An optimized method for fatty acid analysis, including quantification of trans fatty acids, in human adipose tissue by gas-liquid chromatography.
Considering the need for a quick direct method for measurement of the fatty acid composition including trans isomers of human adipose tissue we have developed a procedure using gas-liquid chromatography (GLC) alone, which is thus suitable for validation of fatty acid status in epidemiological studies. Fatty acids ranging in carbon number from 12 to 22 and with 0-6 double bonds were resolved and identified by capillary column GLC with a temperature program starting at 150 degrees C. Following injection, the oven temperature was increased at a rate of 3 degrees C/min to 200 degrees C, then held constant for 25 min, and finally raised at 25 degrees C/min to 225 degrees C. The trans and cis isomers of 18:1 were well separated from each other, as shown by silver-ion thin-layer chromatography. Verification by standards showed that the trans 18:1 isomers with a double bond in position 12 or lower were separated from the cis 18:1 isomers with a double bond in position 6 or higher. As the adipose tissue samples contained only small amounts of the 13t-, 14t- and 15t-18:1 isomers and the 4c- and 5c-18:1 isomers the overlapping was found to be minimal. The GLC method may also be valuable for determining the fatty acid profiles including total trans in other tissues.
Bysted
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Department of Biochemistry and Nutrition, The Technical University of Denmark, Copenhagen. bysted@mimer.be.dtu.dk
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Hølmer
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D000273:Adipose Tissue / Q000737:chemistry*; D000368:Aged; D002849:Chromatography, Gas / Q000379:methods* / Q000592:standards; D002855:Chromatography, Thin Layer / Q000379:methods* / Q000592:standards; D004952:Esters / Q000032:analysis; D005227:Fatty Acids / Q000032:analysis* / Q000737:chemistry; D005260:Female; D006801:Humans; D007536:Isomerism; D008875:Middle Aged; D015203:Reproducibility of Results; D016622:Silver Staining; D013696:Temperature
D004952:Esters; D005227:Fatty Acids