出版社：American Society for Biochemistry and Molecular Biology
摘要：Conventional sample preparation for fatty acid analysis is acomplicated, multiple-step process, and gas chromatography (GC)analysis alone can require >1 h per sample to resolve fattyacid methyl esters (FAMEs). Fast GC analysis was adapted tohuman plasma FAME analysis using a modified polyethylene glycolcolumn with smaller internal diameters, thinner stationary phasefilms, increased carrier gas linear velocity, and faster temperatureramping. Our results indicated that fast GC analyses were comparableto conventional GC in peak resolution. A conventional transesterificationmethod based on Lepage and Roy was simplified to a one-stepmethod with the elimination of the neutralization and centrifugationsteps. A robotics-amenable method was also developed, with lowermethylation temperatures and in an open-tube format using multiplereagent additions. The simplified methods produced results thatwere quantitatively similar and with similar coefficients ofvariation as compared with the original Lepage and Roy method.
The present streamlined methodology is suitable for the directfatty acid analysis of human plasma, is appropriate for researchstudies, and will facilitate large clinical trials and makepossible population studies.Abbreviations: BHT, butylated hydroxytoluene; DHA, docosahexaenoic acid; EPA, eicosapentaenoic acid; FAME, fatty acid methyl ester; FID, flame ionization detector; GC, gas chromatography; HUFA, highly unsaturated fatty acid; RRF, relative response factor
Supplementary key words fatty acid analysis • plasma • transesterification method • robotic chemistry • fast gas chromatographic analysis
