Abstract
In high performance liquid chromatography on a silica gel column, elution of phospholipids (PLs) can be accelerated in the presence of magnesium chloride or calcium chloride in water-containing eluents composed of n-hexane-isopropanol-water. In particular, the addition of these metal chlorides results in markedly reduced retention of the chloline-containing PLs such as phosphatidylcholine, sphingomyelin and lysophosphatidylcholine. The effect on the retention increases with increases in concentration of the metal chlorides and in water content in the eluent systems. Two types of Aquasil SS columns packed with Hypersil-5 of 60 or 120Å pore size showed the same retention time for the PLs. These columns retain PLs more weakly than LiChrosorb Si-60. Seven kinds of phosphoglycerides and sphingolipids (sphingomyelin) can be separated within 40-60 minutes by the use of Aquasil SS column and the isocratic eluent composed of n-hexane-isopropanol-0.2mol dm-3 magnesium chloride (36:48:5, v/v). Five classes of PLs were identified from PL fraction of human platelets by the proposed method.
