A high-performance liquid chromatography tandem mass spectrometry (HPLC–MS/MS) method was developed and fully validated for simultaneous determination of puerarin, 3′-hydroxy puerarin, 6″-O-xylosyl puerarin, 3′-methoxy puerarin, mirificin, puerarin-7-O-glucoside, daidzin, daidzein, daidzein-7-O-glucoside, ginsenoside-Rd, notoginsenoside-R1, ginsenoside-Re, ginsenoside-Rg1, ginsenoside-Rb1 in rat plasma after intravenous injection of 5 mL/kg Gegen-Sanqi compatibility solution (containing Pueraria flavonid 10.8 mg/mL and Panax notoginsenosidum 5.4 mg/mL). After addition of internal standard (IS) baicalin, the analytes and IS were recovered from rat plasma by protein precipitation using acetonitrile containing 0.1% formic acid. Chromatographic separation was performed on a Capcell pak MG C18 column (3.0 mm × 75 mm, 3.0 μm) at 35 °C with a flow rate of 0.75 mL/min. Mass spectrometry was conducted using multiple reaction monitoring in positive mode. The method was linear for all of the analytes over 1000 times concentration range with correlation coefficients greater than 0.99. The precision and accuracy of the LC–MS/MS assay based on the three analytical quality control (QC) levels were well within the acceptance criteria from FDA guidance for bioanalytical method validation. Method recoveries were higher than 75% and the matrix effects were minimal. All analytes were stable under tested conditions. It was the first time to study the pharmacokinetics of daidzein-7-O-glucoside in rat plasma. To the best of our knowledge, this is the first study for simultaneous determination of so many analytes in rat plasma after intravenous injection of Gegen-Sanqi compatibility solution. It was expected that the present work would provide some helpful references for the apprehension of the mechanism of action and further clinical efficacy studies of Gegen-Sanqi herb-pair.

建立了高效液相色谱串联质谱法（HPLC–MS / MS），并已完全验证了同时测定葛根素，3'-羟基葛根素，6” -O-羟基甲苯基葛根素，3'-甲氧基葛根素，mirificin，葛根素7- O-葡萄糖苷，大豆苷，大豆苷，大豆苷7- O静脉注射5 mL / kg Gegen-Sanqi相容性溶液（含葛根黄酮10.8 mg / mL和三七总皂甙5.4）后，在大鼠血浆中测定-葡萄糖苷，人参皂苷-Rd，三七皂苷-R1，人参皂苷-Re，人参皂苷-Rg1，人参皂苷-Rb1毫克/毫升）。加入内标（IS）黄ical苷后，使用含0.1％甲酸的乙腈通过蛋白沉淀从大鼠血浆中回收分析物和IS。在Capcell pak MG C18色谱柱（3.0 mm×75 mm，3.0μm）上于35°C进行色谱分离，流速为0.75 mL / min。使用正反应模式的多反应监测进行质谱分析。对于1000倍浓度范围内的所有分析物，该方法都是线性的，相关系数大于0.99。基于三种分析质量控制（QC）水平的LC-MS / MS分析的准确性和准确性完全符合FDA指导性生物分析方法验证的接受标准。方法的回收率高于75％，基质的影响极小。所有分析物在测试条件下均稳定。这是首次研究黄豆苷元7-的药代动力学大鼠血浆中的O-葡萄糖苷。据我们所知，这是首次在静脉注射Gegen-Sanqi相容性溶液后同时测定大鼠血浆中如此多的分析物的研究。可以预期，目前的工作将为理解葛根-三七草药对的作用机理和进一步的临床疗效研究提供一些有用的参考。