Purification of phospholipids by preparative high pressure liquid chromatography |
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Authors: | W S M Geurts van Kessel M Tieman R A Demel |
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Affiliation: | (1) Laboratory of Biochemistry, State University of Utrecht, University Center “De Uithof”, Padualaan 8, NL-3584 CH Utrecht, The Netherlands |
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Abstract: | A method is described for the purification of a number of phospholipids by preparative high performance liquid chromatography
(HPLC). Purification of digalactosyl-diglyceride from spinach and egg phosphatidylcholine, 1,2-dioleoyl-sn-glycero-3-phosphocholine and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine from its reaction mixture have been resolved. The lipid separation is performed on a polygosil
column and the individual compounds are monitored directly by refractive index detection. Chloroform/methanol mixtures are
used as eluent systems, providing a wide polarity range to separate the classes of lipids. The developed equipment can be
used for columns between 10 and 50 cm long and 4 and 50 mm inner diameter. The flow rate could be varied between 1 and 100
ml/min and applied pressures between 10 and 450 bars. |
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