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Profiling Methods to Identify Cold-Regulated Primary Metabolites Using Gas Chromatography Coupled to Mass Spectrometry

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Plant Cold Acclimation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1166))

Abstract

This book chapter describes the analytical procedures required for the profiling of a metabolite fraction enriched for primary metabolites. The profiling is based on routine gas chromatography coupled to mass spectrometry (GC-MS). The generic profiling method is adapted to plant material, specifically to the analysis of single leaves from plants that were exposed to temperature stress experiments. The described method is modular. The modules include a rapid sampling and metabolic inactivation protocol for samples in a wide size range, a sample extraction procedure, a chemical derivatization step that is required to make the metabolite fraction amenable to gas chromatographic analysis, a routine GC-MS method, and finally the procedures of data processing and data mining. A basic and extendable set of standardizations for metabolite recovery and retention index alignment of the resulting GC-MS chromatograms is included. The method has two applications: (1) the rapid screening for changes of relative metabolite pools sizes under temperature stress and (2) the verification of cold-regulated metabolites by exact quantification using a GC-MS protocol with extended internal and external standardization.

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Author information

Authors and Affiliations

  1. Max-Planck-Institute of Molecular Plant Physiology, Applied Metabolome Analysis Research Group, Am Mühlenberg 1, 14476, Potsdam, Germany

    Frederik Dethloff, Alexander Erban, Isabel Orf, Jessica Alpers, Ines Fehrle, Olga Beine-Golovchuk, Stefanie Schmidt, Jens Schwachtje & Joachim Kopka

Authors
  1. Frederik Dethloff
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  2. Alexander Erban
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  3. Isabel Orf
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  4. Jessica Alpers
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  5. Ines Fehrle
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  6. Olga Beine-Golovchuk
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  7. Stefanie Schmidt
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  8. Jens Schwachtje
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  9. Joachim Kopka
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Corresponding author

Correspondence to Joachim Kopka .

Editor information

Editors and Affiliations

  1. Max-Planck-Institut für Molekulare Pflanzenphysiologie, Potsdam, Germany

    Dirk K. Hincha

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

    Ellen Zuther

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Dethloff, F. et al. (2014). Profiling Methods to Identify Cold-Regulated Primary Metabolites Using Gas Chromatography Coupled to Mass Spectrometry. In: Hincha, D., Zuther, E. (eds) Plant Cold Acclimation. Methods in Molecular Biology, vol 1166. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0844-8_14

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  • DOI: https://doi.org/10.1007/978-1-4939-0844-8_14

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0843-1

  • Online ISBN: 978-1-4939-0844-8

  • eBook Packages: Springer Protocols

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