LC-MS/MS方法同时检测超声提取迷迭香(Rosmarnus officinalis L.)叶片中的4种主要成分

Simultaneous Determination of Four Major Volatile Components in Rosemary (Rosmarnus officinalis L.) Leaves by LC-MS/MS with Ultrasonic-Assisted Extraction

目的:建立一种同时测定迷迭香中迷迭香酸、鼠尾草酸、绿原酸和咖啡酸含量的LC-MS/MS分析方法,并优化超声辅助提取这4种物质的方法。方法:液相条件:采用Agilent Eclipse XDB-C18色谱柱(150mm×4.6mm,5μm),流动相为乙腈和水(含0.1%甲酸)进行梯度洗脱。质谱条件:ESI离子源,MRM负离子扫描方式,检测在不同提取溶剂、料液比、超声时间、pH值及酸种类的超声波辅助提取条件下4种物质的含量。结果:咖啡酸、绿原酸、迷迭香酸和鼠尾草酸在质量浓度范围分别为0.0005～0.5、0.0005～0.5、0.002～0.2、0.01～1mg/mL的条件下线性关系良好,并符合方法学考察要求;在最优提取条件下迷迭香酸含量为4.07mg/g,鼠尾草酸含量为14.50mg/g,绿原酸含量为8.49mg/g,咖啡酸含量为2.92mg/g。结论:该分析方法可快速灵敏地实现同时检测,并优化了这4种化合物的最佳提取条件。

Objective：To establish an LC-MS/MS assay for the simultaneous determination of rosmarinic acid,carnosic acid,chlorogenic acid and caffeic acid in rosemary leaves and to optimize conditions for the ultrasonic extraction of these components.Methods： The chromatographic separation was performed on an Agilent Eclipse XDB-C18 column（150 mm × 4.6 mm,5 μm） using a mobile phase consisting of acetonitrile and 0.1% formic acid in water by gradient elution.The analyztes were monitored by ESI-MS/MS in the negative ion MRM mode.The effects of solvent type,solid/liquid ratio,ultrasonic treatment time,pH and acid type on the extraction of rosmarinic acid,carnosic acid,chlorogenic acid and caffeic acid were investigated.Results： Caffeic acid,chlorogenic acid,rosmarinic acid and carnosic acid showed a good linear relationship over the concentration ranges of 0.0005–0.5,0.0005–0.5,0.002–0.2 and 0.01–1 mg/mL,respectively.Under optimal extraction conditions,the contents of rosmarinic acid,carnosic acid,caffeic acid and chlorogenic acid were respectively,4.07,14.50,2.92 mg/g and 8.49 mg/g as determined by the LCMS/MS assay.Conclusion： This array allows rapid,sensitive and simultaneous determination of osmarinic acid,carnosic acid,chlorogenic acid and caffeic acid in rosemary leaves.Their optimal extraction conditions have been established.