Simultaneous Determination of Gallic Acid, Berberine and Trigonelline in Polyherbal Churna by HPTLC Method

Background: Ayurvedic medicines have a lot of potential because of their comprehensive approach to illness care, yet there is a need for sufficient standardization before they can be used in mainstream medicine. Nevertheless, there is a lack of work efforts directed towards the concurrent assessment of biomarkers in polyherbal churna using HPTLC fingerprinting. The objective of this study is to explore bioactive principles such as Gallic acid, Berberine, and Trigonelline in polyherbal churna using HPTLC fingerprinting. Materials and Methods: The in-house polyherbal churna is a blend of Amla, Methi, and Daruharidra. The formulation underwent a standardization process based on multiple quality control parameters. Using HPTLC technique, pharmacologically active biomarkers including gallic acid, trigonelline, and berberine were successfully identified. The mobile phase used for the HPTLC analysis was a mixture of Toluene: Ethylacetate: Glacial acetic acid: Water in the ratio of 2:3:4:1 v/v/v/v. Results: The method developed exhibited linearity, specificity, reproducibility, robustness, and accuracy, ultimately redefining the characteristics of the approach and cost-effective. The examination confirmed the presence of three biomarkers in the polyherbal churna, with corresponding Rf values of 0.66 for gallic acid, 0.12 for trigonelline, and 0.61 for berberine, respectively. Conclusion: The proposed HPTLC approach, which was developed, utilizes a more recent technique for simultaneously fingerprinting biomarkers from polyherbal formulations containing Amla, Methi, and Daruharidra. The method can be used for standardization of different herbal formulation containing these plant species.