The present study was to develop and validate reverse phase highperformance liquid chromatography (RP-HPLC) method for the estimation of Ritonavir. The RP-HPLC separation was achieved on JASCO HPLC system at 1ml/min flow rate on HiQSilC18 (250 x 4.6 mm, 5 μm) columnby using acetonitrile: 10 mM ammonium acetate buffer (pH 4) (85:15%V/V) as mobile phase at wavelength 239 nm. The data of linear regression analysis indicated a good linear relationship over the range of 5–30 μg/ml concentration with correlation coefficient value of 0.997. The accuracy of the method is indicated by good recovery in the range of 99.43%-100.53%and precision with RSD less than 2%