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Amino Acid Analysis pp 57–68Cite as

Quantification of Isotopologues of Amino Acids by Multiplexed Stable Isotope-Resolved Metabolomics Using Ultrahigh-Resolution Mass Spectrometry Coupled with Direct Infusion

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2030))

Abstract

Stable isotope-resolved metabolomics (SIRM) is increasingly used among researchers for metabolic studies including amino acid metabolism. However, the classical GC- or HPLC-based methods for amino acid quantification do not meet the needs for multiplexed stable isotope-enriched analysis by ultrahigh-resolution Fourier transform mass spectrometry (UHR-FTMS). This is due to insufficient acquisition time during chromatographic separations and large dynamic range in concentrations of analytes, which compromises detection and quantification of the numerous metabolite isotopologues present in crude extracts. This chapter discusses a modified ethyl chloroformate derivatization method to enable rapid quantitative analysis of stable isotope-enriched amino acids using direct infusion ion introduction coupled with UHR-FTMS.

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Acknowledgments

This work was supported in part by the National Institutes of Health grants [5R01ES22191-04, 3R01ES022191-04S1, 1U24DK097215-01A1, and P01 CA163223-01A1].

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Authors and Affiliations

  1. Urologic Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Ye Yang

  2. Department of Toxicology and Cancer Biology, Markey Cancer Center, University of Kentucky, Lexington, KY, USA

    Teresa W.-M. Fan, Andrew N. Lane & Richard M. Higashi

  3. Center for Environmental and Systems Biochemistry, University of Kentucky, Lexington, KY, USA

    Teresa W.-M. Fan, Andrew N. Lane & Richard M. Higashi

Authors
  1. Ye Yang
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  2. Teresa W.-M. Fan
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  3. Andrew N. Lane
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  4. Richard M. Higashi
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Corresponding author

Correspondence to Teresa W.-M. Fan .

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Editors and Affiliations

  1. Office of Policy for Pharmaceutical Quality Office of Pharmaceutical Quality, Center for Drug Evaluation and Research US FDA, Silver Spring, MD, USA

    Michail A. Alterman

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Yang, Y., Fan, T.WM., Lane, A.N., Higashi, R.M. (2019). Quantification of Isotopologues of Amino Acids by Multiplexed Stable Isotope-Resolved Metabolomics Using Ultrahigh-Resolution Mass Spectrometry Coupled with Direct Infusion. In: Alterman, M. (eds) Amino Acid Analysis. Methods in Molecular Biology, vol 2030. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9639-1_6

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  • DOI: https://doi.org/10.1007/978-1-4939-9639-1_6

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