UHPLC-MS/MS法结合化学计量学分析姜味草中的多指标成分

Analysis of Multi-index Components in Herba Micromeriae biflorae by UHPLC-MS/MS Combined with Chemometrics

建立了超高效液相色谱串联质谱(UHPLC-MS/MS)法同时测定姜味草(1)药材中14个活性成分,即新绿原酸、绿原酸、隐绿原酸、咖啡酸、异绿原酸A、芦丁、木犀草苷、异槲皮苷、迷迭香酸、香叶木苷、阿魏酸、野黄芩苷、木犀草素-7-O-β-D-葡萄糖醛酸苷和蒙花苷。色谱柱采用Acquity UPLC BEH C柱(2.1 mm×100 mm,1.7 μm),流动相A为乙腈,B为0.1%甲酸溶液,线性梯度洗脱,流速为0.4 ml/min,柱温为35 ℃,进样量为2 μl。采用电喷雾离子源(ESI),多反应监测模式(MRM)。并结合聚类分析、主成分分析和正交偏最小二乘法-判别分析评价不同批次1药材的差异。结果显示,14个活性成分的线性关系、精密度、稳定性、重复性及加样回收率等均符合要求。化学计量学分析结果表明,云南和四川2个产地的1药材中,14个活性成分存在一定差异,其中迷迭香酸和蒙花苷为主要差异性成分。建立的UHPLC-MS/MS法准确、快速、简单、灵敏度高,为1药材的质量控制提供了参考。

An ultra-high performance liquid chromatography-mass spectrometry(UHPLC-MS/MS) method was established for the simultaneous determination of fourteen active ingredients,including neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,caffeic acid,isochlorogenic acid A,rutin,cynaroside,isoquercitrin,rosmarinic acid,diosmin,ferulic acid,scutellarin,luteolin-7-O-β-D-glucuronide and linarin,in Herba Micromeriae biflorae(1).The Acquity UPLC BEH Ccolumn(2.1 mm×100 mm,1.7 μm) was used,and the analysis was carried out in the linear gradient elution mode with acetonitrile as mobile phase A and 0.1% formic acid solution as mobile phase B.The flow rate was 0.4 ml/min,the column temperature was 35 ℃ and the injection volume was 2 μl.The electron spray ionization(ESI) and multiple reaction monitoring(MRM) mode were used,and combined with cluster analysis,principal component analysis and orthogonal partial least squares-discriminant analysis to evaluate the differences of different batches of 1.The results showed that the linear relationship,precision,stability,repeatability and sample recovery of the fourteen compounds were all met the requirements.The chemometric analysis results showed that there were certain differences of the fourteen compounds in 1 from two origins in Yunnan and Sichuan,among which rosmarinic acid and linarin were the main differential components.The established UHPLC-MS/MS method is accurate,rapid,simple and highly sensitive,which provides a reference for the quality control of 1.