Simultaneous UPLC-TQ-MS/MS determination of six active components in rat plasma: application in the pharmacokinetic study of leaves.

BACKGROUND

(Batal.) Ijinskaja (CP) is a monotypic genus plant, also called sweet tea tree that belongs to the Juglandaceae family, which is mainly distributed in the subtropical highlands in China. Our previous work has verified that CP leaves exhibit a potent hyperglycemic effect by inhibiting pancreatic β cell apoptosis through the regulation of MPAK and Akt signaling pathways. However, the components that contribute to this potential health benefit remain undiscovered.

METHOD

A sensitive, reliable, and validated ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC-TQ-MS/MS) method was developed to simultaneously determine the presence of six active components (neochlorogenic acid, chlorogenic acid, quercetin-3--glucuronide, kaempferol-3--rhamnoside, quercetin, and kaempferol) in rat plasma after a single oral administration (in a dosage of 10.5 g/kg) of an extract of CP leaves to rats. The separation was performed on a Waters ACQUITY BEH C column (50 mm × 2.1 mm, 1.7 μm). The detection was conducted by multiple reaction monitoring (MRM) in negative ionization mode. The two highest abundant MRM transitions without interference were optimized for each analyte. Acetonitrile and formic acid aqueous solution (0.1%) was used as the mobile phase at a flow rate of 0.3 ml/min.

RESULT

The precision, accuracy, and recovery all satisfied the criteria of international guidance (Bioanalytical Method Validation Guidance for Industry, Food and Drug Administration), and the analytes were stable in plasma for all tested conditions. The main pharmacokinetic parameters were calculated by plasma concentration versus time profiles using the pharmacokinetics program.

CONCLUSION

The pharmacokinetic parameters of each compound can facilitate future clinical studies.