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Analysis of Kinetic Labeling of Amino Acids and Organic Acids by GC-MS

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Plant Metabolic Flux Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1090))

Abstract

Plant metabolic pathways and the molecular and atomic fluxes through them can be deduced using stable isotopically labeled substrates. To this end one prerequisite is accurate measurement of the labeling pattern of targeted metabolites. Experiments are generally limited to the use of single-element isotopes, mainly 13C. Here, we summarize the application of gas chromatography-time of flight mass spectrometry (GC-TOF-MS) for metabolic studies using differently labeled elemental isotopes applied to both intact organelles and whole plant tissue. This method allows quantitative evaluation of a broad range of metabolic pathways without the need for laborious (and potentially inaccurate) chemical fractionation procedures commonly used in the estimation of fluxes following incubation in radiolabeled substrates. We focus herein on the determination of isotope labeling in organic and amino acids.

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Acknowledgements

Financial support from the Max-Planck-Society (to WLA and ARF), the Deutsche Forschungsgemeinschaft (grant no. DFG-SFB429 to ARF), and the National Council for Scientific and Technological Development CNPq-Brazil (grant number 472787/2011-0 to WLA) is gratefully acknowledged.

Conflict of interest: The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Departamento de Biologia Vegetal, Universidade Federal de Viçosa, Viçosa, MG, Brazil

    Wagner L. Araújo & Adriano Nunes-Nesi

  2. Max-Planck-Institut für Molekulare Pflanzenphysiologie, Golm, Potsdam, Germany

    Takayuki Tohge & Alisdair R. Fernie

  3. Max-Planck-Institut für Molekulare Pflanzenphysiologie, Potsdam-Golm, Germany

    Toshihiro Obata

Authors
  1. Wagner L. Araújo
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  2. Takayuki Tohge
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  3. Adriano Nunes-Nesi
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  4. Toshihiro Obata
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  5. Alisdair R. Fernie
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Editor information

Editors and Affiliations

  1. INRA Bordeaux Université Bordeaux Segalen, Villenave d'Ornon, France

    Martine Dieuaide-Noubhani

  2. Department of Molecular Genetics, The Ohio State University, Columbus, Ohio, USA

    Ana Paula Alonso

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© 2014 Springer Science+Business Media, New York

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Araújo, W.L., Tohge, T., Nunes-Nesi, A., Obata, T., Fernie, A.R. (2014). Analysis of Kinetic Labeling of Amino Acids and Organic Acids by GC-MS. In: Dieuaide-Noubhani, M., Alonso, A. (eds) Plant Metabolic Flux Analysis. Methods in Molecular Biology, vol 1090. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-688-7_7

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  • DOI: https://doi.org/10.1007/978-1-62703-688-7_7

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-687-0

  • Online ISBN: 978-1-62703-688-7

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