A simple, precise, accurate and robust reverse phase RP-HPLC method was developed and validated for the simultaneous estimation of Emtricitabine, Tenofovir and Dolutegravir in bulk and pharmaceutical dosage form. This method involves a simple isocratic mobile phase Acetonitrile: 0.01N Potassium dihydrogen orthophosphate (40:60) and a chromatographic separation using Ascentis column (C18 150x 4.6mm, 2.7µm) with PDA detection at 265nm, by using WATERS 2695 HPLC system. Diluent is 50:50 v/v of Acetonitrile: 0.01N Potassium dihydrogen orthophosphate. The average retention times of Emtricitabine, Tenofovir and Dolutegravir were found to be 2.243, 2.763 and 3.254 min respectively. The assay of Emtricitabine, Tenofovir and Dolutegravir were performed with tablets and the % assay were found to be 100.01%, 99.57% and 100.64% which shows that the method is useful for routine analysis. The linearity of Emtricitabine, Tenofovir and Dolutegravir were found to be linear with a R2 of 0.999 for all the drugs, this shows that the method is able to produce good sensitivity. The %RSD for method precision for Emtricitabine, Tenofovir and Dolutegravir were 0.7, 1.4 and 0.6, which shows that the method is precise.