Purification of Phosphatidylcholine with High Content of DHA from Squid Illexargentinus by Countercurrent Chromatography

Abstract

Abstract A method for purification by countercurrent chromatography (CCC) of phosphatidylcholine with high content of DHA is described. Phospholipids were extracted from squid Illex argentinus characterized by high performance liquid chromatography and their fatty acids profile determined by gas chromatography. Phosphatidylcholine was primarily represented with 48.3 % (w/w) of total phospholipids and contained 39.7 % of DHA (w/w total fatty acids). The analytical separation of phosphatidylcholine, was achieved on a Kromaton II apparatus, in the systems heptane—ethyl acetate—acetonitrile (1:0.65:1, v/v/v) and isooctane—ethyl acetate—acetonitrile (1:0.65:1, v/v/v), at 550 rpm and 1.5 mL/min flow rate using normal ascending and reversed descending elution modes respectively. The fractions collected were further analysed by high performance thin layer chromatography. Phosphatidylcholine was separated from the other phospholipids of the crude extract and was recovered pure in the lower phase with 95 % yield. Purification at preparative scale was performed on a 1 L column, 550 rpm and 5 mL/min using the same chromatograph. These results prove that CCC constitutes a powerful technique for separation of phospholipids from natural lipidic extracts.