A validated HPLC-MS/MS method for determination of genipin-1-o-glucuronic acid in rat plasma after administration of genipin and its application to a pharmacokinetic study

A sensitive and specific method was developed and validated for the quantitation of one major metabolite of genipin in rats plasma. The major metabolite was isolated from rat bile via semi-preparative HPLC technology and its chemical structure was identified as genipin-1-o-glucuronic acid (GNP-GLU), which was for the first time used as a standard compound for quantitative analysis in rat plasma after administration of genipin. The application of high-performance liquid chromatography-tandem mass spectrometry in negative mode in multiple reaction monitoring mode was investigated. Chromatographic separation was achieved on an Eclipse XDB-C-18 column using a mobile phase consisting of water with 0.1% formic acid (A)-acetonitrile (B). The limit of detecation was 0.214ng/mL and the lower limit of quantification was 0.706ng/mL. The calibration curve was linear from 1.27 to 3810ng/mL for plasma samples, with a correlation coefficient of 0.9924. The intra- and inter-day precisions and accuracy were all within 15%. The recoveries of GNP-GLU and puerarin were above 90.0 and 76.2%, respectively. The highly sensitive method was successfully applied to estimate pharmacokinetic parameters of GNP-GLU following oral and intravenous administration of genipin to rats. Copyright (c) 2013 John Wiley & Sons, Ltd.