UPLC-MS-MS同时测定中药活血益气方KLW中三七皂苷R_1、人参皂苷Rg_1、人参皂苷Rb_1的含量

Simultaneous Determination of Notoginsenoside R_1 ,Ginsenoside Rg_1 and Ginsenoside Rb_1 in Chinese Formula KLW by UPLC-MS-MS

目的:建立一种超高效液相色谱串联质谱分析方法,可同时测定中药活血益气方KLW中的三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1的含量。方法:中药活血益气方KLW采用超声提取,离心取上清液过0.2μm微孔滤膜后测试,采用Waters ACQUITY BEH C18色谱柱(2.1 mm×50 mm,1.7μm),以甲醇和水溶液梯度洗脱,流速0.4 mL.min-1,多反应监测测试方法(MRM)三七皂苷R1选择1 007.2/423.3 m/z;人参皂苷Rg1选择875.2/423.5 m/z;人参皂苷Rb1选择1 183.3/487.3m/z定量。结果:三七皂苷R1、人参皂苷Rg1和人参皂苷Rb1在0.05～2.0 mg.L-1呈现良好的线性关系(r>0.999),平均加样回收率(n=6)分别为97.7%,96.3%,97.1%。结论:方法简便、灵敏、快速、重复性好,适用于同时测定复杂复方KLW中3种皂苷类成分的含量,为该制剂的质量控制和临床药学研究提供了实用的检测方法。

Objective： To develop an ultra performance liquid chromatography tandem mass （UPLC-MS- MS） method for determination of notoginsenoside R1, gensenoside Rgl, and gensenoside Rb, in Chinese formula KLW. Method： The Chinese formula KLW is extracted by ultrasonic and centrifugal, then filtered before test. The ultra performance liquid chromatographic separation was performed on a Waters ACQUITY BEH C18 column （2. 1 mm ×50mm,1.7μm） by gradient elution with methanol-water as the mobile phase at a flow rate of 0. 4 mL · min-1 The analysis of 3 compounds was performed under ESI multiple reaction monitoring （MRM） mode, and the quantification was conducted by external calibration method. The monitoring ions for notoginsenoside RI were m/z 1 007.2/423.3 m/z; gensenoside RgI were 875.2/423.5 m/z; gensenoside Rb1 were 1 183.3/487.3 m/z. Result： The method showed a good linearity in the range of 0.05-2.0 mg .L-1, with the correlation coefficient above 0. 999. The average recoveries （n = 6） of notoginsenoside R1, ginsenoside Rg1 and ginsenoside Rb1 were 97.7%, 96.3%, 97.1% respectively. Conclusion： This method is simple, rapid, accurate and sensitive, can also provide a reliable method for clinical research and can be used for the quality control of Chinese formula KLW.