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Integrated, Nontargeted Ultrahigh Performance Liquid Chromatography/Electrospray Ionization Tandem Mass Spectrometry Platform for the Identification and Relative Quantification of the Small-Molecule Complement of Biological Systems

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Download Hi-Res ImageDownload to MS-PowerPointCite This:Anal. Chem. 2009, 81, 16, 6656-6667
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    Integrated, Nontargeted Ultrahigh Performance Liquid Chromatography/Electrospray Ionization Tandem Mass Spectrometry Platform for the Identification and Relative Quantification of the Small-Molecule Complement of Biological Systems
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    Metabolon, Incorporated, 800 Capitola Drive, Suite 1, Durham, North Carolina 27713
    * To whom correspondence should be addressed. E-mail: [email protected]. Fax: 919-572-1721.
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    Analytical Chemistry

    Cite this: Anal. Chem. 2009, 81, 16, 6656–6667
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    https://doi.org/10.1021/ac901536h
    Published July 22, 2009
    Copyright © 2009 American Chemical Society
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    Abstract

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    To address the challenges associated with metabolomics analyses, such as identification of chemical structures and elimination of experimental artifacts, we developed a platform that integrated the chemical analysis, including identification and relative quantification, data reduction, and quality assurance components of the process. The analytical platform incorporated two separate ultrahigh performance liquid chromatography/tandem mass spectrometry (UHPLC/MS/MS2) injections; one injection was optimized for basic species, and the other was optimized for acidic species. This approach permitted the detection of 339 small molecules, a total instrument analysis time of 24 min (two injections at 12 min each), while maintaining a median process variability of 9%. The resulting MS/MS2 data were searched against an in-house generated authentic standard library that included retention time, molecular weight (m/z), preferred adducts, and in-source fragments as well as their associated MS/MS spectra for all molecules in the library. The library allowed the rapid and high-confidence identification of the experimentally detected molecules based on a multiparameter match without need for additional analyses. This integrated platform enabled the high-throughput collection and relative quantitative analysis of analytical data and identified a large number and broad spectrum of molecules with a high degree of confidence.

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    Analytical Chemistry

    Cite this: Anal. Chem. 2009, 81, 16, 6656–6667
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    https://doi.org/10.1021/ac901536h
    Published July 22, 2009
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