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


Ontology type: schema:Chapter     


Chapter Info

DATE

2014-05-08

AUTHORS

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

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. More... »

PAGES

171-197

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-1-4939-0844-8_14

DOI

http://dx.doi.org/10.1007/978-1-4939-0844-8_14

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1041454340

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/24852636


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