Determination of fatty acids as phenacyl esters in rat adipose tissue and blood vessel walls by high-performance liquid chromatography.

Twenty-two biologically relevant (6:0-22:6) saturated, monounsaturated and polyunsaturated fatty acids were separated by reversed-phase high-performance liquid chromatography after derivatization with phenacyl bromide. An optimal resolution of the critical combinations linolenic-myristic, docosahexaenoic-palmitoleic-arachidonic and palmitic-oleic acids and cis and trans isomers of octadecenoic (n9) and octadecadienoic (n9, 12) acids was achieved by continuous gradient elution with methanol-acetonitrile-water. Elution of mixtures of 6:0-22:1 fatty acids was completed within 80 min at a flow-rate of 1 ml/min. By the use of UV detection at 242 nm the detection limits for short- and long-chain fatty acids were found to be about 0.8 and 12 ng per injection, respectively. Linearity was tested up to 100 ng. The method was applied to the determination of fatty acids in rat adipose tissue and blood vessel walls of animals fed hydrogenated fat diets. The results are comparable to those obtained by gas chromatography and surpass the latter for the resolution of oleic and elaidic acids.