Simultaneous Estimation of Chebulagic acid and Chebulinic acid in Marketed Polyherbal Formulations by HPTLC

A simple, rapid and reliable HPTLC method has been developed for simultaneous estimation of Chebulagic acid and Chebulinic acid in marketed polyherbal formulations. Identification and quantification were performed on 10cm x 10cm, layer thickness 0.2mm, aluminumbacked silica gel 60 F254 HPTLC plates previously washed with methanol. The solvent system consisting of Ethyl Acetate: Toluene: Formic Acid: Methanol (6:1:1:2%v/v) was used as a mobile phase. This system gave compact spots for Chebulagic acid and Chebulinic acid. The spots were scanned at λ=254nm. Calibration plots were established showing the dependence of response on the amount chromatographed. The linear regression analysis data for the calibration plots showed good linear relation with R = 0.998 and 0.9991 with respect to peak area for Chebulagic acid and Chebulinic acid respectively, in concentration range of 420-500ng/spot. The method was validated for precision, recovery, Limit of Detection and Limit of Quantification. The proposed HPTLC method was found to be simple, precise and accurate and can be used for the quality control of the raw materials as well as formulations.