Bioanalytical Method Development and Validation for the Estimation of Saxagliptin in Marketed Tablet Formulation

Abstract

The concepts, importance and application of bioanalytical method development have been discussed for a long time and validation of bioanalytical methods is widely accepted as pivotal before they are taken into routine use. Now it is widely accepted that bioanalysis is an integral part of the clinical diagnosis, biomarker discovery, pharmacokinetic/ pharmacodynamic characterization of a novel chemical entity from the time of its discovery and during various stages of drug development, leading to its market authorization. Bioanalytical methods, based on a variety of physico-chemical and biological techniques such as chromatography, immunoassay and mass spectrometry, must be validated prior to and during use to give confidence in the results generated. This study describes the development of an innovative, rapid, precise, selective and sensitive reverse phase high-performance liquid chromatography method for the quantitative determination of Saxagliptin (SAXA) in human plasma and pharmaceutical dosage form. Extraction of drug from plasma was done by employing optimized liquid-liquid extraction procedure. The sample was analyzed using methanol: acetonitrile in the ratio of 50:50v/v as mobile phase. Chromatographic separation was achieved on Thermo C18 analytical column (250mm×4.6mm i.d., 5.0μm) as stationary phase using isocratic elution (at a flow rate of 1 ml/min). The peak was detected using UV-PDA detector set at 230 nm and the total time for a chromatographic separation was 20 min. The calibration curve obtained was linear (r2 = 0.999) over the concentration range of 5-25μg/ml for SAXA. Method was validated for precision, robustness and recovery.