BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF VILDAGLIPTIN IN RAT PLASMA USING LCMS/MS METHOD

Abstract

A simple, wide range, selective, cost effective method using liquid chromatography tandem mass spectrometry method (LC-MS/MS), needs to be developed for toxicological studies in rat plasma due to unavailability of such kind of method in the industry. A sensitive, accurate and selective method was developed and validated for the quantitation of Vildagliptin (VLD) in rat plasma. Electrospray ionization (ESI) interface in positive ion mode was selected to improve selectivity and the sensitivity required for this application. Additionally, a protein precipitation was performed for sample clean up. The separation was achieved in less than 5 min using a ACE 3 C18 PFP, column and a mobile phase, composed of a mixture of ammonium acetate buffer: acetonitrile (20:80, v/v), in isocratic mode at a flow rate of 0.7 mL/min. Detection was carried out in mass spectrometer at m/z 304.4 (parent) and 154.1 (product) for Vildagliptin and 311.1 (parent) and 161.1 (product) for Vildagliptin D7. The retention time observed was 3.00 to 3.20 min. The standard curve was linear (R2 >0.99) over the concentration range of 7.06 ng/mL to 3023.81 ng/mL. The method has shown tremendous reproducibility, with within- and between batch precision less than 9 %, and accuracy within ± 15% of nominal values and has proved to be highly reliable for the analysis of pre-clinical samples for toxicokinetic studies.