Disopyramide Bioanalytical Method Development and Validation in Rabbit Plasma by Using RP-UPLC Following ICH M10 Guidelines

Abstract

A simple, Accurate, precise method was developed for the estimation of Disopyramide in rabbit plasma was developed and validated. By using precipitation method, 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 recommendation’s limitations.