As it is designed today, the assessment of the risk associated with food contaminants is mainly based on toxicological studies carried out on individual substances, which do not fit the real exposure modalities of consumers to chemicals occurring in food.
In order to meet challenges arised by multi-exposure and low doses in modern food toxicology, it is essential to provide very powerful tools for the determination of trace contaminants within complex matrices in order to look at exposures parameters. The study of their impact on the general metabolism and metabolic networks also necessitates to develop real analytical engineering strategies covering from specific spectrometric approaches for the identification of the contaminant metabolic pathways to emerging global metabonomic approaches.
For that, methodologies to favour are those allowing to test with a high troughput the impact of xenobiotics on a large number of targets, and to monitor the fate of these xenobiotics at the ultra-trace level in the exposed organisms. To be efficient, the research system to set up must be integrative and must target the consumer exposure to xenobiotics, the metabolism of these substances by the organism, their action mode on cellular targets, their effects on organs and functions, and last but not least the global response of the whole organism.
In this context, the analytical platform MetaToul-AXIOM (Analysis of Xenobiotics, Identification & Metabolism) develops its activities for providing tools and methods meeting the needs of research team from TOXALIM and abroad, for the achievement of their research projects. This includes leading edge technological development, mainly based on the use of nuclear magnetic resonance and mass spectrometry, to meet challenges of characterizing the exposure of consumers to low doses of contaminants present in mixtures, and their effects.
The MetaToul-AXIOM platform is part of the MetaToul platform (Toulouse Metabolomics and Fluxomics Centre, https://mth-metatoul.com/) headed by Pr J.C. Portais (INSA Toulouse), and leaned on the Genopole de Toulouse (an organization that coordinates all life sciences research platforms – i.e. Services- in the Région Midi-Pyrénées, see: http://genopole-toulouse.prd.fr/). The MetaToul platform is one of the 4 platforms consituting the national infrastructure for metabolomics and fluxomics MetaboHUB (http://www.metabohub.fr/).
MetaToul is a high level technological platform which provides large-scale facilities (NMR & MS equipments), concepts, tools, and expertise to the scientific community in the emerging field of global analysis of metabolism (metabolomics, fluxomics), to support scientists from public laboratories or private companies in France and abroad whose projects require in-depth investigations of metabolism, its regulation or its engineering. MetaToul federates the technical and scientific potential in metabolomics, fluxomics, and metabolism of 5 different laboratories located in Toulouse, among which TOXALIM is the major actor in the field of food toxicology.
The platform is opened to local, national and European scientific communities (ANR projects, European projects,…). Proposed services include : assistance and expertise for the preparation of projects, complete service (routine analysis), service on demand, access to NMR and MS facilities (restricted to duly authorized users, after relevant training), software and support for data treatment, statistical analysis, assistance for sample preparation, data handling and data interpretation, practical training of students, researchers, engineers and technicians.
MetaToul platform gained the IBiSA certification in 2009 and was recognized the same year as « strategic platform » by the INRA organ for internal recognition “CNOC” (Commission Nationale des Outils Communs).
Within MetaToul platform, AXIOM is in charge of the xenobiotic metabolism activity. It has leading equipment devoted to the development of projects in the field of food toxicology. This mainly includes instruments of mass spectrometry (ion trap GC-MSn, LC-MSn , triple quadripole LC-MS/MS, hybrid Orbitrap technology high resolution LC-MSn), Nuclear Magnetic Resonance (600 MHz NMR equipped with cryo probe), and their associated techniques of sample purification and separation (accelerated solvent extraction ASE, UHPLC systems, on line radioactivity detection…).
Functioning
The MetaToul-AXIOM platform can be sollicited for:
Answering to limited needs which can be the subject of service.
Collaboration or partnership in the frame of the set-up and implementation of research projects.
Access to instruments opened to the scientific community.
In all cases, a preliminary contact must be established by e-mail at metatoul@insa-toulouse.fr in order to estimate the ability of the plateau to meet the request. Functioning modalities are available on the platform websites www.toulouse.inra.fr/axiomm (section functioning) or www.metatoul.fr (section activities / access modalities).
Services
The activities of the MetaToul-AXIOM platform rest on five main skill centers: (i) mass spectrometry, (ii) nuclear magnetic resonance, (iii) purification and separation methods, (iv) bioinformatics, and (v) fine organic synthesis.
The MetaToul-AXIOM platform provides technical and human resources to achieve analyses services for:
- structural analyses by mass spectrometry and NMR of metabolites from the biotransformation of xenobiotics (food contaminants, ...) studied in vitro or in vivo:
Confirmation of compound's structure by UHPLC-HRMS
Confirmation of compound's structure by NMR
- conducting untargeted metabolomics studies by high resolution mass spectrometry and / or NMR from biological fluids or tissues, with or without statistical analysis:
untargeted analyses by UHPLC-HRMS
untargeted metabolomics by NMR
statistical analyses of metabolomics data
tissues extraction using "Fastprep"
- quantitative or qualitative determination of contaminants, metabolites or biomarkers present at the trace level into complex matrices:
Extraction and digestion of DNA into deoxynucleosides for LC-MS quantification of DNA adducts
LC-MS quantification of the DNA adduct dG-PhIP
Extraction of heterocyclic aromatic amines (HCAs) from meat samples
LC-MS quantification of 13 HCAs in meat by UHPLC-APCI-MS/MS
LC-MS quantification of BPA, BPA-Glucuronide and BPA-Sulfate in biological matrices and cells cultures media
LC-MS quantification of urany BPA-Glucuronide
LC-MS quantification of cefquinone in biological matrices and cells cultures media
- access to instruments
training and assistancefor NMR analyses
training and assistancefor quantitative LC-MS analyses (triple quadrupoles)
training and assistance for structural LC-MS analyses (ion traps, high resolution mass spectrometry)
training and assistance for GC-MS analyses
- fine organic synthesis (only for scientists of the Toxalim laboratory)
Research and development
The MetaToul-AXIOM platform is at the crossroads of the activities of several TOXALIM research teams, and thus takes part in several scientific projects developed within TOXALIM. Axes developed by TOXALIM require a leading edge analytical potential and a large part of the MetaToul-AXIOM platform activity is devoted to methodological development essential for the good unwinding of the projects in which MetaToul-AXIOM is involved.
Methodological development under progress
Development of leading edge analytical methodologies for the characterization of DNA adducts by reactive xenobiotic metabolites as markers of exposure and/or toxicity.
Development of quantitative analysis methods for the determination of toxic contaminants and/or their metabolites present at trace level in complex biological matrices.
Development of 1D and 2D NMR techniques for the analysis of complex biological matrices in isotropic (urine, plasma, cells) or anisotropic (tissues by using HR-MAS probe) media.
Development of methods based on high resolution mass spectrometry for the generation of data intended to the modelling of metabolic networks.
Development of semi-targeted profiling methods of specific compounds in complex matrix by high resolution mass spectrometry (metabolites of xenobiotics, DNA adducts, aldehydes, secondary fungal metabolites).
Lipidomics by NMR
Development of statistical tools for (i) integration of data generated from different analytical technics, and (ii) longitudinal data treatment.
Chemicals synthesis of reference compounds including stable isotopes for MS analyses
Participation in research projects
AXIOM participates as partner or service provider to several regional projects (région Midi-Pyrénées "fipronil" et "retardateurs de flamme"), national projects (INVS-AFSSA "biomarqueurs d’exposition à l’acrylamide", AFSSET "retardateurs de flamme", ANR "Caphe", ANR "PlastImpact", ANR "Metaprofile", ANR "Delisus", ANR "ExpomatPest", ANR "Kismet", ANR "Nistec", ANR "Perinatox", ANR "ContrePerf", ANR "Sécuriviande", ANR "Neurodevotox") or european projects (REX "Cascade", IP "ProSafeBeef", IP FP7 "Bridge").
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