Objective: A New method was established for simultaneous estimation of Sertraline and Alprazolam by RP-HPLC method. Methods: Chromatographic separations were carried using HPLC equipped with Auto Sampler and PDA Detector, Inertsil ODS (1.7 x 50 mm, 3mm) column with a mobile phase composition of Methanol: TEA Buffer (65:35 v/v) have been delivered at a flow rate of 1ml/min and the detection was carried out using waters HPLC auto sampler, separation module 2695 HPLC system with PDA detector at wavelength 230 nm. Results: The retention time for Sertraline and Alprazolam were 2.152 and 3.64 minute respectively. The correlation coefficient values in linearity were found to be 0.999 and concentration range 10-50 μg/ml for Sertraline and 20-100 µg/ml for Alprazolam respectively. For accuracy the total recovery was found to be 100.37 % and 100.34 % for Sertraline and Alprazolam. LOD and LOQ for Sertraline 0.9 and 1.2 μg/mL, LOD and LOQ for Alprazolam 2.7 and 3.6 μg/mL. Conclusion: The results of study showed that the proposed RP‐HPLC method is a simple, accurate, precise, rugged, robust, fast and reproducible, which may be useful for the routine estimation of Sertraline and Alprazolam in pharmaceutical dosage form. Keywords: Sertraline, Alprazolam, RP‐HPLC, Simultaneous estimation.