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Papers In Press, published online ahead of print January 1, 2010
J. Lipid Res., doi:10.1194/jlr.D000547

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Journal of Lipid Research, Vol. 51, 216-221, January 2010
Copyright © 2010 by American Society for Biochemistry and Molecular Biology

Methods

High-throughput analysis of fatty acid composition of plasma glycerophospholipids^[S]

Claudia Glaser, Hans Demmelmair and Berthold Koletzko¹

Division of Metabolic Diseases and Nutritional Medicine, Dr. von Hauner Children's Hospital, Ludwig-Maximilians-University of Munich, Lindwurmstr. 4, D-80337 Munich, Germany

¹ To whom correspondence should be addressed. e-mail: office.koletzko{at}med.uni-muenchen.de

Plasma FA composition, a marker of FA status and dietary intake, is associated with health outcomes on a short- and long-term basis. Detailed investigation of the relationships between plasma FA composition and health requires the analysis of large numbers of samples, but manual sample preparation is very cumbersome and time consuming. We developed a high-throughput method for the analysis of FAs in plasma glycerophospholipids (GPs) with increased sensitivity. Sample preparation requires two simple steps: protein precipitation and subsequent base catalyzed methyl ester synthesis. Analysis of GP FAs is performed by gas chromatography. Coefficients of variation for FAs contributing more than 1% to total FAs are below 4%. Compared with the established reference method, results of the new method show good agreement and very good correlations (r > 0.9). The new method reduces the manual workload to about 10% of the reference method. Only 100 µl plasma volume is needed, which allows for the analysis of samples from infants. The method is well suitable for application in large clinical trials and epidemiological studies.

Supplementary key words essential fatty acids • fatty acids • fatty acid methyl esters • high-throughput • glycerophospholipids • phospholipids

Abbreviations: CE, cholesteryl ester; CV, coefficient of variation; FAME, fatty acid methyl ester; GC, gas chromatography; GP, glycerophospholipid; LC-PUFA, long-chain PUFA; PhL, phospholipid; TAG, triacylglycerol

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