Safety Evaluation of Barleria lupulina Lindl. Leaf Extract via Toxicity Assessment and Analysis of Chemical Constituents
Keywords:
Barleria lupulina, Brine shrimp, ICP-MS, UHPLC, QTOF/MSAbstract
Barleria lupulina Lindl. is a perennial herb commonly referred to as the hophead Philippine violet and “Penawar Seribu Bisa” or “Bisa Ular” in Malaysia. It belongs to the Acanthaceae family and holds a significant place in the traditional pharmacopeia of indigenous communities in Malaysia. This study aims to assess the safety profile of the leaf extract, focusing on its elemental composition and potential cytotoxicity. The concentrations of heavy metals were quantified using Inductively Coupled Plasma-Mass Spectrometry (ICP-MS), while cytotoxicity was assessed through the Brine Shrimp Lethality Assay (BSLA). Additionally, Ultra-High-Performance Liquid Chromatography coupled with Quadrupole Time-of-Flight Mass Spectrometry (UHPLC-QTOF/MS) was utilized to characterize the chemical constituents of the extract. Results indicate that the heavy metal concentrations in B. lupulina leaf are within the permissible limits set by the Codex Alimentarius, suggesting they are safe for consumption. The BSLA assessment demonstrated no significant mortality in brine shrimp, even at the highest extract concentration of 1000 μg/mL, further supporting the safety of the extract. UHPLC-QTOF/MS analysis revealed the presence of several compounds—paeonolide, sibiricaxanthone B, scroneoside A, roseoside, and cornuside II—reported for the first time in this species.
Downloads
Metrics
References
Aguilar, N.O. 2001. Barleria lupulina Lindley. In: Plant Resources of South-East Asia No. 12(2): Medicinal and Poisonous Plants 2. van Valkenburg, J.L.C.H. & Bunyapraphatsara, N. (Eds.). Backhuys Publishers, Leiden. p. 527.
Ahmad, W., Alharthy, R.D., Zubair, M., Ahmed, M., Hameed, A. & Rafique, S. 2021. Toxic and heavy metals contamination assessment in soil and water to evaluate human health risk. Scientific Reports, 11(1): 17006. DOI: https://doi.org/10.1038/s41598-021-94616-4
Akoury, E., Baroud, C., El Kantar, S., Hassan, H. & Karam, L. 2022. Determination of heavy metals contamination in thyme products by inductively coupled plasma mass spectrometry. Toxicology Reports, 9: 1962–1967. DOI: https://doi.org/10.1016/j.toxrep.2022.10.014
Balkwill, M.J. & Balkwill, K. 1998. A preliminary analysis of distribution patterns in a large, pantropical genus, Barleria L. (Acanthaceae). Journal of Biogeography, 25(1): 95–110. DOI: https://doi.org/10.1046/j.1365-2699.1998.251120.x
Balqis-Wong, I., Siti Salwa, A.G., Uswatun, H.Z. & Mohd Izuan, E.F. 2025. Toxicological and cosmetic functional properties of microwave-assisted Pereskia bleo (Kunth) DC leaves extract. The International Journal of Engineering and Science, 14(6): 128–143. DOI: https://doi.org/10.9790/1813-1406128143
Bang, H.Y., Kim, Y.K., Kim, H., Baek, E.J., Na, T., Sim, K.S. & Kim, H.J. 2023. Ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry for simultaneous pesticide analysis and method validation in sweet pepper. Molecules, 28(14): 5589. DOI: https://doi.org/10.3390/molecules28145589
Baruah, K., Phong, H.P.P.D., Norouzitallab, P., Defoirdt, T. & Bossier, P. 2015. The gnotobiotic brine shrimp (Artemia franciscana) model system reveals that the phenolic compound pyrogallol protects against infection through its prooxidant activity. Free Radical Biology and Medicine, 89: 593–601. DOI: https://doi.org/10.1016/j.freeradbiomed.2015.10.397
Chaachouay, N. & Zidane, L. 2024. Plant-derived natural products: a source for drug discovery and development. Drugs and Drug Candidates, 3(1): 184–207. DOI: https://doi.org/10.3390/ddc3010011
Chen, L., Jiang, L., Shi, X., Yang, J., Wang, R. & Li, W. 2024. Constituents, pharmacological activities, pharmacokinetic studies, clinical applications, and safety profile on the classical prescription Kaixinsan. Frontiers in Pharmacology, 15: 1338024. DOI: https://doi.org/10.3389/fphar.2024.1338024
Chen, R.J. & Lee, V.R. 2023. Cobalt Toxicity. StatPearls Publishing, Treasure Island.
Choi, S.H., Kim, J.Y., Mi Choi, E., Lee, M.Y., Yang, J.Y., Ho Lee, G., Su Kim, K., Yang, J.S., Russo, R.E., Yoo, J.H., Kang, G.J. & Su Park, K. 2019. Heavy metal determination by inductively coupled plasma–mass spectrometry (ICP-MS) and direct mercury analysis (DMA) and arsenic mapping by femtosecond (fs)–laser ablation (LA) ICP-MS in cereals. Analytical Letters, 52(3): 496–510. DOI: https://doi.org/10.1080/00032719.2018.1471484
Coetzee, P.P., Steffens, F.E., Eiselen, R.J., Augustyn, O.P., Balcaen, L. & Vanhaecke, F. 2005. Multi-element analysis of South African wines by ICP−MS and their classification according to geographical origin. Journal of Agricultural and Food Chemistry, 53(13): 5060–5066. DOI: https://doi.org/10.1021/jf048268n
Darbyshire, I., Fisher, A.E., Kiel, C.A. & McDade, L.A. 2019. Phylogenetic relationships among species of Barleria (Acanthaceae, Lamiales): Molecular data reveal complex patterns of morphological evolution and support a revised classification. Taxon, 68(1): 92–111. DOI: https://doi.org/10.1002/tax.12029
Edo, G.I., Nwachukwu, S.C., Ali, A.B., Yousif, E., Jikah, A.N., Zainulabdeen, K., Ekokotu, H.A., Isoje, E.F., Igbuku, U.A., Opiti, R.A., Akpoghelie, P.O., Owheruo, J.O. & Essaghah, A.E.A. 2025. A review on the composition, extraction and applications of phenolic compounds. Ecological Frontiers, 45(1): 7–23. DOI: https://doi.org/10.1016/j.ecofro.2024.09.008
Frankish, N., de Sousa Menezes, F., Mills, C. & Sheridan, H. 2010. Enhancement of insulin release from the β-cell line INS-1 by an ethanolic extract of Bauhinia variegata and its major constituent roseoside. Planta Medica, 76(10): 995–997. DOI: https://doi.org/10.1055/s-0029-1240868
Gangaram, S., Naidoo, Y., Dewir, Y.H. & El-Hendawy, S. 2021. Phytochemicals and biological activities of Barleria (Acanthaceae). Plants, 11(1): 82. DOI: https://doi.org/10.3390/plants11010082
Garg, A., Agrawal, R. & Deshmukh, R. 2024. Pharmacology of Polygala tenuifolia and its significance in traditional Chinese medicine. Pharmacological Research - Modern Chinese Medicine, 10: 100341. DOI: https://doi.org/10.1016/j.prmcm.2023.100341
Ghisalberti, E.L. 2007. Detection and isolation of bioactive natural products. In: Bioactive Natural Products: Detection, Isolation, and Structural Determination. 2nd Ed. Colegate, S.M. & Molyneux, R.J. (Eds.). CRC Press, Boca Raton. pp. 25–90. DOI: https://doi.org/10.1201/9781420006889-6
Gitlin, M. 2016. Lithium side effects and toxicity: prevalence and management strategies. International Journal of Bipolar Disorders, 4(1): 27. DOI: https://doi.org/10.1186/s40345-016-0068-y
Hamidi, M.R., Jovanova, B. & Panovska, T.K. 2014. Toxicological evaluation of the plant products using Brine Shrimp (Artemia salina L.) model. Macedonian Pharmaceutical Bulletin, 60(1): 9–18. DOI: https://doi.org/10.33320/maced.pharm.bull.2014.60.01.002
Ismail-Suhaimy, N.W., Gani, S.S.A., Zaidan, U.H., Halmi, M.I.E. & Bawon, P. 2021. Optimizing conditions for microwave-assisted extraction of polyphenolic content and antioxidant activity of Barleria lupulina Lindl. Plants, 10(4): 682. DOI: https://doi.org/10.3390/plants10040682
Ito, H., Kobayashi, E., Li, S.H., Hatano, T., Sugita, D., Kubo, N., Shimura, S., Itoh, Y., Tokuda, H., Nishino, H. & Yoshida, T. 2002. Antitumor activity of compounds isolated from leaves of Eriobotrya japonica. Journal of Agricultural and Food Chemistry, 50(8): 2400–2403. DOI: https://doi.org/10.1021/jf011083l
Jaishankar, M., Tseten, T., Anbalagan, N., Mathew, B.B. & Beeregowda, K.N. 2014. Toxicity, mechanism and health effects of some heavy metals. Interdisciplinary Toxicology, 7(2): 60–72. DOI: https://doi.org/10.2478/intox-2014-0009
Järup, L. 2003. Hazards of heavy metal contamination. British Medical Bulletin, 68(1): 167–182. DOI: https://doi.org/10.1093/bmb/ldg032
Karahan, F., Ozyigit, I.I., Saracoglu, I.A., Yalcin, I.E., Ozyigit, A.H. & Ilcim, A. 2020. Heavy metal levels and mineral nutrient status in different parts of various medicinal plants collected from eastern Mediterranean region of Turkey. Biological Trace Element Research, 197(1): 316–329. DOI: https://doi.org/10.1007/s12011-019-01974-2
Kumar, A., P, N., Kumar, M., Jose, A., Tomer, V., Oz, E., Proestos, C., Zeng, M., Elobeid, T., K, S. & Oz, F. 2023. Major Phytochemicals: recent advances in health benefits and extraction method. Molecules, 28(2): 887. DOI: https://doi.org/10.3390/molecules28020887
Lajayer, B.A., Ghorbanpour, M. & Nikabadi, S. 2017. Heavy metals in contaminated environment: destiny of secondary metabolite biosynthesis, oxidative status and phytoextraction in medicinal plants. Ecotoxicology and Environmental Safety, 145: 377–390. DOI: https://doi.org/10.1016/j.ecoenv.2017.07.035
Medić, B., Stojanović, M., Stimec, B.V., Divac, N., Vujović, K.S., Stojanović, R., Čolović, M., Krstić, D. & Prostran, M. 2020. Lithium-pharmacological and toxicological aspects: The current state of the art. Current Medicinal Chemistry, 27(3): 337–351. DOI: https://doi.org/10.2174/0929867325666180904124733
Nguta, J.M., Mbaria, J.M., Gakayu, D.W.P., Gathumbi, K., Kabasa, J.D. & Kiama, S.G. 2012. Evaluation of acute toxicity of crude plant extracts from Kenyan Biodiversity using brine shrimp, Artemia salina. Journal of Emergency Trends in Engineering and Applied Sciences, 3(1): 30–34. DOI: https://doi.org/10.2174/2210289201203010030
Ozyigit, I.I., Yalcin, B., Turan, S., Saracoglu, I.A., Karadeniz, S., Yalcin, I.E. & Demir, G. 2018. Investigation of heavy metal level and mineral nutrient status in widely used medicinal plants’ leaves in Turkey: Insights into health implications. Biological Trace Element Research, 182(2): 387–406. DOI: https://doi.org/10.1007/s12011-017-1070-7
Packer, M. 2016. Cobalt cardiomyopathy: a critical reappraisal in light of a recent resurgence. Circulation: Heart Failure, 9(12): e003604. DOI: https://doi.org/10.1161/CIRCHEARTFAILURE.116.003604
Park, K.R., Lee, J.Y., Cho, M., Hong, J.T. & Yun, H.M. 2021. Paeonolide as a novel regulator of core-binding factor subunit Alpha-1 in bone-forming cells. International Journal of Molecular Sciences, 22(9): 4924. DOI: https://doi.org/10.3390/ijms22094924
Paul, J.K., Azmal, M., Haque, A.S.N.B., Talukder, O.F., Meem, M. & Ghosh, A. 2024. Phytochemical-mediated modulation of signaling pathways: a promising avenue for drug discovery. Advances in Redox Research, 13: 100113. DOI: https://doi.org/10.1016/j.arres.2024.100113
Pröfrock, D. & Prange, A. 2012. Inductively coupled plasma-mass spectrometry (ICP-MS) for quantitative analysis in environmental and life sciences: a review of challenges, solutions, and trends. Applied Spectroscopy, 66(8): 843–868. DOI: https://doi.org/10.1366/12-06681
Quattrocchi, U. 2012. CRC World Dictionary of Medicinal and Poisonous Plants: Common Names, Scientific Names, Eponyms, Synonyms, and Etymology. CRC Press, Boca Raton. 3960 pp.
Rehder, D. 2013. Vanadium. Its role for humans. In: Interrelations between Essential Metal Ions and Human Diseases. Sigel, A., Sigel, H. & Sigel, R.K.O. (Eds.). Springer, Dordrecht. pp. 139–169. DOI: https://doi.org/10.1007/978-94-007-7500-8_5
Rokaya, M.B., Parajuli, B., Bhatta, K.P. & Timsina, B. 2020. Neopicrorhiza scrophulariiflora (Pennell) Hong: a comprehensive review of its traditional uses, phytochemistry, pharmacology and safety. Journal of Ethnopharmacology, 247: 112250. DOI: https://doi.org/10.1016/j.jep.2019.112250
Samuel, A.J.S.J., Kalusalingam, A., Chellappan, D.K., Gopinath, R., Radhamani, S., Husain, H.A., Muruganandham, V. & Promwichit, P. 2010. Ethnomedical survey of plants used by the Orang Asli in Kampung Bawong, Perak, west Malaysia. Journal of Ethnobiology and Ethnomedicine, 6(1): 5. DOI: https://doi.org/10.1186/1746-4269-6-5
Sawarkar, H.A., Pandey, A.K. & Biyani, K.R. 2023. Phytochemicals and therapeutic potentials of Barleria lupulina. Bangladesh Journal of Pharmacology, 18(4): 119–129. DOI: https://doi.org/10.3329/bjp.v18i4.67429
Shahid, M., Dumat, C., Khalid, S., Schreck, E., Xiong, T. & Niazi, N.K. 2017. Foliar heavy metal uptake, toxicity and detoxification in plants: A comparison of foliar and root metal uptake. Journal of Hazardous Materials, 325: 36–58. DOI: https://doi.org/10.1016/j.jhazmat.2016.11.063
Sharp-Tawfik, A.E., Coiner, A.M., MarElia, C.B., Kazantzis, M., Zhang, C. & Burkhardt, B.R. 2019. Compositional analysis and biological characterization of Cornus officinalis on human 1.1 B4 pancreatic β cells. Molecular and Cellular Endocrinology, 494: 110491. DOI: https://doi.org/10.1016/j.mce.2019.110491
Simorangkir, M., Nainggolan, B., Juwitaningsih, T. & Silaban, S. 2021. The toxicity of n-hexane, ethyl acetate and ethanol extracts of sarangbanua (Clerodendrum fragrans Vent Willd) leaves by brine shrimp lethality test (BSLT) method. Journal of Physics: Conference Series, 1811(1): 012053. DOI: https://doi.org/10.1088/1742-6596/1811/1/012053
Sirisidthi, N.K., Chabra, S., Jiraungkoorskul, K. & Jiraungkoorskul, W. 2016. Evaluation of total essential oil and cytotoxic activity of Barleria lupulina Lindl. Asia-Pacific Journal of Molecular Biology and Biotechnology, 24(1): 1–6.
Siti Salwa, A.G., Ezzah, A.M., Uswatun, H.Z. & Mohd Izuan, E.F. 2023. Chemical profile of Hylocereus polyrhizus flesh water-based extract: Assessment of nutritional value, chemical constituents and potential metal toxicity. Universal Journal of Agricultural Research, 11(3): 566–576. DOI: https://doi.org/10.13189/ujar.2023.110307
Tan, H., Lee, H.J., Hillman, P.F., Lee, E.Y., Lee, C., Seo, E.K., Lee, M.J. & Nam, S.J. 2024. The discovery of an anti-inflammatory monoterpenoid, neoroseoside from the Zea mays. Bioorganic & Medicinal Chemistry Letters, 105: 129737. DOI: https://doi.org/10.1016/j.bmcl.2024.129737
Tchounwou, P.B., Yedjou, C.G., Patlolla, A.K. & Sutton, D.J. 2012. Heavy metal toxicity and the environment. In: Molecular, Clinical and Environmental Toxicology. Luch, A. (Ed.). Springer, Basel. pp. 133–164. DOI: https://doi.org/10.1007/978-3-7643-8340-4_6
Urmi, K.F., Mostafa, S., Begum, G. & Hamid, K. 2013. Comparative brine shrimp lethality bioassay of different plant parts of Bauhinia purpurea L. Journal of Pharmaceutical Sciences and Research, 5(10): 190–192.
Vaou, N., Voidarou, C., Rozos, G., Saldari, C., Stavropoulou, E., Vrioni, G. & Tsakris, A. 2025. Unraveling nature’s pharmacy: Transforming medicinal plants into modern therapeutic agents. Pharmaceutics, 17(6): 754. DOI: https://doi.org/10.3390/pharmaceutics17060754
White, S.J.O. & Shine, J.P. 2016. Exposure potential and health impacts of indium and gallium, metals critical to emerging electronics and energy technologies. Current Environmental Health Reports, 3(4): 459–467. DOI: https://doi.org/10.1007/s40572-016-0118-8
Wilschefski, S.C. & Baxter, M.R. 2019. Inductively coupled plasma mass spectrometry: introduction to analytical aspects. The Clinical Biochemist Reviews, 40(3): 115–133. DOI: https://doi.org/10.33176/AACB-19-00024
Witkowska, D., Słowik, J. & Chilicka, K. 2021. Heavy metals and human health: Possible exposure pathways and the competition for protein binding sites. Molecules, 26(19): 6060. DOI: https://doi.org/10.3390/molecules26196060
Yajima, A., Oono, Y., Nakagawa, R., Nukada, T. & Yabuta, G. 2009. A simple synthesis of four stereoisomers of roseoside and their inhibitory activity on leukotriene release from mice bone marrow-derived cultured mast cells. Bioorganic & Medicinal Chemistry, 17(1): 189–194. DOI: https://doi.org/10.1016/j.bmc.2008.11.002
Zulkifli, S.A., Abd Gani, S.S., Zaidan, U.H. & Halmi, M.I.E. 2020. Optimization of total phenolic and flavonoid contents of defatted pitaya (Hylocereus polyrhizus) seed extract and its antioxidant properties. Molecules, 25(4): 787. DOI: https://doi.org/10.3390/molecules25040787
Published
How to Cite
Issue
Section
Any reproduction of figures, tables and illustrations must obtain written permission from the Chief Editor (wicki@ukm.edu.my). No part of the journal may be reproduced without the editor’s permission
