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Determination of red blood cell fatty acid profiles: Rapid and high-confident analysis by chemical ionization-gas chromatography-tandem mass spectrometry B Analytical technologies in the biomedical and life sciences

Schober, Yvonne, Wahl, Hans Günther, Renz, Harald, Nockher, Wolfgang Andreas
Journal of chromatography 2017 v.1040 pp. 1-7
biomarkers, chemical analysis, docosahexaenoic acid, eicosapentaenoic acid, erythrocytes, fatty acid composition, fatty acid methyl esters, gas chromatography-mass spectrometry, human diseases, humans, ionization, ions, linoleic acid, tandem mass spectrometry
Cellular fatty acid (FA) profiles have been acknowledged as biomarkers in various human diseases. Nevertheless, common FA analysis by gas chromatography mass spectrometry (GC–MS) requires long analysis time. Hence, there is a need for feasible methods for high throughput analysis in clinical studies.FA was extracted from red blood cells (RBC) and derivatized to fatty acid methyl esters (FAME). A method using gas chromatography tandem mass spectrometry (GC–MS/MS) with ammonia-induced chemical ionization (CI) was developed for the analysis of FA profiles in human RBC. We compared this method with classical single GC–MS using electron impact ionization (EI). The FA profiles of 703 RBC samples were determined by GC–MS/MS.In contrast to EI ammonia-induced CI resulted in adequate amounts of molecular ions for further fragmentation of FAME. Specific fragments for confident quantification and fragmentation were determined for 45 FA. The GC–MS/MS method has a total run time of 9min compared to typical analysis times of up to 60min in conventional GC–MS. Intra and inter assay variations were <10% for all FA analyzed. Analysis of RBC FA composition revealed an age-dependent increase of the omega-3 eicosapentaenoic and docosahexaenoic acid, and a decline of the omega-6 linoleic acid with a corresponding rise of the omega-3 index.The combination of ammonia-induced CI and tandem mass spectrometry after GC separation allows for high-throughput, robust and confident analysis of FA profiles in the clinical laboratory.