LC-MS/MS versus TLC plus GC methods: Consistency of glycerolipid and fatty acid profiles in microalgae and higher plant cells and effect of a nitrogen starvation
| Autor: | Jouhet, Juliette, Lupette, Josselin, Clerc, Olivier, Magneschi, Leonardo, Bedhomme, Mariette, Collin, Séverine, Roy, Sylvaine, Maréchal, Eric, Rébeillé, Fabrice |
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| Přispěvatelé: | UMR 1417 PCV Laboratoire de Physiologie Cellulaire Végétale, Centre National de la Recherche Scientifique (CNRS), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Institut National de la Recherche Agronomique (INRA), Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Physiologie cellulaire et végétale (LPCV), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), TOTAL-CEA, CEA High Commissioner (Flagship program), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Université Grenoble Alpes (UGA), Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Grenoble Alpes (UGA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Recherche Agronomique (INRA), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes (UGA)-Institut de Recherche Interdisciplinaire de Grenoble (IRIG) |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
Thin-Layer Chromatography
Glycerol Ionization [SDV]Life Sciences [q-bio] lcsh:Medicine Brassica Monomers (Chemistry) plante supérieure Research and Analysis Methods Biochemistry Model Organisms chromatographie à phase gazeuse Plant and Algal Models Macromolecular Structure Analysis Polymer chemistry lcsh:Science Molecular Biology micro-algue Lipid Analysis acide gras lcsh:R Chromatographic Techniques Fatty Acids thin layer chromatography arabidopsis thaliana cellule végétale méthode de quantification Organisms Chemical Reactions Biology and Life Sciences glycerolipide Plants Cell Cultures Lipids Physical sciences Planar Chromatography Chemistry Experimental Organism Systems plant cells lcsh:Q Biological Cultures fatty acid chromatographie en couche mince Research Article |
| Zdroj: | PLoS ONE PLoS ONE, 2017, 12 (8), pp.1-21. ⟨10.1371/journal.pone.0182423⟩ PLoS ONE, Public Library of Science, 2017, 12 (8), pp.1-21. ⟨10.1371/journal.pone.0182423⟩ PLoS ONE, Vol 12, Iss 8, p e0182423 (2017) Plos One 8 (12), 1-21. (2017) |
| ISSN: | 1932-6203 |
| Popis: | Methods to analyze lipidomes have considerably evolved, more and more based on mass spectrometry technics (LC-MS/MS). However, accurate quantifications using these methods require 13C-labeled standards for each lipid, which is not feasible because of the very large number of molecules. Thus, quantifications rely on standard molecules representative of a whole class of lipids, which might lead to false estimations of some molecular species. Here, we determined and compared glycerolipid distributions from three different types of cells, two microalgae (Phaeodactylum tricornutum, Nannochloropsis gaditana) and one higher plant (Arabidopsis thaliana), using either LC-MS/MS or Thin Layer Chromatography coupled with Gas Chromatography (TLC-GC), this last approach relying on the precise quantification of the fatty acids present in each glycerolipid class. Our results showed that the glycerolipid distribution was significantly different depending on the method used. How can one reconcile these two analytical methods? Here we propose that the possible bias with MS data can be circumvented by systematically running in tandem with the sample to be analyzed a lipid extract from a qualified control (QC) of each type of cells, previously analyzed by TLC-GC, and used as an external standard to quantify the MS results. As a case study, we applied this method to compare the impact of a nitrogen deficiency on the three types of cells. |
| Databáze: | OpenAIRE |
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