In the present investigation a more accurate, precise and economically feasible reverse phase liquid chromatographic method was developed and validated for the quantification of simultaneous estimation of Lamivudine and Stavudine in pure and Marketed dosage forms. Quantification of drugs of choice was done with a C18 Kromasil Column using mobile phase of Acetonitrile and Phosphate buffer. The flow rate was adjusted to 1 ml/min and at 257 nm the column eluents were chromatographed. Both the drugs exhibit retention time at 3.3 min and 6.2 min. The established method was linear over a concentration range of 20-100 µg/ml for both the drugs. The established method proved as more reliable and reproducible with a relative standard deviation of less than 2 and have accuracy within the acceptance limits. Marketed formulation assay was determined and proved that formulation was within the percentage purity limits. The analytical method was validated in accordance with International Conference on Harmonization (ICH) and this method can be employed in the quantification of marketed tablet dosage forms also.