HPLC-MS/MS法同时测定百蕊颗粒中紫云英苷和山奈酚的含量

Simultaneous Determination of Astragulin and Kaempferol in Bairui Granule by HPLC-MS/MS

目的:建立同时测定百蕊颗粒中紫云英苷和山奈酚2种有效成分含量的HPLC-MS/MS检测方法。方法:采用Ultimate XB-C_(18)(100 mm×2.1 mm,5μm)色谱柱;流动相为0.1%甲酸水溶液(A)-0.1%甲酸乙腈(B),梯度洗脱,流速0.3 ml·min^(-1);柱温为40℃;采用电喷雾离子源(ESI),以多反应监测方式(MRM)进行定量分析。紫云英苷和山奈酚在负离子模式下定量分析离子对分别为m/z 447.2→m/z 284.0和m/z 285.1→m/z 187.1。结果:紫云英苷和山奈酚的浓度分别在9.86~1 970ng·ml^(-1)(r=0.999 5)和8.73~437 ng·ml^(-1)(r=0.999 6)范围内均呈良好线性关系;紫云英苷和山奈酚低、中、高平均加样回收率分别为99.23%、99.65%、99.23%和98.24%、99.13%、99.69%,RSD分别为1.84%、1.37%、1.21%和1.38%、0.96%、1.47%(n=6)。结论:本文方法简便可靠,重复性好,可同时测定百蕊颗粒中紫云英苷和山奈酚的含量,适用于百蕊颗粒质量控制。

Objective： To develop a quantitative HPLC-MS/MS method for the determination of astragulin and kaempferol in Bairui granule. Methods： A column of Ultimate XB-C C18（ 100 mm × 2. 1 mm,5 μm） was used. The mobile phase was composed of 0. 1%formic acid-water（ A） and 0. 1% formic acid-acetonitrile（ B） with gradient elution. The flow rate was 0. 3 ml·min^（-1）. The detection was performed with multiple teactions monitoring（ MRM） using electrospray ionization（ ESI）. The precursor/product ion transitions were monitored at m/z 447. 2→m/z 284. 0（ anion mode） for astragulin and m/z 285. 1 →m/z 187. 1（ anion mode） for kaempferol.The temperature of column was set at 40℃. Results： A good linearity was obtained with the concentration ranging from 9. 86 to 1 970 ng·ml^-1（ r = 0. 999 5） for astragulin and 8. 73 to 437 ng·ml^-1（ r = 0. 999 6） for kaempferol. The average recovery of the two investigated compounds at low,medium and high concentrations was 99. 23%,99. 65% and 99. 23%,and 98. 24%,99. 13% and99. 69% with the RSD of 1. 84%,1. 37% and 1. 21%,and 1. 38%,0. 96% and 1. 47%,respectively（ n = 6）. Conclusion： The method is rapid,accurate and reliable,which can be used to control the quality of Bairui granule.