Determination and Pharmacokinetics of Glucoraphanin in Rat Plasma by UPLC-MS/MS

Glucoraphanin is a beta-thioglycoside N-hydroxyl sulfate found in broccoli and other cruciferous vegetables. In this work, we used UPLC-MS/MS to detect glucoraphanin in plasma and studied their pharmacokinetics, oral and intravenous, and then calculated absolute bioavailability. Midazolam was used as an internal standard. Plasma samples were processed by protein precipitation using methanol as a precipitant. AUPLC BEH C18 was used for chromatographic separation using mobile phase of methanol 0.1% formic acid with gradient elution. Multiple reaction monitoring (MRM) mode using electrospray ionization (ESI) tandem mass spectrometry in positive ionization was applied for quantitative analysis. Within the range of 2-2000 ng/mL, the linearity of glucoraphanin in rat plasma was acceptable (r>0.995), and the lower limit of quantification (LLOQ) was 2 ng/mL. Accuracy range was between 91.3% and 107.9%, precision was lower than 12%, the recovery was better than 76.7%. The developed UPLCMS/MS method was successfully applied in the pharmacokinetics of glucoraphanin in rats after oral and intravenous administration. The absolute bioavailability of the glucoraphanin was 22.5% in rats.