Development and Validation of Bioanalytical Method for Selected Drugs by Liquid Chromatography Tandem Mass Spectrometry LC MS MS

Abstract

newlineBack ground: Linagliptin is an orally active DPP-4 enzyme inhibitor used to treat newlinetype 2 diabetis. Vitamin K1 (phylloquinone, phytonadione and phytomenadione) is a newlinefat soluble vitamin belongs to vitamin K family used to treat coagulation disorders. newlineThese products were selected for bioanalytical method development and validation newlinedue to unavailability of detailed reports in previous reports. newlineObjectives : (1) To develop a bioanalytical method for Linagliptin and Vitamin K1 in newlinehuman plasma using LC-MS-MS method (2) To validate the developed method on newlineLinagliptin and Vitamin K1 complying with most recent regulations ( e.g: US - FDA). newlineMethodology: Methods for both the drugs was developed while focusing on the newlineselection and optimization of chromatographic and mass spectrometric parameters newlineincluding the selection and optimization of sample extraction method. The most newlinerecent regulatory guidelines on BMV were followed for conducting full method newlinevalidation. Simple, sensitive, rapid, selective and reliable methods were developed newlineusing UFLC-MS/MS for quantification of linagliptin and vitamin K1 isomers in newlinehuman plasma. newlineResults: Both linagliptin and its deuterated IS (linagliptin-D4) was extracted from newlinelow plasma sample volume of 300 µL by simple LLE method. Rapid estimation of newlineanalyte and IS (mean RT of 1.75 and 1.74 min respectively, with a short3.5min run newlinetime per sample) was chromatographically established on Gemini C18 (100 mm × 4.6 newlinemm, 3and#956;) column under simple isocratic elution conditions, using a mixture of newlinemethanol and 10 mM ammonium formate [80:20 (v/v)] which delivered at a flow rate newlineof 0.5 mL/min. Following separation of compounds, protonated parent to product newlineion transitions were monitored for linagliptin (m/z: 473.3 - 420.1) and IS (m/z: 477.5 - newlinexxi newline newline424.3) using triple quadrupole mass spectrometer in MRM mode. The analyte and IS newlinerecovery of not less than 71.0% , precision of and#8804; 8.6 %CV, accuracy between 86.7 % newlineto 95.6 % and linearity (r) of gt 0.99 was established with