HPLC-MS法同时测定益母草及其颗粒剂中盐酸水苏碱与盐酸益母草碱的含量

目的:建立益母草与益母草颗粒中盐酸水苏碱与盐酸益母草碱的含量测定方法。方法:采用LC-MS法同时测定盐酸水苏碱与盐酸益母草碱的含量。使用Waters XBridge Amide色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.1%甲酸为流动相,梯度洗脱,流速1.0 mL·min^-1,分流比为1:4,使用ESI⁺离子源,选择性离子扫描(SIM)下对盐酸水苏碱(m/z 144.0)和盐酸益母草碱(m/z 312.0)进行测定。结果:盐酸水苏碱质量浓度在1.126~281.4 ng·mL^-1线性关系良好(r=0.999 8);在药材及颗粒剂中盐酸水苏碱平均回收率(n=6)分别为99.3%和98.9%,RSD分别为2.2%和2.6%。盐酸益母草碱质量浓度在1.042~260.6 ng·mL^-1性关系良好(r=0.999 5);在药材及颗粒剂中盐酸益母草碱平均回收率(n=6)分别为100.1%和99.1%,RSD分别为2.5%和2.9%。5批益母草中盐酸水苏碱与盐酸益母草碱的含量分别为6.688~12.411、0.686~3.637 mg·g^-1,5批益母草颗粒中盐酸水苏碱与盐酸益母草碱的含量分别为2.067~8.876、0.041~0.231 mg·g^-1。 结论:经过方法学验证,该方法可作为益母草与益母草颗粒的含量测定方法。

Objective:To establish a quantitative method for stachydrine hydrochloride and leonurine hydrochloride determination in Leonuri Herba and Yimucao granules.Methods:The contents of stachydrine hydrochloride and leonurine hydrochloride in Leonuri Herba and Yimucao granules were determined by HPLC-MS.The chromatographic column adopted in this study was Waters XBridge Amide(250 mm×4.6 mm,5 μm).The mobile phase was acetonitrile-0.1% formic acid in gradient mode at the flow rate of 1.0 mL·min^-1,with the split ratio of 1:4.The ESI⁺ source of the mass spectrometer was used in selected ion monitoring(SIM)mode,which was used for the detection of stachydrine hydrochloride(m/z 144.0)and leonurine hydrochloride(m/z 312.0). Results:The linear range of stachydrine hydrochloride was 1.126-281.4 ng·mL^-1(r=0.999 8),the average recovery rates(n=6)of stachydrine hydrochloride in Leonuri Herba and Yimucao granules were 99.3% and 98.9% with RSDs of 2.2% and 2.6%,respectively.The linear range of leonurine hydrochloride was 1.042~ 260.6 ng·mL^-1(r=0.999 5),and the average recovery rates(n=6)of leonurine hydrochloride in Leonuri Herba and Yimucao granules were 100.1% and 99.1% with RSDs of 2.5% and 2.9%,respectively.The contents of stachydrine hydrochloride and leonurine hydrochloride in 5 samples of Leonuri Herba were 6.688-12.411 and 0.686-3.637 mg·g^-1.The contents of stachydrine hydrochloride and leonurine hydrochloride in 5 samples of Yimucao granules were 2.067-8.876 and 0.041-0.231 mg·g^-1.Conclusion:The results of methodology validation showed that the established method can be used for contents determination of stachydrine hydrochloride and leonurine hydrochloride in Leonuri Herba and Yimucao granules.