DEVELOPMENT AND VALIDATION OF UPLC-PDA METHOD FOR QUALITY ASSESSMENT OF BIOMARKERS IN POLYHERBAL ANTI-DIABETIC FORMULATION
DOI:
https://doi.org/10.29121/shodhkosh.v7.i13s.2026.8435Keywords:
Gymnema Sylvestre, Momordica Charantia, Trigonella Foenum-Graecum, Azadirachta Indica, Syzygium Cumini, Simultaneous Analysis, Quality Assessment, UPLC–PDA MethodAbstract [English]
Background/Objectives: It consist of Gymnema sylvestre, Momordica charantia, Trigonella foenum-graecum, Azadirachta indica, Syzygium cumini, has been widely used to treat Diabetes. In the present study, a Ultra-performance liquid chromatography with photodiode array detector (UPLC–PDA) method for the simultaneous quality assessement of the five biomarkers components, i.e., Gymnemic acids from Gymnema sylvestre, Charantin and momordicin from Momordica charantia,4-hydroxyisoleucine and trigonelline from Trigonella foenum-graecum, Quercetin and rutin from various herbs, Gallic acid and ellagic acid as phenolic markers was developed.
The main objectives of this research work to develop and optimize a sensitive and selective UPLC-PDA method for simultaneous determination of key biomarkers in polyherbal anti-diabetic formulations with enhanced resolution and reduced analysis time compared to conventional HPLC methods & to validate the developed UPLC-PDA method according to ICH Q2(R1) guidelines, evaluating all critical validation parameters including specificity, linearity, accuracy, precision, detection limits, quantitation limits, robustness, and system suitability to ensure method reliability and regulatory compliance.
Methods: The developed UPLC–PDA assay for quality assessement of biomarkers for herbal anti diabetic formulation was validated with respect to linearity, limit of detection (LOD), limit of quantification (LOQ), recovery, and precision.
Results: In the regression equation of the calibration curve, the coefficient of determination was (r²) ≥ 0.999 and LOD and LOQ were 0.010–0.022 μg/mL and 0.033–0.070 μg/mL, respectively. Recovery and precision (relative standard deviation) were 98–100% and <2.0%, respectively. In this analytical method, Five compounds were detected and Succesfully validated by UPLC-PDA method.
Conclusions: The development and validation of an analytical method could be used to obtain basic data for the quality assessment of biomarkers in polyherbal anti-diabetic preparation by the use of UPLC-PDA.
References
Ahrén, B., & Pacini, G. (2021). Glucose effectiveness: Lessons from studies on insulin-independent glucose clearance in mice. Journal of Diabetes Investigation, 12(5), 675–685. https://doi.org/10.1111/jdi.13446
Alhamhoom, Y., Ahmed, S. S., Kumar, R., Salahuddin, M., Bharathi, D. R., Ahmed, M. M., Farhana, S. A., & Rahamathulla, M. (2023). Synergistic antihyperglycemic and antihyperlipidemic effect of polyherbal and allopolyherbal formulation. Pharmaceuticals, 16(10), 1368. https://doi.org/10.3390/ph16101368
Al-Qaseemi, S. A., Almulhim, H. A., Almulhim, M. F., & Chaudhry, S. R. (2016). IoT architecture challenges and issues: Lack of standardization. In Proceedings of the 2016 Future Technologies Conference (FTC) (pp. 731–738). IEEE. https://doi.org/10.1109/FTC.2016.7821686
Alves, E., Gurupadayya, B. M., & Prabhakaran, P. (2025). Artificial intelligence in HPLC method development: A critical review of technological integration, limitations, and future directions. Critical Reviews in Analytical Chemistry, 1–43. https://doi.org/10.1080/10408347.2025.2575352
Amaeze, O., Eng, H., Horlbogen, L., Ajaezi, G., & Nworu, C. (2021). Cytochrome P450 enzyme inhibition and herb–drug interaction potential of medicinal plant extracts used for management of diabetes in Nigeria. European Journal of Drug Metabolism and Pharmacokinetics, 46, 437–450. https://doi.org/10.1007/s13318-021-00685-1
Ardalani, H., Hejazi Amiri, F., Hadipanah, A., et al. (2021). Potential antidiabetic phytochemicals in plant roots: A review of in vivo studies. Journal of Diabetes & Metabolic Disorders, 20, 1837–1854. https://doi.org/10.1007/s40200-021-00853-9
Balekundri, A., & Mannur, V. (2020). Quality control of the traditional herbs and herbal products: A review. Future Journal of Pharmaceutical Sciences, 6, 67. https://doi.org/10.1186/s43094-020-00091-5
Bansode, B., & Jungari, S. (2019). Economic burden of diabetic patients in India: A review. Diabetes & Metabolic Syndrome: Clinical Research & Reviews, 13(4), 2469–2472. https://doi.org/10.1016/j.dsx.2019.06.020
Başaran, E., Öztürk, A. A., Şenel, B., Demirel, M., & Sarıca, Ş. (2022). Quercetin, rutin and quercetin–rutin incorporated hydroxypropyl β-cyclodextrin inclusion complexes. European Journal of Pharmaceutical Sciences, 172, 106153. https://doi.org/10.1016/j.ejps.2022.106153
Benchoula, K., Vohra, M. S., Parhar, I. S., et al. (2022). Metabolomics based biomarker identification of anti-diabetes and anti-obesity properties of Malaysian herbs. Metabolomics, 18, 12. https://doi.org/10. 1007/s11306-022-01870-2
Bereman, M. S. (2015). Tools for monitoring system suitability in LC MS/MS centric proteomic experiments. Proteomics, 15(5–6), 891–902. https://doi.org/10.1002/pmic.201400373
Berger, T. A. (2016). Instrument modifications that produced reduced plate heights <2 with sub-2 μm particles and 95% of theoretical efficiency at k = 2 in supercritical fluid chromatography. Journal of Chromatography A, 1444, 129–144. https://doi.org/10.1016/j.chroma.2016.03.021
Bhavna, Ojha, A., & Bhargava, S. (2022). International Council for Harmonisation (ICH) guidelines. In Regulatory affairs in the pharmaceutical industry (pp. 47–74). Academic Press. https://doi.org/10.1016/B978-0-12-822211-9.00008-3
Blessy, M., Patel, R. D., Prajapati, P. N., & Agrawal, Y. K. (2014). Development of forced degradation and stability indicating studies of drugs—A review. Journal of Pharmaceutical Analysis, 4(3), 159–165. https://doi.org/10.1016/j.jpha.2013.09.003
Borman, P., & Elder, D. (2017). Q2(R1) validation of analytical procedures: Text and methodology. In A. Teasdale, D. Elder, & R. W. Nims (Eds.), ICH quality guidelines: An implementation guide (Chap. 5). Wiley. https://doi.org/10.1002/9781118971147.ch5
Burgos, A., Goñi, A., Illarramendi, A., & Bermudez, J. (2010). Real-time detection of apneas on a PDA. IEEE Transactions on Information Technology in Biomedicine, 14(4), 995–1002. https://doi.org/ 10.1109/T ITB.2009.2034975
Caro-Ordieres, T., Marín-Royo, G., Opazo-Ríos, L., Jiménez-Castilla, L., Moreno, J. A., Gómez-Guerrero, C., & Egido, J. (2020). The coming age of flavonoids in the treatment of diabetic complications. Journal of Clinical Medicine, 9(2), 346. https://doi.org/10.3390/jcm9020346
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