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Rapid LC–MS/MS Profiling of Protein Amino Acids and Metabolically Related Compounds for Large-Scale Assessment of Metabolic Phenotypes

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Amino Acid Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 828))

Abstract

Amino acids extracted from a biological matrix can be resolved and measured using a 6-min per sample method through high-performance liquid chromatography with a short C18 column and rapid gradient using the ion-pairing reagent perfluoroheptanoic acid. LC-tandem mass spectrometry with multiple reaction monitoring (MRM) transitions selective for each compound allows simultaneous quantification of the 20 proteinogenic amino acids and 5 metabolically related compounds. Distinct MRM transitions were also established for selective detection of the isomers leucine/isoleucine and threonine/homoserine.

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Acknowledgments

We thank Lijun Chen of the Michigan State University Mass Spectrometry Core and Yan Lu in the Last Laboratory for assistance in various stages of the project. This project was supported by National Science Foundation grant MCB-0519740 (to R.L.L.).

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Authors and Affiliations

  1. Department of Biology and Microbiology, South Dakota State University, Brookings, SD, USA

    Liping Gu

  2. Department of Biochemistry and Molecular Biology, and Department of Chemistry, Michigan State University, East Lansing, MI, USA

    A. Daniel Jones

  3. Departments of Biochemistry and Molecular Biology and Department of Plant Biology, Michigan State University, East Lansing, MI, USA

    Robert L. Last

Authors
  1. Liping Gu
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  2. A. Daniel Jones
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  3. Robert L. Last
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Corresponding author

Correspondence to A. Daniel Jones .

Editor information

Editors and Affiliations

  1. Ctr. Biologics Evaluation & Research, Div. Cellular & Gene Therapies, U.S. Food and Drug Administration, Rockville Pike 8800, Bethesda, 20892, Maryland, USA

    Michail A. Alterman

  2. Functional Genomics Center Zürich, Protein Analysis Group, Universität Zürich, Winterthurerstr. 190, Zürich, 8057, Switzerland

    Peter Hunziker

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Gu, L., Jones, A.D., Last, R.L. (2012). Rapid LC–MS/MS Profiling of Protein Amino Acids and Metabolically Related Compounds for Large-Scale Assessment of Metabolic Phenotypes. In: Alterman, M., Hunziker, P. (eds) Amino Acid Analysis. Methods in Molecular Biology, vol 828. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-445-2_1

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  • DOI: https://doi.org/10.1007/978-1-61779-445-2_1

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-444-5

  • Online ISBN: 978-1-61779-445-2

  • eBook Packages: Springer Protocols

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