化学衍生化辅助高分辨质谱法用于 shotgun 脂质组学分析。

Chemical Derivatization-Aided High Resolution Mass Spectrometry for Shotgun Lipidome Analysis.

机构信息

School of Chemistry, The University of Melbourne, Parkville, VIC, Australia.

Department of Biochemistry and Pharmacology, The University of Melbourne, Parkville, VIC, Australia.

出版信息

Methods Mol Biol. 2021;2306:61-75. doi: 10.1007/978-1-0716-1410-5_5.

DOI:10.1007/978-1-0716-1410-5_5

PMID:33954940

Abstract

Chemical derivatization coupled with nano-electrospray ionization (nESI) and ultra-high resolution accurate mass spectrometry (UHRAMS) is an established approach to overcome isobaric and isomeric mass interference limitations, and improve the analytical performance, of direct-infusion (i.e., "shotgun") lipidome analysis strategies for "sum composition" level identification and quantification of individual lipid species from within complex mixtures. Here, we describe a protocol for sequential functional group selective derivatization of aminophospholipids and O-alk-1'-enyl (i.e., plasmalogen) lipids, that when integrated into a shotgun lipidomics workflow involving deuterium-labeled internal lipid standard addition, monophasic lipid extraction, and nESI-UHRAMS analysis, enables the routine identification and quantification of >500 individual lipid species at the "sum composition" level, across four lipid categories and from >30 lipid classes and subclasses.

摘要

化学衍生化结合纳升电喷雾电离（nESI）和超高分辨率精确质量质谱（UHRAMS）是一种成熟的方法，可以克服同量异位和同型质量干扰的限制，并提高直接进样（即“ shotgun”）脂质组学分析策略的分析性能，用于“总和组成”水平鉴定和对复杂混合物中单个脂质种类进行定量。本文描述了一种对氨基磷脂和 O-alk-1'-烯基（即脑苷脂）脂质进行顺序官能团选择性衍生化的方案，当将其整合到 shotgun 脂质组学工作流程中时，该方案涉及氘标记的内部脂质标准添加、单相脂质提取和 nESI-UHRAMS 分析，可以常规鉴定和定量> 500 种单个脂质种类，涵盖四个脂质类别和> 30 个脂质类别和子类。