Journal
JOURNAL OF LIPID RESEARCH
Volume 49, Issue 5, Pages 1113-1125Publisher
ELSEVIER
DOI: 10.1194/jlr.D800001-JLR200
Keywords
lipidomics; metabolomics; RAW264.7; MCF7; lignoceroyl-CoA; nervonoyl-CoA; cerotoyl-CoA
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Fatty acyl-CoAs participate in numerous cellular processes. This article describes a method for the quantitation of subpicomole amounts of long chain and very-long-chain fatty acyl-CoAs by reverse-phase LC combined with electrospray ionization tandem mass spectrometry in positive ion mode with odd-chain-length fatty acyl-CoAs as internal standards. This method is applicable to a wide range of species [at least myristoyl- (C14:0-) to cerotoyl(C26:0-) CoA] in modest numbers of cells in culture (similar to 10(6)- 107), with analyses of RAW264.7 cells and MCF7 cells given as examples. Analysis of these cells revealed large differences in fatty acyl-CoA amounts (12 +/- 1.0 pmol/10(6) RAW264.7 cells vs. 80.4 +/- 6.1 pmol/106 MCF7 cells) and subspecies distribution. Very-long-chain fatty acyl-CoAs with alkyl chain lengths > C20 constitute <10% of the total fatty acyl-CoAs of RAW264.7 cells versus >50% for MCF7 cells, which somewhat astonishingly contain approximately as much C24:0- and C26:0-CoAs as C16:0- and C18:0-CoAs and essentially equal amounts of C26:1- and C18:1-CoAs. This simple and robust method should facilitate the inclusion of this family of compounds in lipidomics and metabolomics studies.
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