二醇基硅胶固相萃取-多维质谱对乌鳢组织脂质组学分析  

Si O2/diol Based Solid-phase Extraction and Multi-dimensional Mass Spectrometry for Lipidomics Study of Channa argus

作　　者：沈清[1] 金仁耀[1] 冯俊丽[1] 薛静[1] 陈康[1] 戴志远[1] SHEN Qing;JIN Renyao;FENG Junli;XUE Jing;CHEN Kang;DAI Zhiyuan(Institute of Seafood, Zhejiang Gongshang University, Hangzhou 310012, China)

机构地区：[1]浙江工商大学海洋食品研究院,浙江杭州310012

出　　处：《食品科学》2017年第2期154-158,共5页Food Science

基　　金：国家国际科技合作专项(2014DFA32880);国家自然科学基金青年科学基金项目(31601542);浙江省自然科学基金项目(LQ16C200001)

摘　　要：建立以二醇基键合硅胶(Si O2/diol)为填料的固相萃取(solid-phase extraction,SPE)和基于多维度串联质谱的鸟枪法分离纯化并测定乌鳢肌肉组织中的磷脂。利用Folch法和SPE法提取并纯化磷脂,比较3种不同填料SPE柱的萃取效果,优化SPE参数,最佳条件:上样量100?μL,淋洗液p H值为2,洗脱液为体积分数50%乙腈溶液1 m L。所得提取物以直接进样的方法流动注射入电喷雾离子源,经三重四极杆质谱的母离子扫描和中性丢失扫描模式,实现对磷脂分子种类的鉴定和定量。同时,通过多维度串联质谱分析,得到各个磷脂分子中2条脂肪酸链长度和不饱和度信息。实验结果显示,乌鳢样品中成功检出磷脂酰胆碱、磷脂酰乙醇胺、磷脂酰肌醇和磷脂酰丝氨酸4类磷脂,共计67种磷脂分子,且大部分磷脂均含有不饱和脂肪酸链。本方法简单高效,分析准确快捷,结果稳定。In this study, a method using SiO2/diol based solid-phase extraction (SPE) and multi-dimensional mass spectrometry-based shotgun lipidomics was developed for studying the phospholipids in muscle tissue of Channa argus.Crude lipids were extracted by the Folch method. Phospholipids were obtained by SiO2/diol-SPE, and it’s the extraction efficiency was compared with that of two other commercial SPE cartridges. The conditions for SiO2/diol-SPE were optimized as follows: loading volume, 100 μL; washing solvent pH 2, and 1 mL of 50% acetonitrile as eluent. The eluate was directly injected into the electrospray ionization (ESI) source. The phospholipid molecular species were identified and quantified using the precursor-ion scan and neutral loss scan modes, and the structures of two fatty acid chains in each species were confirmed by multi-dimensional mass spectrometry. The results showed that a total of 67 phospholipid molecular specieswere detected, including 18 phosphatidylcholines (PC), 21 phosphatidylethanoamines (PE), 12 phosphatidylinositols (PI),and 16 phosphatidylserines (PS). The phospholipids in C. argus contained abundant unsaturated fatty acyl chains. This analytical method proved to be simple, efficient, and reliable.

关 键 词：鸟枪法脂质组学 多维质谱 二醇基硅胶 固相萃取 乌鳢 

分 类 号：S912[农业科学—水产科学]