A Unique Method for Simultaneous HPLC Analysis of Gallic Acid, Glycyrrhizic Acid and E-Guggulsterone in a Polyherbal Formulation

Background: The therapeutic effects of herbal medicines are exerted due to the presence of different phytoconstituents and the effects are further potentiated when compatible herbals are formulated together. High Performance Liquid Chromatography (HPLC) is an important analytical tool towards standardization of polyherbal formulation. Complex nature, co- existence with other constituents trace quantities of active phytoconstituents and their simultaneous estimation are challenges encountered in HPLC analysis of polyherbal formulation. Simultaneous estimation of active biomarkers in polyherbal formulation by HPLC provides a unique method for verifying the quality and efficacy of polyherbal formulation. Materials and Methods: Present research work is about inventing an unique, selective and sensitive HPLC (High Performance Liquid Chromatography) method for simultaneous estimation of Gallic acid, Glycyrrhizic acid and E-Guggulsterone from a polyherbal formulation containing thirteen Ethnobotanical herbs, reported to have Cardioprotective effects, marketed as Collasyn tablets by Dr. Palep Research Labs Ltd. in Indian market. Reverse-phase column (C18, 250 mM X4.6 mM i.d., 5μm) was used to analyze the marker compounds Gallic acid, Glycyrrhizic acid and E-Guggulsterone at λmax 224 nm. The mobile phase was composed of acetonitrile and 0.05% OPA (Ortho-phosphoric acid) in water (50:50) with a flow rate of 1 mL/min. Results: A unique HPLC method is developed for simultaneous estimation of active markers-Gallic acid, Glycyrrhizic acid and E-Guggulsterone from the polyherbal formulation. Quantitative analysis shows Gallic acid, Glycyrrhizic acid and E-Guggulsterone to be 11.47%, 1.05% and 0.63% respectively in the polyherbal formulation. The developed method is validated in accordance with the International Council for Harmonisation of Technical Requirements for Pharmaceuticals guidelines. The method shows good linearity in a relatively wide range of Concentration of 50-150 mcg/mL. The method is also found to be precise, accurate and robust. Conclusion: This unique and validated method for simultaneous estimation of cardioprotective biomarkers Gallic acid, Glycyrrhizic acid and E-Guggulsterone from polyherbal formulation, marketed as Collasyn tablets by Dr. Palep Research Labs Ltd. is important for its quality control. It helps in harnessing the complete plant potential for therapeutics. Thus, this approach to standardizing polyherbal formulations will be instrumental in verifying the quality and efficacy of the polyherbal extract. The simultaneous estimation of constituents from plants presents a significant advancement in phytochemical analysis, offering a comprehensive understanding of plant properties and their potential applications. The methodologies employed demonstrate high efficiency and accuracy, allowing for the effective quantification of multiple compounds in a single analytical run. This approach not only streamlines the process of phytochemical profiling but also enhances the ability to explore the therapeutic potentials of various plant species.