The estimation of unsaturated fatty acids by reversed-phase partition chromatography

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Abstract
The separation of representative unsaturated acids on a standard reversed-phase column was investigated. A number of conclusions are drawn regarding the relationship between structure and elution rate. A method is devised for analysis of milligram quantities of fatty acid mixtures containing both saturated and unsaturated acids. The most widely distributed unsaturated fatty acids are separable from each other by the present chromatographic technique but not from certain saturated fatty acids which are eluted at similar rates. The latter are estimated by a 2d chromatogram following oxidation by alkaline permanganate, which removes the unsaturated material. The amounts of unsaturated fatty acids are then determined by difference. The method was tested on standard fatty acid mixtures, and on fatty acids from a number of natural oils. It gives good agreement with other techniques.