A method for purification by countercurrent chromatography (CCC) of phosphatidylcholine with high content of DHA is described. Phospholipids were extracted from squidIllex argentinuscharacterized 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.