An Untargeted Metabolomic Workflow to Improve Structural Characterization of Metabolites

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• 作者：Igor Nikolskiy ; Nathaniel G. Mahieu ; Ying-Jr Chen ; Ralf Tautenhahn ; Gary J. Patti
• 刊名：Analytical Chemistry
• 出版年：2013
• 出版时间：August 20, 2013
• 年：2013
• 卷：85
• 期：16
• 页码：7713-7719
• 全文大小：332K
• 年卷期：v.85,no.16(August 20, 2013)
• ISSN：1520-6882

文摘

Mass spectrometry-based metabolomics relies on MS² data for structural characterization of metabolites. To obtain the high-quality MS² data necessary to support metabolite identifications, ions of interest must be purely isolated for fragmentation. Here, we show that metabolomic MS² data are frequently characterized by contaminating ions that prevent structural identification. Although using narrow-isolation windows can minimize contaminating MS² fragments, even narrow windows are not always selective enough, and they can complicate data analysis by removing isotopic patterns from MS² spectra. Moreover, narrow windows can significantly reduce sensitivity. In this work, we introduce a novel, two-part approach for performing metabolomic identifications that addresses these issues. First, we collect MS² scans with less stringent isolation settings to obtain improved sensitivity at the expense of specificity. Then, by evaluating MS² fragment intensities as a function of retention time and precursor mass targeted for MS² analysis, we obtain deconvolved MS² spectra that are consistent with pure standards and can therefore be used for metabolite identification. The value of our approach is highlighted with metabolic extracts from brain, liver, astrocytes, as well as nerve tissue, and performance is evaluated by using pure metabolite standards in combination with simulations based on raw MS² data from the METLIN metabolite database. A R package implementing the algorithms used in our workflow is available on our laboratory website (wustl.edu/decoms2.php" class="extLink">http://pattilab.wustl.edu/decoms2.php).