Objective: In order to evaluate the reliability and feasibility of pueraria reference extractive substance
(RES) used in biological sample, the pharmacokinetics of 3
–hydroxy puerarin (3
-HP), puerarin, 3
–
methoxy puerarin (3
-MP), and daidzein-8-C-apiosyl-(1-6)-glucoside (DAG) in beagle plasma following
oral administration of Yufeng Ningxin Tablet were quantitated.
Methods: A reliable and sensitive high-performance liquid chromatography-tandem triple quadrupole
mass spectrometry (HPLC-QQQ-MS/MS) method developed with chromatographic separation was operated on a Merck C18 column, and acetonitrile-5 mmol/L ammonium was used as mobile phase in gradient
elution. The plasma samples were deproteinized by acetone, detected by triple quadrupole mass spectrometry with an electrospray ionization interface, and quantified using selected ion monitoring mode.
The pharmacokinetic parameters were calculated by Winnonlin 4.1.
Results: The calibration curves of the reference extractive substance and standard substance methods
were linear over the ranges 0.0417–11.3309 μg/mL and 0.0394–10.0000 μg/mL. The intra-day and interday precision of the two methods at three concentrations were less than 13.63%, and the average recoveries of 3
-HP, puerarin, 3
-MP, and DAG were more than 70.67%. The RSD of the mean plasma concentrations of the analytes calculated by the two methods was less than 5%, and cos (ϑ) = =1.000. Among the
analytes, puerarin showed the highest blood concentration [(940 ± 185) ng/mL] and the longest retention
time [(5 ± 1) h] in the dog’s bodies