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Liquid chromatography high‐resolution mass spectrometry for fatty acid profiling
Author(s) -
Bromke Mariusz A.,
Hochmuth Anton,
Tohge Takayuki,
Fernie Alisdair R.,
Giavalisco Patrick,
Burgos Asdrubal,
Willmitzer Lothar,
Brotman Yariv
Publication year - 2015
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/tpj.12739
Subject(s) - chemistry , chromatography , derivatization , mass spectrometry , gas chromatography–mass spectrometry , fatty acid , gas chromatography , liquid chromatography–mass spectrometry , fatty acid methyl ester , resolution (logic) , biochemistry , artificial intelligence , computer science , biodiesel , catalysis
Summary Quantification of fatty acids has been crucial to elucidate lipid biosynthesis pathways in plants. To date, fatty acid identification and quantification has relied mainly on gas chromatography ( GC ) coupled to flame ionization detection ( FID ) or mass spectrometry ( MS ), which requires the derivatization of samples and the use of chemical standards for annotation. Here we present an alternative method based on a simple procedure for the hydrolysis of lipids, so that fatty acids can be quantified by liquid chromatography mass spectrometry ( LC ‐ MS ) analysis. Proper peak annotation of the fatty acids in the LC ‐ MS ‐based methods has been achieved by LC ‐ MS measurements of authentic standard compounds and elemental formula annotation supported by 13 C isotope‐labeled Arabidopsis. As a proof of concept, we have compared the analysis by LC ‐ MS and GC ‐ FID of two previously characterized Arabidopsis thaliana knock‐out mutants for FAD 6 and FAD 7 desaturase genes. These results are discussed in light of lipidomic profiles obtained from the same samples. In addition, we performed untargeted LC ‐ MS analysis to determine the fatty acid content of two diatom species. Our results indicate that both LC ‐ MS and GC ‐ FID analyses are comparable, but that because of higher sensitivity and selectivity the LC ‐ MS ‐based method allows for a broader coverage and determination of novel fatty acids.