Simultaneous determination of calycosin, prim-O-glucosylcimifugin, and paeoniflorin in rat plasma by HPLC-MS/MS: application in the pharmacokinetic analysis of HQCF.

ABSTRACT

OBJECTIVE: This study aimed to develop and validate a high-performance liquid chromatography-tandem mass spectrometry method to simultaneously determine three bioactive components of the Huangqi Chifeng decoction (HQCF) in rat plasma. METHODS: Taxol was used as an internal standard in the developed method. Chromatographic separation was performed on a C18 column using a gradient elution with 0.1% formic acid in acetonitrile (v/v) and 0.1% formic acid in water (v/v) as the mobile phases at a flow rate of 0.4 mL·minute-1. All compounds were monitored via selected reaction monitoring with an electrospray ionization source. RESULTS: The lower limits of quantification of paeoniflorin, calycosin, and prim-O-glucosylcimifugin were 15.0, 0.75, and 0.75 ng·mL-1, respectively. The calibration curves indicated optimal linearity (r > 0.99) across the concentration ranges. The specificity, precision, accuracy, recovery, matrix effect, and stability of the method were validated. This method was successfully applied in a pharmacokinetics study of the three compounds in rat plasma. CONCLUSION: The pharmacokinetics results provide insights into the mechanisms of HQCF in vivo and its future clinical application.