Bioanalytical method Development and Validation for Estimation of Olanzapine in K3edta Human Plasma by Liquid Chromatography–Tandem Mass Spectrometry and Application to a Pharmacokinetic Study

Abstract

A rapid and sensitive LC–MS/MS method for the bioanalytical method development and validation for estimation of olanzapine in k₃EDTA human plasma using Olanzapine D₃ as internal standard has been developed and validated. The analytes and IS were extracted from plasma by solid phase extraction using Oasis HLB 1cc (30mg) Extraction Cartridge and separated on a Cosmosil, 5µm, C18 150*4. 6 mmcolumn using a 10 mM ammonium formate in Water: Acetonitrile (10: 90) at a flow rate of 1.0 mL/min. Detection involved an API-4000 LC–MS/MS with electrospray ionization in the positive ion mode and multiple-reaction monitoring for analysis. The method was validated according to FDA guidelines and shown to provide intra- and inter-day precision and accuracy within acceptable limits in a run time of only 4.0 min. The method was successfully applied to a pharmacokinetic study involving a single oral administration of a combination tablet to human male volunteers