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Quehenberger, Oswald; Armando, Aaron M.; Dennis, Edward A.
Biochimica and biophysica acta. Molecular and cell biology of lipids, 11/2011, Volume: 1811, Issue: 11Journal Article
Historically considered to be simple membrane components serving as structural elements and energy storing entities, fatty acids are now increasingly recognized as potent signaling molecules involved in many metabolic processes. Quantitative determination of fatty acids and exploration of fatty acid profiles have become common place in lipid analysis. We present here a reliable and sensitive method for comprehensive analysis of free fatty acids and fatty acid composition of complex lipids in biological material. The separation and quantitation of fatty acids are achieved by capillary gas chromatography. The analytical method uses pentafluorobenzyl bromide derivatization and negative chemical ionization gas chromatography–mass spectrometry. The chromatographic procedure provides base line separation between saturated and unsaturated fatty acids of different chain lengths as well as between most positional isomers. Fatty acids are extracted in the presence of isotope-labeled internal standards for high quantitation accuracy. Mass spectrometer conditions are optimized for broad detection capacity and sensitivity capable of measuring trace amounts of fatty acids in complex biological samples. This article is part of a Special Issue entitled Lipodomics and Imaging Mass Spectrometry. ► We review quantitative analyses of fatty acids in biological material. ► Chromatographic separation of fatty acids is achieved by capillary GC. ► Mass spectrometry using soft ionization techniques enables sensitive detection. ► Isotope dilution method facilitates accurate and precise quantitation.
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