New Analytical method was developed for the estimation of Lamivudine and Dolutegravir in drug product by liquid chromatography. The chromatographic separation was achieved on C18 column (Eclipse XDB-Phenyl 250*4.6mm) at ambient temperature .The separation achieved employing a mobile phase consists of 0.1%v/v Trifluoro acetic acid in water: Methanol (300:700). The flow rate was 1.0ml/ minute and ultra violet detector at 260m. The average retention time for Lamivudine and Dolutegravir found to be 2.412 min and 3.263 min. the proposed method was validated for selectivity, precision, linearity and accuracy. All validation parameters were within the acceptable range. The assay methods were found to be linear from 300.0 – 900.0?g/ml for Lamivudine and 50.0 -150.0?g/ml of Dolutegravir.