超高效液相质谱法测定增强免疫类中药保健食品中多种功能成分含量

Deteetion of multiple functional components in health food containing Chinese materia medica for enhancing immunity by ultra performance liquid chromatography-tandem mass spectrometry

目的:采用超高效液相色谱-串联质谱法(Ultra Performance Liquid Chromatography-tandem mass spectrometry,UPLC-MS/MS)建立了同时测定增强免疫类中药保健食品中葛根素、红景天苷、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、金丝桃苷等功效成分的含量测定方法。方法:样品通过甲醇超声提取,色谱柱分离,水-0.1%甲酸乙腈梯度洗脱,采用负离子多反应监测(multiple reaction monitoring, MRM)模式进行定量定性分析。结果:6种目标化合物在13 min内完成分离且1~100 ng/mL范围内均呈现良好的线性关系(r>0.9979),0.55、0.60和0.65μg/kg添加水平的平均回收率为83.0%~107.0%,实验室内变异系数为0.74%~3.71%(n=7),方法检出限为0.06~0.44μg/kg。结论:该方法快速准确,灵敏度高,专属性强,适用于同时测定中药保健食品中葛根素、红景天苷、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1、金丝桃苷等具有增强免疫力功效成分的含量,可为具有增强免疫力中药保健食品的质量控制提供依据。

Objective:To establish a method for the simultaneous determination of puerarin,salidroside,ginsenoside Rgl,ginsenoside Re,ginsenoside Rbl,and hypericin in health food containing Chinese materia medica for enhancing immunity by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).Method:Samples were extracted by methanol ultrasonically,separated by chromatographic column,and eluted with a gradient of water-0.1%formic acid acetonitrile.Quantitative and qualitative analyses were performed in multiple reaction monitoring(MRM)mode using an electrospray negative ion source.Result:The 6 target compounds were separated within 13 minutes and showed good linearity in the range of 1 to 100 ng·mL^(-1)(r>0.9979),and the average recoveries of spiked levels of 0.55,0.60 and 0.65μg·kg^(-1) ranged from 83.0%to 107.0%.The coefficient of variation(CV)ranged from 0.74%to 3.71%(n=7),and the method detection limit(MDL)ranged from 0.06 to 0.44μg kg.Conclusion:The method is fast,accurate,sensitive and specific.It is suitable for the simultaneous determination of the contents of puerarin,salidroside,ginsenoside Rgl,ginsenoside Re,ginsenoside Rbl,hypericin with immunity enhancing effects in health food containing Chinese materia medica.The method can provide the basis for the quality control of the health food containing Chinese materia medica for enhancing immunity.