Separation and quantitation of water soluble cellular metabolites by hydrophilic interaction chromatography-tandem mass spectrometry
Separation and quantitation of water soluble cellular metabolites by hydrophilic interaction chromatography-tandem mass spectrometry
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DOI：
10.1016/j.chroma.2006.05.019
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发表时间：
2006-08-25
影响因子：
4.1
通讯作者：
Rabinowitz, Joshua D.
中科院分区：
文献类型：
--
作者：
Bajad, Sunil U.;Lu, Wenyun;Rabinowitz, Joshua D.
代谢组学中一个关键的未满足的需求是有效定量大量已知细胞代谢物的能力。在这里，我们提出了一个液相色谱（LC）-电喷雾串联质谱（ESI-MS/MS）的方法可靠的测量141代谢物，包括中心碳，氨基酸和核苷酸代谢的成分。所选的LC方法（使用氨基柱的亲水相互作用色谱法）可有效分离使用标准反相色谱法无法保留的高水溶性代谢物。通过在三重四极杆仪器上扫描许多选定的反应监测事件来实现MS/MS检测。当应用于[C-12]-与[C-13]葡萄糖中生长的大肠杆菌提取物时，该方法显示了79种不同代谢产物的适当C-12-和C-13-峰。(c)2006 Elsevier B. V.保留所有权利。
A key unmet need in metabolomics is the ability to efficiently quantify a large number of known cellular metabolites. Here we present a liquid chromatography (LC)-electrospray ionization tandem mass spectrometry (ESI-MS/MS) method for reliable measurement of 141 metabolites, including components of central carbon, amino acid, and nucleotide metabolism. The selected LC approach, hydrophilic interaction chromatography with an amino column, effectively separates highly water soluble metabolites that fail to retain using standard reversed-phase chromatography. MS/MS detection is achieved by scanning through numerous selected reaction monitoring events on a triple quadrupole instrument. When applied to extracts of Escherichia coli grown in [C-12]- versus [C-13]glucose, the method reveals appropriate C-12- and C-13-peaks for 79 different metabolites. (c) 2006 Elsevier B.V. All rights reserved.