Phytochemical Profiling and Antioxidant Activity Assessment of Ethanolic Extract of Phyllanthus niruri
Abstract
Introduction: Phyllanthus niruri, recognized as Stonebreaker due to its medicinal value, has been used for centuries in traditional medicine. This plant exhibits significant antioxidant activity due to various bioactive components such as alkaloids, phenols, and flavonoids. Aim: This study aimed at screening for bioactive components and investigating its antioxidant activity. Methods: The aerial parts of plant were harvested, dried and grounded into fine powder. 10 grams of the powdered plant material was soaked in 100ml of ethanol to prepare ethanolic extract. A qualitative examination was conducted to identify the phytochemicals in the sample. DPPH and ABTS assays were performed to determine antioxidant activity. Absorbance was recorded at 517 nm for DPPH and 734 nm for ABTS. In both assays gallic acid was used as reference standard. Results: Many bioactive compounds such as flavonoids, alkaloids, phenols, triterpenoids, tannins, saponin, steroids, glycosides and carbohydrates were identified in plant extract. The plant extract demonstrated notable antioxidant activity showing an IC₅₀ value of 3.12 ± 0.26 μg/ml in the DPPH assay, which was less than gallic acid (IC₅₀ 1.51 ± 0.02 μg/ml). In ABTS assay, IC₅₀ value of plant extract was 19.98±0.09 μg/ml, which is higher than gallic acid (5.69 ± 0.05 μg/ml). Conclusion: Our results revealed potential antioxidant activity of Phyllanthus niruri, which is useful to treat several clinical conditions like inflammation and cellular damage by oxidative stress.
Keywords:
Antioxidants, Free Radical Scavenging, Oxidative Stress, Phyllanthus niruri, Phytochemicals
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References
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Mostofa, R., Ahmed, S., Begum, M. M., Sohanur Rahman, M., Begum, T., Ahmed, S. U., ... & Begum, R. (2017). Evaluation of anti-inflammatory and gastric anti-ulcer activity of Phyllanthus niruri L. (Euphorbiaceae) leaves in experimental rats. BMC Complementary and Alternative Medicine, 17(1), 267. https://doi.org/10.1186/s12906-017-1771-7
Muhammad, A. A., Ibrahim, R. B., Aminu, C., Abbas, A. Y., Kabiru, A., & Bisallah, C. I. (2020). Activity of aqueous extract of Phyllanthus niruri Linn in 2,4-dinitrophenylhydrazine induced anaemic rats. Journal of Pharmacy and Pharmacology Research, 4(3), 79–95. https://doi.org/10.26502/fjppr.034
Nawfetrias, W., Nurhangga, E., Reninta, R., Chotimah, S., Bidara, I. S., Maretta, D., Akmal, S., & Faizal, A. (2023, October). Metabolite profiling of the medicinal herb Phyllanthus niruri L. under drought stress. IOP Conference Series: Earth and Environmental Science, 1255(1), 012046. https://doi.org/10.1088/1755-1315/1255/1/012046
Noreen, H., Semmar, N., Farman, M., & McCullagh, J. S. (2017). Measurement of total phenolic content and antioxidant activity of aerial parts of medicinal plant Coronopus didymus. Asian Pacific Journal of Tropical Medicine, 10(8), 792–801. https://doi.org/10.1016/j.apjtm.2017.07.024
Pineda, E., Napalan, R., Obañana, R., Pablico, J., Pasaporte, J., Parreño, G., Pimentel, R., Polong, J., Parreño-de Guzman, E., Mendoza, M., & Pakingking, R. (2025). Phytochemical constituents, antioxidant and antibacterial activity of the Philippine Sampa-sampalukan (Phyllanthus niruri) leaf extract against selected human bacterial pathogens. Journal of Biological Studies, 8(1), 34–47. https://doi.org/10.62400/jbs.v8i1.13543
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Shaikh, J. R., & Patil, M. (2020). Qualitative tests for preliminary phytochemical screening: An overview. International Journal of Chemical Studies, 8(2), 603-608. https://doi.org/10.22271/chemi.2020.v8.i2i.8834
Singh, A., Chitra, V., & Ilango, K. (2024). Network pharmacology exploration to reveal molecular insights of Phyllanthus niruri in non-alcoholic fatty liver: In vitro and in silico evidence. Pharmacia, 71, 1-16. https://doi.org/10.3897/pharmacia.71.e110280
Singh, M. K., & Ahmad, M. S. (2020). Phytochemical profile of Phyllanthus niruri L. and evaluation of its potent bioactive compounds. Pharmacognosy Communications, 13, 1545–1551. https://doi.org/10.21786/bbrc/13.3/82
Singh, T., Kumar, R., & Singh, J. K. (2016). Acute toxicity study of Phyllanthus niruri and its effect on the cyto-architectural structure of nephrocytes in Swiss albino mice Mus-musculus. Pharmacognosy Journal, 8(1), 77-80. https://doi.org/10.5530/pj.2016.1.17
Sirajudeen, J., Ahamath, J. M., Vahith, R. A., Manivel, V., & Elamparithi, R. (2017). Phytochemical screening and antimicrobial activity of Phyllanthus niruri. Journal of Advanced Applied Scientific Research, 1(7). https://doi.org/10.46947/joaasr17201743
Skibska, B., Kochan, E., Stanczak, A., Lipert, A., & Skibska, A. (2023). Antioxidant and anti-inflammatory effects of α-lipoic acid on lipopolysaccharide-induced oxidative stress in rat kidney. Archivum Immunologiae et Therapiae Experimentalis, 71(1), 16. https://doi.org/10.1007/s00005-023-00682-z
Tang, J., Li, R., Wu, B., Tang, J., Kan, H., Zhao, P., Luo, X., & Liu, Y. (2024). Secondary metabolites with antioxidant and antimicrobial activities from Camellia fascicularis. Current Issues in Molecular Biology, 46(7), 6769–6782. https://doi.org/10.3390/cimb46070404
Tiwana, G., Cock, I. E., & Cheesman, M. J. (2024). Phyllanthus niruri Linn.: Antibacterial activity, phytochemistry, and enhanced antibiotic combinatorial strategies. Antibiotics, 13(7), 654. https://doi.org/10.3390/antibiotics13070654
Trivadila, T., Iswantini, D., Rahminiwati, M., Rafi, M., Salsabila, A. P., Sianipar, R. N. R., ... & Murni, A. (2025). Herbal Immunostimulants and Their Phytochemicals: Exploring Morinda citrifolia, Echinacea purpurea, and Phyllanthus niruri. Plants, 14(6), 897. https://doi.org/10.3390/plants14060897
Vishal., Dahiya, D. P., Sankhyan, A., & Sharma, A. (2025). An updated review on Phyllanthus niruri: Its phytochemistry, pharmacological activity, medicinal use and traditional use. International Journal of Pharmaceutical Sciences, 3(3), 2022-2027. https://doi.org/10.5281/zenodo.15062486
Wulandari, E. T., Soetrisno, S., Purwanto, B., Reviono, R., Wasita, B., & Laqif, A. (2024). Impacts of Phyllanthus niruri extract on biomarker levels, macrophage count, and lesion area in an endometriotic rat model. Narra Journal, 4(3), e1002. https://doi.org/10.52225/narra.v4i3.1002
Zonooz, S. R., Hasani, M., Morvaridzadeh, M., Pizarro, A. B., Heydari, H., Yosaee, S., Setayesh, L., & Heshmati, J. (2021). Effect of alpha-lipoic acid on oxidative stress parameters: A systematic review and meta-analysis. Journal of Functional Foods, 87, 104774. https://doi.org/10.1016/j.jff.2021.104774
Abdel-Sattar, O. E., Allam, R. M., Al-Abd, A. M., El-Halawany, A. M., El-Desoky, A. M., Mohamed, S. O., Sleem, A. A., Singab, A. N. B., & Meselhy, M. R. (2023). Hypophyllanthin and phyllanthin from Phyllanthus niruri synergize doxorubicin anticancer properties against resistant breast cancer cells. ACS Omega, 8(31), 28563–28576. https://doi.org/10.1021/acsomega.3c03169
Al Zarzour, R. H., Ahmad, M., Asmawi, M. Z., Kaur, G., Saeed, M. A. A., Al-Mansoub, M. A., Ibrahim, I. A., & Yam, M. F. (2017). Phyllanthus niruri standardized extract alleviates the progression of non-alcoholic fatty liver disease and decreases atherosclerotic risk in Sprague–Dawley rats. Nutrients, 9(7), 766. https://doi.org/10.3390/nu9070766
Arif, J., Nasution, A. N., Nasution, S. W., Widowati, W., Dewi, N. S. M., & Kusuma, H. S. W. (2024). Antioxidant activities and potential wound healing effects of meniran (Phyllanthus niruri) extract gel. Folia Medica Indonesiana, 60(1), 69-77. https://doi.org/10.20473/fmi.v60i1.42396
Beidokhti, M. N., Andersen, M. V., Eid, H. M., Villavicencio, M. L. S., Staerk, D., Haddad, P. S., & Jäger, A. K. (2017). Investigation of antidiabetic potential of Phyllanthus niruri L. using assays for α-glucosidase, muscle glucose transport, liver glucose production, and adipogenesis. Biochemical and Biophysical Research Communications, 493(1), 869–874. https://doi.org/10.1016/j.bbrc.2017.09.080
Ezzat, M. I., Okba, M. M., Ahmed, S. H., El-Banna, H. A., Prince, A., Mohamed, S. O., & Ezzat, S. M. (2020). In-depth hepatoprotective mechanistic study of Phyllanthus niruri: In vitro and in vivo studies and its chemical characterization. PLoS One, 15(1), e0226185. https://doi.org/10.1371/journal.pone.0226185
Giribabu, N., Rao, P. V., Kumar, K. P., Muniandy, S., Swapna Rekha, S., & Salleh, N. (2014). Aqueous extract of Phyllanthus niruri leaves displays in vitro antioxidant activity and prevents the elevation of oxidative stress in the kidney of streptozotocin‐induced diabetic male rats. Evidence‐Based Complementary and Alternative Medicine, 2014(1), 834815. https://doi.org/10.1155/2014/834815
Hilal, B., Khan, M. M., & Fariduddin, Q. (2024). Recent advancements in deciphering the therapeutic properties of plant secondary metabolites: phenolics, terpenes, and alkaloids. Plant Physiology and Biochemistry, 211, 108674. https://doi.org/10.1016/j.plaphy.2024.108674
Himawati, S., Rosdayanti, H., Reninta, R., Devy, L., Bidara, I. S., Roswanjaya, Y. P., ... & Taula’bi’, D. (2024). Impact of cultivation, harvesting time, and extraction time on flavonoid content of Phyllanthus niruri L. In AIP Conference Proceedings (Vol. 2957, No. 1, p. 040032). AIP Publishing LLC. https://doi.org/10.1063/5.0184244
Ilhami, F. B., Cahyaningrum, S. E., Wardana, A. P., Gultom, N. S., Subekti, H., Rahmawati, A., & Puspitarini, S. (2025). Meniran (Phyllanthus niruri L.) embedded zeolitic imidazolate framework (ZIF-8) nanoparticle for cancer chemotherapy: Supported molecular docking analysis. RSC Advances, 15(1), 223–230. https://doi.org/10.1039/D3RA07293A
Krishnamurthy, H. K., Pereira, M., Rajavelu, I., Jayaraman, V., Krishna, K., Wang, T., ... & Rajasekaran, J. J. (2024). Oxidative stress: fundamentals and advances in quantification techniques. Frontiers in Chemistry, 12, 1470458. https://doi.org/10.3389/fchem.2024.1470458
Kumar, P., Kumar, D., Pal, S., & Singh, S. (2025). Plant secondary metabolites in defense against phytopathogens: Mechanisms, biosynthesis, and applications. Physiological and Molecular Plant Pathology, 138,102639. https://doi.org/10.1016/j.pmpp.2025.102639
Lee, N. Y., Khoo, W. K., Adnan, M. A., Mahalingam, T. P., Fernandez, A. R., & Jeevaratnam, K. (2016). The pharmacological potential of Phyllanthus niruri. Journal of Pharmacy and Pharmacology, 68(8), 953-969. https://doi.org/10.1111/jphp.12565
Lin, B., Xu, P., Zheng, J., Deng, X., Ye, Q., Huang, Z., & Wang, N. (2022). Effects and mechanisms of natural alkaloids for prevention and treatment of osteoporosis. Frontiers in Pharmacology, 13, 1014173. https://doi.org/10.3389/fphar.2022.1014173
Markiewicz-Górka, I., Pawlas, K., Jaremków, A., Januszewska, L., Pawłowski, P., & Pawlas, N (2019). Alleviating effect of α-lipoic acid and magnesium on cadmium-induced inflammatory processes, oxidative stress and bone metabolism disorders in Wistar rats. International Journal of Environmental Research and Public Health, 16(22), 4483. https://doi.org/10.3390/ijerph16224483
Mohammed, M. S., Osman, W. J., Garelnabi, E. A., Osman, Z., Osman, B., Khalid, H. S., & Mohamed, M. A. (2014). Secondary metabolites as anti-inflammatory agents. The Journal of Phytopharmacology, 3(4), 275–285. https://doi.org/10.31254/phyto.2014.3409
Mostofa, R., Ahmed, S., Begum, M. M., Sohanur Rahman, M., Begum, T., Ahmed, S. U., ... & Begum, R. (2017). Evaluation of anti-inflammatory and gastric anti-ulcer activity of Phyllanthus niruri L. (Euphorbiaceae) leaves in experimental rats. BMC Complementary and Alternative Medicine, 17(1), 267. https://doi.org/10.1186/s12906-017-1771-7
Muhammad, A. A., Ibrahim, R. B., Aminu, C., Abbas, A. Y., Kabiru, A., & Bisallah, C. I. (2020). Activity of aqueous extract of Phyllanthus niruri Linn in 2,4-dinitrophenylhydrazine induced anaemic rats. Journal of Pharmacy and Pharmacology Research, 4(3), 79–95. https://doi.org/10.26502/fjppr.034
Nawfetrias, W., Nurhangga, E., Reninta, R., Chotimah, S., Bidara, I. S., Maretta, D., Akmal, S., & Faizal, A. (2023, October). Metabolite profiling of the medicinal herb Phyllanthus niruri L. under drought stress. IOP Conference Series: Earth and Environmental Science, 1255(1), 012046. https://doi.org/10.1088/1755-1315/1255/1/012046
Noreen, H., Semmar, N., Farman, M., & McCullagh, J. S. (2017). Measurement of total phenolic content and antioxidant activity of aerial parts of medicinal plant Coronopus didymus. Asian Pacific Journal of Tropical Medicine, 10(8), 792–801. https://doi.org/10.1016/j.apjtm.2017.07.024
Pineda, E., Napalan, R., Obañana, R., Pablico, J., Pasaporte, J., Parreño, G., Pimentel, R., Polong, J., Parreño-de Guzman, E., Mendoza, M., & Pakingking, R. (2025). Phytochemical constituents, antioxidant and antibacterial activity of the Philippine Sampa-sampalukan (Phyllanthus niruri) leaf extract against selected human bacterial pathogens. Journal of Biological Studies, 8(1), 34–47. https://doi.org/10.62400/jbs.v8i1.13543
Rajput, A., Sharma, R., & Bharti, R. (2022). Pharmacological activities and toxicities of alkaloids on human health. Materials Today: Proceedings, 48, 1407–1415. https://doi.org/10.1016/j.matpr.2021.09.189
Roy, A., Khan, A., Ahmad, I., Alghamdi, S., Rajab, B. S., Babalghith, A. O., ... & Islam, M. R. (2022). Flavonoids a bioactive compound from medicinal plants and its therapeutic applications. BioMed Research International, 2022(1), 5445291. https://doi.org/10.1155/2022/5445291
Shaikh, J. R., & Patil, M. (2020). Qualitative tests for preliminary phytochemical screening: An overview. International Journal of Chemical Studies, 8(2), 603-608. https://doi.org/10.22271/chemi.2020.v8.i2i.8834
Singh, A., Chitra, V., & Ilango, K. (2024). Network pharmacology exploration to reveal molecular insights of Phyllanthus niruri in non-alcoholic fatty liver: In vitro and in silico evidence. Pharmacia, 71, 1-16. https://doi.org/10.3897/pharmacia.71.e110280
Singh, M. K., & Ahmad, M. S. (2020). Phytochemical profile of Phyllanthus niruri L. and evaluation of its potent bioactive compounds. Pharmacognosy Communications, 13, 1545–1551. https://doi.org/10.21786/bbrc/13.3/82
Singh, T., Kumar, R., & Singh, J. K. (2016). Acute toxicity study of Phyllanthus niruri and its effect on the cyto-architectural structure of nephrocytes in Swiss albino mice Mus-musculus. Pharmacognosy Journal, 8(1), 77-80. https://doi.org/10.5530/pj.2016.1.17
Sirajudeen, J., Ahamath, J. M., Vahith, R. A., Manivel, V., & Elamparithi, R. (2017). Phytochemical screening and antimicrobial activity of Phyllanthus niruri. Journal of Advanced Applied Scientific Research, 1(7). https://doi.org/10.46947/joaasr17201743
Skibska, B., Kochan, E., Stanczak, A., Lipert, A., & Skibska, A. (2023). Antioxidant and anti-inflammatory effects of α-lipoic acid on lipopolysaccharide-induced oxidative stress in rat kidney. Archivum Immunologiae et Therapiae Experimentalis, 71(1), 16. https://doi.org/10.1007/s00005-023-00682-z
Tang, J., Li, R., Wu, B., Tang, J., Kan, H., Zhao, P., Luo, X., & Liu, Y. (2024). Secondary metabolites with antioxidant and antimicrobial activities from Camellia fascicularis. Current Issues in Molecular Biology, 46(7), 6769–6782. https://doi.org/10.3390/cimb46070404
Tiwana, G., Cock, I. E., & Cheesman, M. J. (2024). Phyllanthus niruri Linn.: Antibacterial activity, phytochemistry, and enhanced antibiotic combinatorial strategies. Antibiotics, 13(7), 654. https://doi.org/10.3390/antibiotics13070654
Trivadila, T., Iswantini, D., Rahminiwati, M., Rafi, M., Salsabila, A. P., Sianipar, R. N. R., ... & Murni, A. (2025). Herbal Immunostimulants and Their Phytochemicals: Exploring Morinda citrifolia, Echinacea purpurea, and Phyllanthus niruri. Plants, 14(6), 897. https://doi.org/10.3390/plants14060897
Vishal., Dahiya, D. P., Sankhyan, A., & Sharma, A. (2025). An updated review on Phyllanthus niruri: Its phytochemistry, pharmacological activity, medicinal use and traditional use. International Journal of Pharmaceutical Sciences, 3(3), 2022-2027. https://doi.org/10.5281/zenodo.15062486
Wulandari, E. T., Soetrisno, S., Purwanto, B., Reviono, R., Wasita, B., & Laqif, A. (2024). Impacts of Phyllanthus niruri extract on biomarker levels, macrophage count, and lesion area in an endometriotic rat model. Narra Journal, 4(3), e1002. https://doi.org/10.52225/narra.v4i3.1002
Zonooz, S. R., Hasani, M., Morvaridzadeh, M., Pizarro, A. B., Heydari, H., Yosaee, S., Setayesh, L., & Heshmati, J. (2021). Effect of alpha-lipoic acid on oxidative stress parameters: A systematic review and meta-analysis. Journal of Functional Foods, 87, 104774. https://doi.org/10.1016/j.jff.2021.104774
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Devi, N., Kumar, S., Bindu, N., Gnanavel, A. and Karthick, S. (2025) “Phytochemical Profiling and Antioxidant Activity Assessment of Ethanolic Extract of Phyllanthus niruri”, International Journal of Advancement in Life Sciences Research, 8(4), pp. 128-137. doi: https://doi.org/10.31632/ijalsr.2025.v08i04.010.
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