Isolation and Purification of Lipids by Centrifugal Partition Chromatography

Department of Chemistry, Villanova University, 노리스타운, Pennsylvania, United States
Journal of liquid chromatography 11/1990; 13(18):3603-3614. DOI: 10.1080/01483919008049558


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

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    ABSTRACT: Summary The present study was undertaken in order to characterize then to purify fatty acids from marine phytoplankton. From a crude mixture of fatty acid methyl esters it was possible to isolate by countercurrent chromatography a mixture of four polyunsaturated fatty acid methyl ester identified as being hexadecatrienoic acid methyl ester, octadecatetraenoic acid methyl ester, eicosapentaenoic acid methyl ester and docosahexaenoic acid methyl ester by gas chromatography coupled with mass spectrometry in electron impact and in positive-ion chemical ionization mode. The four polyunsaturated fatty acids are in different ratios in mixtures from the two microorganisms:Skeletonema costatum andIsochrysis galbana.
    No preview · Article · Jul 1994 · Chromatographia
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    ABSTRACT: Hexadecatrienoic acid (C16:3), octadecatetraenoic acid (C18:4), eicosapentaenoic acid (C20:5) and docosahexaenoic acid (C22:6) were purified in two steps by counter-current chromatography (CCC). The selected liquid phases were heptane-acetonitrile-water for the first step and heptane-methanol-water for the second. The upper non-polar phase was kept stationary in the coil of the instrument by a rotational force field. The separation was followed by UV detection at 210 nm and by analysing the eluted fractions by gas chromatography after esterification. The purity of the fatty acids therefore purified was checked by capillary gas chromatograpy. CCC may be an alternative to HPLC for the preparative-scale purification of such compounds.
    No preview · Article · Jun 1995 · Journal of Chromatography A
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    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.
    No preview · Article · Jun 1996 · Journal of Liquid Chromatography & Related Technologies
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