyme-2 (ece-2) plasmin angiotensin-1 converting enzyme (ace) aminopeptidase a angiotensin converting enzyme-2 (ace-2) mitochondrial presequence protease (prep) insulin-degrading enzyme (ide) acyl peptide hydrolase (apeh) matrix metalloproteinase-2 (mmp-2) cathepsin b matrix metalloproteinase-9 (mmp-9) cathepsin d beta secretase 1 (bace 1) beta secretase 2 (bace 2) 26s proteasome. source: adopted from (żukowska et al., 2023) pa ge 24 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 4(1) 16-28, 2025 conclusion it is concluded that amyloid-beta (aβ) oligomers have important role in alzheimer’s disease (ad) pathogenesis, neuroinflammation and cognitive decline through liquid-liquid phase separation (llps) and dynamic aggregation. this study highlighted progress in targeting aβ oligomers with metal complexes, small molecules and immunotherapies, which make stable nontoxic forms and control aggregation, or regulate degradation. main challenges include the heterogeneity in structure and the desire for blood-brain barrier-permeable therapeutics. 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(2023). molecular basis of selective amyloid‐β degrading enzymes in alzheimer’s disease. the febs journal, 291(14), 2999-3029. https://doi.org/10.1111/ febs.16939 pa ge 1 pa ge 86 american journal of chemistry and pharmacy (ajcp) potent antioxidant agents: dithiocarbamates of ω-substituted (2-naphthyloxy) alkanes sadaf zaidi1*, devdutt chaturvedi2, nitin srivastava1, manisha shukla3 volume 2 issue 2, year 2023 issn: 2834-0116 (online) doi: https://doi.org/10.54536/ajcp.v2i2.1686 https://journals.e-palli.com/home/index.php/ajcp article information abstract received: may 25, 2023 accepted: june 10, 2023 published: july 13, 2023 a series of dithiocarbamates of ω-substituted (2-naphthyloxy) alkanes (4-48) was tested for antioxidant activity by radicals 2,2-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (abts) assay, dpph assay (1,1-diphenyl-2-picryl-hydrazyl), o2 . (net) assay and roo. (trap) assay against curcumin and vitamin c as standard drugs. most of these compounds have shown promising activities, such compounds are 11, 12, 13, 25, 26, 27, 28, 41, 42, and 43. the series was synthesized by the condensation reaction of 2-(2-chloro-alkoxy)-naphthalene with different types of aliphatic, alicyclic, aromatic, heterocyclic primary, as well as secondary amines to develop dithiocarbamates of ω-substituted (2-naphthyloxy) alkanes. keywords amines, antioxidant, dithiocarbamates, condensation, catalyst 1 department of applied chemistry, amity school of applied sciences, amity university uttar pradesh (auup), lucknow campus, lucknow-226028, u. p. india 2 department of chemistry, school of physical & material sciences, mahatma gandhi central university (mgcu), motihari, bihar, india 3 department of chemistry, babu banarasi das national technology & management, lucknow-227105, u. p. india * corresponding author’s e-mail: sadaf.zaidi00@gmail.com introduction dithiocarbamates have procured a very special position in various areas of organic chemistry such as pharmaceuticals (shaw, 2008) intermediates in organic synthesis (halls, 1969) peptide chemistry (greene & wurtz, 2007) and combinatorial chemistry linkage (mayer et al, 1997) organic dithiocarbamates are also used to synthesize structurally varied biologically potent molecules such as antimalarial(yang liu y et al , 2011) anticholinergics, (ozkanli f et al 2010) antimicrobial (ozkanli f et al 2010) antimitotic (bacharaju k et al 2012) antitubercular (horita y et al 2011) antifungal (zou y et al 2014) , anticancer (cao sl et al 2010) antioxidant (zahram m a h et al 2008) , antiprotozoal (coro j et al 2006), antileprosy (marakov v et al 2006) antifolates (cao sl et al 2006) antitubulin(hou x et al 2011), antialzheimer (mohsin ua anti-hiv (he xy et al 2013) antipoliferative (cao sl et al 2013), and anticontraceptives (jangir s et al 2014) active agents (figure 1). a wide range of dithiocarbamates has been found to act as starting material to synthesize figure 1: structurally diverse biologically potent dithiocarbamates pa ge 87 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 86-98, 2023 structurally diverse biological strong synthetic molecules or intermediates such as isothiocyanates (liu p et al 2013), thiourea (halimjani az et al 2009) cynamide (jamir j et al 2012), dithiobenzophene (kienle m et al 2010), glycosides (aucagne v et al 2005) amide (kumar nk et al 2010) dicarboxylates (khalizadeh ma et al 2010) benzimidazole (das p et al 2008), carbamate (tandel sk et al 1993) pyran ( charati fr et al 2012) flavonoids (bahrin lj et al 2012), etc. in recent years, much focus has been made upon the antioxidant activity of dithiocarbamates, keeping in view the high utility of pyrrolidine dithiocarbamate (ptdc) 2 as an inhibitor of nuclear factor-kappa b (moellering d et al 1999) and diethyl dithiocarbamate (dtdc) 3 is widely used both in basic and clinical research (zhu bz et al 2002). other dithiocarbamate compounds like thalidomide sulphur analog 1 (zahran mah et al 2008) and aliphatic amines 4 (orlinski mm et al 1998) were also found to exhibit pronounced antioxidant activity (figure 1&2). based upon our on-going research work of drug designing figure 2: structurally diverse antioxidant dithiocarbamates figure 3: prototype i (comp. no. 4-48) scheme 1: reaction procedure for the synthesis of prototype i: a anhyd. k2co3, dry acetone, reflux, 12-15 h, 98%; b triton b, cs2, dry dmso, amine, 20-30 min. and synthesizing of semisynthetic/natural/synthetic molecules (zaidi s et al 2019), we became inquisitive to explore the “antioxidant activity of dithiocarbamates of ω-substituted (2-naphthyloxy) alkanes (prototype i)”. result and discussion chemistry as already reported, a series of of dithiocarbamates of ω-substituted (2-naphthyloxy) alkanes (4–48) scheme 1 is produced by using different types of alicyclic, aromatic, aliphatic, heterocyclic primary as well as secondary amines. it was noticed that the final production of prototype i (4-48) dithiocarbamates depends on electron releasing impact of the amines such as phenyl ethyl, cyclohexane, n-methyl piperazine, piperidine, and pyrrolidine amine and table 1 illustrates that phenyl propyl amine has a highest yield than primary amines. pa ge 88 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 86-98, 2023 table 1: synthesis designed dithiocarbamates of prototype i (comp. no. 4-48) comp. no. n r1 r2 time (min.) yield (%) 4 1 c4h9 h 35 93 5 1 c6h11 h 30 94 6 1 c8h15 h 40 90 7 1 c10h19 h 40 93 8 1 r1=r2= pyrrolidine 25 96 9 1 r1=r2= piperidine 30 98 10 1 r1=r2= n methyl piperazine 20 98 11 1 r1=r2= morpholine 25 90 12 1 r1=r2= toluidine 25 92 13 1 r1=r2= anisidine 30 92 14 1 r1=r2= cyclohexane 25 95 15 1 ph(ch2) h 30 82 16 1 ph(ch2ch2) h 30 97 17 1 ph(ch2ch2ch2) h 25 95 18 1 r¬1=r2= dibutyl 35 80 19 2 c4h9 h 35 93 20 2 c6h11 h 30 94 21 2 c8h15 h 40 90 22 2 c10h19 h 40 93 23 2 r1=r2= pyrrolidine 25 96 24 2 r1=r2= piperidine 30 98 25 2 r1=r2= n methyl piperazine 20 98 26 2 r1=r2= morpholine 25 90 27 2 r1=r2= toludine 25 92 28 2 r1=r2= anisidine 30 92 29 2 r1=r2= cyclohexane 25 95 30 2 ph(ch2) h 30 82 31 2 ph(ch2ch2) h 30 97 32 2 ph(ch2ch2ch2) h 25 95 33 2 r1=r2= dibutyl 35 80 34 3 c4h9 h 35 93 35 3 c6h11 h 30 94 36 3 c8h15 h 40 90 37 3 c10h19 h 40 93 38 3 r1=r2= pyrrolidine 25 96 39 3 r1=r2= piperidine 30 98 40 3 r1=r2= n methyl piperazine 20 98 41 3 r1=r2= morpholine 20 98 42 3 r1=r2= toludine 25 90 43 3 r1=r2= anisidine 25 92 44 3 r1=r2= cyclohexane 30 92 45 3 ph(ch2) h 25 95 46 3 ph(ch2ch2) h 30 97 47 3 ph(ch2ch2ch2) h 25 95 48 3 r1=r2= dibutyl 35 80 pa ge 89 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 86-98, 2023 biological evaluation trap and net radical scavenging (kienle m et al 2010), dpph (aucagne v et al 2005), and abts assays (kumar nk et al 2010) in vitro were done to examine the series of compounds for antioxidant activities. table 2 illustrates that the results have been normalized using ic50. by varying the alkyl chain and its connected amines, wide range of compounds are produced, the sar of these compounds may be determined. it has been shown that compounds with the three-carbon chain are more potent than those with the two-carbon or four-carbon chain. the 25, 26, 27 and 28 compounds have a greater potency than other compounds due to the three-carbon chain bound to them. the hydrophilicity is responsible for the increased potency of the three-carbon chain. when analysing the effects of different amines groups, we observed that compounds as 12, 13, 26, 27, 28, 41, 42 and 43 having aromatic amines like anisidine and toludine show comparable value to control drugs curcumin and vc. compounds having substituted heterocyclic amines (10, 11, 25, 26, 40 and 41), generated promising results. better results were obtained by substitution of aromatic amines like benzyl amine (15, 30 & 45) when compared with phenyl propyl amine (17, 32 and 47) and phenyl ethyl (16, 31 and 46). table 2: antioxidant activities of prototype i (comp. 4-48) compound dpph ic50 (µm) abts ic50 (µm) trap ic50 (µm) net ic50 (µm) 4 42.78±0.65 88.45±0.67 95.35±0.34 156.76±1.12 5 50.12±0.45 90.54±1.15 98.12±1.12 175.54±0.67 6 30.21±0.65 68.15±0.25 78.67±1.12 126.65±0.67 7 >4820 >4820 >4820 >4820 8 29.21±0.65 62.15±0.25 72.67±1.12 125.65±0.67 9 31.89±0.90 64.45±0.55 74.35±0.85 122.56±0.89 10 28.09±0.48 60.65±0.55 76.54±0.98 120.54±0.45 11 25.45±0.98 85.89±0.78 74.67±0.85 112.45±0.58 12 24.78±0.25 58.34±0.45 74.32±0.98 110.25±0.65 13 22.89±1.05 55.98±1.76 70.56±0.78 105.32±0.89 14 >4820 >4820 >4820 >4820 15 33.35±1.43 66.56±0.43 86.45±0.67 130.24±0.65 16 35.68±0.47 68.24±0.56 88.25±0.65 138.75±0.34 17 38.65±0.90 70.89±0.35 102.34±0.54 142.89±0.67 18 >4820 >4820 >4820 >4820 19 25.45±0.83 82.32±0.78 85.15±0.65 142.32±0.56 20 28.09±0.98 88.09±0.45 88.45±1.35 155.24±0.45 21 25.24±0.90 66.01±0.34 82.45±1.21 120.22±1.34 22 61.25±1.21 175.45±0.65 178.32±0.48 204.32±0.45 23 24.24±0.90 61.01±0.34 80.45±1.21 110.22±1.34 24 26.89±1.05 62.56±1.02 83.62±1.43 118.56±1.25 25 22.65±0.47 57.56±2.01 68.45±0.98 105.24±1.24 26 30.56±2.01 76.98±1.35 85.65±1.28 133.22±1.21 27 15.24±1.85 54.34±0.35 60.65±1.05 92.45±1.89 28 18.56±2.01 54.15±1.47 61.12±1.85 96.78±1.85 29 >4820 >4820 >4820 >4820 30 28.89±0.45 68.98±1.76 76.01±0.78 120.25±1.76 31 32.28±2.06 72.45±2.16 85.65±1.21 125.89±1.32 32 34.45±0.56 88.34±0.18 98.32±1.35 134.24±1.25 33 >4820 >4820 >4820 >4820 34 40.22±0.34 78.58±0.25 86.45±0.56 135.75±0.56 35 45.67±0.52 84.45±0.65 90.12±0.45 140.45±0.65 36 26.14±0.34 62.65±1.12 75.34±0.65 118.34±0.45 37 >4820 >4820 >4820 >4820 38 28.14±0.34 61.65±1.12 76.34±0.65 115.34±0.45 pa ge 90 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 86-98, 2023 39 30.56±0.87 63.25±0.54 78.45±1.12 120.45±0.56 40 25.94±0.65 60.15±0.55 74.34±0.56 110.45±0.46 41 35.18±1.21 68.32±0.85 81.24±0.85 126.54±0.65 42 24.45±0.89 58.34±0.54 70.34±0.98 105.65±0.56 43 21.89±0.75 56.45±0.75 68.54±0.65 102.34±0.45 44 >4820 >4820 >4820 128.89±0.45 45 32.34±0.45 65.32±0.85 80.32±0.65 132.45±0.98 46 34.67±1.12 66.35±0.55 88.45±0.24 145.67±0.56 47 38.55±0.35 68.34±0.54 110.56±0.85 >4820 48 >4820 >4820 >4820 >4820 vc 43.41±1.80 104.47±3.10 103.22±2.22 165.±1.45 curcumin 12.63±0.50 49.55±2.15 55.22±2.10 88.91±1.25 experimental all of the synthesized compounds were detected by melting point, 1h nmr, 13c nmr, and hrms. chemicals have been acquired from fluka, aldrich, and merck chemical firms. bomem mb-104–ftir spectrophotometer has recorded the ir spectra of 4000200 cm-1 range, while ac-300f was used to scan nmrs, nmr (300mhz) is carried out with cdcl3 and tms and other deuterated solvents as an internal standard. “carlo-erba ea 1110-cnno-s” analyser performed the elemental analysis and accepted favourably the measured values. synthetic process for ω-substituted 2-napthyloxy haloalkanes typical procedure measured amount of β-naphthol 1 was poured in dry acetone and later anhydrous k2co3 (10 eqv.) was added into it. for alkylation, 1-bromo-3-chloro propane 2 (2.5 eqv.) was also included and the reaction mixture was allowed to reflux for 12-15h. the progress of reaction has been recorded by tlc and a new less polar spot appeared on the tlc which indicated the formation of a product. the filtrate of the reaction was extracted thrice with ethyl acetate. the organic layer which afforded the required compound 3 was separated, dried over anhydrous na2so4. different spectroscopic, as well as analytical methods, have verified compound 3. general procedure for dithiocarbamates of prototype i (4-48) synthesis measured amount of required amine was dissolved in dry dmso. to this triton-b, as well as cs2, were also introduced in the reaction mixture drop by drop along with constant stirring for around 15 min. the reaction mixture was later mixed with compound 3, which was stirred for around 20-40 minutes. the formation of desired product was monitored by tlc. at the completion of the reaction, the resultant mixture was extracted thrice by using ethyl acetate. the organic layer has been isolated as well as dried over anhydrous na2so4 which gave the product, i.e., the prototype i (compound no. 4-48). experimental methods synthesis a general method to prepare ω naphthyloxy halo alkanes (3a–c) in dry acetone (200 ml), the mixture was refluxed for around 12–15 hours comprising of β-naphthol 1 (20 grams, 0.14 mol), anhydrous k2co3 (in excess amount of 100 g) and bromochloroalkane 2 (0.14 mol). the reaction mixture was purified, and the filtrate was condensed to obtain the oily compound crystallized by using benzenehexane, giving the pure desired compound which is colourless crystals. 2-(2-naphthyloxy)-1-chloroethane (3a) yield: 27.5 g (96%); mp: 94 0c; ir (kbr, cm-1): ν = 1455 (ar), 1508 (ar), 1585 (ar), 2878 (ch), 2927 (ch); 1h nmr (400 mhz, cdcl3): δ = 3.81 (t, 2h, ch2cl), 4.26 (t, 2h, och2), 6.97–7.64 (m, 7h, ar–h); 13c nmr (100 mhz, cdcl3): δ = 45.3, 75.1, 105.8, 118.6, 123.6, 126.4, 129.5, 134.5, 157.7 ppm; mass (eims): m/z = 206; analysis: c12h11clo, calcd: c, 69.74; h, 5.36; obsd: c, 70.04; h, 5.66%. 3-(2-naphthyloxy)-1-chloropropane (3b) yield: 29.7 g (97%); mp: 980c; ir (kbr, cm-1): ν = 1461 (ar), 1512 (ar), 1596 (ar), 2855 (ch), 2940 (ch) cm-1; 1h nmr (400 mhz, cdcl3): δ = 2.27– 2.33 (m, 2h, ch2), 3.80 (t, 2h, ch2cl), 4.25 (t, 2h, och2), 7.12–7.77 (m, 7h, ar–h) ppm; mass (eims): m/z = 220; analysis: c13h13clo, calcd: c, 70.75; h, 5.94; obsd: c, 70.79; h, 6.21%. 4-(2-naphthyloxy)-1-chlorobutane (3c) yield: 32 g (98%); mp: 1120c; ir (kbr, cm-1): ν = 1464 (ar), 1510 (ar), 1599 (ar), 2887 (ch), 2941 (ch); 1h nmr (cdcl3): δ = 2.15–2.20 (m, 4h, ch2ch2), 3.79 (t, 2h, ch2cl), 4.24 (t, 2h, och2), 7.13–7.78 (m, 7h, ar– h) ppm; mass (eims): m/z = 234; analysis:c14h15clo, a general method to prepare dithiocarbamates of ω-substituted (2-naphthyloxy) alkanes a mixture of dry dmso of 35 ml along with desired amines (0.6 ml, 5 m mole) comprising of carbon pa ge 91 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 86-98, 2023 disulphide (in excess amount 3 ml) and triton-b (0.9 ml, 4 m mole) system. at room temperature, the reaction has been agitated for 1 h and finally (2-naphthyloxy)1-chloroalkanes (0.5 grams, 2 m mole) was added. the reaction was allowed to proceed until they were complete (2 hours) under tlc monitoring. the final reaction mixture was poured into 50 ml of distilled water and extraction was done thrice by using ethyl acetate. the organic layer was segregated as well as dried over anhydrous sodium sulphate, then condensed to produce the ω-substituted dithiocarbamate (2-naphthyloxy) alkanes (4-48). the compound was generated as a yellow solid. butyl-dithiocarbamic acid-2-(napthalen-2-yloxy)ethyl ester (4) yield: 0.73 g (93.5 %); m.p.: 106°c; ir (kbr, νmax, cm-1): 661 (c-s), 1114 (c=s), 1454 (ar), 1511 (ar), 1612 (ar), 2864 (ch), 2936 (ch), 3390 (nh); 1 h nmr (cdcl3): δ = 0.920.96 (t, 3h, ch3), 1.30-1.34 (m, 2h, ch2ch3), 1.531.56 (m, 2h, ch2.ch2.ch3), 2.0 (bs, h, nh), 2.62-2.64 (m, 2h, nhch2), 3.28-3.32 (t, 2h, ch2-s-c=s), 4.71-4.74 (t, 2h, ch2-o-naphthyl), 6.97-7.64 (m, 7h, ar-h of naphthyloxy); mass: m/e 319; analysis: c17h21nos2, calcd. (%): c, 63.91, h 6.63, n, 4.38; obsd. (%): c, 64.19, h, 6.49, n, 4.24. butyl-dithiocarbamic acid-3-(naphthalene-2-yloxy) propyl ester (5) yield: 0.73 g (93.5 %); m.p.: 106°c; ir (kbr, νmax, cm-1): 661 (c-s), 1115 (c=s), 1454 (ar), 1511 (ar), 1610 (ar), 2864 (ch), 2935 (ch), 3390 (nh); 1 h nmr (cdcl3): δ = 0.93-0.96 (t, 3h, ch3), 1.33-1.35 (m, 2h, ch2ch3), 1.531.55 (m, 2h, ch2.ch2.ch3), 2.0 (bs, h, nh), 2.62-2.65 (m, 2h, nhch2), 2.84-2.86 (t, 2h, ch2-s-c=s), 2.35-2.39 (m, 2h, ch2ch2ch2) 4.01-4.05 (t, 2h, ch2-onaphthyl), 6.96-7.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 333; analysis: c18h23nos2, calcd. (%): c, 65.91, h 6.83, n, 4.38, s, 19.15, obsd. (%): c, 65.82, h, 6.95, n, 4.20, s, 19.23 %: o, 4.80. butyl-dithiocarbamic acid-4-(naphthalene-2-yloxy) butyl ester (6) yield: 0.73 g (93.5 %); m.p.: 106 °c; ir (kbr, νmax, cm1): 661 (c-s), 1114 (c=s), 1454 (ar), 1511 (ar), 1612 (ar), 2864 (ch), 2936 (ch), 3390 (nh); 1 h nmr (cdcl3): δ = 0.93-0.96 (t, 3h, ch3), 1.33-1.35 (m, 2h, ch2ch3), 1.531.55 (m, 2h, ch2.ch2.ch3), 2.0 (bs, h, nh), 2.62-2.66 (m, 2h, nhch2), 2.85-2.87 (t, 2h, ch2s-c=s), 1.92-1.96 (m, 2h, ch2ch2ch2), 1.68-1.71 (m, 2h, ch2ch2ch2), 4.00-4.03 (t, 2h, ch2-o-naphthyl), 6.96-7.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 347.14; analysis: c19h25nos2, calcd. (%): c, 65.60, h 7.22, n, 4.58; s, 18.42 %: o, 4.58. obsd. (%): c, 65.66, h, 7.25, n, 4.60, s, 18.45 %: o, 4.60. hexyl-dithiocarbamic acid 2 -(napthalen-2-yloxy)ethyl ester (7) yield: 0.8 g (96.4 %); m.p.: 119 °c; ir (kbr, νmax, cm-1): 665 (c-s), 1116 (c=s), 1475 (ar), 1514 (ar), 1602 (ar), 2875 (ch), 2936 (ch), 3394 (nh); 1 h nmr (cdcl3): δ = 0.920.95 (t, 3h, ch3), 1.27-1.29 (m, 4h, ch2ch2ch2ch3 of hexyl group), 1.31-1.35 (m, 2h, ch2ch3 of hexyl group), 1.52-1.56 (m, 2h, ch2.ch2.ch3 of hexyl group), 2.0 (bs, h, nh), 2.36-2.40 (m, 2h, naphthyl-o-ch2ch2ch2-s-c=s), 2.63-2.65 (m, 2h, nhch2), 3.24-3.29 (t, 2h, ch2-s-c=s), 4.68-4.73 (t, 2h, ch2-o-naphthyl), 6.96-7.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 347.14; analysis: c19h25nos2, calcd. (%): c, 66.44, h 7.53, n, 3.87, 18.45 %: o, 4.60. obsd. (%): c, 65.66, h, 7.25, n, 4.03, s, 18.45 %: o, 4.60. hexyl-dithiocarbamic acid-3-(naphthalene-2-yloxy) propyl ester (8) yield: 0.8 g (96.4 %); m.p.: 119°c; ir (kbr, νmax, cm-1): 662 (c-s), 1116 (c=s), 1475 (ar), 1514 (ar), 1602 (ar), 2875 (ch), 2936 (ch), 3394 (nh); 1 h nmr (cdcl3): δ = 0.920.94 (t, 3h, ch3), 1.27-1.29 (m, 4h, ch2ch2ch2ch3 of hexyl group), 1.31-1.35 (m, 2h, ch2ch3 of hexyl group), 1.52-1.58 (m, 2h, ch2.ch2.ch3 of hexyl group), 2.0 (bs, h, nh), 2.34-2.42 (m, 2h, naphthyl-o-ch2ch2ch2-s-c=s), 2.63-2.66 (m, 2h, nhch2), 2.81-2.86 (t, 2h, ch2-s-c=s), 4.02-4.05 (t, 2h, ch2-o-naphthyl), 6.97-7.64 (m, 7h, ar-h of naphthyloxy); mass: m/e 361; analysis: c20h27nos2, calcd. (%): c, 66.44, h 7.53, n, 3.87, obsd. (%): c, 66.75, h, 7.38, n, 3.71. hexyl-dithiocarbamic acid-4-(naphthalene-2-yloxy) butyl ester (9) yield: 0.72 g (98 %); m.p.: 129 °c; ir (kbr, νmax, cm-1): 669 (c-s), 1116 (c=s), 1475 (ar), 1514 (ar), 1610 (ar), 2875 (ch), 2936 (ch), 3410 (nh); 1 h nmr (cdcl3): δ = 0.92-0.96 (t, 3h, ch3), 1.25-1.29 (m, 4h, ch2ch2ch2ch3 of hexyl group), 1.30-1.34 (m, 2h, ch2ch3 of hexyl group), 1.53-1.56 (m, 2h, ch2ch2ch2.ch2.ch3 of hexyl group), 1.711.73 (m, 2h, naphthyl-o-ch2ch2), 1.94-1.96 (m, 2h, sch2ch2), 2.0 (bs, h, nh), 2.62-2.64 (m, 2h, nhch2), 2.822.86 (t, 2h, ch2-s-c=s), 4.024.06 (t, 2h, ch2-o-naphthyl), 6.95-7.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 375; analysis: c21h29nos2, calcd. (%): c, 67.15, h 7.78, n, 3.73, obsd. (%): c, 67.59, h, 7.56, n, 3.51. octyl-dithiocarbamic acid-2-(napthalen-2-yloxy) ethyl ester (10) yield: 0.85 g, (96.2 %); m.p.: 172 °c; ir (kbr, νmax, cm-1): 667 (c-s), 1120 (c=s), 1475 (ar), 1521 (ar), 1612 (ar), 2886 (ch), 2941 (ch), 3399 (nh); 1 h nmr (cdcl3): δ = 0.92-0.94 (t, 3h, ch3), 1.27-1.29 (m, 8h, ch2ch2ch2ch2ch2ch3 of octyl group), 1.32-1.34 (m, 2h, ch2ch3 of octyl group), 1.53-1.56 (m, 2h, ch2ch2n of n-octyl group), 2.0 (bs, h, nh), 2.62-2.64 (m, 2h, nhch2), 3.25-3.29 (t, 2h, ch2-sc=s), 4.01-4.04 (t, 2h, ch2-o-naphthyl), 6.95-7.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 375.17; analysis: c21h29nos2, calcd. (%): c, 67.15, h, 7.78, n, 3.73; o, 4.22; s, 17.04. obsd. (%): c, 67.15, h, 7.78, n, 3.73, o, 4.26; s, 17.07. pa ge 92 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 86-98, 2023 octyl-dithiocarbamic acid -3-(naphthalene-2-yloxy) propyl ester (11) yield: 0.85 g, (96.2 %); m.p.: 172 °c; ir (kbr, νmax, cm1): 667 (c-s), 1120 (c=s), 1472 (ar), 1521 (ar), 1612 (ar), 2884 (ch), 2941 (ch), 3399 (nh); 1 h nmr (cdcl3): δ = 0.92-0.94 (t, 3h, ch3), 1.27-1.29 (m, 8h, ch2ch2ch2ch2 ch2ch3 of octyl group), 1.30-1.32 (m, 2h, ch2ch3 of octyl group), 1.53-1.56 (m, 2h, ch2ch2n of noctyl group), 2.0 (bs, h, nh), 2.38-2.42 (m, 2h, naphthyloch2ch2ch2-s-c=s), 2.63-2.66 (m, 2h, nhch2), 2.83-2.87 (t, 2h, ch2-s-c=s), 4.01-4.04 (t, 2h, ch2-onaphthyl), 6.987.66 (m, 7h, ar-h of naphthyloxy); mass: m/e 389; analysis: c22h31nos2, calcd. (%): c, 67.62, h, 8.02, n, 3.59; obsd. (%): c, 67.89, h, 7.90, n, 3.44. octyl-dithiocarbamic acid -4-(naphthalene-2-yloxy) butyl ester (12) yield: 0.86 g (94 %); m.p.: 156 °c; ir (kbr, νmax, cm1): 664 (c-s), 1108 (c=s), 1463 (ar), 1514 (ar), 1602 (ar), 2862 (ch), 2926 (ch), 3392 (nh); 1 h nmr (cdcl3): δ = 0.930.95 (t, 3h, ch3), 1.27-1.29 (m, 8h, ch2ch2ch2ch2ch2ch3 of octyl group), 1.30-1.34 (m, 2h, ch2ch3 of octyl group), 1.52-1.57 (m, 2h, ch2. ch2.n), 2.0 (bs, h, nh), 2.63-2.67 (m, 2h, nhch2), 3.26-3.32 (t, 2h, ch2-s-c=s), 4.71-4.74 (t, 2h, ch2-onaphthyl), 6.96-7.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 375; analysis: c21h29nos2, calcd. (%): c, 67.15, h 7.78, n, 3.73, obsd. (%): c, 67.54, h, 7.56, n, 3.56. decyl-dithiocarbamic acid-2-(napthalen-2-yloxy)ethyl ester (13) yield: 0.86 g (94 %); m.p.: 156 °c; ir (kbr, νmax, cm1): 663 (c-s), 1108 (c=s), 1465 (ar), 1511 (ar), 1605 (ar), 2862 (ch), 2926 (ch), 3392 (nh); 1 h nmr (cdcl3): δ = 0.92-0.94 (t, 3h, ch3), 1.27-1.29 (m, 10h, ch2ch2ch2ch2ch2ch2ch2ch3 of decyl group), 1.311.35 (m, 2h, ch2ch3 of decyl group), 1.53-1.56 (m, 2h, ch2ch2n of n-decyl group), 2.01 (bs, h, nh), 2.63-2.65 (m, 2h, nhch2), 3.253.29 (t, 2h, ch2-sc=s), 4.68-4.71 (t, 2h, ch2-o-naphthyl), 6.97-7.64 (m, 7h, ar-h of naphthyloxy); mass: m/e 403.25; analysis: c23h33nos2, calcd. (%): c, 68.40, h 8.20, n, 3.90, o, 3.92, s, 15.86; obsd. (%): c, 68.44, h, 8.24, n, 3.96. o, 3.96, s, 15.89. decyl-dithiocarbamic acid-3-(naphthalene-2-yloxy) propyl ester (14) yield: 0.86 g (94 %); m.p.: 156 °c; ir (kbr, νmax, cm1): 664 (c-s), 1109 (c=s), 1462 (ar), 1513 (ar), 1602 (ar), 2863 (ch), 2925 (ch), 3392 (nh); 1 h nmr (cdcl3): δ = 0.92-0.95 (t, 3h, ch3), 1.27-1.29 (m, 10h, ch2ch2ch2ch2ch2ch2ch2ch3 of decyl group), 1.311.35 (m, 2h, ch2ch3 of decyl group), 1.53-1.56 (m, 2h, ch2ch2n of n-decyl group), 2.0 (bs, h, nh), 2.62-2.66 (m, 2h, nhch2), 2.84-2.87 (t, 2h, ch2-s-c=s), 2.34-2.38 (m, 2h, ch2ch2), 4.68-4.71 (t, 2h, ch2-o-naphthyl), 6.96-7.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 417.67; analysis: c24h35nos2, calcd. (%): c, 69, h 8.40, n, 3.35, o, 3.80, s, 15.31. o, 3.83, s, 15.35. obsd. (%): c, 69.02, h, 8.45, n, 3.35, o, 3.83, s, 15.35. decyl-dithiocarbamic acid-4-(naphthalene-2-yloxy) butyl ester (15) yield: 0.86 g (94 %); m.p.: 156 °c; ir (kbr, νmax, cm1): 663 (c-s), 1107 (c=s), 1462 (ar), 1510 (ar), 1606 (ar), 2865 (ch), 2926 (ch), 3392 (nh); 1 h nmr (cdcl3): δ = 0.92-0.94 (t, 3h, ch3), 1.27-1.29 (m, 10h, ch2ch2ch2ch2ch2ch2ch2ch3 of decyl group), 1.30-1.34 (m, 2h, ch2ch3 of decyl group), 1.52-1.58 (m, 2h, ch2ch2n of n-decyl group), 2.0 (bs, h, nh), 2.62-2.66 (m, 2h, nhch2), 2.842.87 (t, 2h, ch2-sc=s), 1.93-1.95 (m, 2h, ch2ch2ch2), 1.67-1.71 (m, 2h, ch2ch2ch2),4.68-4.71 (t, 2h, ch2-onaphthyl), 6.977.64 (m, 7h, ar-h of naphthyloxy); mass: m/ e 431.70; analysis: c25h37nos2, calcd. (%): c, 69.52, h 8.63, n, 3.21, o, 3.70, s, 14.82. obsd. (%): c, 69.56, h, 8.64, n, 3.24, o, 3.71, s, 14.86. pyrollidine-dithiocarbamic acid-2-(napthalen-2yloxy) ethyl ester (16) yield: 0.62 g (80.8 %); m.p.: 79 °c; ir (kbr, νmax, cm-1): 657 (c-s), 1106 (c=s), 1454 (ar), 1502 (ar), 1600 (ar), 2863 (ch), 2925 (ch); 1 h nmr (cdcl3): δ = 1.58-1.60 (m, 4h, ch2 of pyrrolidine ring), 2.8 (t, 4h, ch2n of pyrrolidine ring (2h, ch2-s-c=s), 4.71-4.73 (t, 2h, ch2o-naphthyl), 6.96-7.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 317; analysis: c17h19nos2, calcd. (%): c, 64.32, h, 6.03, n, 4.41, obsd. (%): c, 63.85, h, 6.29, n, 4.64. pyrollidine-dithiocarbamic acid-3-(napthalen-2yloxy) propyl ester (17) yield: 0.63 g (83.2 %); m.p.: 86 °c; ir (kbr, νmax, cm1): 672 (c-s), 1124 (c=s), 1475 (ar), 1524 (ar), 1605 (ar), 2885 (ch), 2926 (ch); 1 h nmr (cdcl3): δ = 1.57-1.61 (m, 4h, ch2 of pyrolidine ring), 2.35-2.38 (m, 2h, naphthyl-o-ch2ch2ch2-s-c=s), 2.8 (t, 4h, ch2n of pyrolidine ring), 2.82-2.86 (t, 2h, ch2-s-c=s), 4.024.04 (t, 2h, ch2-o-naphthyl), 6.96-7.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 331; analysis: c18h21nos2, calcd. (%): c, 65.22, h, 6.39, n, 4.23, obsd. (%): c, 65.63, h, 6.12, n, 4.01. pyrollidine dithiocarbamic acid 4 (napthalen 2 yloxy) butyl ester (18) yield: 0.64 g (86.5 %); m.p.: 95 °c; ir (kbr, νmax, cm-1): 679 (c-s), 1125 (c=s), 1484 (ar), 1525 (ar), 1610 (ar), 2885 (ch), 2947 (ch); 1 h nmr (cdcl3): δ = 1.56-1.60 (m, 4h, ch2 of pyrolidine ring), 1.71-1.72 (m, 2h, naphthyloch2ch2), 1.95-1.98 (m, 2h, s-ch2ch2), 2.8 (t, 4h, ch2n of pyrolidine ring), 2.84-2.88 (t, 2h, ch2-s-c=s), 4.024.05 (t, 2h, ch2-o-naphthyl), 6.97-7.64 (m, 7h, ar-h of naphthyloxy); mass: m/e 345; analysis: c19h23nos2, calcd. (%): c, 66.09, h, 6.57, n, 4.15, obsd. (%): c, 66.57, h, 6.32, n, 3.86. pa ge 93 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 86-98, 2023 piperidine-1-dithiocarbamic acid-2-(napthalen-2yloxy) ethyl ester (19) yield: 0.66 g (82.5 %); m.p.: 89 °c; ir (kbr, νmax, cm-1): 660 (c-s), 1110 (c=s), 1461 (ar), 1509 (ar), 1605 (ar), 2859 (ch), 2926 (ch); 1 h nmr (cdcl3): δ = 1.481.50 (m, 6h, ch2 of piperidine ring), 2.7 (t, 4h, ch2n of piperidine ring), 3.28-3.30 (t, 2h, ch2-s-c=s), 4.714.73 (t, 2h, ch2-o-naphthyl), 6.95-7.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 331; analysis: c18h21nos2, calcd. (%): c, 65.22, h 6.39, n, 4.23, obsd. (%): c, 65.73, h, 6.13, n, 3.98. piperidine-1-dithiocarbamic acid-3-(napthalen-2-yloxy) propyl ester (20) yield: 0.66 g (82.5 %); m.p.: 89 °c; ir (kbr, νmax, cm-1): 662 (c-s), 1112 (c=s), 1461 (ar), 1509 (ar), 1605 (ar), 2859 (ch), 2927 (ch); 1 h nmr (cdcl3): δ = 1.48-1.50 (m, 6h, ch2 of piperidine ring), 2.7 (t, 4h, ch2n of piperidine ring), 2.82-2.86 (t, 2h, ch2-s-c=s), 2.35-2.38 (m, 2h, ch2ch2), 4.01-4.04 (t, 2h, ch2-o-naphthyl), 6.957.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 392.54; analysis: c19h24n2o2s3, calcd. (%): c, 58.22, h 6.20, n, 7.23, obsd. (%): c, 58.14.73, h, 6.16, n, 7.14, o, 12.23, s, 16.13. piperidine-1-dithiocarbamic acid-4-(napthalen-2yloxy) butyl ester (21) yield: 0.66 g (82.5 %); m.p.: 89 °c; ir (kbr, νmax, cm-1): 660 (c-s), 1110 (c=s), 1461 (ar), 1509 (ar), 1605 (ar), 2859 (ch), 2925 (ch); 1 h nmr (cdcl3): δ = 1.48-1.50 (m, 6h, ch2 of piperidine ring), 2.7 (t, 4h, ch2n of piperidine ring), 2.83-2.86 (t, 2h, ch2-s-c=s), 1.92-1.96 (m, 2h, ch2ch2), 1.68-1.71 (m, 2h, ch2ch2), 4.00-4.02 (t, 2h, ch2-o-naphthyl), 6.96-7.65 (m, 7h, ar-h of naphthyloxy); mass: m/e 406.56; analysis: c20h26n2o3s2, calcd. (%): c, 59.12, h 6.49, n, 6.82, o, 11.82, s, 15.75. obsd. (%): c, 59.08, h, 6.45, n, 6.89, o, 11.89, s, 15.79. 4methyl piperazine-dithiocarbamic acid 2 (napthalen2 yloxy) ethyl ester (22) yield: 0.66 g (82.5 %); m.p.: 89 °c; ir (kbr, νmax, cm1): 662 (c-s), 1112 (c=s), 1461 (ar), 1509 (ar), 1605 (ar), 2859 (ch), 2926 (ch); 1 h nmr (cdcl3): δ = 2.24-2.27 (s, 3h, ch3 of methyl piperazine ring), 2.442.48 (t, ch2n of piperazine ring), 2.62-2.65 (t, ch2n of piperazine ring), 2.0 (bs, h, nh), 3.25-3.29 (t, 2h, ch2s-c=s), 4.684.71 (t, 2h, ch2-o-naphthyl), 6.96-7.65 (m, 7h, ar-h of naphthyloxy); mass: m/e 361; analysis: c18h23n3os2, calcd. (%): c, 59.84, h 6.39, n, 11.60, o, 4.40, s, 17.72 % obsd. (%): c, 59.80, h, 6.41, n, 11.62 % o, 4.43, s, 17.74. 4methyl piperazine dithiocarbamic acid 3 (napthalen2-yloxy)propyl ester (23) yield: 0.66 g (82.5 %); m.p.: 89 °c; ir (kbr, νmax, cm-1): 660 (c-s), 1110 (c=s), 1461 (ar), 1509 (ar), 1605 (ar), 2859 (ch), 2926 (ch); 1 h nmr (cdcl3): δ = 2.24-2.27 (s, 3h, ch3 of methyl piperazine ring), 2.44-2.48 (t, ch2n of piperazine ring), 2.62-2.65 (t, ch2n of piperazine ring), 2.0 (bs, h, nh), 2.84-2.87 (t, 2h, ch2-sc=s), 2.342.38 (m, 2h, ch2ch2), 4.68-4.71 (t, 2h, ch2-onaphthyl), 6.96-7.63 (m, 7h, ar-h of naphthyloxy); mass: m/e 375.55; analysis: c19h25n3os2, calcd. (%): c, 60.72, h 6.69, n, 11.23, o, 4.22, s, 17.05 % obsd. (%): c, 60.76, h, 6.71, n, 11.19, o, 4.26, s, 17.08. 4 methyl piperazine dithiocarbamic acid 4 (napthalen2-yloxy)butyl ester (24) yield: 0.66 g (82.5 %); m.p.: 89 °c; ir (kbr, νmax, cm1): 662 (c-s), 1112 (c=s), 1461 (ar), 1509 (ar), 1605 (ar), 2859 (ch), 2925 (ch); 1 h nmr (cdcl3): δ = 2.24-2.27 (s, 3h, ch3 of methyl piperazine ring), 2.442.48 (t, ch2n of piperazine ring), 2.62-2.65 (t, ch2n of piperazine ring), 2.0 (bs, h, nh), 2.84-2.86 (t, 2h, ch2-sc=s), 1.941.96 (m, 2h, ch2ch2ch2), 1.68-1.71 (m, 2h, ch2ch2ch2), 4.68-4.71 (t, 2h, ch2-o-naphthyl), 6.967.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 389.58; analysis: c20h27n3os2, calcd. (%): c, 61.64, h 6.93, n, 10.73, o, 4.08, s, 16.42 % obsd. (%): c, 61.66, h, 6.99, n, 10.79, o, 4.11, s, 16.46. morpholine 4 dithiocarbamic acid 2 (napthalen-2 yloxy)ethyl ester (25) yield: 0.66 g (82.5 %); m.p.: 89 °c; ir (kbr, νmax, cm-1): 660 (c-s), 1110 (c=s), 1461 (ar), 1509 (ar), 1605 (ar), 2859 (ch), 2926 (ch); 1 h nmr (cdcl3): δ = 3.62-3.67 (t, 2h, ch2 of morphaline ring), 2.34-2.37 (t, ch2n of morphaline ring), 2.35-2.40 (s, ch2scs), 2.84-2.86 (t, 2h, ch2-s-c=s), 2.34-2.38 (m, 2h, ch2ch2), 4.68-4.71 (t, 2h, ch2-o-naphthyl), 6.95-7.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 382.50; analysis: c17h22n2o4s2, calcd. (%): c, 53.35, h 5.79, n, 7.30, o, 16.70, s, 16.72 obsd. (%): c, 53.38, h, 5.80, n, 7.32, o, 16.73, s, 16.77. morpholine 4 dithiocarbamic acid 3 (napthalen-2 yloxy)propyl ester (26) yield: 0.66 g (82.5 %); m.p.: 89 °c; ir (kbr, νmax, cm-1): 662 (c-s), 1112 (c=s), 1461 (ar), 1509 (ar), 1605 (ar), 2859 (ch), 2926 (ch); 1 h nmr (cdcl3): δ = 3.62-3.67 (t, 2h, ch2 of morphaline ring), 2.34-2.37 (t, ch2n of morphaline ring), 2.35-2.40 (s, ch2-s-c=s), 1.95-1.99 (m, 4h, ch2ch2), 4.68-4.71 (t, 2h, ch2-o-naphthyl), 6.967.65 (m, 7h, ar-h of naphthyloxy); mass: m/e 396.52; analysis: c18h24n2o4s2, calcd. (%): c, 45.50, h 6.09, n, 7.10, o, 16.10, s, 16.15. obsd. (%): c, 45.52, h, 6.10, n, 7.06 % o, 16.14, s, 16.17. morpholine 4 dithiocarbamic acid 4 (napthalen-2 yloxy)butyl ester (27) yield: 0.66 g (82.5 %); m.p.: 89 °c; ir (kbr, νmax, cm-1): 660 (c-s), 1110 (c=s), 1461 (ar), 1509 (ar), 1605 (ar), 2859 (ch), 2926 (ch); 1 h nmr (cdcl3): δ = 3.623.67 (t, 2h, ch2 of morphaline ring), 2.34-2.37 (t, ch2n of morphaline ring), 2.35-2.40 (s, ch2scs), 3.25-3.28 (t, 2h, ch2-s-c=s), 1.94-1.96 (m, 2h, ch2ch2ch2), 1.68-1.71 (m, 2h, ch2ch2ch2), 4.68-4.71 (t, 2h, ch2pa ge 94 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 86-98, 2023 o-naphthyl), 6.967.63 (m, 7h, ar-h of naphthyloxy); mass: m/e 410.55; analysis: c19h26n2o4s2, calcd. (%): c, 55.55, h 6.39, n, 6.30, o, 15.55, s, 15.60. obsd. (%): c, 55.58, h, 6.35, n, 6.85, o, 15.59, s, 15.62. p -tolyl dithiocarbamic acid-2-(napthalen-2-yloxy) ethyl ester (28) yield: 0.76 g (88.4 %); m.p.: 137 °c; ir (kbr, νmax, cm1): 660 (c-s), 1111 (c=s), 1454 (ar), 1502 (ar), 1602 (ar), 2851 (ch), 2928 (ch), 3388 (nh); 1 h nmr (cdcl3): δ = 2.34 (s, 3h, ch3), 3.28-3.30 (t, 2h, ch2-s-c=s), 4.0 (bs, h, nh), 4.70-4.72 (t, 2h, ch2-o-naphthyl), 6.35-7.62 (m, 11h, ar-h of naphthyloxy and phenyl ring); mass: m/e 353; analysis: c20h19nos2, calcd. (%): c, 67.89, h, 5.45, n, 3.99, obsd. (%): c, 67.63, h, 5.58, n, 4.12. p-tolyl-dithiocarbamic acid-3-(napthalen-2-yloxy) propyl ester (29) yield: 0.76 g (88.4 %); m.p.: 137 °c; ir (kbr, νmax, cm-1): 662 (c-s), 1111 (c=s), 1454 (ar), 1502 (ar), 1601 (ar), 2851 (ch), 2928 (ch), 3388 (nh); 1 h nmr (cdcl3): δ = 2.34 (s, 3h, ch3), 3.28-3.30 (t, 2h, ch2-s-c=s), 4.0 (bs, h, nh), 2.35-2.40 (s, ch2-s-c=s), 1.95-1.99 (m, 4h, ch2ch2), 4.68-4.71 (t, 2h, ch2-o-naphthyl), 6.34-7.64 (m, 11h, ar-h of naphthyloxy and phenyl ring); mass: m/e 367.53; analysis: c21h21nos2, calcd. (%): c, 68.60, h, 5.72, n, 3.80, o, 4.31, s, 17.45 % obsd. (%): c, 68.63, h, 5.76, n, 3.81 % o, 4.35, s, 17.45. p -tolyl dithiocarbamic acid-4-(napthalen-2-yloxy) butyl ester (30) yield: 0.76 g (88.4 %); m.p.: 137 °c; ir (kbr, νmax, cm-1): 662 (c-s), 1111 (c=s), 1454 (ar), 1502 (ar), 1601 (ar), 2851 (ch), 2928 (ch), 3388 (nh); 1 h nmr (cdcl3): δ = 2.34 (s, 3h, ch3), 3.28-3.30 (t, 2h, ch2-s-c=s), 4.0 (bs, h, nh), 2.35-2.40 (s, ch2scs), 3.25-3.27 (t, 2h, ch2s-c=s), 1.94-1.96 (m, 2h, ch2ch2ch2), 1.68-1.71 (m, 2h, ch2ch2ch2), 4.71-4.73 (t, 2h, ch2-o-naphthyl), 6.35-7.63 (m, 11h, ar-h of naphthyloxy and phenyl ring); mass: m/e 381.55; analysis: c22h23nos2, calcd. (%): c, 69.20, h, 6.06, n, 3.62, o, 4.15, s, 16.79. obsd. (%): c, 69.25, h, 6.08, n, 3.67, o, 4.19, s, 16.81. (4 methoxy 4 phenyl) dithiocarbamic acid 2 (napthalen2-yloxy)ethyl ester (31) yield: 0.8 g (89.2 %); m.p.: 117 °c; ir (kbr, νmax, cm-1): 659 (c-s), 1106 (c=s), 1455 (ar), 1502 (ar), 1600 (ar), 2854 (ch), 2926 (ch), 3389 (nh); 1 h nmr (cdcl3): δ = 3.28-3.30 (t, 2h, ch2-s-c=s), 3.72 (s, 3h, och3), 4.0 (bs, h, nh), 4.70-4.72 (t, 2h, ch2-o-naphthyl), 6.35-7.64 (m, 11h, ar-h of naphthyloxy and phenyl ring); mass: m/e 369; analysis: c20h19no2s2, calcd. (%): c, 65.01, h, 5.18, n, 3.79, obsd. (%): c, 65.47, h, 5.03, n, 3.48. (4 methoxy 4 phenyl) dithiocarbamic acid3 (napthalen2-yloxy)propyl ester (32) yield: 0.82 g (93.8 %); m.p.: 139 °c; ir (kbr, νmax, cm1): 668 (c-s), 1117 (c=s), 1472 (ar), 1524 (ar), 1614 (ar), 2876 (ch), 2938 (ch), 3396 (nh); 1 h nmr (cdcl3): δ = 2.38-2.42 (m, 2h, naphthyl-och2ch2ch2-s-c=s), 2.84-2.88 (t, 2h, ch2-s-c=s), 3.74 (s, 3h, och3), 4.0 (bs, h, nh), 4.02-4.05 (t, 2h, ch2-onaphthyl), 6.34-7.65 (m, 11h, ar-h of naphthyloxy and phenyl ring); mass: m/e 383; analysis: c21h21no2s2, calcd. (%): c, 65.76, h 5.52, n, 3.65, obsd. (%): c, 65.27, h, 5.85, n, 3.81. (4-methoxy4 phenyl) dithiocarbamic acid 4 (napthalen2-yloxy)butyl ester (33) yield: 0.83 g (94.5 %); m.p.: 126 °c; ir (kbr, νmax, cm1): 681 (c-s), 1126 (c=s), 1484 (ar), 1523 (ar), 1610 (ar), 2885 (ch), 2936 (ch), 3407 (nh); 1 h nmr (cdcl3): δ = 1.71-1.74 (m, 2h, naphthyl-o-ch2ch2), 1.94-1.96 (m, 2h, s-ch2ch2), 2.01 (bs, h, nh), 2.82-2.86 (t, 2h, ch2-s-c=s), 3.72 (s, 3h, och3),3.92-3.94 (d, 2h, ch2 of benzylic proton),4.01-4.04 (t, 2h, ch2-o-naphthyl), 6.657.62 (m, 11h, ar-h of naphthyloxy and phenyl ring); mass: m/e 411; analysis: c23h25no2s2, calcd. (%): c, 67.12, h, 6.12, n, 3.40, obsd. (%): c, 67.67, h, 6.40, n, 3.67. cyclohexyl dithiocarbamic acid 2 (napthalen 2 yloxy)ethyl ester (34) yeild: 0.714 g (85.5 %), m.p; 112 °c; ir (kbr, νmax, cm1): 658 (c-s), 1103 (c=s), 1454 (ar), 1502 (ar), 1600 (ar), 2851 (ch), 2926 (ch), 3373 (nh); 1 h nmr (cdcl3): δ = 1.41-1.45 (m, 6h,ch2 of cyclohexyl ring), 1.621.64 (m, 4h, ch2 of cyclohexyl ring), 2.0 (bs, h, nh), 2.54 2.58 (m, h, tertiary h of cyclohexyl ring), 3.26-3.29 (t, 2h, ch2-s-c=s), 4.71-4.74 (t, 2h, ch2-o-naphthyl), 6.98-7.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 345; analysis: c19h23nos2, calcd. (%): c, 66.05, h, 6.71, n, 4.05, obsd. (%): c, 65.65, h, 6.97, n, 4.23. cyclohexyl dithiocarbamic acid 3 (napthalen-2 yloxy)propyl ester (35) yield: 0.714 g (85.5 %); m.p.: 112 °c; ir (kbr, νmax, cm1): 658 (c-s), 1103 (c=s), 1454 (ar), 1502 (ar), 1600 (ar), 2851 (ch), 2927 (ch), 3373 (nh); 1 h nmr (cdcl3): δ = 1.44-1.48 (m, 6h,ch2 of cyclohexyl ring), 1.63-1.66 (m, 4h, ch2 of cyclohexyl ring), 2.0 (bs, h, nh), 2.542.59 (m, h, tertiary h of cyclohexyl ring), 3.28-3.30 (t, 2h, ch2-s-c=s), 4.71-4.73 (t, 2h, ch2-o-naphthyl), 6.957.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 345; analysis: c19h23nos2, calcd. (%): c, 66.05, h, 6.71, n, 4.05, obsd. (%): c, 65.65, h, 6.97, n, 4.23. cyclohexyl dithiocarbamic acid 4 (napthalen-2 yloxy)butyl ester (36) yield: 0.75 g (94.5 %); m.p.: 126 °c; ir (kbr, νmax, cm-1 ): 668 (c-s), 1121 (c=s), 1469 (ar), 1523 (ar), 1617 (ar), 2879 (ch), 2937 (ch), 3408 (nh); 1 h nmr (cdcl3): δ = 1.42-1.44 (m, 6h, ch2 of cyclohexyl ring), 1.65 1.68 (m, 4h, ch2 of cylohexyl ring), 1.71-1.73 (m, 2h, naphthyl-o-ch2ch2), 1.94-1.96 (m, 2h, s-ch2ch2), 2.0 (bs, h, nh), 2.54-2.57 (m, h, tert. ch of cylohexyl ring), 2.842.88 (t, 2h, ch2-s-c=s), 4.02-4.06 (t, 2h, ch2-opa ge 95 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 86-98, 2023 naphthyl), 6.98-7.62 (m, 7h, ar-h of naphthyloxy); mass: m/e 373; analysis: c21h27nos2, calcd. (%): c, 67.52, h, 7.28, n, 3.75, obsd. (%): c, 67.84, h, 7.12, n, 3.59. benzyl dithiocarbamic acid -2-(napthalen-2-yloxy) ethyl ester (37) yield: 0.75 g (87.3 %); m.p.: 101 °c; ir (kbr, νmax, cm1): 662 (c-s), 1112 (c=s), 1464 (ar), 1512 (ar), 1603 (ar), 2865 (ch), 2926 (ch), 3385 (nh); 1 h nmr (cdcl3): δ = 2.0 (bs, h, nh), 3.26-3.32 (t, 2h, ch2-s-c=s), 3.923.94 (d, 2h, benzylic proton), 4.71-4.73 (t, 2h, ch2-onaphthyl), 6.98-7.65 (m, 12h, ar-h of naphthyloxy); mass: m/e 353; analysis: c20h19nos2, calcd. (%): c, 67.95, h 5.42, n, 3.96, obsd. (%): c, 67.63, h, 5.58, n, 4.12. benzyl dithiocarbamic acid -3-(napthalen-2-yloxy) propyl ester (38) yield: 0.75 g (89.8 %); m.p.: 109 °c; ir (kbr, νmax, cm1): 668 (c-s), 1114 (c=s), 1474 (ar), 1514 (ar), 1612 (ar), 2863 (ch), 2926 (ch), 3398 (nh); 1 h nmr (cdcl3): δ = 2.0 (bs, h, nh), 2.38-2.42 (m, 2h, naphthyloch2ch2ch2-s-c=s), 2.82-2.86 (t, 2h, ch2-s-c=s), 3.91-3.93 (d, 2h, ch2 of benzylic hydrogens), 4.014.04 (t, 2h, ch2-onaphthyl), 6.96-7.62 (m, 12h, ar-h of naphthyloxy); mass: m/e 367; analysis: c21h21nos2, calcd. (%): c, 68.63, h, 5.76, n, 3.81, obsd. (%): c, 68.27, h, 5.94, n, 4.02. benzyl dithiocarbamic acid -4-(napthalen-2-yloxy) butyl ester (39) yield: 0.75 g (92.8 %); m.p.: 119 °c; ir (kbr, νmax, cm1): 673 (c-s), 1126 (c=s), 1484 (ar), 1529 (ar), 1612 (ar), 2873 (ch), 2936 (ch), 3398 (nh); 1 h nmr (cdcl3): δ = 1.71-1.74 (m, 2h, naphthyl-o-ch2ch2), 1.96-1.99 (m, 2h, s-ch2ch2), 2.0 (bs, h, nh), 2.84-2.88 (t, 2h, ch2s-c=s), 3.91-3.94 (d, 2h, ch2 of benzylic proton), 4.014.04 (t, 2h, ch2-o-naphthyl), 6.96-7.65 (m, 12h, ar-h of naphthyloxy); mass: m/e 381; analysis: c22h23nos2, calcd. (%): c, 69.25, h 6.08, n, 3.67, obsd. (%): c, 69.67, h, 5.87, n, 3.46. phenyl ethyl dithiocarbamic acid 2 (napthalen-2 yloxy)ethyl ester (40) yield: 0.78 g (88.2 %); m.p.: 146 °c; ir (kbr, νmax, cm1): 661 (c-s), 1112 (c=s), 1464 (ar), 1514 (ar), 1605 (ar), 2865 (ch), 2923 (ch), 3376 (nh); 1 h nmr (cdcl3): δ = 2.0 (bs, h, nh), 2.80-2.82 (t, 2h, phch2), 2.962.98 (m, 2h, nhch2ch2ph), 3.28-3.31 (t, 2h, ch2-s-c=s), 4.71-4.73 (t, 2h, ch2-o-naphthyl), 6.98-7.62 (m, 12h, ar-h of naphthyloxy and phenyl ring); mass: m/e 367; analysis: c21h21nos2, calcd. (%): c, 68.63, h, 5.76, n, 3.81, obsd. (%): c, 68.19, h, 6.06, n, 3.95. phenyl ethyl dithiocarbamic acid 3 (napthalen-2 yloxy)propyl ester (41) yield: 0.8 g (91.4 %); m.p.: 172 °c; ir (kbr, νmax, cm-1): 668 (c-s), 1126 (c=s), 1478 (ar), 1519 (ar), 1614 (ar), 2878 (ch), 2933 (ch), 3396 (nh); 1 h nmr (cdcl3): δ = 2.0 (bs, h, nh), 2.35-2.41 (m, 2h, naphthyloch2ch2ch2-s-c=s), 2.80-2.82 (t, 2h, phch2), 2.842.86 (t, 2h, ch2-s-c=s), 2.96-3.02 (m, 2h, ch2nh), 4.02-4.06 (t, 2h, ch2-o-naphthyl), 6.95-7.62 (m, 12h, ar-h of naphthyloxy and phenyl group); mass: m/e 381; analysis: c22h23nos2, calcd. (%): c, 69.25, h, 6.08, n, 3.67, obsd. (%): c, 68.87, h, 6.29, n, 3.89. phenyl ethyl dithiocarbamic acid 4 -(napthalen-2 yloxy)butyl ester (42) yield: 0.8 g (94.8 %); m.p.: 179 °c; ir (kbr, νmax, cm-1): 679 (c-s), 1149 (c=s), 1487 (ar), 1533 (ar), 1622 (ar), 2884 (ch), 2944 (ch), 3438 (nh); 1 h nmr (cdcl3): δ = 1.72-1.74 (m, 2h, naphthyl-o-ch2ch2), 1.961.97 (m, 2h, s-ch2ch2), 2.01 (bs, h, nh),2.80-2.82 (t, 2h, phch2), 2.86-2.88 (t, 2h, ch2-s-c=s), 2.96-3.00 (m, 2h, ch2nh), 4.02-4.06 (t, 2h, ch2-o-naphthyl), 6.98-7.62 (m, 12h, ar-h of naphthyloxy and phenyl ring); mass: m/e 395; analysis: c23h25nos2, calcd. (%): c, 69.93, h, 6.37, n, 3.54, obsd. (%): c, 69.57, h, 6.55, n, 3.72. phenyl propyl dithiocarbamic acid 2 -(napthalen-2 yloxy)ethyl ester (43) yield: 0.84 g (90.2 %); m.p.: 119 °c; ir (kbr, νmax, cm1): 668 (c-s), 1114 (c=s), 1462 (ar), 1514 (ar), 1600 (ar), 2862 (ch), 2925 (ch), 3388 (nh); 1 h nmr (cdcl3): δ = 1.86-1.88 (m, 2h, phch2ch2ch2nh), 2.0 (bs, h, nh), 2.54-2.56 (t, 2h, phch2), 2.65-2.64 (m, 2h, nhch2ch2ch2ph), 3.28-3.31 (t, 2h, ch2-s-c=s), 4.72-4.74 (t, 2h, ch2-o-naphthyl), 6.95-7.62 (m, 12h, ar-h of naphthyloxy and phenyl ring); mass: m/e 381; analysis: c22h23nos2, calcd. (%): c, 69.25, h 6.08, n, 3.67, obsd. (%): c, 69.66, h, 5.99, n, 3.35. phenyl propyl dithiocarbamic acid 3 (napthalen-2 yloxy)propyl ester (44) yield: 0.84 g (93.2 %); m.p.: 135 °c; ir (kbr, νmax, cm-1 ): 682 (c-s), 1129 (c=s), 1481 (ar), 1533 (ar), 1626 (ar), 2884 (ch), 2936 (ch), 3416 (nh); 1 h nmr (cdcl3): δ = 1.86-1.89 (m, 2h, ph.ch2ch2ch2), 2.0 (bs, h, nh), 2.38-2.42 (m, 2h, naphthyl-o-ch2ch2ch2-sc=s), 2.52-2.55 (t, 2h, phch2), 2.62-2.64 (m, 2h, nhch2), 2.84-2.88 (t, 2h, ch2-s-c=s), 4.02-4.05 (t, 2h, ch2onaphthyl), 6.98-7.65 (m, 12h, ar-h of naphthyloxy and phenyl group); mass: m/e 395; analysis: c23h25nos2, calcd. (%): c, 69.83, h 6.37, n, 3.54, obsd. (%): c, 69.34, h, 6.66, n, 3.74. phenyl propyl dithiocarbamic acid 4-(napthalen-2 yloxy)butyl ester (45) yield: 0.85 g (97.6 %); m.p.: 154 °c; ir (kbr, νmax, cm1): 692 (c-s), 1139 (c=s), 1486 (ar), 1539 (ar), 1628 (ar), 2882 (ch), 2948 (ch), 3427 (nh); 1 h nmr (cdcl3): δ = 1.71-1.73 (m, 2h, naphthyl-o-ch2ch2), 1.861.88 (m, 2h, phch2ch2ch2nh), 1.96-1.99 (m, 2h, sch2ch2), 2.02 (bs, h, nh), 2.54-2.56 (t, 2h, phch2), 2.642.68 (m,2h,phch2ch2ch2-n), 2.84-2.86 pa ge 96 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 86-98, 2023 (t, 2h, ch2-s-c=s), 2.98-3.01 (m, 2h, ch2nh), 4.024.06 (t, 2h, ch2-onaphthyl), 6.98-7.62 (m, 12h, ar-h of naphthyloxy and phenyl ring); mass: m/e 409; analysis: c24h27nos2, calcd. (%): c, 70.37, h, 6.64, n, 3.42, obsd. (%): c, 69.95, h, 6.86, n, 3.62. di sec butyl dithiocarbamic acid-2-(napthalen-2 yloxy)ethyl ester (46) yield: 0.84 g (90.2 %); m.p.: 119 °c; ir (kbr, νmax, cm1): 668 (c-s), 1115 (c=s), 1462 (ar), 1514 (ar), 1601 (ar), 2865 (ch), 2926 (ch), 3388 (nh); 1 h nmr (cdcl3): δ = 1.08-1.10 (d, ch3), 2.75-2.79 (m, 8h, ch3ch2ch3), 1.381.41 (m, 6h, ch3chch3), 0.94-0.96 (t, 2h,ch3chch2), 3.283.30 (t, 2h, ch2-s-c=s), 4.714.73 (t, 2h, ch2-o-naphthyl), 6.95-7.63 (m, 12h, ar-h of naphthyloxy and phenyl ring); mass: m/e 375.59; analysis: c21h29nos2, calcd. (%): c, 67.10, h 7.75, n, 3.70, o, 4.22, s, 17.05. obsd. (%): c, 67.15, h, 7.78, n, 3.73, o, 4.26, s, 17.07. di sec butyl dithiocarbamic acid 3-(napthalen-2 yloxy) propyl ester (47) yield: 0.84 g (90.2 %); m.p.: 119 °c; ir (kbr, νmax, cm1): 668 (c-s), 1114 (c=s), 1462 (ar), 1514 (ar), 1600 (ar), 2862 (ch), 2925 (ch), 3388 (nh); 1 h nmr (cdcl3): δ = 1.08-1.10 (d, ch3), 2.75-2.79 (m, 8h, ch3ch2ch3), 1.38-1.41 (m, 6h, ch3chch3), 0.94-0.96 (t, 2h,ch3chch2), 3.28-3.30 (t, 2h, ch2-s-c=s), 2.352.39(m, 2h, ch2ch2ch2), 4.72-4.74 (t, 2h, ch2-onaphthyl), 6.967.62 (m, 12h, ar-h of naphthyloxy and phenyl ring); mass: m/e 389.62; analysis: c22h31nos2, calcd. (%): c, 67.80, h 8.01, n, 3.60, o, 4. 10, s, 16.42. obsd. (%): c, 67.82, h, 8.02, n, 3.59, o, 4.11, s, 16.46. di sec butyl dithiocarbamic acid 4 -(napthalen-2 yloxy)butyl ester (48) yield: 0.84 g (90.2 %); m.p.: 119 °c; ir (kbr, νmax, cm-1): 668 (c-s), 1114 (c=s), 1465 (ar), 1514 (ar), 1600 (ar), 2865 (ch), 2927 (ch), 3388 (nh); 1 h nmr (cdcl3): δ = 1.08-1.10 (d, ch3), 2.75-2.79 (m, 8h, ch3ch2ch3), 1.381.41 (m, 6h, ch3chch3), 0.94-0.96 (t, 2h,ch3chch2), 3.273.31 (t, 2h, ch2-sc=s), 1.92-1.96 (m, 2h, ch2ch2ch2), 1.68-1.71 (m, 2h, ch2ch2ch2), 4.71-4.73 (t, 2h, ch2-onaphthyl), 6.977.64 (m, 12h, ar-h of naphthyloxy and phenyl ring); mass: m/e 403.64; analysis: c23h33nos2, calcd. (%): c, 68.41, h 8.20, n, 3.44, o, 3.92, s, 15.87. obsd. (%): c, 68.44, h, 8.24, n, 3.47, o, 3.96, s, 15.89. biological testing dpph radical scavenging activity of the experiment the initial absorbance of the dpph in ethanol (concentration = 0.04 mm) was measured at 517 nm and was maintained constant throughout the period of assay. all the sample compounds were dissolved and eventually further diluted in 80% etoh. different test concentrations (2.5, 5, 10, 20 and 40 mg/ ml) or carrier solvent alone was added to 2 ml of ethanolic dpph solution with three replicates each. the change in absorbance at 517 nm was measured with time and free radical scavenging activity was calculated as inhibition using following equation: percentage dpph radical scavenging activity = 1 [as/ ac] x 100, where as: absorbance of the dpph solution containing samples. ac: absorbance of the control solution without sample but with dpph. the experiment was also conducted using vitamin c as a reference antioxidant. abts radical scavenging activity of the experiment(trap) in this method, an antioxidant was added to a pre-formed abts radical solution and after a fixed time period the remaining abts.+ was quantified spectrophotometrically at 734 nm. abts.+ was produced by reacting abts with oxidant solution (k2s2o8) at the volume ratio of 1:1, stored in the dark at room temperature for 16 h. the abts.+ solution was diluted to give an absorbance of 0.750 ± 0.025 at 734 nm in 80% etoh. all the compounds tested were dissolved and eventually further diluted in 80% etoh. 200 ml different test concentrations (2.5, 5, 10, 20 and 40 mg/ml) or carrier solvent alone was added to 4 ml of ethanolic dpph solution with three replicates each. the absorbance was recorded as time goes on after mixing and the percentage of radical scavenging was calculated for each concentration relative to a blank containing no scavenger. the extent of decolorization was calculated as percentage reduction of absorbance. percentage abts radical scavenging activity = 1 [as/ ac] x 100, where as: absorbance of the abts solution containing samples. ac: absorbance of the control solution without sample but with abts. the experiment was also conducted using vitamin c as a reference antioxidant. roo_radical scavenging activity of the experiment (trap) in this method, 0.02 g abts and 0.27 g of 2,20-azobis (2-methylpropionamidine) dihydrochloride (aaph) was dissolved in acetate buffer solution (ph = 4.3) and transferred to the 500ml volumetric flask. the solution was treated with 45 0c water bath for an hour, and then cooled to room temperature. 200 ml different concentrations (2.5, 5,10, 20, and 40 mg/ml) or carrier solvent alone was added to 4 ml of ethanolic roo. solution with three replicates each. the changes in absorbance at 734 nm were recorded. percentage roo. radical scavenging activity = 1 [as/ ac] x 100, where as: absorbance of the roo. solution containing samples. ac: absorbance of the control solution without sample but with roo.. the experiment was also conducted using vitamin c as a reference antioxidant. pa ge 97 https://journals.e-palli.com/home/index.php/ajcp am. j. chem. pharm. 2(2) 86-98, 2023 o2-. radical scavenging activity of the experiment (net) 0.04 g ko2 and 0.3 g 18-crown-6 were dissolved with 100 ml anhydrous dmso to create o2-. radical. 1.2 g nitrotetrazolium blue chloride (net) was dissolved in 100ml of anhydrous dmso to be used for determining absorbance. 100 ml of different test concentrations (2.5, 5, 10, 20, and 40 mg/ ml) or carrier solvent alone was added to 200 ml of ethanolic o2-. solution with three replicates each react for 5 min and then added with 200 ml of net solution reacted for 5 min. to determine the absorbance of 680 nm through following equation:percentage o2-. radical scavenging activity = 1 [as/ac] x 100, where as: absorbance of the o2-. solution containing samples. ac: absorbance of the control solution without sample but with o2-.. the experiment was also conducted using vitamin c as a reference antioxidant. conclusion we have synthesized a series of dithiocarbamates of ω-substituted (2-naphthyloxy) alkanes. antioxidant activity of these hybrid compounds were studied by radicals 1,1-diphenyl-2-picryl-hydrazyl (dpph) assay, 2,2-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) (abts) assay, roo. 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