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Direct Analysis of Underivatized Amino Acids in Plant Extracts by LC-MS-MS

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Book cover Amino Acid Analysis

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

Abstract

In this chapter, we describe a method for quantification of 20 proteinogenic amino acids as well as 13 15N-labeled amino acids by liquid chromatography–mass spectrometry without the need for derivatization and use of organic solvents. Analysis of the underivatized amino acids is performed by liquid chromatography–electrospray ionization-tandem mass spectrometry (LC-ESI-MS-MS) in the positive ESI mode. Separation is achieved on a strong cation exchange (SCX) column (Luna 5 μm SCX 100 Å) with 30 mM ammonium acetate in water (A) and 5% acetic acid in water (B). Quantification is accomplished by use of d5-phenylalanine as internal standard achieving limits of detection of 0.1–3.0 μM. The method was successfully applied for the determination of proteinogenic and 15N-labeled amino acids in plant extracts.

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Acknowledgments

The authors are grateful to Matthias Gehre (Helmholtz Centre for Environmental Research – UFZ Leipzig, Germany) for providing the 15N-labeled amino acids. We also like to thank Bernd Kastenholz for his assistance in the optimization of the leaf extraction procedure.

Author information

Authors and Affiliations

  1. Institute of Bio- and Geosciences, Forschungszentrum Jülich GmbH, ICG-3, Jülich, Germany

    Björn Thiele

  2. IBG-1: Biotechnology, Jülich, Germany

    Nadine Stein

  3. IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, Jülich, Germany

    Marco Oldiges

  4. Central Division of Analytical Chemistry/BioSpec, Jülich, Germany

    Diana Hofmann

Authors
  1. Björn Thiele
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  2. Nadine Stein
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  3. Marco Oldiges
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  4. Diana Hofmann
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Corresponding author

Correspondence to Björn Thiele .

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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Thiele, B., Stein, N., Oldiges, M., Hofmann, D. (2012). Direct Analysis of Underivatized Amino Acids in Plant Extracts by LC-MS-MS. 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_25

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  • DOI: https://doi.org/10.1007/978-1-61779-445-2_25

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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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