Determination of Diosmetin-7-o-beta-d-Glucoside in Rat Plasma by UPLC-MS/MS

In this study, we used UPLC-MS/MS to determine diosmetin-7-o-beta-d-glucoside in rat plasma and investigated its pharmacokinetics in rats. Six rats were given diosmetin-7-o-beta-d-glucoside (5 mg/kg) by intravenous (i.v.) administration. The blood (150 mu L) was withdrawn from the caudal vein after administration. Diazepam was used as an internal standard (IS), and a one-step acetonitrile precipitation method was used to process the plasma samples. Chromatographic separation was achieved using a UPLC BEH C18 column using a mobile phase of acetonitrile-0.1% formic acid with gradient elution. Electrospray ionization (ESI) tandem mass spectrometry in multiple reaction monitoring (MRM) mode with positive ionization was applied, 463.1 -> 301.0 for diosmetin-7-o-beta-d-glucoside, m/z 285.1 -> 193.0 for diazepam (IS). Infra-day and inter-day precision of diosmetin-7-o-beta-d-glucoside in rat plasma were less than 14%. The method was successfully applied in the phannacokinetics of diosmetin-7-o-beta-d-glucoside in rats after intravenous administration. The t(1/2) of diosmetin-7-o-beta-d-glucoside is 1.4 +/- 0.4 h, which indicates the quick elimination.