Biology, Medicine, & Natural Product Chemistry ISSN 2089-6514 (paper) Volume 14, Number 1, April 2025 | Pages: 567-573 | DOI: 10.14421/biomedich.2025.141.567-573 ISSN 2540-9328 (online) Phytochemistry, Toxicity, and Antimalaria Activity of Mangrove Plant from Muara Badak Beach, East Kalimantan, Indonesia Usman1,*, Muh. Amir Masruhim1, Agung Rahmadani1, Erwin2 1Master Study Program of Chemistry Education and Undergraduate Study Program of Chemistry Education, Faculty of Teacher Training and Education, Mulawarman University, Samarinda, Indonesia. 2Department of Chemistry, Faculty of Mathematics and Natural Sciences, Mulawarman University, Samarinda, Indonesia Jl. Muara Pahu, Kampus Gn. Kelua, Samarinda, Kalimantan Timur, Indonesia. Corresponding author* sainusman@ymail.com Manuscript received: 06 February, 2025. Revision accepted: 27 June, 2025. Published: 26 August, 2025. Abstract This study aimed to determine the secondary metabolite content, toxicity, and antimalarial activity of methanol extracts of mangrove leaves Avicennia marina, Rhizophora mucronata, and Sonneratia caseolaris. Mangrove leaf extraction by maceration using methanol solvent, then qualitative phytochemical test (color test), toxicity test by BSLT method using Artemia salina, and antimalarial activity test using Plasmodium falciparum strain 3D7 with Giemsa staining microscopic method. The content of secondary metabolite compounds of methanol extracts of the three mangrove leaf species are alkaloids, flavonoids, saponins, phenolic compounds, steroids, triterpenoids and tannins. The toxicity of methanol extract of R. mucronata mangrove leaves against A. salina is included in the strong category. Meanwhile, the toxicity of S. caseolaris is included in the moderate category and A. marina is in the weak category. Mangrove species R. mucronata and S. caseolaris have good activity against P. falciparum 3D7 with IC50 values < 30 μg/ml, while A. marina shows poor activity against P. falciparum 3D7 with IC50 values > 50 μg/ml. Other tissue parts of these three mangrove species still need further exploration for their bioactivity against A. salina and P. falciparum 3D7. Keywords: Mangrove plant; Phytochemical; Toxicity; Antimalarial. INTRODUCTION Mangrove forests are plant communities located at the confluence of sea and land in tropical and subtropical regions, especially at latitudes 25°N. Various species of mangrove plants can grow and thrive in areas of high salt content, tides, extreme temperatures, strong winds, anaerobic and muddy, and sandy soils with morphological adaptations to their environment (Aljaghthmi et al., 2018; Bibi et al., 2018). Indonesia is one of the countries in Asia with the largest mangrove forest with the highest mangrove species diversity in the world. This condition is supported by the geographical location, geological history and typology of the islands, as well as the unique oceanographic characteristics of Indonesia Mangrove forests function to protect coastal areas from damage due to sea water abrasion, keep the coastline stable, as a breeding ground for marine biota such as fish, crabs, and shrimp. Mangrove plants also have various medicinal properties so that mangrove plants are widely used by people in coastal areas as traditional medicines (Joandani et al., 2019; Tefarani et al., 2019). People in coastal areas have long known and used mangrove plants as traditional medicines, especially to treat; skin diseases, rheumatism, blisters, arthritis, bleeding, asthma, sore throat, eye problems, stomach pain, infections, diabetes, HIV, hepatitis, smallpox, ulcers, diarrhea, malaria, astringent, aphrodisiac, antiulcer, antitumor, snake bite treatment, anti-aging drugs and anticancer. Mangrove plants are utilized in traditional medicine because they contain active chemical compounds. (Lopez et al., 2018; Lalitha et al., 2021; Syamsul E.S., 2022). Active chemical compounds that are thought to act as drugs to fight various diseases are compounds of the alkaloid group, steroids, triterpenes, phenolics compounds compounds, flavonoids, terpenoids, stilbenes, carotenoids, anthocyanins, anthocyanidins, inositol, saponins, long chain alcohols, tannins, amino acids, benzoquinones, coumarins, quinins, chalcones, lipid compounds, phorbol esters, rotenone, polyphenols, benzofurans, limonoids, sulfur procyanidins, gibberellins and xyloccensins (Kartikaningsih et al., 2024; Usman et al., 2023). Several Indonesian mangrove species such as S. caseolaris, A. marina, R. mucronata, and R. apiculata have pharmacological activity as potent abiomicrobials. https://doi.org/10.14421/biomedich.2025.141.567-573 mailto:sainusman@ymail.com 568 Biology, Medicine, & Natural Product Chemistry 14 (1), 2025: 567-573 A. marina, has activity as an anticancer. A. corniculatum, A. aureum, A. alba, and R. mucronata showed cytotoxic activity against WiDr cancer cells. A. illicifolius, A. marina and E. agallocha showed signifi cant analgesic activity. Other plant species from the family Meliaceae, Rubiaceae, Piperaceae, Acanthaceae, Myrtaceae, and Myrsinaceae, were shown to have antimalarial activity. Then the study reported that Meliaceae and Rhizophorae have antimalarial activity (Audah et al., 2022; Arbiastutie et al., 2022). This means that the content of secondary metabolite compounds of mangrove plants A. marina, R. mucronata, and S. caseolaris originating from the coastal beach of Muara Badak District, Kutai Kartanegara Regency, East Kalimantan, has the potential to find bioactive compounds as natural antimalarial alternative medicinal ingredients. Figure 1. Mangrove plant species A. marina, R. mucronata, and S. Caseolaris. MATERIALS AND METHODS Materials The materials used in this study were the mangrove leaf samples of A. marina, R. mucronata, and S. caseolaris species. Phytochemical test reagents include alkaloids, flavonoids, phenolics compounds compounds, steroids, triterpenoids, saponins, and tannins. Methanol solvent, H2SO4(p) solution, HNO3(p) solution, HCl(p) solution, and DMSO solution, Artemia salina shrimp larvae and P. falciparum strain 3D7. The instruments used were glassware generally used in the laboratory, analytical balances, vials, aluminum foil, test tube, micro*pipette, measuring flask, volume pipette, aerator, study lamp, mikroskop, and rotary evaporator. Preparation of sample The samples used in this study were mangrove species of A. marina, R. mucronata, and S. caseolaris. Samples were taken from the coastal beach, Muara Badak District, Kutai Kartanegara, East Kalimantan. Then, the sample was washed and dried by air drying without direct sunlight. The dried samples were mashed using a blender until they became powder. After obtaining the sample in powder form, the sample was macerated. The sample powder was weighed as much as 500 g, then put into a beaker and added methanol solvent to cover the entire surface of the sample and then covered with aluminum foil. After 24 h, the results of the first maceration can be taken and then macerated again on the sample dregs up to 3 x 24 h. The methanol extract obtained from the first to third maceration was collected and then concentrated using a rotary evaporator to obtain a concentrated methanol extract of A. marina, R. mucronata and S. caseolaris mangrove leaves (Mitra et al., 2021). Phytochemical test A Phytochemical test is a qualitative test carried out by observing the occurrence of color changes after the extract is added to the reagent. Phytochemical tests are carried out to determine the presence or absence of secondary metabolite compounds contained in a sample. In this study, phytochemical tests were carried out to determine the secondary metabolites contained in A. marina, R. mucronata, and S. caseolaris mangrove leaves (Harahap et al., 2021; Akasia et al., 2021). Alkaloid test The alkaloid test was carried out by mixing 10 mL of A. marina, R. mucronata, and S. caseolaris mangrove, which was heated, cooled and then filtered. 2 drops of the filtrate were added to the drip plate, 2 drops of Meyer's reagent, Dragendroff's reagent, and Wagner's reagent were added. Observe the color changes that occur. A positive indicator of the alkaloid test on Mayer's reagent is the formation of a white precipitate. The alkaloid test is said to be positive in the Wagner test if there is a brownish/brick red precipitate, and a positive test on the Dragendrof reagent is said to be positive if there is a brownish/brick red/red-orange precipitate. Phenolics compound test The phenolics compounds test was carried out by adding 2 drops of A. marina, R. mucronata, and S. caseolaris mangrove filtrate to a drip plate, adding 1% FeCl3 solution, then observing the color changes. A positive Usman et al. – Phytochemistry, Toxicity, and Antimalaria Activity of Mangrove Plant … 569 indicator of the phenol test is the formation of a blue- black color. Saponin test The Saponin test was carried out by mixing 2 drops of A. marina, R. mucronata, and S. caseolaris mangrove filtrate into a drip plate. Then 2 mL of distilled water was added, then shaken until a stable foam was formed, then 1 drop of 2N HCl was added. A positive indicator of the saponin test is the formation of a stable foam. Flavonoid test Flavonoid test was carried out by mixing 2 drops of A. marina, R. mucronata, and S. caseolaris mangrove filtrate with 5 mL of methanol, then adding a few drops of concentrated HCl and 1.5 g of magnesium powder. A positive indicator of the flavonoid test is the formation of a red color. Steroid and terpenoid test Triterpenoid and Steroid tests were carried out by adding 2 drops of the filtrate to the drip plate, then adding one drop of acetic anhydride and one concentrated sulfuric acid (Liebermann Burchard reagent). A positive indicator of the terpenoid test is the formation of a red or purple color and a positive steroid if the solution is blue or green. Toxicity test against A. salina The extracts of A. marina, R. mucronata, and S. caseolaris mangrove were prepared as 1000 mg/L mother liquor by dissolving 3.5 g of sample in 50 mL of methanol. The mother liquor was then made into a solution with various concentrations of 125 mg/L, 250 mg/L and 500 mg/L (Khasanah et al., 2020). The next step in the toxicity test using the BSLT method is hatching shrimp larvae. The hatching of eggs is carried out in an artificial aquarium container in the form of a tube, which is assisted by a 5 W incandescent lamp to stimulate the growth of larvae. The medium used to incubate shrimp larvae is artificial seawater, a mixture of distilled water and pure salt. The oxygen level needed during hatching must be more than 3 mg/L, therefore the artificial seawater media must be aired with an aerator. 0.5 mg/mL of yeast was added as a source of nutrition for A. salina. Within 24-36 h, usually the eggs have hatched into larvae called naupli. Active nauplii aged for 48 h are used as test animals in the experiment. Vials were provided for each group according to the concentration level and were repeated 2 times. The parent solution of A. marina, R. mucronata, and S. caseolaris mangrove was added to the vial according to the concentration level. The vial containing the test solution was dried until the solvent had evaporated for several days at room temperature in a desiccator so that only the scale remained from the sample extract and no longer smelled of solvent. Then added 2 drops of 1% DMSO to dissolve the sample. After the sample was dissolved with DMSO, 1 mL of artificial seawater was added, then 10 A. salina L shrimp larvae aged 48 h were added to the vial. One drop of yeast (0.6 mg/mL) was put into each vial as food for A. salina, then artificial seawater was added to the volume limit of 10 mL. The standard criterion for assessing the mortality of shrimp larvae is if the shrimp larvae do not show movement for a few seconds of observation. The manual method is to observe the larvae in the vial (Kurniawan et al., 2021). Antimalarial test Samples of methanol extract of A. marina, R. mucronata, and S. caseolaris mangrove leaves as much as 10 mg were dissolved in 1000 mL l % DMSO solution to make a test solution. The resulting test solution was then made into various concentrations by diluting it, so that the obtained concentrations of 100 μg/mL, 50 μg/mL, 10 μg/mL, 1 μg/mL and 0.1 μg/mL, and 0.01 μg/mL. Prepared test parasites that will be used have been synchronized as a ring stage with parasitemia ± 1% (hematocrit 5%). A test well (well 96) was prepared, 2 μL of the test solution was added with various concentrations. After that, 198 μL of the parasite was added at each concentration of each test solution. The test was repeated twice (Duplo). The test well is then put into the chamber and given mixed gas as 5% O2, 5% CO2, and 90% N2. Incubated for 2 x 24 h at 37°C. Then, after 2 x 24 h, the cultures were harvested and a thin blood film was prepared with 20% gymnasia staining. Then, data analysis was performed by counting the number of infected erythrocytes per 1000 normal erythrocytes under a microscope. The data is then used to determine the percent growth and percent inhibition. After obtaining the percent inhibition value, the data were analyzed using the SPSS version 20 probit analysis program to obtain the IC50 value, which is the concentration of the test material that can inhibit the growth of parasites by 50% (Wardani et al., 2020). RESULTS AND DISCUSSION Result Phytochemical of Mangrove Plants Based on the results of phytochemical tests of methanol extracts of mangrove plant leaves (A. marina, R. mucronata, and S. caseolaris) is known to contain secondary metabolite compounds such as alkaloids, flavonoids, phenol compounds, steroids, triterpenoids, saponins, and tannins. The complete results of the phytochemical test of methanol extracts of the three mangrove leaf species are presented in Table 1. 570 Biology, Medicine, & Natural Product Chemistry 14 (1), 2025: 567-573 Table 1. Phytochemical screening of the third mangrove species. Secondary Metabolite Compounds Methanol Extract of Mangrove Leaf A. marina R. mucronata S. caseolaris Alkalaoids - + + Flavonoids - + + Saponins + + + Fenolic compound + + + Steroids - + + Triterpenoids + - - Tannins + + - Toxicity against A. salina Toxicity tests in this study used the Brine Shrimp Lethality Test (BSLT) method to determine the toxicity of methanol extracts from mangrove leaves (A. marina, R. mucronata, and S. caseolaris) against A. salina. The toxicity of methanol extracts of the three mangrove leaf species was determined by calculating IC50 values using SAS probit analysis. LC50 (Lethal Concentration 50%) value is the dose value of a compound that can kill 50% of test animals. The results of calculating the LC50 value of the three methanol extracts of mangrove leaves are presented in Table 2. Table 2. The LC50 value and toxicity level of mangrove species A. marina, R. mucronata and S. saseolaris. Methanol Extract of Mangrove Leaf LC50 Value (ppm) Category Toxicity A. marina 256,132 Low R. mucronata 48,165 Strong S. caseolaris 104,96 Moderate Antimalarial Activity The antimalarial test of methanol extracts of mangrove leaves of A. marina, R. mucronata, and S. caseolaris was carried out in vitro with Giemsa staining test method. This test uses P. falciparum strain 3D7. Antimalarial test data were obtained using SPSS probit analysis to determine the IC50 value of the methanol extract of mangrove leaves of A. marina, R. mucronata, and S. caseolaris. The IC50 values obtained are presented in Table 3 and Figure 2. Table 3, The IC50 values of mangrove A. marina, R. mucronata and S. sasseolaris. Methanol Extract of Mangrove Leaf Treatment of Concentration Variation (μg/ml) IC50 (μg/ml) (μg/ml) 100 50 10 1 0,1 0,01 A. marina 52,93 45,93 40,23 22,80 10,91 2,36 57,341 R. mucronate 73,04 52,28 37,13 17,59 12,05 6,76 24,118 S. casseolaris 100 63,76 36,32 14,66 9,85 0,24 21,975 Figure 2. IC50 values (antimalarial activity) of methanol extracts of mangrove leaves of A. marina, R. mucronata and S. caseolaris. Discussion Phytochemical According to Table 1. It is known that the methanol extract of A. marina mangrove leaves contains secondary metabolite compounds; saponins, phenolics compounds, triterpenoids, and tannins. R. mucronata leaf extract contains compounds; alkaloids, flavonoids, saponins, phenolics compounds, steroids, and tannins. While S. caseolaris mangrove leaf extract contains alkaloid compounds, flavonoids., saponins, phenolics compounds, and steroids. In the research of Audah et al., (2022) reported that S. caseolaris mangrove leaf extract contains compounds; steroids, flavonoids, and tannins. A. marina leaf extract is; steroids, flavonoids, saponins, and tannins. And R. mucronata leaf extracts are steroids, flavonoids, saponins, and tannins. Then Muhaimin, et al., (2019), stated that the content of secondary metabolite compounds in methanol extracts of S. caseolaris leaves are alkaloids, flavonoids, phenolics compounds, steroids, tannins, quinones, and glycosides. According to Akasia et al., (2021) the secondary metabolite compounds contained in mangrove extracts of S. caseolaris are flavonoids, saponins, phenolics compoundss and steroids. In other studies, it has been reported that the secondary metabolite compounds of R. mucronata are alkaloids, flavonoids, lipids, inositol, triterpene, phenolics compoundss compounds, and tannins (Hardoko et al., 2016; Taniguchi et al., 2018). It was reported that the methanol extract of R. mucronata stem bark has successfully isolated six compounds, including cinchonain Ib, breynioside B, polystachyol, β-sitosterol 3-O-β-D-glucopyranoside, β-sitosterol 3-O-β-D-(6'-O- palmitoyl) glucopyranoside, and β-sitosterol 3-O-β-D- (6'-O-stearoyl) glucopyranoside (Linh et al., 2020). 57,341 24,118 21,975 0 10 20 30 40 50 60 70 A. marina R. mucronate S. caseolaris N il ai I C 5 0 ( μ g /m l Methanol Extract of Mangrove Leaf Usman et al. – Phytochemistry, Toxicity, and Antimalaria Activity of Mangrove Plant … 571 The difference in the content of phytochemical compounds in the three mangrove plant species may be due to the location of their habitat, where each location of mangrove plant habitat has a different pH value so that it can affect the content of secondary metabolite compounds contained in mangrove plants. Then the solvent used during extraction also affects the content of the compounds obtained (Akasia et al, 2021). Toxicity According to the data presented in Table 2, it shows that the methanol extract of R. mucronata mangrove leaves has the lowest LC50 value of 48,165 ppm, this shows that the methanol extract of R. mucronata mangrove leaves is toxic to A. salina with a strong category. While the toxicity of the methanol extract of S. caseolaris mangrove leaves has an LC50 value of 104.96 ppm with a moderate category, then the toxicity of the methanol extract of A. marina mangrove leaves has an LC50 value of 256.132 ppm with a weak category. Meyer et al., (1982) stated that the toxicity level of plant extracts can be determined by looking at the LC50 value. If the LC50 value is smaller than 1000 ppm, it is considered toxic, whereas if the LC50 value is higher than 1000 ppm, it is considered non-toxic. Under the Meyer category, it can be declared that the methanol extract of mangrove leaves A. marina, R. mucronata, and S. caseolaris, are toxic to A. salina with an LC50 value of < 1000 ppm. Audah et al. (2022), reported that the water extract of S. caseolaris leaves, ethanol extract of A. marina mangrove leaves, and n-hexane extract of R. mucronata are toxic to A. salina, each with an LC50 value of 229.77 ppm and 160.43 ppm is toxic to A. salina with an LC50 value of 488.93 ppm. Bokshi B. (2020), reported that the ethyl acetate fraction of the stems and the carbon tetrachloride fraction of the leaves of S. caseolaris mangrove showed very strong toxicity properties against A. salina, with LC50 values of 25.0±0.05 and 25.0±0.07 μg/ml, respectively. According to Rozirwan et al. (2022), it was reported that A. marina mangrove leaf extract contains bioactive compounds such as alkaloids, saponins, flavonoids, steroids and steroids which are toxic to A. salina which can be developed for the pharmaceutical field. Based on the Brine Shrimp Lethality Test (BSLT), ethanol, ethyl acetate, and n- hexane extracts of R. mucronata mangrove are classified as toxic to A. salina, and the secondary metabolite compounds found in the ethanol extract are terpenoids, alkaloids, ketones, phenols, turpentine oil, essential oils, unsaturated fats, and camphor (Zulfahmi I et al., 2024) The bioactivity of medicinal plants can be detected by the content of phytochemical compounds in plants. From the results of the phytochemical test, it is known that the methanol extract of R. mucronata mangrove leaves contains alkaloids and flavonoids, where the two compounds are known to be toxic because they can work as respiratory tract poisons and even alkaloids can cause stomach poisoning, thus inhibiting the ability of organisms to eat. At the same time, alkaloid compounds can also block taste receptors in the mouth area of A. salina larvae, so that larvae do not get taste stimuli and are unable to recognize their food and as a result A. salina larvae die of starvation (Davis et al., 2019). The three mangrove leaf extracts in this study were positive for saponin compounds. Saponin compounds can affect the life of A. salina larvae, because the glycosides contained in saponins can bind oxygen in water, so that the oxygen content in the solution is reduced and as a result the larvae will die from lack of oxygen (Khasanah et al., 2020). While the methanol extract of mangrove leaves A. marina does not contain alkaloids and flavonoids, so the level of toxicity is lower than mangrove R. mucronata and S. caseolaris. Antimalarial Activity According to the results obtained (as presented in Table 3 and Figure 3) methanol extracts of mangrove leaves A. marina, R. mucronata, and S. caseolaris can inhibit the growth of P. falciparum strain 3D7 respectively with IC50 values of 57.34; 24.12; and 21.97 µg/mL. According to Gessler, the level of antimalarial activity with in vitro methods can be categorized into 3 groups, namely very good if the IC50 value is < 10 µg/mL, a fairly good category if the IC50 value is between 10 - 50 µg/mL, and less active of category if the IC50 value is > 50 µg/mL (Gessler et al., 1994). Depending on the IC50 value, A. marina mangrove leaf extract has antimalarial activity with less active of category because the IC50 value > 50 µg/mL. Meanwhile, methanol extracts of mangrove leaves of R. mucronata and S. caseolaris showed fairly good antimalarial activity with IC50 values between 10- 50 µg/mL. R. mucronata mangrove plants contain saponin compounds, which can act as an inhibitory mechanism against malaria by forming complexes with cell membranes through hydrogen bonds, thereby damaging the permeability of malaria cell walls and causing death. Saponins are also substances that can hemolyzes blood. Therefore, saponins can hemolyzes malaria cell membranes in the same way as red blood cell membranes (Juniharti et al., 2021; Gunawan*et al., 2018). R. mucronata leaf extract also contains alkaloid compounds, where these alkaloid compounds can prevent parasite growth in the blood and prevent the formation of peptidoglycan so as to prevent the formation of cell walls due to cell breakdown. The mechanism of action of alkaloids as antimalarial drugs is to prevent the detoxification of hemiparasites in the food vacuole. Likewise, the leaves of mangrove S. caseolaris also contain saponin compounds, so they act as substances that can hemolyzes blood. The mechanism of flavonoid compounds as antimalarial agents is to block the nutrient channels needed by parasites by forming specific membranes (Uzor P. F, 2020; Variani et al., 2021). However, A. marina mangrove leaf extract does not contain alkaloids and flavonoids, so it does not have 572 Biology, Medicine, & Natural Product Chemistry 14 (1), 2025: 567-573 antimalarial activity and has no potential as a malaria herbal medicine. The accumulation of phytochemical compounds contained in the extracts of mangrove leaves R. mucronata and S. caseolaris can provide antimalarial activity by increasing red blood cell oxidation or inhibiting protein synthesis, thus becoming a factor that influences the antimalarial potential of mangrove plants R. mucronata and S. caseolaris (Ahmed et al., 2010). R. mucronata extract is also known to have antidiabetic, antioxidant, and tissue-enhancing effects that are potentially compared to A. marina (Al-Jaghthmi et al., 2020). In another study it was reported that ethanol extract derived from S. alba leaves showed significant antiplasmodial efficacy in reducing parasitemia levels, inducing significant inhibition percentages, and inhibiting Plasmodium growth. Thus, S. alba leaves have the potential to be a promising natural source of antimalarial agents because ethanol extract derived from S. alba leaves has diverse phytochemical constituents that are efficacious as antiplasmodial. (Muhaimin M et al., 2024). Traditional medicine using mangrove skin, bark, fruit, and flowers from the species Lumnitzera littorea, Rhizophora mucronata, Scyphiphora hydrophyllacea, Sonneratia alba, Xylocarpus granatum, and Xylocarpus moluccensis is a popular method used as a malaria drug in people's social life. (Tamalene M.N. et al., 2021). CONCLUSION The secondary metabolite compounds contained in the methanol extract of mangrove leaves R. mucronata and S. caseolaris have antimalarial activity against P. falciparum starin 3D7 with a fairly good category including the IC50 value between 10 - 50 µg/ml, hence the two species of mangrove have potential to be developed as a natural antimalarial herbal medicine. The mangrove species A. marina, R. mucronata and S. caseolaris are toxic to A. salina with a strong category (LC50 value < 1000 ppm), Hence that the third species of mangrove can be examined by cancer cells to determine its potential as a natural anticancer medicinal material. Acknowledgement: We acknowledge, Institute of Tropical Disease (ITD) Airlangga University, Chemistry Education Laboratory, and Pharmacy Laboratory of Mulawarman University for all the support to carry out work. Conflict of Interest: The authors declare that they have no conflict of interests. REFERENCES Ahmed R., Moushumi S. J., Ahmed H., Ali M. 2010. Serum glucose and lipid profiles in rats following administration of Sonneratia caseolaris (L.) Engl. (Sonneratiaceae) leaf powder in diet. Adv Nat App Sci. 4(2), 171-173. https://www.researchgate.net/publication/289417859 Akasia A. I., Putra I. D. N. N., and Putra I. N. G. 2021. Phytochemical Screening of Mangrove Leaf Extracts of Rhizopora mucronata and Rhizopora apiculata collected from Mangrove Area of Tuban Village, Bali. Journal of Marine Research and Technology. 4, 16-22. https://doi.org/10.24843/JMRT.2021.v04.i01.p03 Al-Jaghthmi O., Heba H., Zeid I. A. 2018. Bioactive Compounds Extracted from Mangrove Plants (Avicennia marina, and Rhizophora mucronata): an Overview. Pathophysiology. DOI: https://doi.org/10.1016/j.pathophys.2018.09.002 Al-Jaghthmi O.H.A., Zeid I.E.M..E.A., Al-Ghamdi K.M.S., Heba H.M., Ahmad M.S. 2020. Antihyperglycemic, Antioxidant and Antiapoptotic Effect of Rhizophora Mucronata and Avicennia Marina in Streptozotocin-induced iabetic Rats. Original Paper, Md Ach. 2020 E; 74(6): 421-427 421. doi: 10.5455/medarh.2020.74.421-427 Arbiastutie Y., Diba F., and Masriani M. 2022. Cytotoxicity Activity of Several Medicinal Plants Grow in Mangrove Forest against Human’s Cervical (HELA), Breast (T47D), and Colorectal (WiDr) Cancer Cell Lines. International Journal of Nutrition, Pharmacology, Neurological Diseases. 12, 46-50. Doi: 10.4103/ijnpnd.ijnpnd_57_21 Audah K. A., Ettin J., Darmadi J., Azizah N. N., Anisa A. S., Hermawan T. D. F., Tjampakasari C. R., Heryanto R., Ismail I. S., and Batubara I. 2022. Indonesian Mangrove Sonneratia caseolaris Leaves Ethanol Extract Is a Potential Super Antioxidant and Anti Methicillin-Resistant Staphylococcus aureus Drug. Molecules. 27, 1-18. https://doi.org/10.3390/molecules27238369 Bibi S. N., Fawzi M. M., Gokhan Z., Rajesh J., Nadeem N., Kannan R. R. R., Albuquerque R. D. D. G., and Pandian S. K. 2018. Ethnopharmacology, Phytochemistry, and Global Distribution of Mangroves A Comprehensive Review. Mar. Drugs, 17, 1-82. https://doi.org/10.3390/md17040231 Bokshi B., Zilani Md.N.H., Hossain H., Ahmed Md.I., Anisuzzman M., Biswas N.N., and Sadhu S.K. 2020. Bioactivities of Sonneratia Caseolaris (Linn) Leaf and Stem Using Different Solvent Systems. Biomedical Journal of Scientific & Technical Research. 31(5), 24578-24582. DOI: 10.26717/BJSTR.2020.31.005175 Chen L., Zan Q., Shen J., Liao W. 2009. Litter dynamics and forest structure of the introduced Sonneratia caseolaris mangrove forest in Shenzhen, China. Estuarine, Coastal and Shelf Sci. 85 (2009) 241-246. DOI:10.1016/j.ecss.2009.08.007 Davis, Veronica, Maarisit W., Karauwan F., and Untu S. 2019. Toxicity Test of Ethanol Extract of Gossypium Hirsutum Cotton Leaf against Artemia Salina Shrimp Larvae with Brine Shrimp Lethality Test (BSLT) Method. Biofarmasetikal Tropis. 2, 71–77. https://doi.org/10.55724/jbiofartrop.v2i1.41 Gessler M. C., Nkunya M. H. H., Mwasumbi L. B., Heinrich M., and Tanner M. 1994. Screening Tanzanian Medicinal Plants for Antimalarial Activity. Acta Tropica. 56 (2019) 65–77. DOI: 10.1016/0001-706x(94)90041-8 Gunawan Desdy Hendra. 2018. Reduction of Saponin Compounds in Aloe Vera Gel by Boiling and Steaming. Journal of Food Technology. 9(1), 41-44. DOI: https://doi.org/10.35891/tp.v9i1.938 https://www.researchgate.net/publication/289417859 https://doi.org/10.24843/JMRT.2021.v04.i01.p03 https://doi.org/10.1016/j.pathophys.2018.09.002 https://doi.org/10.3390/md17040231 https://ui.adsabs.harvard.edu/link_gateway/2009ECSS...85..241C/doi:10.1016/j.ecss.2009.08.007 https://doi.org/10.55724/jbiofartrop.v2i1.41 https://doi.org/10.1016/0001-706x(94)90041-8 https://doi.org/10.35891/tp.v9i1.938 Usman et al. – Phytochemistry, Toxicity, and Antimalaria Activity of Mangrove Plant … 573 Harahap I. S., Halimatussakdiah, Amna U. 2021. Phytochemical Screening of Lemon Citrus Leaf Extract (Citrus limon L.) from Langsa City, Aceh. Quimica: Journal of Science and Applied Chemistry. 3(1), 19-23. DOI:10.33059/jq.v3i1.3492 Hardoko, Sasmito B. B., Puspitasari Y. E. 2016. Antidiabetic and antioxidant activities of tannin extract of Rhizophora mucronata leaves. Journal of Chemical and Pharmaceutical Research. 8(3), 143-148 Joandani G. K., Pribadi R., and Suryono C. A. 2019. Kajian Potensi Pengembangan Ekowisata Sebagai Upaya Konservasi Mangrove Di Desa Pasar Banggi, Kabupaten Rembang. Journal of Marine Research. 8(1), 117-126. https://ejournal3.undip.ac.id/index.php/jmr Juniharti A. E., Aulena D. N., Yantih N., Miftahurrohmah N., Kumala S. 2021. Produk Esktraseluler Isolat Kapang Endofi t C.1.1 dan C.3.3 dari Ranting Cempaka Kuning (Michelia champaca L.) sebagai Antimikroba. Jurnal Ilmu Kefarmasian Indonesia. 19(1), 131-138. DOI: https://doi.org/10.35814/jifi.v19i1.985 Kartikaningsih H., Iranawati F., Harlan L.I., Fauziyah J.N., Fathoni H.I., Koentjoro M.P., 2024.Toxicity of Aqueous Extracts of the Leaves of Sonneratia caseolaris Grown in Ujung Pangkah, Gresik, East Java. Trop J Nat Prod Res, 8(5):7213-7220. DOI: https://doi.org/10.26538/tjnpr/v8i5.24 Khasanah N. W., Karyadi B., Sundaryono A. 2020. Uji Fitokimia dan Toksisitas Ekstrak Umbi Hydnophytum Sp. terhadap Artemia salina Leach. PENDIPA Journal of Science Education. 4(1), 47-53. https://doi.org/10.33369/pendipa.4.1.47-53 Kurniawan H., and Rofiqa M. 2021. Uji Toksisitas Ekstrak Etanol Daun Ekor Kucing (Acalypha hispida Burm.f.) Dengan Metode Brine Shrimp Lethality Test (BSLT). Journal Syifa Sciences and Clinical Research. 3(2), 52-62 Lalitha P., Parthiban A., Sachithanandam V., Purvaja R., and Ramesh R.2021. Antibacterial and Antioxidant Potential of GC-MS Analysis of Crude Ethyl Acetate Extract from The Tropical Mangrove Plant Avicennia officinalis L. South African Journal of Botany. 142, 149–155. https://doi.org/10.1016/j.sajb.2021.06.023 Linh K. T. P., Quan N. H., Chien N. V., Trung N. Q., Thong V. H., Tuyen N. V., Thao N .P. 2020. Secondary Metabolites From The Stem Barks of Rhizophora mucronata L. Vietnam Journal of Science and Technology. 58(6), 653-664. DOI: https://doi.org/10.15625/2525-2518/0/0/14783 Lopez D., Cherigo L., de Sedas A., Spadafora C., Martinez L. A. 2018. Evaluation of antiparasitic, anticancer, antimicrobial and hypoglycemic properties of organic extracts from Panamanian mangrove plants. Asian Pacific Journal of Tropical Medicine. 11(1), 32-39. doi: 10.4103/1995-7645.223531 Meyer B. N., Ferrigny N. R., Putnam J. E., Jacobsen L. B., Nicols D. E., and Mc Laughlin J. L. 1982. Brine Shrimp, A Covenient General Bioassay for Active Plant Contituent. Journal of Medical Plant Research. 45 (1982) 31-34. doi: 10.1055/s- 2007-971236 Muhaimin M., Latief M., Putri R. D., Chaerunisaa A. Y., Aditama A. Y., Pravitasari N. E., Siregar J. E. 2019. Antiplasmodial Activity of Methanolic Leaf Extract of Mangrove Plants against Plasmodium berghei. Pharmacogn J. 11, 929-935. DOI:10.5530/pj.2019.11.148 Muhaimin M, Latifah N, Chaerunisaa A.Y, Subarnas A, Susilawati Y, Hirzan R. 2024. Antiplasmodial Activity of Ethanol Extract of Sonneratia alba Leaves. Trop J Nat Prod Res, 8(4):6884- 6890. https://doi.org/10.26538/tjnpr/v8i4.19 Rozirwana, Nugrohob R.Y., Hendria M., Fauziyaha, Putria W.A.E., Agussalima A. 2022. Phytochemical profile and toxicity of extracts from the leaf of Avicennia marina (Forssk.) Vierh. collected in mangrove areas affected by port activities. South African Journal of Botany 150 (2022) 903-919. https://doi.org/10.1016/j.sajb.2022.08.037. Syamsul E.S., Umar S, Wahyuni F.S., Martien R., Hamidi D. 2022. Anti-aging Activity, In Silico Modeling and Molecular Docking from Sonneratia Caseolaris Maced J Med Sci. 2022 Aug 27; 10(A):1471-1477. https://doi.org/10.3889/oamjms.2022.10558. Tamalene M.N, Sen U.K, Bhakat R.K, Vianti E, Bahtiar, Suparman/ 2021. Utilization of mangrove plants as a source of Malaria medicine in North Maluku Province, Indonesia. Asian Journal of Ethnobiology. 4(2), 86-92. DOI: 10.13057/asianjethnobiol/y040203 Taniguchi K., Funasaki M., Kishida A., Sadhu S. K., Ahmed F., Ishibashi M., Ohsaki A. 2018. Two new coumarins and a new xanthone from the leaves of Rhizophora mucronate. Bioorganic & Medicinal Chemistry Letters. 28 (6) 1063-1066. https://doi.org/10.1016/j.bmcl.2018.02.022 Tefarani R., Martuti N. K. T., and Ngabekti S. 2019. Mangrove Species Diversity and Zoning in the Mangunharjo Subdistrict, Tugu District, Semarang City. Life Science. 8(1), 41–53. https://doi.org/10.15294/lifesci.v8i1.29989 Usman, Amir M.M., Erika F., Nurdin M., Kuncoro H. 2019. Antidiabetic activity of leaf extract from three types of mangrove originating from sambera coastal region Indonesia. Res. J. Pharm. Technol. 12, 1707–1712. doi: 10.5958/0974- 360X.2019.00284.1 Usman U, Masruhim M.A, Kusumingtyas P, Erwin E. and Bulan D.E. 2023. Antioxidant and Antidiabetic from Rhizophora mucronata Derived from Sambera Beach, East Kalimantan, Indonesia. Trop J Nat Prod Res, October 2023; 7(10):4921- 4926. http://www.doi.org/10.26538/tjnpr/v7i10.31 Uzor Philip F. 2020. Alkaloids from Plants with Antimalarial Activity: A Review of Recent Studies. Evidence Based Complementary and Alternative Medicine. 8749083 (2020) 1- 17. https://doi.org/10.1155/2020/8749083 Variani Y. A., Setyaningrum E., Handayani K., Nukmal N., Arifiyanto A. 2021. Bioactive Compound Analysis of Secondary Metabolite Extract of Serratia marcescens strain MBC1. Indonesia Journal of Chemical Analysis. 4(2), 64-71. DOI: 10.20885/ijca.vol4.iss2.art3 Wardani A. K., Wahid A. R., Astuti Y. 2020. In Vitro Antimalarial Activity Assay of Ashitaba Leaf Ethanol Extract (Angelica keiskei). Pharmacy Education (2021) 21(2) 27 - 30. https://doi.org/10.46542/pe.2021.212.2730 Wirandita N.A., Hendri M., and Aryawati R. 2023. Antioxidant Activity and Toxicity Test on Mangrove (Rhizophora Apiculata) from Banyuasin Waters, South Sumatra. Oceanography & Fisheries Open access Journal. 16(3), 001- 008. DOI: 10.19080/OFOAJ.2023.16.555936 Zhao C., and Qin C. Z. 2022. Identifying large-area mangrove distribution based on remote sensing: A binary classification approach considering subclasses of non-mangroves. International Journal of Applied Earth Observation and Geoinformation. 108 (2022) 1–17. https://doi.org/10.1016/j.jag.2022.102750 https://doi.org/10.35814/jifi.v19i1.985 https://doi.org/10.26538/tjnpr/v8i5.24 https://doi.org/10.33369/pendipa.4.1.47-53 https://doi.org/10.1016/j.sajb.2021.06.023 https://doi.org/10.15625/2525-2518/0/0/14783 http://dx.doi.org/10.5530/pj.2019.11.148 https://doi.org/10.26538/tjnpr/v8i4.19 https://doi.org/10.1016/j.sajb.2022.08.037 https://doi.org/10.3889/oamjms.2022.10558 https://doi.org/10.1016/j.bmcl.2018.02.022 https://doi.org/10.15294/lifesci.v8i1.29989 http://www.doi.org/10.26538/tjnpr/v7i10.31 https://doi.org/10.1155/2020/8749083 http://dx.doi.org/10.20885/ijca.vol4.iss2.art3 https://doi.org/10.46542/pe.2021.212.2730 https://doi.org/10.1016/j.jag.2022.102750 THIS PAGE INTENTIONALLY LEFT BLANK