Acacia catechu have been in use in Indian traditional medicine, ‘Ayurveda’ since antiquity. However, there has been no attempt to standardize the herbal extract as the main ingredient in terms of its active principle or marker compound. Chromatographic fingerprint symbolize the active chemical constituents of herbal medicines for desired therapeutic action. This study presents a simple, rapid and selective HPTLC method for the quantitative estimation of rutin and quercetin from ethanolic leaf extract of Acacia catechu, precoated HPTLC silica gel 60 F254 as stationary phase and mobile phase for Formic Acid: water: Ethyl acetate [10:10:80 v/v]. detection and quantification were performed densitometrically at 366nm for rutin and quercetin.