Simultaneous Determination of Six Bioactive Components of Jiawei Baiteng Extracts in Plasma by HPLC-TQ-MS/MS and Their Pharmacokinetics in Rats / 中国药学杂志

ABSTRACT

OBJECTIVE: To establish an HPLC-TQ-MS/MS assay for simultaneous determination of berberine, plantamajoside, saikosaponin a, tetrahydropalmatine, paeoniflorin and amygdalin in rat plasma and investigate the pharmacokinetics of Jiawei Baiteng extracts in rats. METHODS: The separation was achieved on an Agilent Poroshell 120 EC-C18 column(3.0 mm × 100 mm, 2.7 μm) at 35℃. The mobile phase was consisted of 0.05% formic acid aqueous solution and acetonitrile containing 0.05% formic acid, eluting with a gradient procedure. Mass spectrometry was performed in the multiple reaction monitoring (MRM) mode, with programmed ionization modes witching and time segment scanning. The ion reactions for quantification were as follows m/z 335.9→m/z 319.9 (berberine, ESI+), m/z 639.1 → m/z 160.9 (plantamajoside, ESI-), m/z 779.3 → m/z 617.4 (saikosaponin a, ESI-), m/z 356.0→m/z 192.0(tetrahydropalmatine, ESI+), m/z 449.1→m/z 121.1(paeoniflorin, ESI-), m/z 456.1→m/z 323.1(amygdalin, ESI-), m/z 323.9→m/z 126.9(gliclazide, internal standard, ESI+) and m/z 321.9→m/z 170.1 (gliclazide, internal stand- ard, ESI-). Blood samples were collected in heparinized tubes via the retinal venous plexus from each rat after a single oral dose of Jiawei Baiteng extracts(3.1 g·kg-1). The plasma samples were pretreated by methanol precipitation to remove protein components, and then analyzed by HPLC-TQ-MS/MS. The pharmacokinetic parameters of the bioactive components of Jiawei Baitengex tracts in rats were calculated by DAS (Drug and Statistics for Windows) software(Version 2.0). RESULTS: The methodological test showed that the linear concentration ranges of berberine, plantamajoside, saikosaponin a, tetrahydropalmatine, paeoniflorin and amygdalin were 0.59-292.50, 0.68-168.75, 6.05-1 512.50, 0.68-337.50, 6.70-1675.00 and 5.60-1400.00ng·mL-1, respectively. The limits of quantification (LOQs) of the six analytes were 0.59, 0.68, 6.05, 0.68, 6.70 and 5.60 ng·mL-1, respectively. The HPLC-TQ-MS/MS method was also validated with good precision, recovery and stability, which conformed to the analytical standards of biological samples. CONCLUSION: The established method is proved to be sensitive, simple and reliable, which is suitable for the pharmacokinetic study of Jiawei Baiteng extracts.