非靶向代谢组学分析方案：从样品制备到数据处理

A Protocol for Untargeted Metabolomic Analysis: From Sample Preparation to Data Processing.

作者信息

Souza Amanda L, Patti Gary J

机构信息

Life Science Mass Spectrometry Division, Thermo Fisher Scientific, San Jose, CA, USA.

Department of Chemistry, Washington University in St. Louis, Saint Louis, MO, USA.

出版信息

Methods Mol Biol. 2021;2276:357-382. doi: 10.1007/978-1-0716-1266-8_27.

Untargeted metabolomics has rapidly become a profiling method of choice in many areas of research, including mitochondrial biology. Most commonly, untargeted metabolomics is performed with liquid chromatography/mass spectrometry because it enables measurement of a relatively wide range of physiochemically diverse molecules. Specifically, to assess energy pathways that are associated with mitochondrial metabolism, hydrophilic interaction liquid chromatography (HILIC) is often applied before analysis with a high-resolution accurate mass instrument. The workflow produces large, complex data files that are impractical to analyze manually. Here, we present a protocol to perform untargeted metabolomics on biofluids such as plasma, urine, and cerebral spinal fluid with a HILIC separation and an Orbitrap mass spectrometer. Our protocol describes each step of the analysis in detail, from preparation of solvents for chromatography to selecting parameters during data processing.

非靶向代谢组学已迅速成为包括线粒体生物学在内的许多研究领域中首选的分析方法。最常见的是，非靶向代谢组学采用液相色谱/质谱联用技术进行，因为它能够测量范围相对广泛的物理化学性质各异的分子。具体而言，为了评估与线粒体代谢相关的能量途径，在使用高分辨率精确质量仪器进行分析之前，通常会应用亲水作用液相色谱（HILIC）。该工作流程会生成大型、复杂的数据文件，手动分析不切实际。在此，我们展示了一种使用HILIC分离和轨道阱质谱仪对血浆、尿液和脑脊液等生物流体进行非靶向代谢组学分析的方案。我们的方案详细描述了分析的每个步骤，从色谱溶剂的制备到数据处理过程中的参数选择。