甜茶叶中甜茶苷的UPLC-MS/MS分析

Determination of Rubusoside from Rubus Suavissimus S.Lee Leaves by UPLC-MS/MS

采用超高效液相色谱-质谱联用法测定广西特产甜茶叶中的主要甜味成分甜茶苷的含量。样品经前处理后以Acquity UPLC BEHC18柱为色谱柱,流动相为乙腈(含体积分数为0.1%甲酸)-蒸馏水(含体积分数0.1%甲酸),梯度条件为10min乙腈相体积由20%变化到65%,柱温35℃,进样量5μL,流速为0.15mL/min。在质谱仪上以电喷雾电离正离子模式多离子反应监测(MRM)方式进行定量分析,监测的离子为m/Z 643→319。甜茶苷回归方程的线性关系良好,线性范围0.1—10.0μg/mL,r=0.9996,方法加标回收率为96.6%,检出限为0.04μg/mL;定量下限为0.1μg/mL。该法操作简便快速、灵敏度高、重复性好,适用于甜茶叶中甜茶苷的测定。

A method for determination of rubusoside in sweet tea leaves was established by ultra performance liquid chromatography electrospray ionization mass spectrometry（UPLC-ESI MS/MS）.After pretreatment procedure,sample was separated on a Acquity UPLC BEH C18 column with A acetonitrile（with 0.1% formic acid,V/V） and B distilled water（with 0.1% formic acid,V/V）as mobile phase.The gradient elution was from 0 to 10 min followed a linear change from 20% to 65%（V/V）for phase A with injection volume of 5μL at a flow rate of 0.15 mL/min.The column temperature was set at 35℃.Detection was performed by multiple reaction monitoring（MRM） on mass spectrometer.Electrospray ionization source was applied and operated in the positive ion mode（ESI＋）,and m/Z 643→319 was selecting as detecting ion.A good linear calibration was obtained in the range of 0.1—10.0μg/mL with correlation coefficient of 0.9996.The average addition standard recovery was 96.6%.The LOD and LOQ were 0.04μg/mL and 0.1μg/mL.The proposed method is specific,sensitive and reproducible enough to be used for the determination of rubusoside in sweet tea leaves.