Abstract
The development of gas-liquid chromatography (GLC) made possible the rapid separation of fatty acid esters on a micro scale. However the hitherto published analyses by GLC have been limited to vegetable oils and terrestrial animal fats. The reason is that the separation of polyenes from other fatty acids is not always clean, which complicates the analysis of other oils such as fish oils comprising the great variety of saturated and unsaturated acids and polyenes. So further improvement has been required in the technique. The present study was undertaken to establish the analytical method of fatty acids in fish oil by GLC. The procedures employed for this purpose are as follows: GLC is carried out on succinate polyester of diethylene glycol, one of the most effective stationary liquid phase for separating fatty acids. Fish oil is methylated, hydrogenated and brominated, bromides being removed, and then each of these samples is subjected to GLC. At first, carbon number composition of fatty acids is determined from the hydrogenated sample, secondly the presence of saturated acids and monoenes is confirmed from the brominated sample, and at last fatty acids are identified on the basis of retention time of chromatogram of methylated sample, reference being made to these results and the presence of polyenes revealed by an alkaliisomerization ultraviolet spectrophotometry. The percentage composition of oil is determined from weights of peaks, the results are the means of duplicate determinations. The fatty acid composition of fish oil was compared by GLC and spectrophotometry. GLC gave somewhat lower values for the slow-moving polyenes and higher values for the fastmoving saturated acids than did spectrophotometry. GLC was not successful in the detection of very small amounts of unsaturated acids which were estimated to be below 1% by spectrophotometry. Odd-numbered acids were demonstrated in fish oil by GLC. In general, the GLC results and spectrophotometric values were in agreement. It is concluded from the present data that analytical results obtained by GLC are reliable.
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