Simple, sensitive high performance thin layer chromatography method for the estimation of gallic acid and quercetin in in-house polyherbal blend has been developed and validated. Methanolic solution of herbal blend comprising of Emblica officinalis, Camellia sinensis and Garcinia cambogia was used for analysis. The sepa-ration was performed on TLC aluminum plates precoated with silica gel G60 F254 and toluene : ethyl ace-tate : formic acid (5:1.5:1 v/v/v) at 254 nm scanning wavelength. The system gave well resolved peaks for gallic acid and quercetin at Rf 0.14 and Rf 0.29 respectively. The method validated as per ICH Q2R1 guide-lines which shows regression co-efficient 0.9939 for gallic acid and 0.9988 for quercetin in range of 2–6 μg/ml. Recovery of gallic acid and quercetin was found in range of 98–102 % which confirms the accuracy of method. Precision study (interday & intraday) showed that the relative standard deviation is less than 2 %, showing method is well precise. Proposed validated HPTLC method is simple, precise, specific, robust and accurate, and could find application in routine quality-control analysis. The method was used for quantitative estimation of gallic acid and quercetin in the polyherbal blend and was found as 1.648 % w/w and 3.165 % w/w respectively.
A sensitive High-performance thin-layer chromatography method was developed for simultaneous estimation of berberine, gallic acid and ursolic acid in a polyherbal blend and validated as per ICH guidelines.Polyherbal blend was prepared using widely recommended herbal plants for platelet augmentation activity viz. Carica papaya, Berberis aristata, Ocimum Sanctum and Tinospora Cordifolia. The optimized separation was obtained with TLC aluminum plates pre-coated with silica gel G60 F254 as a stationary phase and a solvent system containing Toluene : Ethyl acetate : Methanol : Formic acid (3:3:0.2:0.1 v/v/v/v). Berberine and gallic acid were found to demonstrate linearity in the range of 1μg/band – 6μg/band and ursolic acid in the range of 20 μg/band – 100 μg/band with the regression coefficient in acceptable limits. The method was also found to be specific and precise one. The accuracy of the developed method at 80 %, 100 % and 120 % levels was found to be within limits and % RSD was found to be less than 2. LOD and LOQ of all three standards were also determined. Blend was also quantified for the amount of berberine, gallic acid and ursolic acid presence and was found to be 37.8 μg, 46.1 μg and 108 μg per mg, respectively.
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