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24, chemin de Borde Rouge -Auzeville - CS52627 31326 Castanet Tolosan cedex - France

Last update: May 2021

Metabolism and Mass Spectrometry

Team members ( Click here):

Publications ( Click here):

Scientific context

The objective of the human nutrition research carried out in Clermont-Ferrand is to improve human diet particularly during the aging period. Metabolic studies have been of major interest as they integrate all post-genome phenomena. Metabolomics is an essential tool to understand the functionality of living organisms at various levels of biomolecular organization from the cell to the whole organism. This new high-throughput approach permits to apprehend biocomplexity by describing the small molecules (or metabolites) formed or modified by a stimuli, in that case a nutritional challenge. Metabolomics studies as developed by this analytical platform imply mass spectrometry analyses of biological samples.

Presentation

The team is the major part of a larger platform (Platform of "Exploration of Metabolism", PFEM) which is a federative technological structure. The investigation of metabolome is major topic of PFEM, which possess supplemental facilities to transcriptome and proteome analysis. This platform was labelled as an emerging strategic platform in 2008 by INRAE (CNOC) and it obtained the French IBiSA certification in 2010.
As part of a platform, the team is a transverse structure dedicated to metabolism studies, including fingerprinting, profiling, absolute quantification and finally flux analysis. It provides analytical services and method developments. Its purposes are:
- to give researchers technical equipments and related skills dedicated to metabolism studies
- to give researchers an opportunity to consider new approaches ensuring an optimal holding of scientifi c methodologies and data analysis.
- to give a technical assistance to users
- to propose academic and technical training

Analytical approaches

Units in Clermont-Ferrand are well recognized using stable isotopes metabolite tracers and the team has a long experience in fluxomics. Activities include the analysis of organic compounds ranging from the characterization and the quantitative determination of small molecules present in biological fluids and tissues, to the measurement of stable isotopes enrichment of amino acids or fatty acids, which allows calculation of their metabolic rates in vivo after infusion of these isotopes.
The team develops also the metabonomic approach, which permits to characterize with great details the so-called metabolome, i.e. the metabolites present in a given biological sample and to study its modulation according to physiological factors. Activities allow the determination of metabolic fingerprints of urine and plasma samples and the identification of interesting biomarkers.

Equipments and skills

This facility is equipped with about 10 different instruments in mass spectrometry and isotopic mass spectrometry.
• GC-MS: dedicated to quantifi cation or fluxomic studies (high isotopic enrichments)
• GC-C-IRMS: dedicated to fluxomic studies (very low isotopic enrichments) and metabolomics (fecal water metabolic profiling)
• LC-QqQ: dedicated to quantification or fluxomic studies
• LC-ToF: dedicated to metabolomic studies (urine and plasma metabolic profiling)
• LTQ Orbitrap: dedicated to metabolomic studies (biomarker identification)
The laboratory offers an access to the following instruments
Metabolic Profiler (LC / RMN / ToF): dedicated to metabolomic studies (installed at B. Pascal University, Clermont-Ferrand, France)

Research

The methodological developments concern both sample preparation, analysis of metabolites, their identification and the signal processing. The fluxomic developments concern mainly the quantification of isotopic enrichment by MS / MS analysis in order to increase the sensitivity and the specificity of the measures.
The developments in metabolomics concern the validation of analytical methods in various biological matrices, as well as the optimization of the analytical workflow and the automation of data processing and biomarker identification.