High-Throughput Metabolomics: Isocratic and Gradient Mass Spectrometry-Based Methods
High-Throughput Metabolomics: Isocratic and Gradient Mass Spectrometry-Based Methods
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DOI：
10.1007/978-1-4939-9236-2_2
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发表时间：
2019-01-01
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影响因子：
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通讯作者：
D'Alessandro, Angelo
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作者：
Nemkov, Travis;Reisz, Julie A.;D'Alessandro, Angelo
代谢组学在过去的十年中已经成为一种非常有吸引力和影响力的技术，用于各种生物学应用中的表型水平分析。最近，高通量分析技术的双重发展沿着高分辨率质谱仪灵敏度的显著提高，使得每天能够对数百个独特样品进行常规分析。我们之前已经报道了一个强大的3分钟等度代谢组学平台，用于定量氨基酸以及中心碳和氮代谢的关键途径。在这项工作的基础上，我们在这里描述了一个5分钟的反相梯度，然后是全球性的，非靶向的亲水性代谢组分析。除了观察在3分钟运行中测量的那些代谢物之外，这里使用较长梯度运行还允许覆盖极性较低的化合物，例如脂肪酸和酰基肉毒碱，这两者都是线粒体和脂质代谢中的关键参与者，而不会显著牺牲通量。
Metabolomics has emerged in the past decade as a highly attractive and impactful technique for phenotype-level profiling in diverse biological applications. Most recently, the dual developments of high-throughput analytical techniques along with dramatically increased sensitivity of high-resolution mass spectrometers have enabled the routine analysis of hundreds of unique samples per day. We have previously reported a robust 3 min isocratic metabolomics platform for the quantification of amino acids and the key pathways of central carbon and nitrogen metabolism. Building on this work, we describe here a 5 min reverse phase gradient followed by global, untargeted profiling of the hydrophilic metabolome. In addition to observing those metabolites measured in the 3 min run, the use of the longer gradient run here also allows for coverage of less polar compounds such as fatty acids and acylcarnitines, both key players in mitochondrial and lipid metabolism, without a significant sacrifice in throughput.