Validated UPLC–MS/MS method for determination of hordenine in rat plasma and its application to pharmacokinetic study

Abstract

Hordenine is an active compound found in several foods, herbs and beer. In this work, a sensitive and selective UPLC–MS/MS method for determination of hordenine in rat plasma was developed. After addition of caulophylline as internal standard (IS), protein precipitation by acetonitrile–methanol (9:1, v/v) was used as sample preparation. Chromatographic separation was achieved on a UPLC BEH HILIC (2.1mm×100mm, 1.7μm) with acetonitrile (containing 10mM ammonium formate) and water (containing 0.1% formic acid and 10mM ammonium formate) as mobile phase with gradient elution. An electrospray ionization source was applied and operated in positive ion mode; multiple reaction monitoring (MRM) mode was used for quantification using target fragment ions m/z 166.1→121.0 for hordenine and m/z 205.1→58.0 for IS. Calibration plots were linear over the range of 2–2000ng/mL for hordenine in rat plasma. Mean recoveries of hordenine in rat plasma were in the range of 80.4–87.3%. RSD of intra-day and inter-day precision were both <8%. The accuracy of the method ranged from 97.0% to 107.7%. The method was successfully applied to pharmacokinetic study of hordenine after oral and intravenous administration.