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Multiplexed Profiling and Data Processing Methods to Identify Temperature-Regulated Primary Metabolites Using Gas Chromatography Coupled to Mass Spectrometry.
Methods Mol Biol. 2020; 2156:203-239.MM

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 plant material that was exposed to temperature stress. The method can be combined with stable isotope labeling and tracing experiments and is equally applicable to preparations of plant material and microbial photosynthetic organisms. The described methods are modular and can be multiplexed, that is, the same sample or a paired identical backup sample can be analyzed sequentially by more than one of the described procedures. The modules include rapid sampling and metabolic inactivation protocols for samples in a wide weight range, sample extraction procedures, chemical derivatization steps that are required to make the metabolite fraction amenable to gas chromatographic analysis, routine GC-MS methods, and 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 methods have two applications: (1) The rapid screening for changes of relative metabolite pools sizes under temperature stress and (2) the verification by exact quantification using GC-MS protocols that are extended by internal and external standardization.

Authors+Show Affiliations

Applied Metabolome Analysis Research Group, Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany.Applied Metabolome Analysis Research Group, Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany.Applied Metabolome Analysis Research Group, Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany.Applied Metabolome Analysis Research Group, Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany. Proteomics and Biomarkers, Max Planck Institute of Psychiatry, München, Germany.Applied Metabolome Analysis Research Group, Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany. Owlstone Medical Ltd, Cambridge, UK.Applied Metabolome Analysis Research Group, Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany.Applied Metabolome Analysis Research Group, Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany.Applied Metabolome Analysis Research Group, Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany. Nuclear Pore Complex and Ribosome Assembly, Biochemie-Zentrum, Universität Heidelberg, Heidelberg, Germany.Applied Metabolome Analysis Research Group, Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany. kopka@mpimp-golm.mpg.de.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32607984

Citation

Erban, Alexander, et al. "Multiplexed Profiling and Data Processing Methods to Identify Temperature-Regulated Primary Metabolites Using Gas Chromatography Coupled to Mass Spectrometry." Methods in Molecular Biology (Clifton, N.J.), vol. 2156, 2020, pp. 203-239.
Erban A, Martinez-Seidel F, Rajarathinam Y, et al. Multiplexed Profiling and Data Processing Methods to Identify Temperature-Regulated Primary Metabolites Using Gas Chromatography Coupled to Mass Spectrometry. Methods Mol Biol. 2020;2156:203-239.
Erban, A., Martinez-Seidel, F., Rajarathinam, Y., Dethloff, F., Orf, I., Fehrle, I., Alpers, J., Beine-Golovchuk, O., & Kopka, J. (2020). Multiplexed Profiling and Data Processing Methods to Identify Temperature-Regulated Primary Metabolites Using Gas Chromatography Coupled to Mass Spectrometry. Methods in Molecular Biology (Clifton, N.J.), 2156, 203-239. https://doi.org/10.1007/978-1-0716-0660-5_15
Erban A, et al. Multiplexed Profiling and Data Processing Methods to Identify Temperature-Regulated Primary Metabolites Using Gas Chromatography Coupled to Mass Spectrometry. Methods Mol Biol. 2020;2156:203-239. PubMed PMID: 32607984.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Multiplexed Profiling and Data Processing Methods to Identify Temperature-Regulated Primary Metabolites Using Gas Chromatography Coupled to Mass Spectrometry. AU - Erban,Alexander, AU - Martinez-Seidel,Federico, AU - Rajarathinam,Yogeswari, AU - Dethloff,Frederik, AU - Orf,Isabel, AU - Fehrle,Ines, AU - Alpers,Jessica, AU - Beine-Golovchuk,Olga, AU - Kopka,Joachim, PY - 2020/7/2/entrez PY - 2020/7/2/pubmed PY - 2020/7/2/medline KW - Absolute quantification KW - GC-MS KW - Gas chromatography KW - Metabolism KW - Metabolite profiling KW - Metabolomics KW - Relative quantification KW - Stable isotope labeling KW - TOF-MS KW - Time-of-flight mass spectrometry SP - 203 EP - 239 JF - Methods in molecular biology (Clifton, N.J.) JO - Methods Mol. Biol. VL - 2156 N2 - 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 plant material that was exposed to temperature stress. The method can be combined with stable isotope labeling and tracing experiments and is equally applicable to preparations of plant material and microbial photosynthetic organisms. The described methods are modular and can be multiplexed, that is, the same sample or a paired identical backup sample can be analyzed sequentially by more than one of the described procedures. The modules include rapid sampling and metabolic inactivation protocols for samples in a wide weight range, sample extraction procedures, chemical derivatization steps that are required to make the metabolite fraction amenable to gas chromatographic analysis, routine GC-MS methods, and 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 methods have two applications: (1) The rapid screening for changes of relative metabolite pools sizes under temperature stress and (2) the verification by exact quantification using GC-MS protocols that are extended by internal and external standardization. SN - 1940-6029 UR - https://www.unboundmedicine.com/medline/citation/32607984/Multiplexed_Profiling_and_Data_Processing_Methods_to_Identify_Temperature-Regulated_Primary_Metabolites_Using_Gas_Chromatography_Coupled_to_Mass_Spectrometry L2 - https://dx.doi.org/10.1007/978-1-0716-0660-5_15 DB - PRIME DP - Unbound Medicine ER -
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