PURIFICATION OF FISH-OIL ETHYL-ESTERS BY HIGH-SPEED COUNTERCURRENT CHROMATOGRAPHY USING NONAQUEOUS SOLVENT SYSTEMS

High-speed countercurrent chromatography (HSCCC) was applied to purifi cation of fish oil ethyl esters (FOEE). The CCC separations were perfo rmed using non-aqueous two-phase solvent systems in two steps: The fir st purification was carried out with a hexane-acetonitrile system wher e 30 mL of crude FOEE was cleaned in 140 min yielding 23.1 mL of refin ed FOEE. The product was colorless and free of saturated fatty acid es ters. The second and final purification was performed with a hexane-di chloromethane-acetonitrile (5:1:4, v/v/v) where 1 g of the refined FOE E was resolved into three fractions, ethyl hexadecenoate, ethyl octade cenoate and the third composed of 56.4% of ethyl docosahexaenoate and 39.3% of ethyl eicosapentenoate. The separation was completed in 145 m in. However, polyunsaturated fatty acids are not resolved with this so lvent system due to their low partition coefficients.