SIMULTANEOUS DETERMINATION OF FLAVONOIDS IN POLYHERBAL FORMULATION BY A VALIDATED HPTLC METHOD

A rapid, sensitive, selective and robust quantitative densitometric high-performance thin-layer chromatographic method is developed and validated for separation and quantification of flavonoids namely rutin, quercetin, kaempferol and myricetin in herbal formulations. The separation is performed on silica gel 60F₂₅₄ high-performance thin-layer chromatography plates using phase cyclohexane: ethyl acetate: formic acid 12:9:0.4 (v/v/v) as the mobile phase. A precise and accurate quantification is performed using linear regression analysis by plotting the peak area vs concentration in range of 200–1000 ng/band with regression coefficient, R² from 0.997 to 0.998 for selected flavonoids. The developed method is validated in terms of accuracy, recovery and inter- and intraday study as per International Conference on Harmonisation (ICH) guidelines. The method is found sensitive, accurate and reproducible. Therefore, it can be recommended for marker-based standardization and quality assurance. The present research work is an attempt for simultaneous quantification of rutin, quercetin, kaempferol and myricetin in a single mobile phase. The validated HPTLC method can be further applied to evaluate the quality of any plant reported to possess these phytochemicals.