Estradiol (ESD) is widely used in post climacteric replacement therapy. Most of the methods used for quantitation are expensive and time consuming. A rapid, selective and precise stability indicating high performance thin layer chromatography method was developed and validated for the estimation of ESD in bulk and pharmaceutical dosage forms. The method employed TLC aluminium plate precoated with silica gel 60F254 as the stationary phase. The solvent system employed consisted of chloroform-acetone-isopropyl alcohol-glacial acetic acid (9:1:0.4:0.1, v/v/v/v). Such a complex system was essential to obtain a dense and compact spot of the drug at an Rf value of 0.40±0.02. The drug on intentional degradation gave two products with Rf values of 0.52±0.01 and 0.58±0.01 respectively. Spectrodensitometric scanning-integration was performed on a Camag system using a wavelength of 286 nm. The polynomial regression data for the calibration plots exhibited good linear relationship (r=0.9947) over a concentration range of 1-8 μg. Recovery studies were also performed at three experimental levels. The recovery data reveals that the RSD for intra-day and inter-day analysis was found to be 1.27% and 1.75%, respectively. The proposed method was found to be stability indicating. Statistical analysis proves that the method is precise, accurate and reproducible, hence can be employed for the routine analysis of the drug. Copyright (C) 2000 Elsevier Science B.V.