A new, simple, precise, rapid, selective and stability reversed-phase high performance liquid chromatographic (RP-HPLC) method has been developed and validated for the simultaneous quantification of Perphenazine and Amitriptyline in pure form and its pharmaceutical dosage form. The method is based on Phenomenex Gemini C18 (4.6×250mm) 5µ column. The separation is achieved using isocratic elution by Methanol: TEA Buffer in the ratio of 65:35% v/v, pumped at flow rate 1.0mL/min and UV detection at 230nm. The column is maintained at 40°C throughout the analysis. The total run time is about 6min. The method is validated for specificity, accuracy, precision and linearity, robustness and ruggedness, system suitability, limit of detection and limit of quantitation as per International conference of harmonization (ICH) Guidelines. The method is accurate and linear for quantification of Perphenazine, Amitriptyline between 10 - 50µg/mL and 20 - 100µg/mL respectively. Further, satisfactory results are also established in terms of mean percent- age recovery (100.37% for Perphenazine and 100.34% for Amitriptyline, intra-day and inter-day precision (<2%) and robustness. The advantages of this method are good resolution with sharper peaks and sufficient precision. The results indicate that the method is suitable for the routine quality control testing of marketed tablet formulations.