Lopes Aline Soriano, Cruz Elisa Castañeda Santa, Sussulini Alessandra, Klassen Aline
Department of Exact and Earth Sciences, Federal University of São Paulo (UNIFESP), Diadema, SP, Brazil.
Laboratory of Bioanalytics and Integrated Omics (LaBIOmics), University of Campinas (UNICAMP), Campinas, SP, Brazil.
Adv Exp Med Biol. 2017;965:77-98. doi: 10.1007/978-3-319-47656-8_4.
Amongst all omics sciences, there is no doubt that metabolomics is undergoing the most important growth in the last decade. The advances in analytical techniques and data analysis tools are the main factors that make possible the development and establishment of metabolomics as a significant research field in systems biology. As metabolomic analysis demands high sensitivity for detecting metabolites present in low concentrations in biological samples, high-resolution power for identifying the metabolites and wide dynamic range to detect metabolites with variable concentrations in complex matrices, mass spectrometry is being the most extensively used analytical technique for fulfilling these requirements. Mass spectrometry alone can be used in a metabolomic analysis; however, some issues such as ion suppression may difficultate the quantification/identification of metabolites with lower concentrations or some metabolite classes that do not ionise as well as others. The best choice is coupling separation techniques, such as gas or liquid chromatography, to mass spectrometry, in order to improve the sensitivity and resolution power of the analysis, besides obtaining extra information (retention time) that facilitates the identification of the metabolites, especially when considering untargeted metabolomic strategies. In this chapter, the main aspects of mass spectrometry (MS), liquid chromatography (LC) and gas chromatography (GC) are discussed, and recent clinical applications of LC-MS and GC-MS are also presented.
在所有组学科学中,毫无疑问,代谢组学在过去十年中经历了最重要的发展。分析技术和数据分析工具的进步是使代谢组学得以发展并确立为系统生物学中一个重要研究领域的主要因素。由于代谢组学分析需要高灵敏度来检测生物样品中低浓度存在的代谢物,需要高分辨率来鉴定代谢物,以及需要宽动态范围来检测复杂基质中不同浓度的代谢物,质谱法正成为满足这些要求使用最广泛的分析技术。单独的质谱法可用于代谢组学分析;然而,一些问题,如离子抑制,可能会使低浓度代谢物或某些不易离子化的代谢物类别的定量/鉴定变得困难。最佳选择是将气相或液相色谱等分离技术与质谱联用,以提高分析的灵敏度和分辨率,此外还能获得有助于鉴定代谢物的额外信息(保留时间),尤其是在考虑非靶向代谢组学策略时。在本章中,将讨论质谱(MS)、液相色谱(LC)和气相色谱(GC)的主要方面,并介绍LC-MS和GC-MS最近的临床应用。