HPLC Quantification, Antibacterial and Antioxidant Activity of Alkaloids from Stephania glandulifera Miers
DOI:
https://doi.org/10.3126/jist.v30i2.61714Keywords:
Antibacterial activity, Antioxidant property, Bioactive compounds, High-performance liquid chromatography, Stephania glandulifera MiersAbstract
Stephania glandulifera Miers has been extensively used for many years as a traditional medicine; mainly roots, caudex, stem and leaves are used in the treatment of various diseases in the eastern world. This study aimed to standardize the caudex of S. glandulifera through the development and validation of a high-performance liquid chromatography method for the quantification of its major compounds. Among the five previously isolated compounds, tetrahydropalmatine (1), palmatine (2), stepholidine (4), and stepharine (5) were successfully quantified for the first time, yielding concentrations of 20.6 mg, 20.4285 mg, 19.962 mg, and 62.262 mg per gram of plant respectively. Chromatographic separation was achieved using an Agilent, Eclipse plus C18 5 µm reversed-phase column with a methanol-water gradient system with a flow rate of 1mL/ min detecting tetrahydropalmatine at 290 nm and other compounds at 190 nm. The antibacterial activity of all the four compounds along with stepharanine (3) was evaluated, with palmatine (2) showing significant efficacy against Staphylococcus aureus ATCC 25925 and vancomycin-resistant Staphylococcus aureus (MIC=0.3225 mg/mL, MBC=0.625 mg/mL). Additionally, stepharanine (3) and stepharine (5) demonstrated notable antioxidant activity (IC50=0.1384 mg/mL and 0.2944 mg/mL, respectively). These highlight stepharine, stepharanine, and palmatine as promising candidates for future drug development. Future research should explore their pharmacological potential and clinical applicability
Downloads
References
Ainsworth, E.A & Gillespie, KM. (2007). Estimation of total phenolic content and other oxidation substrates in plant tissues using Folin-Ciocalteu reagent, Nature Protocols, 2, 875–877.
Bhujbal, S., Rupenthal, I. D., & Agarwal, P. (2024). Development and validation of a stability-indicating HPLC method for assay of tonabersat in pharmaceutical formulations. Methods, 231, 178–185.
Clinical and Laboratory Standards Institute, Performance standards for antimicrobial susceptibility testing. 30th ed., CLSI supplement M100 Clinical and Laboratory Standards Institute, Wayne, PA, 2020.
Dary, C., Bun, S.-S., Herbette, G., Mabrouki, F., Bun, H., Kim, S., Jabbour, F., Hul, S., Baghdikian, B., & Ollivier, E. (2017). Chemical profiling of the tuber of Stephania cambodica Gagnep. (Menispermaceae) and analytical control by UHPLC-DAD. Natural Product Research, 31(7), 802–809.
Desgrouas, C., Taudon, N., Bun, S. S., Baghdikian, B., Bory, S., Parzy, D., & Ollivier, E. (2014). Ethnobotany, phytochemistry and pharmacology of Stephania rotunda Lour, Journal of Ethnopharmacology, 154, 537–563.
De Wet H., Struwig M., & Van W. B. E. (2014). Taxonomic notes on the genus Stephania (Menispermaceae) in southern Africa, South African Journal of Botany, 95, 146– 151.
Dhungel, J., Marasini, B. P., Manandhar, E., Rathnayaka R. K., Samarakoon S. R., & Shyaula S. L. (2023). Cytotoxic activity of alkaloids isolated from Stephania glandulifera Miers, Journal of Biologically Active Products from Nature, 13, 118-128.
Elsheikh M., Ahmed S., Funston S., Dunlop P., McGaw M., Marchant R & Banat IM. (2016). Resazurin- based 96-well plate microdilution method for the determination of minimum inhibitory concentration of biosurfactants. Biotechnology Letters, 38, 1015–1019.
Forman, L.L. (1988). A synopsis of Thai Menispermaceae, Kew Bulletin, 43, 369–407.
Hao, D.C., Xiao, P.G., Ma, H.Y., Peng, Y., & He, C.N. (2015). Mining chemodiversity from biodiversity: Pharmacophylogeny of Ranunculales medicinal plants, Chinese Journal of Natural Medicines, 13, 507-520.
Hussein, J. (2025). Principles and applications of high-performance liquid chromatography (HPLC): A review. Biomedical & Pharmacology Journal, 18(2), 1085–1089.
Kongkiatpaiboon, S., Duangdee, N., Prateeptongkum, S., Tayana, N., & Inthakusol, W. (2017). Simultaneous HPLC analysis of crebanine, dicentrine, stephanine and tetrahydropalmatine in Stephania venosa. Revista Brasileira de Farmacognosia, 27(6), 691–697.
Long, J., Song, J., Zhong, L., Liao, Y., Liu, L & Li, X. (2019). Palmatine: A review of its pharmacology, toxicity and pharmacokinetics, Biochimie, 162, 176–184.
Qiang-min X. I. E., Hui-fang T., Ji-qiang C. & Ru-lian B. (2002) Pharmacological actions of tetrandrine in inflammatory pulmonary diseases. Acta Pharmacology Sin 23(12): 1107–1113.
Semwal, D. K., Badoni, R., Semwal, R., Kothiyal, S. K., Singh, G. J. P & Rawat, U. (2010a). The genus Stephania (Menispermaceae): Chemical and pharmacological perspectives, Journal of Ethnopharmacology, 132, 369–383.
Semwal D. K. & Semwal R. B. (2015) Efficacy and safety of Stephania glabra: An alkaloid rich traditional medicinal plant. Natural Product Research 29(5): 396–410.
Siddiqui, M. R., AlOthman, Z. A., & Rahman, N. (2017). Analytical techniques in pharmaceutical analysis: A review. Arabian Journal of Chemistry, 10n, 1409–1421.
Singh, A. R., Singh, T. D., Kom, L. E., Rovei, L., Thokchom, D. S., & Rawat, A. K. (2019). Evaluation of hypoglycemic and antioxidant activities of ‘Koubru Yai’ (Stephania rotunda Engl.): An important medicinal plant used in traditional medicine of Manipur, Journal of Pharmacognosy and Phytochemistry, 8, 1364-1369.
Wang, S. T, Qian, W., Q., He, P., Feng, M. Q., Kang, Y., Wang, Y.Q & Huang, J. M. (2019). Two new glycoalkaloids from Stephania succifera, Phytochemistry Letters, 34, 99–102.
Xiaoli, S., Xu, L., & Ming Z. (2015). Chemical constituents and biological activities of Stephania yunnanensis, Biomedical Research, 26, 715-720.
Zhu, H., Wen, L., Geng, Y., Wang, X., J. I. A, W., Wang, D., & Yan, Y. (2016). Antioxidant activity of alkaloids in Caulis Mahoniae, Shandong Science, 29, 24-28.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2025 Institute of Science and Technology, T.U.

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
The views and interpretations in this journal are those of the author(s). They are not attributable to the Institute of Science and Technology, T.U. and do not imply the expression of any opinion concerning the legal status of any country, territory, city, area of its authorities, or concerning the delimitation of its frontiers of boundaries.
The copyright of the articles is held by the Institute of Science and Technology, T.U.