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The chloroform was pooled and dried under nitrogen and subjected to methylation. To monitor the recovery rate, the fatty acid C23:0 was added to the fatty acid ester samples fatty acid ester (usually 1 μg added to 2 mg tissue sample) as an internal standard. Fatty acid methyl esters were prepared by methods similar to those described previously [5,6] using BF3/methanol reagent (14% Boron Trifluoride). Lipid sample was mixed with 1 ml hexane in 16 ml glass tubes with Teflon-lined caps. BF3/MeOH fatty acid ester reagent (1 ml) was added and the mixture was heated at 90-110°C in a metal block or a sand bath for 1 hour, cooled to room temperature and methyl esters extracted in the hexane phase after addition of 1 ml H2O. Samples were allowed to stand for 20-30 min, and then the upper hexane layer was removed and concentrated under nitrogen. Fatty acid methyl esters were analyzed by gas chromatography using a fully automated HP5890 system equipped with a flame-ionization detector, as described previously [7] The chromatography utilized an Omegawax 250 capillary column (30 m × 0.25
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