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A Protocol for Untargeted Metabolomic Analysis: From Sample Preparation to Data Processing

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Mitochondrial Medicine

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2276))

Abstract

Untargeted metabolomics has rapidly become a profiling method of choice in many areas of research, including mitochondrial biology. Most commonly, untargeted metabolomics is performed with liquid chromatography/mass spectrometry because it enables measurement of a relatively wide range of physiochemically diverse molecules. Specifically, to assess energy pathways that are associated with mitochondrial metabolism, hydrophilic interaction liquid chromatography (HILIC) is often applied before analysis with a high-resolution accurate mass instrument. The workflow produces large, complex data files that are impractical to analyze manually. Here, we present a protocol to perform untargeted metabolomics on biofluids such as plasma, urine, and cerebral spinal fluid with a HILIC separation and an Orbitrap mass spectrometer. Our protocol describes each step of the analysis in detail, from preparation of solvents for chromatography to selecting parameters during data processing.

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Acknowledgments

The authors would like to thank Dr. Clary Clish (Broad Institute of Harvard and MIT) for discussion and insights that helped develop this protocol.

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Authors and Affiliations

  1. Life Science Mass Spectrometry Division, Thermo Fisher Scientific, San Jose, CA, USA

    Amanda L. Souza

  2. Department of Chemistry, Washington University in St. Louis, Saint Louis, MO, USA

    Gary J. Patti

  3. Department of Medicine, Washington University in St. Louis, Saint Louis, MO, USA

    Gary J. Patti

Authors
  1. Amanda L. Souza
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  2. Gary J. Patti
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Corresponding author

Correspondence to Amanda L. Souza .

Editor information

Editors and Affiliations

  1. Department of Pharmaceutical Sciences, Midwestern University, Glendale, AZ, USA

    Volkmar Weissig

  2. Cochin Hospital, Cochin Institute, INSERM U1016, PARIS, France

    Marvin Edeas

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Souza, A.L., Patti, G.J. (2021). A Protocol for Untargeted Metabolomic Analysis: From Sample Preparation to Data Processing. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine . Methods in Molecular Biology, vol 2276. Springer, New York, NY. https://doi.org/10.1007/978-1-0716-1266-8_27

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  • DOI: https://doi.org/10.1007/978-1-0716-1266-8_27

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-0716-1265-1

  • Online ISBN: 978-1-0716-1266-8

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