ISOLATION AND PURIFICATION OF LIPIDS BY CENTRIFUGAL PARTITION CHROMATOGRAPHY

Methodologies for the separation of various lipid classes including free fatty acids (FA) and fatty acid ethyl esters (FAEE), monoglycerides (MG), diglycerides (DG), triglycerides (TG), phosphoglycerides (PG), and glycosphingolipids (GSL) by Centrifugal Partition Chromatography are presented. The fatty acid ethyl esters, Hexadecanoate, Octadecanoate, cis-9-Octadecenoate; and cis,cis-9,12-Octadecadienoate, and all-cis-9,12,15-Octadecatrienoate, were separated at 800 rpm; flow rate of 2.0 ml/min; using n-hexane/acetonitrile (1:1, v/v) as the solvent system and normal ascending and reversed descending elution modes respectively. The fatty acid ethyl esters, all-cis-5,8,11,14,17-Icosapentaenoate and all-cis-4,7,10,13,16,19-Docosahexaenoate, were separated at 600 rpm; flow rate of 3.3 ml/min; using n-hexane/methanol/water (1/0.9/0.1, v/v/v) as the solvent system and normal ascending mode. FA and MG; and DG and TG were separated at 600 rpm; 10 ml/min; using n-hexane/methanol/water (1:1:0.05, v/v/v) as the solvent system and reversed descending and normal ascending modes respectively. PE and PC were separated at 600 rpm; flow rate of 5.0 ml/min; using heptane/ethanol (1:1, v/v) as the solvent system and normal ascending and reversed descending modes respectively. The peracetylated GalCer, SF, GB4; and GM1, GD1a, and GT1b, were partially resolved at 800 rpm; flow rate of 4.0 ml/min; using heptane/acetonitrile/water (5:4:1, v/v/v) as the solvent system and normal ascending and reversed descending modes, respectively. These results indicate that CPC constitutes a powerful technique for the separation of large amounts of the aforementioned lipids. Its applications should prove most useful in the purification of these lipids and in faciliating their commercial availability.