Sustainable And Economical in-Vitro Bio Analytical Method Development and Validation of Azelnidipine in Rabbit Plasma Using RP-UPLC Follows ICH Guidelines M10

Abstract

A simple, Accurate, precise method was developed for the simultaneous estimation of Azelnidipine in Rabbit plasma was developed and validated. By using Centrifugation technique, the sample preparation was prepared. Chromatogram run through Std ACQUITY UPLC HSS C18 Column, 1.8 µm, and 2.1 mm X 50 mm, Mobile phase containing Acetonitrile: Buffer Potassium dihydrogen Phosphate taken in the ratio 70:30 v/v was pumped through column at a flow rate of 0.3 ml/min. KH₂PO₄.in this method was buffer. For the separation of Azelnidipine Internal Standard [IS] used is Nicardipine. The Temperature was maintained at 30°C. Optimized wavelength selected was 257 nm.  Retention time of Azelnidipine and Internal Standard were found to be 0.972 min and 0.603 min. The standard curve was linear (R2 >0.999) over the concentration range of 2 – 80 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.