HPTLC method has been developed for determination of gallic acid and piperine in appetomax®, a polyherbal ayurvedic proprietary liquid oral formulation manufactured and marketed by Ocean Life care Pvt. Ltd., Pune, containing extracts form ten plant materials. Gallic acid (GA) and piperine (PI) are important compounds present in the formulation and were selected as marker analytes. Different volumes of sample were spotted for estimation of the GA (2 μl) and PI (40 μl) at 300 nm by densitometry using silica gel 60F254 as stationary phase. Mobile phase used consisted of toluene: methanol: acetic acid 8:1.5:0.5 (v/v/v), Rf values of GA and PI were 0.16±0.02 and 0.55±0.02 respectively. Linearity range was 80-280 ng/band and 60-260 ng/band and coefficient of determination was 0.996 and 0.992 for GA and PI respectively. Limits of detection and quantitation were recorded as 7 and 21 ng/band for GA and 6.67 and 20.21 ng/band for PI. The developed high performance thin layer chromatography method was validated for accuracy, precision, robustness, specificity as per ICH guidelines. Thus the methods can be used for the routine estimation of GA and PI in Ayurvedic proprietary liquid oral formulation.
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