Workflow methodology for rat brain metabolome exploration using NMR, LC–MS and GC–MS analytical platforms
| Autor: | Christian R. Andres, Blandine Madji Hounoum, Lydie Nadal-Desbarats, Frédéric Montigny, Sylvie Mavel, Laurent Galineau, Binta Diémé, Antoine Lefèvre, Patrick Emond, Hélène Blasco |
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| Přispěvatelé: | Imagerie et cerveau (iBrain - Inserm U1253 - UNIV Tours ), Université de Tours-Institut National de la Santé et de la Recherche Médicale (INSERM), Plateforme Scientifique et Technique 'Analyses des Systèmes Biologiques' (PST-ASB), Université Francois Rabelais [Tours], Université de Tours (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM) |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
[SDV]Life Sciences [q-bio] Metabolite Clinical Biochemistry Analytical chemistry Pharmaceutical Science Mass spectrometry 01 natural sciences Gas Chromatography-Mass Spectrometry Mass Spectrometry Workflow Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound Metabolomics Liquid chromatography–mass spectrometry Drug Discovery Metabolome Animals Derivatization ComputingMilieux_MISCELLANEOUS Spectroscopy Chromatography 010401 analytical chemistry Brain Reproducibility of Results Reversed-phase chromatography Rats 0104 chemical sciences 030104 developmental biology chemistry Gas chromatography–mass spectrometry Chromatography Liquid |
| Zdroj: | Journal of Pharmaceutical and Biomedical Analysis Journal of Pharmaceutical and Biomedical Analysis, Elsevier, 2017, 142, pp.270-278. ⟨10.1016/j.jpba.2017.03.068⟩ |
| ISSN: | 0731-7085 |
| DOI: | 10.1016/j.jpba.2017.03.068 |
| Popis: | We developed a multi-platform approach for the metabolome exploration of rat brain tissue, using liquid chromatography coupled with mass spectrometry (LC-MS), nuclear magnetic resonance spectroscopy (NMR) and gas-chromatography coupled with mass spectrometry (GC-MS). The critical steps for metabolite exploration of cerebral tissues are tissue lysis and metabolites extraction. We first evaluated the impact of freeze-drying compared to wet tissue metabolites extraction using NMR and LC-MS with a reversed phase liquid chromatography. Then, we compared four metabolite extraction methods Based on the number of metabolites extracted, their intensity and their coefficient of variation (%CV), the most reproducible protocol (one-step extraction with acetonitrile on lyophilized material) was chosen to further evaluate the impact of sample mass on method performance (3, 6, and 9mg were essayed). GC-MS analysis was also investigated by analyzing four different methoximation/silylation derivatization combinations. The optimal analytical protocols were proposed to establish the reliability required to realize untargeted brain tissue metabolomics exploration. The most reliable workflow was then exemplified by analyzing three rat brain regions (cerebellum, frontal and parietal cortices, n=12) by 1H NMR, LC-MS and GC-MS, allowing their clustering based on their metabolic profiles. We present here an example of development of methodology that should be done before running analysis campaigns. |
| Databáze: | OpenAIRE |
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