Microsoft Word - 11423 NSB El Mekkaoui 2023.03.16.docx Received: 29 Dec 2022. Received in revised form: 02 Feb 2023. Accepted: 10 Mar 2023. Published online: 16 Mar 2023. From Volume 13, Issue 1, 2021, Notulae Scientia Biologicae journal uses article numbers in place of the traditional method of continuous pagination through the volume. The journal will continue to appear quarterly, as before, with four annual numbers. SHSTSHSTSHSTSHST Horticulture and ForestryHorticulture and ForestryHorticulture and ForestryHorticulture and Forestry Society of TransylvaniaSociety of TransylvaniaSociety of TransylvaniaSociety of Transylvania El Mekkaoui A et al. (2023) Notulae Scientia BiologicaeNotulae Scientia BiologicaeNotulae Scientia BiologicaeNotulae Scientia Biologicae Volume 15, Issue 1, Article number 11423 DOI:10.15835/nsb15111423 Research ArticleResearch ArticleResearch ArticleResearch Article.... NSBNSBNSBNSB Notulae Scientia Notulae Scientia Notulae Scientia Notulae Scientia BiologicaeBiologicaeBiologicaeBiologicae Phytochemical studies and Phytochemical studies and Phytochemical studies and Phytochemical studies and in vitroin vitroin vitroin vitro evaluation of the antioxidant evaluation of the antioxidant evaluation of the antioxidant evaluation of the antioxidant activity of some medicinal and aromatic plants from Moroccoactivity of some medicinal and aromatic plants from Moroccoactivity of some medicinal and aromatic plants from Moroccoactivity of some medicinal and aromatic plants from Morocco Anas EL MEKKAOUI1,2*, Mohamed KHAMAR2, Chaimae SLIMANI3, Abderrahman NOUNAH2, Essediya CHERKAOUI2, Fatima BENRADI2, Chaimae RAIS1 1National Agency for Medicinal and Aromatic Plants, Laboratory of Botany, P.O. Box 159 Taounate, 34025, 10, Morocco; raischaimae18@gmail.com 2Mohammed V University of Rabat, Higher School of Technology, Laboratory of Civil and Environmental Engineering (LGCE), P.O. Box 227, Salé, Morocco; anasitsugo@gmail.com (*corresponding author); mohamed.khamar@um5.ac.ma; abderrahman.nounah@um5.ac.ma; essediya.cherkaoui@um5.ac.ma; fatima.benradi@um5.ac.ma 3Sidi Mohamed Ben Abdellah University, Faculty of Sciences and Technologies, Department of Biology, Laboratory of Functional Ecology and Environmental Engineering, P.O. Box 2202, route d’Imouzzer, Fez, Morocco; chaimae.slimani94@gmail.com AbstractAbstractAbstractAbstract The present work was carried out to evaluate the phenolic compounds and the antioxidant activities of some solvent extract (methanol, hydroethanol and aqueous) of several Moroccan medicinal plants known for their high antioxidant properties. The extracts were obtained by sonication, then, the total phenolics and flavonoids compounds were determined using Folin-Ciocalteu and Aluminium chloride. Afterwards, the Total Antioxidant Capacity and DPPH scavenging methods were performed. Results of phytochemical analysis showed that the total phenolics content were the highest in the hydroethanolic extract of Arbutus unedo with 160.76 mg GAE g-1DM, and the flavonoids content were the highest for the hydroethanolic extracts of Inula viscosa with 489.77 mg QE g-1 DM. Also, it can be noted that Arbutus unedo, Argania spinosa, and Myrtus communis exhibited the most potent antioxidant activity respectively with 0.026; 0.043; 0.036 mg ml-1. Keywords:Keywords:Keywords:Keywords: antioxidant activity; medicinal plants; Morocco; oxidative stress; phenolic compounds; radical scavenging IntroductionIntroductionIntroductionIntroduction Oxidative stress is a phenomenon that reflects an imbalance between the production of reactive oxygen species (ROS) called oxidants, and their elimination by protective mechanisms called antioxidant systems, which can detoxify the reactive intermediates, or repair the resulting damage, causing toxic effects through the production of peroxides and free radicals (Pizzino et al., 2017). In addition, some reactive oxidant species act as cellular messengers in redox signaling that can cause disruptions in normal cell signaling mechanisms (Milkovic et al., 2019). https://www.notulaebiologicae.ro/index.php/nsb/index El Mekkaoui A et al. (2023). Not Sci Biol 15(1):11423 2 Oxidative stress is thought to be involved in the development of atherosclerosis, neurodegenerative diseases such as Alzheimer's and Parkinson's, cancer, diabetes mellitus, and inflammatory diseases, as well as psychological diseases or aging processes (Forman and Zhang, 2021). The participation of oxidative stress, which is connected to the formation of reactive oxygen and nitrogen species by all aerobic organisms, including free radicals, is a common factor in the pathogenesis of the majority of chronic diseases (Reza et al., 2010). These reactive molecular species have a major impact on intra- and extra cellular signaling, and can start harmful metabolic processes. A sophisticated antioxidant defense has been established to counteract this damage, and dietary antioxidants play an important role in this defense (Chib et al., 2020). Antioxidants prevent oxidative stress in biological systems. Because of the unpaired electron in their structure, antioxidants are able to neutralize free radicals or radical ions. They have the effect to remove the radical ions produced as a result of oxidation from the system without damaging the biological system (Çalişkan and Cengiz Çalişkan, 2021). Medicinal and aromatic plants (MAP) are inexhaustible natural sources of bioactive compounds, including antioxidants (Lourenço et al., 2019). In light of its geographical location, Morocco is said to have a large variety of MAP, with over 4,200 species and subspecies, of which 22% are endemic (Rankou et al., 2013). The aim of our work is to exploit the potential of Moroccan medicinal plants in order to highlight antioxidant activity which is closely related to the content of phenolic compounds (Aryal et al., 2019) and demonstrate their possible use as alternative therapies against oxidative diseases. Materials and MethodsMaterials and MethodsMaterials and MethodsMaterials and Methods Plant material The plant material used for the present study consists of seven medicinal and aromatic plants that were selected according to their known medicinal effects, their uses in traditional medicine, and the endemic character for some plants. The data related to these plants are represented in Table 1. The leaves of these MAPs were sorted and dried for 72 h at 35 °C. Thereafter, they were crushed and sieved to 300 μm to obtain a fine homogeneous powder, that will be used for extracts. Table 1Table 1Table 1Table 1. Medicinal and aromatic plants studied Species Family GPS data Marrubium vulgare L. Lamiaceae Douar Sahel Boutaher (34°30.4718'N, 4°47.8572'O) Inula viscosa L. Asteraceae Douar Sahel Boutaher (34°30.1870'N, 4°46.9117'O) Retama monosperma L. Fabaceae Settat (33°0.4556'N, 7°34.9844'O) Myrtus communis L. Myrtaceae ANPMA (34°29.8993'N, 4°48.1756'O) Arbutus unedo L. Ericaceae Tamesnit Ratba (34°44.6810'N, 4°53.3705'O) Argania spinosa L. Sapotaceae Agadir (30°26.1546'N, 9°27.3926'O) Cannabis sativa Cannabinaceae Douar Rkaiba (34°43.9322'N, 4°52.0309'O) Preparation of the extract Three extracts were prepared for each plant: a methanol, hydroethanol (20/80) and aqueous extract. To do this, 40 mg of dry matter was added to 10 ml of solvent, the extraction was made by ultrasound at 35 Khz, the whole was then centrifuged for 30 min at 3000 rpm. The supernatant was recovered and stored at 4 °C. El Mekkaoui A et al. (2023). Not Sci Biol 15(1):11423 3 Total Phenolic Content (TPC) The Folin-Ciocalteu technique was used to determine the total phenolic content of these MAPs (Cheng and Li, 2004). Indeed, 200 μL of the extract was mixed with 1.5 mL of 10% Folin-Ciocalteu reagent. After 5 minutes, 1.5 mL of 5% sodium carbonate was added. The absorbance was measured at 725 nm after 2 hours of incubation. The concentration of total polyphenols was calculated based on a previous calibration curve performed with standard gallic acid. Results are expressed as mg gallic acid equivalents per gram dry matter (mg GAE g-1 DM). Total Flavonoids Contents (TFC) The flavonoid contents were assessed in accordance with the method used Barros et al., 2011. Therefore, 0.3 mL of 5% NaNO2 were added to 1 mL of extract. After 5 minutes, 0.3 mL of 10% AlCl3 were added. Then, 2 mL of NaOH 1 M were added, and the mixture's volume was subsequently raised to 10 mL using distilled water. The absorbance was measured at 510 nm. Total flavonoids were calculated from a standard curve made with quercetin. Results are expressed as milligrams of quercetin equivalents per gram of dry matter (mg QE g-1 DM) Evaluation of the antioxidant activity DPPH assay The purpose of this method was to determine which of the prepared extracts had the greatest antioxidant activity against DPPH (2,2'-diphenyl-1-picrylhydrazyl). It is important to note that DPPH is a stable free radical, and in the presence of antioxidants, the characteristic purple colour of DPPH changes to yellow, and the absorbance is measured at 517 nm.... A) B) Figure 1. Figure 1. Figure 1. Figure 1. Reaction of an antioxidant with DPPH (Molyneux, 2004); (A) Diphenylpicrylhydrazyl + Antioxidant-OH (Purple color); (B) Diphenylpicrylhydrazyl + Antioxidant-O (Yellow color) The antioxidant activity was tested according to the method described by Brand-Williams et al. (1995). This procedure consists in preparing dilutions of the order of 1/2, 1/4, 1/8 and 1/16 from a stock solution of 4 mg ml-1 of the studied extract. Then, 1 ml of each dilution was added to 1 ml of DPPH (0.004%). Ascorbic acid was used as a reference molecule. The absorbance was measured at 517 nm by spectrophotometer, and the antioxidant activity was calculated according to the following formula: Antioxidant activity %� � Abs DPPH � ��� �� �ℎ� ��� !"� Abs DPPH # 100 • Abs DPPH: Absorbance of the solution of DPPH • Abs of the extract: value of the absorbance after the addition of the extract The regression curve of this activity allowed to determine the concentration that corresponds to 50% inhibition (The half maximal inhibitory concentration IC50505050: Concentration of the tested sample or ascorbic acid necessary to reduce 50% of the DPPH radical). A low IC50505050 value indicates a high capacity of the extract to act as a DPPH scavenger (Cheng and Li, 2004). El Mekkaoui A et al. (2023). Not Sci Biol 15(1):11423 4 Total antioxidant capacity (TAC) The total antioxidant capacity was measured using the protocol described by Prieto (Prieto et al., 1999). In practice, 200 μL of extract were mixed with 3 mL of reagent solution (6M sulfuric acid, 280 mM sodium phosphate and 40 mM aluminum molybdate). The incubation was done at 95 °C for 90 min. After cooling, the absorbance was measured at 695 nm. The total antioxidant capacity was expressed as milligram ascorbic acid equivalent / gram dry matter (mg AAE g-1 DM). Statistical analysis The data obtained were subjected to analysis of variance (ANOVA). Statistical analysis was based on Two-way ANOVA followed by Tukey's Honestly Significant Difference Test. The data were processed with the software "SYS-TAT 12". A test of comparison of the means was done each time there was a significant effect of factor studied by the ANOVA Results Results Results Results Total phenolic content Figure 2 displays the results of the total phenolic contents. They ranged from 7.90 to 160.76 mg GAE g- 1 DM. The highest concentration of phenols was observed in the hydroethanol extract for A. unedo. On the other hand, the aqueous extract of M. vulgare had the lowest content. The analysis of variance for total phenol content showed a highly significant difference between both the species and the solvents used in the present experimentation (df = 12, F-ratio = 10.843, p <0.001). Figure 2Figure 2Figure 2Figure 2. Total phenolic content of the extracts 0 20 40 60 80 100 120 140 160 180 A. unedo I. viscoa R. monosperma A. spinosa C. sativa M. communis M. vulgare Methanol extract Hydroethanol extract Aqueous extract mg GAE g-1 DM El Mekkaoui A et al. (2023). Not Sci Biol 15(1):11423 5 Total flavonoids contents The results of flavonoid contents were showed in Figure 3. Therefore, the concentration of flavonoids in the extract ranged from 36.01 to 489.77 mg QE g-1 DM. Analysis of the results allowed us to highlight that the highest content was the ethanol extract of I. viscosa. However, the aqueous extract of R. monosperma, had the lowest concentration. Statistically a highly significant difference was observed between the different species studied and the three extraction solvents used (dl = 12, F-ratio = 23.251, p <0.001). Figure 3.Figure 3.Figure 3.Figure 3. Total flavonoid content of the extracts Evaluation of antioxidant activity DPPH method From the results, we found that all the extracts were able to reduce DPPH free radicals. Table 2 summarizes the IC50505050 values of the examined extracts and ascorbic acid. A. unedo recorded the lowest IC50505050 values for the three extraction solvents (methanol, hydroethanol, and aqueous extracts) with respectively 0.033; 0.026; and 0.034 mg ml-1, reflecting its potent antioxidant activity approaching that of ascorbic acid (IC50505050 = 0.01 mg ml-1). On the other hand, R. monosperma had the lowest anti-radical activity (IC50505050 = 0.957; 0.873 and 0.860 mg ml-1). The analysis of variance for IC50505050 showed a highly significant difference between the extracts and solvents used (dl = 12, F-ration = 101.246, p <0.001). 0 100 200 300 400 500 600 A. unedo I. viscosa R. monosperma A. spinosa C. sativa M. communis M. vulgare Methanol Extract Hydroethanol extract Aqueous extract El Mekkaoui A et al. (2023). Not Sci Biol 15(1):11423 6 Table 2.Table 2.Table 2.Table 2. IC50505050 values (mg ml-1) of tested extracts and ascorbic acid MAPs Extract IC50 (mg ml-1) Methanol Hydroethanol Aqueous A. unedo 0.033 ± 0.0004 0.026 ± 0.0006 0.034 ± 0.0007 I. viscosa 0.065 ± 0.0001 0.057 ± 0.0004 0.094 ± 0.0007 R. monosperma 0.957 ± 0.0101 0.873 ± 0.0045 0.86 ± 0.0495 A. spinosa 0.048 ± 0.0001 0.043 ± 0.0012 0.046 ± 0.0011 C. sativa 0.666 ± 0.0175 0.467 ± 0.0040 1.032 ± 0.0175 M. communis 0.037 ± 0.0002 0.036 ± 0.0002 0.038 ± 0.0001 M. vulgare 0.346 ± 0.0017 0.329 ± 0.0009 0.635 ± 0.0081 Ascorbic acid 0.01 ± 0.0001 Total antioxidant capacity The results of the total antioxidant capacity are showed in Figure 4. The values obtained are expressed in terms of mg equivalent of ascorbic acid per gram of dry matter. According to these results, the evaluation of the total antioxidant capacity of the extracts showed a variability relative to the solvents. We found that the hydroethanol extracts present the highest antioxidant capacity, with 267.37 and 269.32 mg AAE g-1 DM respectively for A. unedo and A. spinosa. In contrast, the lowest concentration belongs to R. monosperma for the aqueous extract, with a value of 29.37 mg AAE g-1 DM. The statistical analysis of the total antioxidant capacity shows a highly significant difference between the studied plants and the different solvents used (dl = 12, F-ratio = 15.293, p <0.001). Figure 4Figure 4Figure 4Figure 4. Total antioxidant capacity of the extracts 0 50 100 150 200 250 300 A. unedo I. viscosa R. monosperma A. spinosa C. sativa M. communis M. vulgare Methanol extract Hydroethanol Extract Aqueous extract mg AAE g-1 DM El Mekkaoui A et al. (2023). Not Sci Biol 15(1):11423 7 DiscussionDiscussionDiscussionDiscussion Organic solvent extraction is the most commonly used method for the preparation of plant extracts (Zhang et al., 2018), due to the modification of the solvent influencing the yield/composition of the isolated molecules, it also offers a selective method for the extraction of bioactive compounds (Kapadia et al., 2022). In terms of antioxidant activity, our results indicated that hydroethanol extracts have the strongest antioxidant power, a finding consistent with the study conducted by Özbek et al. (2020). For the phenolic contents of Arbutus unedo, our results ranged from 116.67 to 160.76 mg GAE g-1 DM, which are higher compared to those reported by Ait lhaj et al. (2022) with a highest result of 107.67 mg GAE g-1 DM and Habachi et al. (2022) with a highest result of 86 mg GAE g-1 DM, who used different extraction methods and solvents for the leaves. The best IC50505050 value in our study for the antioxidant activity was 0.026 mg ml-1 obtained in the hydroethanol extract, which is higher than the value reported by Bebek Markovinović et al. (2022) of 0.076 mg ml-1, who used the same solvent. In the case of I. viscosa, the phenolic contents were found to range from 36.82 to 78.41 mg GAE g-1 DM, consistent with the results of Aydar et al. (2022) who obtained a value of 54.39 mg GAE g-1 DM using a combination of ultrasonic and microwave extraction. Our best IC50505050 value was 0.057 mg ml-1 obtained in the hydroethanol extract, which is lower than the result of Yıldırım et al. (2022) who obtained 0.014 mg ml-1, while using chloroform as solvent. For R. monosperma, the phenolic contents were found to range from 24.7 to 32.44 mg GAE g-1 DM, with an IC50505050 value of 0.86 mg ml-1. These results are lower compared to those reported by Selaimia et al. (2020) of 155.61 mg GAE g-1 DM and 0.118 mg ml-1, they used a diethyl ether solvent and a maceration method. For A. spinosa, we obtained for phenolic contents values between 99.02-106.21 mg GAE g-1 DM, which is higher than what was reported by Afrokh et al. (2023) with 47.75 mg GAE g-1 DM. On the other side, our IC50505050 value of 0.043 mg ml-1 obtained in the hydroethanol extract is consistent with the IC50505050 obtained in the same study of 0.048 mg ml-1. M. vulgare, resulted in values between 7.9-13.43 mg GAE g-1 DM for the phenolic contents, which is lower than what reported Afrokh et al. (2023) with a value of 29.25 mg GAE g-1 DM, whereas our IC50505050 of 0.329 mg ml-1 is higher than their result of 2.42 mg ml-1. For C. sativa, the phenolic contents were found to range from 14.07 to 27.88 mg GAE g-1 DM, which is in agreement with the findings of Aazza (2021) who reported a value of 19.07 mg GAE g-1 DM. The best IC50505050 value obtained was 0.467 mg ml-1, which is higher compared to the results reported by Benkirane et al. (2023) at 1.83 mg ml-1. Finally, for the phenolic contents of M. Communis the results were between 89.47-100.3 mg GAE g-1 DM, which is higher compared to the results reported by Hazrati et al. (2022) at 66.52 mg GAE g-1 DM using a 24-hour hydro-ethanol maceration extraction method. The best IC50505050 value in our study was 0.036 mg ml-1 (hydroethanol extract), which is consistent with the findings of Yangui et al. (2021) of 0.038 mg ml-1 where they used a hydromethanol solvent. The difference in phenol contents and antioxidant activity can be explained by the polarity of the solvent used in the extraction, knowing that the high solubility of phenols in polar solvents facilitates a better yield in extracts obtained from them (Alara et al., 2021). This difference may also be due to several factors such as climate, soil, harvest period, storage condition, environmental factors (temperature, pH), or the extraction method (Ben Ahmed et al., 2017; Sun et al., 2019; Ngoune Liliane and Shelton Charles, 2020; Zeroual et al., 2021; Zhang et al., 2022). Furthermore, it has been established that antioxidant activity is positively correlated with the structure of phenols (Osman et al., 2020). Generally, phenols with a high number of hydroxyl groups present the highest antioxidant activity (Heim et al., 2002) and that is due to their power to give more atoms to stabilize the free El Mekkaoui A et al. (2023). Not Sci Biol 15(1):11423 8 radicals (Torres de Pinedo et al., 2007). Thus, the antioxidant effect is not only dose-dependent but also structure-dependent (Rodríguez-Bernaldo de Quirós et al., 2010). Therefore, a mixture of these plants could be considered and could accentuate even more their antioxidant effects, allowing them to be considered as a possible alternative to remedy oxidative diseases. ConclusionsConclusionsConclusionsConclusions During this work, we were able to highlight the medical potential of these plants. The quantitative analysis of the methanol, ethanol and aqueous extracts showed the presence of flavonoid and polyphenols. Furthermore, we were able to show that these plants have a good antioxidant activity especially A. unedo, A. spinosa and M. communis. Additionally, these plants compounds have significant therapeutic effects with few side effects, their natural antioxidants may offer an alternative to conventional treatments for oxidative stress and could be considerate strong candidates for the prevention of free radical diseases like cancer, aging process, cardiovascular disease, and diabetes. It would be important to extend the range of this study parameters as well as the isolation, characterization, and identification of active compounds to valorise more these plants. Authors’ ContributionsAuthors’ ContributionsAuthors’ ContributionsAuthors’ Contributions Data curation: AEM, MK; Methodology: AEM, CS, CR; Supervision: MK, CR, AN, EC, FB; Writing - original draft: AEM; Writing - review and editing: AEM, CS. All authors read and approved the final manuscript. Ethical approvalEthical approvalEthical approvalEthical approval (for researches involving animals or humans) Not applicable. AcknowledgementsAcknowledgementsAcknowledgementsAcknowledgements This research received no specific grant from any funding agency in the public, commercial, or not-for- profit sectors. Conflict of InterestsConflict of InterestsConflict of InterestsConflict of Interests The authors declare that there are no conflicts of interest related to this article. ReferencesReferencesReferencesReferences Aazza S (2021). Application of multivariate optimization for phenolic compounds and antioxidants extraction from Moroccan Cannabis sativa waste. Journal of Chemistry 2021:1-11. https://doi.org/10.1155/2021/9738656 Afrokh M, Tahrouch S, EL Mehrach K, Fahmi F, Ait Bihi M, Weber-Ravn H, … Tabyaoui M (2023). Ethnobotanical, phytochemical and antioxidant study of fifty aromatic and medicinal plants. Chemical Data Collections 43:100984. https://doi.org/10.1016/j.cdc.2022.100984 El Mekkaoui A et al. (2023). Not Sci Biol 15(1):11423 9 Ait lhaj Z, Taghzouti K, Bouyahya A, Diria G, Bakhy K, Bchitou R (2022). Phenolic composition and antioxidant activity of leaves of strawberry tree (Arbutus unedo l.) populations from Morocco. Phytotherapy 20:192-204. https://doi.org/10.3166/phyto-2021-0296 Alara OR, Abdurahman NH, Ukaegbu CI (2021). Extraction of phenolic compounds: A review. Current Research in Food Science 4:200-214. https://doi.org/10.1016/j.crfs.2021.03.011 Aryal S, Baniya MK, Danekhu K, Kunwar P, Gurung R, Koirala N (2019). Total phenolic content, flavonoid content and antioxidant potential of wild vegetables from western Nepal. Plants 8(4):96. https://doi.org/10.3390/plants8040096 Aydar AY, Aydın T, Yılmaz T, Kothakota A, Claudia Terezia S, Florin Leontin C, Pandiselvam R (2022). Investigation on the influence of ultrasonic pretreatment on color, quality and antioxidant attributes of microwave dried Inula viscosa (L.). Ultrasonics Sonochemistry 90:106184. https://doi.org/10.1016/j.ultsonch.2022.106184 Barros L, Carvalho AM, Ferreira ICFR (2011). Exotic fruits as a source of important phytochemicals: Improving the traditional use of Rosa canina fruits in Portugal. Food Research International 44(7):2233-2236. https://doi.org/10.1016/j.foodres.2010.10.005 Bebek Markovinović A, Brčić Karačonji I, Jurica K, Lasić D, Skendrović Babojelić M, Duralija B, … Bursać Kovačević D (2022). Strawberry tree fruits and leaves (Arbutus unedo l.) as raw material for sustainable functional food processing: a review. Horticulturae 8(10):881. https://doi.org/10.3390/horticulturae8100881 Ben Ahmed Z, Yousfi M, Viaene J, Dejaegher B, Demeyer K, Mangelings D, vander Heyden Y (2017). Seasonal, gender and regional variations in total phenolic, flavonoid, and condensed tannins contents and in antioxidant properties from Pistacia atlantica ssp. leaves. Pharmaceutical Biology 55(1):1185-1194. https://doi.org/10.1080/13880209.2017.1291690 Benkirane C, Mansouri F, Ben Moumen A, Taaifi Y, Melhaoui R, Caid HS, … Abid M (2023). Phenolic profiles of non- industrial hemp (Cannabis sativa L.) seed varieties collected from four different Moroccan regions. International Journal of Food Science Technology 58(3):1367-1381. https://doi.org/10.1111/ijfs.16298 Brand-Williams W, Cuvelier ME, Berset C (1995). Use of a free radical method to evaluate antioxidant activity. Food Science and Technology 28(1):25-30. https://doi.org/10.1016/S0023-6438(95)80008-5 Çalişkan B, Cengiz Çalişkan A (2021). Antioxidant and oxidative stress. in antioxidants - benefits, sources, mechanisms of action. IntechOpen. https://doi.org/10.5772/intechopen.96643 Cheng Z, Li Y (2004). Reducing power: the measure of antioxidant activities of reductant compounds? Redox Report 9(4):213-217. https://doi.org/10.1179/135100004225005994 Chib A, Gupta N, Bhat A, Anjum N, Yadav G (2020). Role of antioxidants in food. International Journal of Chemical Studies 8(1):2354-2361. https://doi.org/10.22271/chemi.2020.v8.i1aj.8621 Forman HJ, Zhang H (2021). Targeting oxidative stress in disease: promise and limitations of antioxidant therapy. Nature Reviews Drug Discovery 20(9):689-709. https://doi.org/10.1038/s41573-021-00233-1 Habachi E, Rebey IB, Dakhlaoui S, Hammami M, Sawsen S, Msaada K, … Bourgou S (2022). Arbutus unedo: innovative source of antioxidant, anti-inflammatory and anti-tyrosinase phenolics for novel cosmeceuticals. Cosmetics 9(6):143. https://doi.org/10.3390/cosmetics9060143 Hazrati S, Hosseini SJ, Ebadi MT, Nicola S (2022). Evolution of phytochemical variation in myrtle (Myrtus communis l.) organs during different phenological stages. Horticulturae 8(9):757. https://doi.org/10.3390/horticulturae8090757 Heim KE, Tagliaferro AR, Bobilya DJ (2002). Flavonoid antioxidants: chemistry, metabolism and structure-activity relationships. The Journal of Nutritional Biochemistry 13(10):572-584. https://doi.org/10.1016/S0955- 2863(02)00208-5 Kapadia P, Newell AS, Cunningham J, Roberts MR, Hardy JG (2022) Extraction of high-value chemicals from plants for technical and medical applications. International Journal of Molecular Sciences 23(18):10334. https://doi.org/10.3390/ijms231810334 Lourenço SC, Moldão-Martins M, Alves VD (2019). antioxidants of natural plant origins: from sources to food industry applications. Molecules 24(22):4132. https://doi.org/10.3390/molecules24224132 Milkovic L, Cipak Gasparovic A, Cindric M, Mouthuy PA, Zarkovic N (2019). Short overview of ROS as cell function regulators and their implications in therapy concepts. Cells 8(8):793. https://doi.org/10.3390/cells8080793 El Mekkaoui A et al. (2023). Not Sci Biol 15(1):11423 10 Molyneux P (2004). The use of the stable free radical diphenylpicryl-hydrazyl (DPPH) for estimating antioxidant activity. Songklanakarin Journal of Science and Technology 26(2):211-219. Ngoune Liliane T, Shelton Charles M (2020). Factors affecting yield of crops. in agronomy - climate change and food security. IntechOpen. https://dx.doi.org/10.5772/intechopen.90672 Osman MA, Mahmoud GI, Shoman SS (2020). Correlation between total phenols content, antioxidant power and cytotoxicity. Biointerface Research in Applied Chemistry 11(3):10640-10653. https://doi.org/10.33263/briac113.1064010653 Özbek HN, Halahlih F, Göğüş F, Koçak Yanık D, Azaizeh H (2020). Pistachio (Pistacia vera L.) hull as a potential source of phenolic compounds: evaluation of ethanol–water binary solvent extraction on antioxidant activity and phenolic content of pistachio hull extracts. Waste and Biomass Valorization 11(5):2101-2110. https://doi.org/10.1007/s12649-018-0512-6 Pizzino G, Irrera N, Cucinotta M, Pallio G, Mannino F, Arcoraci V, … Bitto A (2017). Oxidative stress: harms and benefits for human health. Oxidative Medicine and Cellular Longevity 2017:8416763. https://doi.org/10.1155/2017/8416763 Prieto P, Pineda M, Aguilar M (1999). Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of Vitamin E1. Analytical Biochemistry 269(2):337-341. https://doi.org/10.1006/abio.1999.4019 Rankou H, Culham A, Jury SL, Christenhusz MJM (2013). The endemic flora of Morocco. Phytotaxa 78(1):1-27. https://doi.org/10.11646/phytotaxa.78.1.1 Reza M, Ardekani S, Khanavi M, Hadjiakhoondi A (2010). Comparison of antioxidant activity and total phenol contents of some date seed varieties from Iran. Iranian Journal of Pharmaceutical Research 9(2):141-146. Rodríguez-Bernaldo de Quirós A, Lage-Yusty MA, López-Hernández J (2010). Determination of phenolic compounds in macroalgae for human consumption. Food Chemistry 121(2):634-638. https://doi.org/10.1016/j.foodchem.2009.12.078 Selaimia A, Azouz M, Chouikh A, Zga N, Besbes N (2020). Phytochemical study, antioxidant and antimicrobial activities of flavonoids and diethyl ether extracts from leaves and seeds of medicinal plant of Algeria flora: Retama monosperma (l.) boiss. PONTE International Scientific Researches Journal 76(4):42-52. https://doi.org/10.21506/j.ponte.2020.4.4 Sun A, Chi X, Yang X, Feng J, Li Y, Zhou J (2019). Applications and prospects of ultrasound-assisted extraction in Chinese herbal medicine. Open Access Journal of Biomedical Science 1(1):5-15. https://doi.org/10.38125/OAJBS.000103 Torres de Pinedo A, Peñalver P, Morales JC (2007). Synthesis and evaluation of new phenolic-based antioxidants: Structure–activity relationship. Food Chemistry 103(1):55-61. https://doi.org/10.1016/j.foodchem.2006.07.026 Yangui I, Younsi F, Ghali W, Boussaid M, Messaoud C (2021). Phytochemicals, antioxidant and anti-proliferative activities of Myrtus communis L. genotypes from Tunisia. South African Journal of Botany 137:35-45. https://doi.org/10.1016/j.sajb.2020.09.040 Yıldırım A, Şen A, Tuysuz M, Birteksöz Tan AS, Şenkardeş İ, Bitiş L (2022). In vitro investigation of antimicrobial, enzyme inhibitory and free radical scavenging activities of Inula salicina L. International Journal of Agriculture Environment and Food Sciences 6(3):389-395. https://doi.org/10.31015/jaefs.2022.3.7 Zeroual A, Sakar EH, Mahjoubi F, Chaouch M, Chaqroune A, Taleb M (2021). Effects of extraction technique and solvent on phytochemicals, antioxidant, and antimicrobial activities of cultivated and wild Rosemary (Rosmarinus officinalis L.) from Taounate region (northern Morocco). Biointerface Research in Applied Chemistry 12(6):8441-8452. https://doi.org/10.33263/BRIAC126.84418452 Zhang QW, Lin LG, Ye WC (2018). Techniques for extraction and isolation of natural products: A comprehensive review. Chinese Medicine 13(1):20. https://doi.org/10.1186/s13020-018-0177-x Zhang Y, Cai P, Cheng G, Zhang Y (2022). A brief review of phenolic compounds identified from plants: their extraction, analysis, and biological activity. Natural Product Communications 17(1):1-14. https://doi.org/10.1177/1934578X211069721 El Mekkaoui A et al. (2023). Not Sci Biol 15(1):11423 11 The journal offers free, immediate, and unrestricted access to peer-reviewed research and scholarly work. Users are allowed to read, download, copy, distribute, print, search, or link to the full texts of the articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author. License License License License ---- Articles published in Notulae Scientia BiologicaeNotulae Scientia BiologicaeNotulae Scientia BiologicaeNotulae Scientia Biologicae are Open-Access, distributed under the terms and conditions of the Creative Commons Attribution (CC BY 4.0) License. © Articles by the authors; Licensee SMTCT, Cluj-Napoca, Romania. The journal allows the author(s) to hold the copyright/to retain publishing rights without restriction. 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