Objective: A Simple, accurate, specific and rugged reverse phase liquid chromatographic method was developed and validated for the simultaneous estimation of Lamivudine, Tenofovir, and Dolutegravir in bulk and tablet dosage form.Method: A reverse phase gradient program has been developed to separate the all four active ingredients using 0.1% triflouro acetic acid, acetonitrile was used as mobile phase. A gradient programing has been developed and validated, on a reverse phase C18 column (150 X4.6 mm, 3µ) with a flow rate of 0.9 mL/min by monitoring at 258 nm of wavelength.Results: The mean retention times of Lamivudine, Tenofovir, and Dolutegravir were found to be 3.06, 9.37 and 10.08 min respectively. Linearity of Lamivudine, Tenofovir, and Dolutegravir was found to be 10-50 µg/mL, 10-50 µg/mL and 1-10 µg/mL respectively. The accuracy of the proposed method was determined by performing recovery studies and was found to be between 98-102%. The repeatability testing for both sample and standard solutions was found as %RSD<2.0% which is within the acceptable limits showing that the method is precise as well. The LOD and LOQ were found to be 0.18 and 0.53 µg/ml for Lamivudine, 0.18 and 0.53 µg/ml for Tenofovir, 0.08 and 0.25 µg/ml for Dolutegravir respectively.Conclusion: The proposed method was validated in terms of linearity, range, accuracy, precision, specificity, robustness and stability studies and the method is successfully applied for the estimation of lamivudine, tenofovir, and dolutegravir in combined tablet dosage form.