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Direct Analysis of Underivatized Amino Acids in Plant Extracts by LC-MS/MS (Improved Method)

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Amino Acid Analysis

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

Abstract

In this chapter we describe a method for quantification of 20 proteinogenic amino acids by liquid chromatography-mass spectrometry which affords neither derivatization nor the 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 μ SCX 100 Å) with 5% acetic acid in water (A) and 75 mM ammonium acetate in water (B). Quantification is accomplished by use of d2-phenylalanine as internal standard achieving limits of detection of 5–50 nM. The method was successfully applied for the determination of proteinogenic amino acids in plant extracts.

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Acknowledgment

The authors 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, IBG-2: Plant Sciences, Forschungszentrum Jülich GmbH, 52425 Jülich, Germany

    Björn Thiele

  2. Institute of Bio- and Geosciences, IBG-3: Agrosphere, Forschungszentrum Jülich GmbH, 52425 Jülich, Germany

    Björn Thiele & Diana Hofmann

  3. Central Institute for Engineering, Electronics and Analytics, ZEA-3: Analytics, Forschungszentrum Jülich GmbH, 52425 Jülich, Germany

    Michelle Hupert & Beatrix Santiago-Schübel

  4. Institute of Bio- and Geosciences, IBG-1: Biotechnology, Forschungszentrum Jülich GmbH, 52425 Jülich, Germany

    Marco Oldiges

Authors
  1. Björn Thiele
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  2. Michelle Hupert
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  3. Beatrix Santiago-Schübel
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  4. Marco Oldiges
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  5. Diana Hofmann
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Corresponding author

Correspondence to Björn Thiele .

Editor information

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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Thiele, B., Hupert, M., Santiago-Schübel, B., Oldiges, M., Hofmann, D. (2019). Direct Analysis of Underivatized Amino Acids in Plant Extracts by LC-MS/MS (Improved Method). 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_30

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  • DOI: https://doi.org/10.1007/978-1-4939-9639-1_30

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9638-4

  • Online ISBN: 978-1-4939-9639-1

  • eBook Packages: Springer Protocols

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