Fatty acid analysis by high resolution gas chromatography and mass spectrometry for clinical and experimental applications

Abstract

Quantitative fatty acid profiles analyzed via fatty acid methyl esters (FAME) are among the most common metabolite panels and fit into the category of omics techniques. Recently, preparation and analysis methods for high throughput clinical analysis have become routine, and novel methods for structure analysis enable rapid identification of unknowns and confounded peaks. Observation of one hundred FAME in a single mixture is common with high resolution capillary gas chromatography columns. Structural analysis of FAME requires high resolution gas chromatography with specialized tandem mass spectrometry to obtain fragments indicative of structure. Covalent adduct chemical ionization provides unambiguous double bond positions, whereas electron ionization with fragmentation of the molecular ion identifies branch points. Quantitative analysis requires response calibration using external standards and/or isotopically labeled internal standards with mass spectrometry detection. Modern high throughput methods enable routine analysis of well behaved clinical samples. Careful attention to structure analysis using recent methods avoids biases due to interfering or mischaracterized fatty acids.