Bio- Method Development And Validation Of Ketotifen Fumarate In Rabbit Plasma Using RP-HPLC

Abstract

A simple, Accurate, precise method was developed for the simultaneous estimation of Ketotifen Fumarate in human plasma was developed and validated. By using Protein Precipitation, the sample preparation was prepared. Chromatogram was run through Discovery C₁₈ (150x 4.6 mm, 5m) Mobile phase containing Buffer Na₂HPO₄:  Methanol taken in the ratio 55:45 was pumped through column at a flow rate of 1.0ml/min. Buffer used Sodium Phosphate Buffer in this method was buffer. For the separation of Ketotifen Fumarate Internal Standard [IS] used is Remogliflozin. The Temperature was maintained at 30°C. Optimized wavelength selected was 222.0nm.  Retention time of Ketotifen Fumarate and Internal Standard were found to be 2.980 min and 2.344 min. The standard curve was linear (R2 >0.995) over the concentration range of 9-360 ng/ml. According to ICH guidelines, each analytical validation parameter was determined. As accuracy, precision, recovery, and other validation parameters were all within the guidelines' constraints, the bioanalytical technique created approach was selective, robust, and reliable. Without any interference from plasma, the peaks generated for the target substance and the internal standard were adequately separated from one another and had a sufficient tailing factor. Therapeutic drug monitoring (TDM), bioequivalence research, pharmacokinetics studies, toxicology, and biological investigations might all greatly benefit from the technique.