Neurology Department, Columbia University Medical Center, New York, NY, USA.
Metabolomics Core Facility, European Molecular Biology Lab, Heidelberg, Germany.
Methods Mol Biol. 2021;2275:379-391. doi: 10.1007/978-1-0716-1262-0_24.
Untargeted lipidomics profiling by liquid chromatography -mass spectrometry (LC-MS) allows researchers to observe the occurrences of lipids in a biological sample without showing intentional bias to any specific class of lipids and allows retrospective reanalysis of data collected. Typically, and in the specific method described, a general extraction method followed by LC separation is used to achieve nonspecific class coverage of the lipidome prior to high resolution accurate mass (HRAM) MS detection . Here we describe a workflow including the isolation of mitochondria from liver tissue, followed by mitochondrial lipid extraction and the LC-MS conditions used for data acquisition. We also highlight how, in this method, all ion fragmentation can be used to identify species of lower abundances, often missed by data dependent fragmentation techniques. Here we describe the isolation of mitochondria from liver tissue, followed by mitochondrial lipid extraction and the LC-MS conditions used for data acquisition.
采用液相色谱-质谱(LC-MS)的非靶向脂质组学分析可以让研究人员在观察生物样本中脂质存在情况时避免对任何特定脂质类别产生有意的偏见,并允许对已收集的数据进行回溯性重新分析。通常,在描述的特定方法中,在使用高分辨精确质量(HRAM)MS 检测之前,采用一般提取方法和 LC 分离来实现脂质组的非特异性类别覆盖。在此,我们描述了一个工作流程,包括从肝组织中分离线粒体,然后进行线粒体脂质提取以及用于数据采集的 LC-MS 条件。我们还强调了在这种方法中,如何使用所有离子碎裂来鉴定低丰度的物种,这些物种通常会被基于数据的碎裂技术所遗漏。