一种测定植物甾醇的高灵敏度衍生化HPLC-MS/MS方法  

A Highly Sensitive Method for Quantitative Analysis of Phytosterols by HPLC-MS/MS with Pre-Column Derivatization

作　　者：孙琪[1] 陈帆[1] 

机构地区：[1]温州大学化学与材料工程学院,浙江温州325035

出　　处：《分析测试技术与仪器》2016年第3期169-178,共10页Analysis and Testing Technology and Instruments

基　　金：国家自然科学基金面上项目(21272178)资助~~

摘　　要：建立以丹磺酰肼为柱前衍生试剂,采用 Agilent Poroshell 120 EC-C18（50 mm×3. 0 mm,2. 7 μm）反相色谱柱,甲醇-水（体积比为99 ∶ 1）为流动相,流速为 0. 3 mL/ min,进样量为10 μL,电喷雾电离源,选择反应监测（ SRM）方式正离子检测,测定豆甾醇、 β-谷甾醇、岩藻甾醇、麦角甾醇的柱前衍生化高效液相色谱串联质谱法. 同时分析测定羊栖菜、海带、紫菜中的甾醇种类及含量. 以胆固醇为内标物,采用内标工作曲线法定量,各甾醇线性范围在0. 05 ~ 100 μg / mL 之间,相关系数在0. 990 3 ~ 0. 998 1 之间,检出限（LODs）在0. 2 ~ 20 ng / mL 之间,定量限（LOQs）在0. 667 ~ 66. 7 ng / mL 之间,甾醇类物质灵敏度比高效液相色谱法提高25 ~ 500 倍.A sensitive liquid chromatography-electrospray ionization-tandem mass spectrometry （LC-ESI-MS / MS） methodfor the determination of phytosterols in sargassum fusiforme has been developed and validated. Four kinds of phytosterols werederivatived by dansyl hydrazine and separated on a reversed-phase Agilent Poroshell 120 EC-C18 column（50 mm×3. 0 mm,2. 7 μm） with a mobile phase of methanol-water （99 ∶ 1,V ∶ V） at a flow rate of 0. 3 mL/ min, and the injection volume was10 μL. An electrospray ionization source was applied and operated in the positive ion mode, confirmed by the selectedreaction monitoring （SRM） mode using an internal standard method. Phytosterols showed a wide linearity in 0. 05 ~ 100 μg /mL, with correlation coefficients between 0. 990 3 and 0. 998 1. The limits of detection of phytosterols were between 0. 2 and20 ng / mL. This simple, selective and sensitive method can be successfully used for the determinations of different types ofmatrices, and has a 25 ~ 500 fold increase in sensitivity than that in high performance liquid chromatography.

关 键 词：植物甾醇 高效液相色谱串联质谱法 柱前衍生化 选择反应监测 

分 类 号：O657.3[理学—分析化学]