Development of a selective and sensitive LC-MS/MS method for the quantification of alpha-asarone in mouse plasma and its application to pharmacokinetic studies

Abstract

A simple, sensitive and selective liquid chromatography-tandem mass spectrometric method was developed and validated for the quantification of alpha-asarone in mouse plasma with its application to pharmacokinetic studies. An electrospray ionization (ESI) with multiple reaction monitoring (MRM) mode was used to monitor the precursor-product ion transitions of 209.1 > 193.9 m/z for alpha-asarone and 157.8 > 114.0 miz for allantoin. Chromatographic separation was acquired on a Sepax BR-C18 (5 mu m, 120 A 1.0 x 100 mm) column with an isocratic mobile phase consisting of methanol and 0.1% formic acid (80:20, v/v). The developed bioanalytical method was successfully validated according to the United "States Food and Drug Administration (US FDA) guidelines for linearity, selectivity, accuracy, precision, recovery, matrix effect, and stability. The validated method was successfully applied to a pharmacoldnetics study of alpha-asarone along with a combination of pharmacokinetic techniques, including small-volume serial blood sampling in mice, reducing drug doses and the number of animals used, using a simple protein precipitation method and less solvent consumption will enable its use in further bioequivalence studies. (C) 2018 Elsevier B.V. All rights reserved.