High performance liquid chromatography of fatty acids as naphthacyl derivatives

A procedure for the separation of fatty acids, after their derivatization a s fatty acid naphthacyl esters, by reverse-phase high performance liquid ch romatography is described. A reproducible resolution (50 min), shorter than former HPLC analyses, of a standard mixture of fatty acid naphthacyl ester s (25 fatty acids; C7:0 - C22:6, n - 3), was achieved by a ternary elution gradient of methanol-acetonitrile-water. Compared with gas chromatography. HPLC analysis of fatty acid naphthacyl esters showed similar percentages of molar distribution of long-chain fatty acids (greater than or equal to 14 carbons). The separation was monotered by UV absorbance at 246 nm. which wa s highly sensitive (the detection limit was about 0.1 ng), did not destroy the fatty acid naphthacyl esters and thus allowed individual recovery for f urther analysis (specific radioactivity determination or positional isomer identification). This preparative HPLC method provides, therefore, a useful process for the study of fatty acid metabolism in biological systems.