A simple, efficient, and robust stability-indicating RP-HPLC method has been developed and validated to measure Lamivudine, Zidovudine and Nevirapine at a single wavelength (268 nm) in order to assay. The samples were eluted in an isocratic method using an inertsil ODS column (4.6 mm × 250 mm with a particle size of 5 μ) with a mobile phase consisting of 10 mM Ammonium acetate buffer (with a pH adjusted to 3.8 using Acetic acid): acetonitrile (60:40, v/v), acetonitrile and water (1:1, v/v) using as diluent, through ambient temperature delivered at a flow rate 1.2 mL/mins. A good linear response was obtained in the range from 15-75 μg/mL, 30-150 μg/mL and 20-100 μg/mL of Lamivudine, Zidovudine and Nevirapine respectively. The LODs for Lamivudine, Zidovudine and Nevirapine were found to be 0.315, 0.405 and 0.600 μg/mL, respectively and the LOQs for Lamivudine, Zidovudine and Nevirapine were 0.945, 1.080 and 2.100 μg/mL respectively. The method was quantitatively evaluated in terms of accuracy (recovery), linearity, precision, selectivity and robustness in accordance with standard guidelines. The method is simple, suitable and conducive for analyzing Lamivudine, Nevirapine and Zidovudine in bulk and in pharmaceutical formulations.