A rapid, simple, sensitive, accurate, and precise UV spectrophotometric method has been developed for the simultaneous estimation of anti-retroviral agents lamivudine, didanosine and efavirenz in pharmaceutical dosage form. The absorption maxima of the drugs were found to be 271, 250 and 247 nm for lamivudine, didanosine and efavirenz respectively. Lamivudine, didanosine and efavirenz obeyed Beer’s law in the concentration range of 10-100 µg/ml, 10-100 µg/ml and 10-70 µg/ml respectively. The percentage recovery was within the range of 98% - 101%, indicating that insignificant interference from the other ingredients in the formulation. The above method was validated in terms of linearity, accuracy, precision, Limit of Detection (LOD), Limit of Quantification (LOQ) etc. in accordance with ICH guide lines. The developed method was free from interferences due to excipients present in tablets. The method was rapid, simple and suitable for routine quality control analysis.