Journal of pharmaceutical and biomedical analysis

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

PMID 25880244


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 (2.1 mm × 100 mm, 1.7 μm) with acetonitrile (containing 10mM ammonium formate) and water (containing 0.1% formic acid and 10 mM 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 2-2000 ng/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.