An accurate, highly sensitive, precise, simple, efficient and reproducible, isocratic Reversed Phase-High Performance Liquid Chromatography (RP-HPLC) method was developed and validated for the quantitative determination of anastrozole in pharmaceutical tablet dosage forms. RP-HPLC method was developed by using Welchrom C18 Column (4.6 X 250mm, 5μm), Shimadzu LC-20AT Prominence Liquid Chromatograph. The mobile phase composed of 10mM Phosphate buffer (pH-3.0, adjusted with triethylamine): acetonitrile (50:50 v/v). The flow rate was set to 1.0 mL/min with the responses measured at 215 nm using Shimadzu SPD-20A Prominence UV-Vis detector. The retention time of anastrozole was found to be 6.143 min. Linearity was established for anastrozole HCl in the range of 2-10 μg/mLwith correlation coefficient 0.999. The percentage recovery was found to be 99.84 % to 100.2%. Validation parameters such as specificity, linearity, precision, accuracy, robustness, limit of detection (LOD) and limit of quantitation (LOQ) were evaluated for the method according to the International Conference on Harmonization (ICH) Q2 R1 guidelines. The developed method was successfully applied for the quantification of bulk and active pharmaceutical ingredient present in tablet dosage form.