Regarding plasma fatty acid profile, the analysis was carried out blind to the subject status. Plasma was stored at −80 °C until used for analysis. Lipids were extracted with different chloroform/methanol mixtures according to Folch [33 (link)] and fractionated by HPLC-ELSD and analyzed as previously describe [34 (link), 35 (link)].
Fatty acid composition of whole plasma was determined by gas-chromatographic analysis. The fatty acid methylesters, obtained after trans-derivatization with sodium methoxide in methanol 3.33 % w/v, were injected into gas chromatograph (Agilent Technologies 6850 Series II) equipped with a flame ionization detector (FID) under the following experimental conditions: capillary column: AT Silar length 30 m, film thickness 0.25 μm, Gas carrier: helium, temperature: injector 250 °C, detector 275 °C, oven 50 °C for 2 min, rate of 10 °C min-1 until 200 °C for 20 min. A standard mixture containing methyl ester fatty acids was injected for calibration.
Fatty acid composition of whole plasma was determined by gas-chromatographic analysis. The fatty acid methylesters, obtained after trans-derivatization with sodium methoxide in methanol 3.33 % w/v, were injected into gas chromatograph (Agilent Technologies 6850 Series II) equipped with a flame ionization detector (FID) under the following experimental conditions: capillary column: AT Silar length 30 m, film thickness 0.25 μm, Gas carrier: helium, temperature: injector 250 °C, detector 275 °C, oven 50 °C for 2 min, rate of 10 °C min-1 until 200 °C for 20 min. A standard mixture containing methyl ester fatty acids was injected for calibration.
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