Validation of the method for the simultaneous estimation of bioactive marker gallic acid and quercetin in Abutilon indicum by HPTLC

ObjectiveTo establish and validate an simultaneous estimation of the two biomarker compounds gallic acid (GA) and quercetin (QE) from methanolic extract of Abutilon indicum (AI).MethodsChromatography was performed on aluminium foil-backed silica gel 60 F254 HPTLC plates with the binary mobile phase toluene-ethyl acetate-formic acid (5:4:1, v/v/v). Ultraviolet detection was performed densitometrically at the maximum absorbance wavelength, 270nm. The method was validated for precision, recovery, robustness, specificity, and detection and quantification limits, in accordance with ICH guidelines.ResultsThe system was found to give compact spots for GA and QE (Rf value of 0.31 and 0.50, respectively). The limit of detection (23 and 41 ng band-1) limit of quantification (69 and 123 ng band-1), recovery (99.4–99.9 and 98.7–99.4%), and precision (≤1.98 and 1.97) were satisfactory for gallic acid and quercetin respectively. Linearity range for GA and QE were 100-1000 (r2= 0.9991) and 150–900 ng band-1 (r2 = 0.9956) and the contents estimated as 0.69% ± 0.01% and 0.57% ± 0.01% w/w respectively.ConclusionsThis simple, precise and accurate method gave good resolution from other constituents present in the extract. The method has been successfully applied in the analysis and routine quality control of herbal material and formulations containing AI.