A simple, precise, selective and rapid reverse phase high-performance liquid chromatographic method was developed and validated for the simultaneous estimation of Tenofovir and Lamivudine in bulk and combined dosage form. The mobile phase used was mixture of phosphate buffer (6.5 mM) adjusted to pH 2.5 with orthophosphoric acid and acetonitrile (50:50 v/v). The Inertsil C18 column (15 cm x 4.6 mm, 5 µm) was used and drugs were detected by UV detector at 260 nm. The retention time of Lamivudine and Tenofovir were found to be 2.04 and 3.54 min respectively. The method was linear in the concentration range of 60-140 μg/ml and 180-420 μg/ml with correlation coefficient (r2 ) of 0.998 and 0.999 for Lamivudine and Tenofovir respectively. The method was validated according to ICH guidelines with respect to accuracy, precision, specificity and can be used to determine drug content of marketed formulation.