||A simple and fast method was developed for simultaneous quantitative determination of two biologically active flavonoid compounds i.e. quercetin and rutin in bark of Anogeissus latifolia using High-Performance Thin-layer Chromatography. The separation was performed on TLC aluminium plates precoated with silica gel 60 F254. Good separation was achieved in the mobile phase of Ethyl acetate: Formic acid: Glacial acetic acid: Water (100:11:11:26, v/v) and densitometric determination of these compounds was carried out at 366 nm in reflection/absorbance mode. The rutin and quercetin content of hydroalcohol bark extract of Anogeissus latifolia were found to be 0.1617% w/w and 1.875%w/w respectively. The linear regression analysis data for the calibration plots showed a good linear relationship with r=0.9997 and r= 0.9942 for rutin and quercetin, respectively. The average recovery of rutin and quercetin was 99.98 % and 100.11%, respectively indicating the excellent reproducibility. Statistical analysis of the data showed that the method is reproducible. The present method is being reported first time and may be used for routine quality control of the bark of Anogeissus latifolia. This HPTLC method was found to be simple and convenient for rapid screening of active compounds and quantification of the investigated flavonoids in Anogeissus latifolia.