Toxicity of Extracts and Fractions of Bangkal Leaves (Nauclea subdita (Korth.) Steud.) and Genjer (Limnocharis flava (L.) Buch) on the Bioindicator Artemia salina Leach

Arsyik Ibrahim; Sealti Puji Salea; Sri Windarti; M. Rahmad Ramadhan; Hidfdzur Rashif Rija'i; M. Arifuddin; Laode Rijai

doi:10.70392/jpns.v3i1.47

Authors

Arsyik Ibrahim Mulawarman University Author
Sealti Puji Salea Mulawarman University Author
Sri Windarti Mulawarman University Author
M. Rahmad Ramadhan Mulawarman University Author
Hidfdzur Rashif Rija'i Mulawarman University Author
M. Arifuddin Mulawarman University Author
Laode Rijai Mulawarman University Author

DOI:

https://doi.org/10.70392/jpns.v3i1.47

Keywords:

Bangkal, Genjer, Metabolites, IC50, Artemia salina

Abstract

A study was conducted on the secondary metabolite content and toxicity testing of Artemia salina leach larvae using extracts from Bangkal leaves (Nauclea subdita (Korth.) Steud.) and Genjer plants (Limnocharis flava (L.) Buch). The objective of this study is twofold: firstly, to ascertain the compounds present in Bangkal leaf extract and Genjer plant extract, and secondly, to assess the toxicity levels of the extracts and fractions from Bangkal leaves and Genjer plants on Artemia salina Leach larvae. In this study, the extraction method employed was maceration with 96% ethanol as the solvent. The crude 96% ethanol extract was fractionated in stages using n-hexane, ethyl acetate, and n-butanol solvents based on polarity levels. The toxicity testing was conducted using the Brine Shrimp Lethality Test (BSLT) method, with Artemia salina Leach larvae serving as the bioindicator. A range of test concentrations were utilised, with each concentration having three replicates. The mortality of Artemia salina was observed after a 24-hour period and was analysed using the Reed and Muench method to determine the LC₅₀ value. The results of the study demonstrated that the secondary metabolites present in the 96% ethanol extract of bangkal leaves com-prised the following groups of compounds: steroids, and triterpenoids, phenols, quinones, and saponins. The toxicity test results of the bangkal leaf extract and fractions yielded LC₅₀ values of 418.79 ppm for the 96% ethanol extract; 716.14 ppm for the n-hexane fraction; 263.03 ppm for the ethyl acetate fraction; and 70,79 ppm for the extract n-buthanol fraction. Meanwhile, the toxicity test results of the genjer plant extract and fractions obtained an LC₅₀ value of 490.23 ppm for the 96% ethanol extract; n-hexane fraction 551.31 ppm; ethyl acetate fraction 425.01 ppm; and n-butanol fraction extract 46.02 ppm. These results indicate that the n-butanol fraction of bangkal leaves and genjer plants have stronger cytotoxic activity than other fraction extracts.

Author Biography

Arsyik Ibrahim, Mulawarman University

Laboratorium Riset dan Pengembangan Kefarmasian “FARMAKA TROPIS”, Fakultas Farmasi

References

Harborne, A.J. Phytochemical methods: A guide to modern techniques of plant analysis, 1998, 2nd ed., Chapman and Hall Ltd, London, Great Britain.

Manjang, Y. Research on Natural Material Chemistry for Plant Preservation and Development Through Agro-Tourism Plants, 2006, Director General of Higher Education of the Ministry of National Education, Padang.

Anonymous. Indonesia has 7,500 medicinal plants in the Indonesian Institute of Sciences. Available online: URL http://lipi.go.id/berita/single/Indonesia-Miliki-7500Tanaman-Obat/11540 (accessed Desember 24, 2025).

Soendjoto, M.A., Riefani, M.K. Bangkal (Nauclea sp.), a wetland plant, is the material for the cool face powder. Wetland Conservation News, 2023, 13–18.

Abbah, J., Amos, S., Chindo, B., Ngazal, I., Vongtau, H.O., Adzu, B., Farida, T., Gamaniel, K.S., Pharmacological evi-dence favouring the use of Nauclea latifolia in malaria ethnopharmacy: Effects against nociception, inflammation, and py-rexia in rats and mice. Journal of Ethnopharmacology, 2010, 127(1), 85–90. doi:10.1016/j.jep.2009.09.045.

Traore, F., Gasquet, M., Laget, M., Guiraud, H., Di Giorgio, C., Azas, N., Doumbo, O., Timon-David, P. Toxicity and genotoxicity of antimalarial alkaloid-rich extracts derived from Mitragyna inermis O. Kuntze and Nauclea latifolia. Phyto-therapy Research, 2000, 14(8), 608–611. Doi: 10.1002/1099-1573(200012)14:8<608:AIDPTR667> 3.0.CO; 2-D.

Wardhani, R.A.A., Akhyar, O. Phytochemical Screening, Antioxidant and Antibacterial Activity Test of Propionibacte-rium acnes Ethanol Extract of Bark and Leaves of Scarlet Plant (Nuclea subdita) Screening. Periodical Scientific Journal of Chemical Science and Applications, 2018, 12, 64–75. doi: http://dx.doi.org/10.20527/jstk.v12i2.4956.

El-Mahmood, A.M., Doughari, J.H., Chanji, F.J. In vitro antibacterial activities of crude extracts of Nauclea latifolia and Daniella oliveri. Scientific Research and Essays, 2008, 3(3), 102–105.

Liu, W., Di Giorgio, C., Lamidi, M., Elias, R., Ollivier, E., De Méo, M.P. Genotoxic and clastogenic activity of saponins extracted from Nauclea bark as assessed by the micronucleus and the comet assays in Chinese Hamster Ovary cells. Journal of Ethnopharmacology, 2011, 137(1),176–183. doi: 10.1016/j.jep.2011.05.005.

Rahmi, N, Salim, R, Miyono,and Ikhwan, M, The Effect of Solvents and Extraction Methods on Antibacterial and Free Radical Scavenging Activities from Bangkal (Nauclea subdita) Bark Extracts. Jurnal Penelitian Hasil Hutan, 2021, 39 (1), 13-26.

Brata, A, Azizah, L, Muin, D. Effectiveness of the Use of Bengkal Leaf Infusion (Nauclea orientalis L.) As an antidiabetic against mice. Bahana Journal of Public Health, 2023, 7(2), 75-81

Perkasa, A.Y., Petropoulos, S. Genjer Yellow Velvetleaf used as an indigenous vegetable in Indonesia, Anatolian Journal of Botany, 2020, 4(1), 76–79. doi: 10.30616/ajb.710777.

Ade, F., Kurnia, A. Identification and Isolation of Flavonoid Compounds of Ethanol Extract of Genjer Leaves (Limnocharis flava (L.) Buchenau). Journal of the National Pharmaceutical Society, 2021, 1(1), 1-9.

Abriyani, E., Fikayuniar, L, Silvi, M.A, and Wichandar, A. Phytochemical Screening and Antioxidant Test of Limnocharis flava L Flower Extractby DPPH Method. Journal of Buana Farma, 2022, 2(2), 56-59.

Kristiani, S, Billi, J, Efendi, H. Determination of Total Flavonoid Levels and Antioxidant Activity Test of Genjer Leaf and Stem Extract (Limnocharis flava) by ABTS Method. Journal of Borneo Scholars, 2023, 7(1), 11-21. DOI: https://doi.org/10.36805/jbf.v2i2.392

Socajiwa, P. Antioxidant Activity Test of Genjer Plant Extract (Limnocharis Flava) Using the DPPH Method, 2022. Thesis of the Bachelor of Pharmacy Study Program, Faculty of Health Sciences, Dr. Soebandi University Jember. p.1-51

Jacoeb, A.M., Abdullah, A., Rusydi, R. Microscopic characteristics and bioactive components of genjer plant (Limnocharis flava) from Situ Gede, Bogor. Journal of Aquatic Resources, 2010, 4(2),1-8.

Narwanti, I. and Hamida, I.A., Antioxidant Activity Test of N-Hexane Fraction, Chloroform, and Ethyl Acetate of Ethanol Extract of Limnocharis flava with DPPH Method. Journal of Indonesian Pharmaceutical Personnel, 2018, 1(2), 251–259.

Rachman, V.S., Qomariyah, A., Najib, A., Antioxidant Test on Genjer Extract (Limnocharis flava) and Hand Soap from Genjer Leaves. Journal of Chemical Engineering Innovation. 2024, 9(4), 295-303

Ooh, K.F, Ong, H.C, Wong, F.C, Chai, T. HPLC profiling of phenolic acids and flavonoids and evaluation of an-ti-lipoxygenase and antioxidant activities of aquatic vegetable Limnocharis flava. Acta Poloniae Pharmaceutica Drug Re-search, 2015, 72(5), 973-979

Utami, N.F., Nurdayanty, S.M., Sutanto, U.S. Effect of Various Extraction Methods on the Determination of Flavonoid Levels of Ethanol Extract of Iler Leaf (Plectranthus scutellarioides). Phytopharmaceuticals: J. Pharmaceutical Sciences, 2020, 10(1),76–83. DOI: 10.33751/jf.v10i1.2069.

Payung, R, Gunawan, E, Pratiwi, R.D. Cytotoxic Activity Test of Matoa Leaf Fraction (Pometia pinnata) by Brine Shrimp Lethality Test (BSLT) Method. J Biol Papua, 2021, 13, 83–91.

Parlin, D.A, Nasution, M.P, Nasution, H.M, Daulay, A.S. Phytochemical Screening and Cytotoxicity Test of Matoa Leaf Ethanol Extract (Pometia pinnata) using the BSLT Method. Journal of Pharmacy, Science and Health, 2022, 2(1), 38–48. DOI: https://doi.org/10.32696/fjfsk.v2i1.1372.

Marliza, H, Oktaviani, D. Cytotoxic Test of Ethanol Extract of Kemunu Leaves (Colacasia gigantea Hook.f) by Brine Shrimp Lethality Test (BSLT) Method. Bencoolen J Pharm, 2021, 1, 38–45.

Lu, F.C. Basic, Basic, Target Organ, and Risk Assessment. 2nd edition. Koensoenmardiyah Translation, IKIP Semarang Press, Semarang, 1995, 265-287

Meyer, L., Ferrigni. Brine Shrimp Coveniens General Bioassay for Active Constituents. Planta Medica 1, 1982, 4, 31-34

Ibrahim, A., Bulan, A.S., Ramadhan, M.R., Bone, M., Rashif, H.R., Rusman, A., Arifuddin, M., Junaidin, Riki, Rijai, L, Secondary Metabolites and Cytotoxicity of Kitolod Leaf Extract (Hippobroma longiflora L.) Against Shrimp Larvae (Artemia salina Leach). J Riseta Naturafarm, 2025, 2(1), 33-42. https://doi.org/10.70392/jrn.v2i1.3342.

Saifudin, A. Standardization of Natural Medicinal Ingredients. 2011, Yogyakarta: Graha Ilmu.

Nur, T., Rusman, A., Hikmawan, B.D., Almaida, M., Herman, H., Ibrahim, A. Secondary Metabolites and Cytotoxic Bioactivity Tests of Chopstick Fruit Extract (Brucea javanica (L.) Merr) against Artemia salina Leach. J Researchta Natura-farm, 2024, 1(1), 22–30. DOI: https://doi.org/10.70392/368zcy93.

Ahmad, I., Ibrahim, A. Bioactivity of Methanol Extract and N-Hexane Fraction of Sungkai Leaf (Peronema canescens Jack) Against Shrimp Larvae Artemia salina Leach. Jurnal Sains dan Kesehatan, 2015, 1(3), 114–9. DOI: https://doi.org/10.25026/ jsk.v1i3.27.

Colegate, S.M. Molyneux, and Russel. J. Bioactive Natural Products. 1993, CRC Press, United States of America.

McLaughlin, J.L, Roggers, L.L. The Use of Biological Assays to Evaluate Botanicals. Drug Information Journal, 1998, 32, 513 – 524.

Sutrisno, R.B. Thin Layer Chromatography Reagents, 1993, Edition 1, Print 1, Pancasila University, Jakarta, Indonesia.

Monica, C., Zamzani, I., Nashihah, S. Antibacterial Activity of Ethanol Extract 96% of Bangkal Leaves (Nauclea subdita) Against Staphylococcus aureus Bacteria. Journal of Indonesian Pharmaceutical Personnel, 2024, 7(2), 33-43. doi: 10.36387/jifi.v7i2.2129

Gadouche, L., Alsoufi, A.L., Pacholska, D., Skotarek, A., Pączkowski, C.,and Szakiel, A. Triterpenoid and Steroid Con-tent of Lipophilic Extracts of Selected Medicinal Plants of the Mediterranean Region. Molecules J, 2023. 28(2), 1-10. Doi: 10.3390/molecules28020697.

Wardana, A.P. Elucidation of Isolated Compound Structures from Gowok (Shzygius polycephalum) Chloroform Tract Skin Extracts and Antioxidant Activity Test. 2016, Skripsi Department of Chemistry FMIPA, Surabaya University.

Suhendi, A., Sjahid, R, Hanwar, D. Isolation and Identification of Flavonoids from Dewandaru (Eugenia uniflora L.) Leaf. J.Pharmacon, 2011, 12(2), 73–81.

Ibrahim, A, Siswandono, S, Prajogo, B.E.W. Anticancer Activity of Peronema canescens Jack Leaves Extracts Against Human Cells: HT-29 and HeLa In Vitro. Research J. Pharm. and Tech., 2022, 15(10), 4739 – 4745. DOI: 10.52711/0974-360X.2022.00796.

Haryati, N.A., Erwin, C.S. Toxicity Test and Antibacterial Activity of Red Leaf Extract (Syzygium mytifolium Walp) against Staphylococcus aureus and Escherichia coli Bacteria. J. Chemistry Mulawarman, 2015, 13(1), 35-39.

Murtiningsih, T. M., Supriningrum, R., and Nurhasnawati, H. Identification and Determination of Saponin Content from Extract of Embelia borneensis Bark. Journal Sains dan Teknologi, 2023. 9(3), 117-120. Doi: 10.31602/jst.v9i3.12499.

Marliana, S.D, Saleh, C. Phytochemical Test and Antibacterial Activity of Ethanol Rough Extract, n-Hexane, Ethyl Ace-tate, and Methanol Fraction from Water Pumpkin Fruit (Lagenaria siceraria (Morliana). J. Mulawarman Chemistry, 2011, 8(2), 39-63.

Robinson, T. High Plant Organic Content. ITB Bandung, 1995, Bandung, Indonesia, 71–285

Aprilyanie, I, Handayani, V, Syarif, R.A. Toxicity Test of Fruit Skin Extract of Kaffir Lime Plant (Citrus hystrix DC) using the Brine Shrimp Lethality Test (BSLT) Method. Makassar Natural Product Journal, 2023, 1(1),1–9.

Nastiti, M., Erwin, Kusuma, I.W. Phytochemical Screening and Toxicity Test on Applied Leaves (Artocarpus elasticus) by Brine Shrimp Lethality Test (BSLT) Method. Pros Semin Nas Kim, 2017, 60, 69–73.

Rajabi, S, Ramazani, A. M, Naji, T. Artemia salina as a Model Organism in Toxicity Assessment of Nanoparticles. Journal of Pharmaceutical Sciences, 2015, 23(20), 1-6. Doi 10.1186/s40199-015-0105-x.

Bareta, A.R., Widiastuti E.L., Nurcahyani, N. Cytotoxicity Test of Taurine and Brown Macroalgae Ethanol Extract using BSLT (Brine Shrimp Lethality Test) method], Berita Biologi, 2023, 22(2), 133-157. Doi: 10.55981/beritabiologi.2023.660.