Abstract

METHOD DEVELOPMENT AND VALIDATION FOR ESTIMATION OF OLANZPINE IN HUMAN PLASMA BY HYPHENATED TECHNIQUE LIQUID CHROMATOGRAPHY MASS- SPECTROMETRY

Pancholi Nidhi*, Sharma C.S., Kamble P.R., Kamble R.K., Chauhan C.S.

ABSTRACT

A simple, sensitive and rapid liquid chromatography/electro spray ionization tandem mass spectrometry (LC-ESI-MS/MS) method was developed and validated for the quantification of olanzapine, atypical antipsychotic drug, in human plasma using olanzapine D3 as internal standard (IS). Following liquid-liquid extraction, the analytes were separated using an isocratic mobile phase on a reverse phase C18 column and analyzed by MS in the multiple reaction monitoring mode using the respective [M+H]+ ions, m/z 313.2/256.2 for olanzapine and m/z 384.2/253.2 for the IS. The assay exhibited a linear dynamic range of 0.200-20.00 ng/mL for olanzapine in human plasma. The lower limit of quantification was 0.200ng/ml. Acceptable precision and accuracy was obtained for concentrations over the standard curve range. The recovery was assessed by comparing the standards spiked before and after extraction at four concentration levels (0.560, 2.800, 7.280, 14.000 ng∙mL−1). A run time of 3.20 min for each sample made it possible to analyze more than 400 human plasma samples per day. The validated method has been successfully used to analyze human plasma samples for application in pharmacokinetic, bioavailability or bioequivalence studies.

Keywords: Olanzapine, Liquid-liquid extraction, Antipsychotic drug.