Expanding coverage of the metabolome for global metabolite profiling.
Expanding coverage of the metabolome for global metabolite profiling.
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DOI：
10.1021/ac102981k
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发表时间：
2011-03-15
影响因子：
7.4
通讯作者：
Siuzdak, Gary
中科院分区：
文献类型：
--
作者：
Yanes, Oscar;Tautenhahn, Ralf;Patti, Gary J.;Siuzdak, Gary
基于质谱的代谢组学是对天然存在的小分子（统称为代谢组）的综合研究。鉴于内源性代谢物的结构多样性和化学性质，生物提取和色谱方法会使质谱法检测到的代谢物的数量、性质和浓度产生偏差，这对全球非靶向研究构成了挑战。在这项工作中，我们使用大肠杆菌细菌细胞，探索溶剂极性，温度和pH值的影响，同时提取极性和非极性代谢产物。此外，我们探索了涉及不同固定相和移动的相的色谱条件，这些条件优化了内源性代谢物提取物以及合成标准品混合物的分离和电离。我们的研究结果表明，热极性溶剂是最有效的同时提取亲水性和疏水性代谢产物。此外，在移动的相中的氟化铵显著地增加了负电喷雾电离模式中的电离效率，如通过检测到的特征数量和平均特征强度分别增加2.5倍和5.7倍所证明的。与乙酸铵或甲酸富集的移动的相相比，氟化铵灵敏度的提高导致数据库中代谢物命中数的3.5倍，并允许鉴定参与基本细胞途径的独特代谢物。
Mass spectrometry-based metabolomics is the comprehensive study of naturally occurring small molecules collectively known as the metabolome. Given the vast structural diversity and chemical properties of endogenous metabolites, biological extraction and chromatography methods bias the number, property, and concentration of metabolites detected by mass spectrometry and creates a challenge for global untargeted studies. In this work, we used Escherichia coli bacterial cells to explore the influence of solvent polarity, temperature, and pH in extracting polar and non-polar metabolites simultaneously. In addition, we explored chromatographic conditions involving different stationary and mobile phases that optimize the separation and ionization of endogenous metabolite extracts as well as a mixture of synthetic standards. Our results reveal that hot polar solvents are the most efficient in extracting both hydrophilic and hydrophobic metabolites simultaneously. In addition, ammonium fluoride in the mobile phase substantially increased ionization efficiency in negative electrospray ionization mode as demonstrated by 2.5-fold and 5.7-fold increases in the number of features detected and the average feature intensity, respectively. The improvement in sensitivity with ammonium fluoride resulted in 3.5 times as many metabolite hits in databases compared to ammonium acetate or formic acid enriched mobile phases and allowed for the identification of unique metabolites involved in fundamental cellular pathways.
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影响因子：
7.4
作者：
Masson, Perrine;Alves, Alexessander Couto;Want, Elizabeth J.
通讯作者：
Want, Elizabeth J.
影响因子：
4.1
作者：
Pesek, Joseph J.;Matyska, Maria T.;Sana, Theodore R.
通讯作者：
Sana, Theodore R.
DOI：
10.1016/j.jchromb.2008.04.031
发表时间：
2008-08-15
影响因子：
3
作者：
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通讯作者：
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影响因子：
4.1
作者：
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通讯作者：
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影响因子：
4.1
作者：
Luo, Bing;Groenke, Karsten;Oldiges, Marco
通讯作者：
Oldiges, Marco