Pharmacokinetic study of puerarin of Zige freeze-dried powder in cerebral ischemia rats and normal rats / 中国药学杂志

ABSTRACT

OBJECTIVE: To investigate the pharmacokinetics of puerarin of Zige freeze-dried powder in the plasma of the normal ruts and the model rats with middle cerebral artery occlusion(MACO). METHODS: A high-performance liquid chromatography-tan-dem mass spectrometry (HPLC-MS/MS) method was developed and validated for quantification of puerarin in rat plasma after injection administration of Zige freeze-dried powder. The plasma samples were pretreated with methanol for protein precipitation and the supernatants were analysis on a Diamonsil C18 column (4.6 mm × 150 mm, 5 μm) with the mobile phase consisting of 0.1% formic acid in methanol and 10 mmol · L-1 ammonium acetate in water (8020) at a flow rate of 0. 6 mL · min-1 to separate puerarin and genistein (internal standard, IS). A triple-quadrupole tandem mass spectrometer under a multiple reaction-monitoring (MRM) mode to determine puerarin(m/z 415.1 → 295.1) and genistein(m/z 269.2 → 133.1). The Zige freeze-dried powder was administered to rats in 26.7 mg · kg-1 doses via intravenous (iv) injection, the plasma sample was collected at each time point (3, 5, 10, 30 min, 1, 2, 3 and 5 h post-administration). The pharmacokinetic analyses of puerarin in rat plasma were performed via the proprietary DAS 2.1 computer software package. RESULTS: The calibration curves had good linear in 0.02-100 μg · mL-1 with r=0.9954. The intra- and inter day precisions were found to be less than 13.2%, the accuracy was ranged in -7.3%-10.5%, and the extraction recoveries of the puerarin from the plasma was 86.3%. The contents of puerarin arrived the max values with (56.5 ± 3.4) and (62.3 ± 14.0) μg · mL-1, the t1/2 were (0.311 ± 0.133) and (0.755 ± 0.128) h, the values of AUG0-t, were (17.1 ± 1.5) and (20.0 ± 6.8) μg · h · mL-1, the values of MRT0-t, were (0.269 ± 0.044) and (0.360 ± 0.045) h in the plasma of the normal and the model respectively. Compared with the normal, the t1/2, MRT0-t, of the model were significantly longer (P < 0.05). CONCLUSION: The developed and validated method is specifically, rapid and sensitive, and has been successfully applied to research the pharmacokinetics of puerarin in rats plasma after iv zige freeze-dried powder. The pharmacokinetic; parameters showed that the puerarin in the Zige freeze-dried powder has better biological availability, slower elimination and longer resistance time in the model rats, which indicate the reduce of the rate of metabolism in the model and is beneficial to the clinical pharmacokinetic studies after iv administration of Zige freeze-dried powder.