1. 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
- Author
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Jouhet, Juliette, Lupette, Josselin, Clerc, Olivier, Magneschi, Leonardo, Bedhomme, Mariette, Collin, Séverine, Roy, Sylvaine, Maréchal, Eric, Rébeillé, Fabrice, 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), and 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)
- Subjects
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 - Abstract
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.
- Published
- 2017
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