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

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, quantif...

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Bibliographic Details
Published in:PloS one 2017-08, Vol.12 (8), p.e0182423-e0182423
Main Authors: Jouhet, Juliette, Lupette, Josselin, Clerc, Olivier, Magneschi, Leonardo, Bedhomme, Mariette, Collin, Séverine, Roy, Sylvaine, Maréchal, Eric, Rébeillé, Fabrice
Format: Article
Language:English
Subjects:
Algae
Analysis
Analytical methods
Aquatic microorganisms
Arabidopsis - growth & development
Arabidopsis - metabolism
Arabidopsis thaliana
Biodiesel fuels
Biology
Biology and Life Sciences
Case studies
Chromatography
Chromatography, Liquid - methods
Chromatography, Thin Layer - methods
Comparative analysis
Fatty acids
Fatty Acids - analysis
Fatty Acids - metabolism
Gas chromatography
Gas Chromatography-Mass Spectrometry - methods
Glycerol
Glycolipids - analysis
Glycolipids - metabolism
Ions
Life Sciences
Lipids
Liquid chromatography
Mass spectrometry
Mass spectroscopy
Methods
Microalgae
Microalgae - growth & development
Microalgae - metabolism
Nitrogen - deficiency
Physical sciences
Physiological aspects
Plant cells
Research and Analysis Methods
Scientific imaging
Starvation
Tandem Mass Spectrometry - methods
Thin layer chromatography
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Summary: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.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0182423