School of Chemistry, The University of Melbourne, Parkville, VIC, Australia.
Department of Biochemistry and Pharmacology, The University of Melbourne, Parkville, VIC, Australia.
Methods Mol Biol. 2021;2306:61-75. doi: 10.1007/978-1-0716-1410-5_5.
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.
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。
