A simple, Accurate, precise method was developed for the simultaneous estimation of Disopyramide in human plasma was developed and validated. By using solvent phase extraction [SPE] the sample preparation was prepared. Chromatogram was run through Std Hibar C18 (100 x 2.1 mm, 2m) Mobile phase containing Buffer Ammonium Acetate: Methanol taken in the ratio 60:40 was pumped through column at a flow rate of 0.2ml/min. For the separation of Disopyramide, Internal Standard [IS] used was Darunavir. The Temperature was maintained at 30°C. Optimized wavelength selected was 215nm. Retention time of Disopyramide and Internal Standard were found to be 1.281 min and 1.535 min. The standard curve was linear (R2 >0.995) over the concentration range of 0.15-6 ng/ml. All the analytical validation parameters were determined as per ICH guidelines The bioanalytical method developed approach was selective, robust, and reliable, as accuracy, precision, recovery, and other validation parameters were all within the recommendations limitations. The peaks produced for the drug of interest and the internal standard were well separated from one another without any plasma interferences, and the peaks were symmetrical with an adequate tailing factor. The method has the potential to be very beneficial in therapeutic drug monitoring (TDM), bioequivalence research, pharmacokinetics studies, toxicology, and biomedical investigations.