A rapid, isocratic method for phospholipid separation by high-performance liquid chromatography.
A rapid, isocratic method for separating the most prevalent phospholipids by high-performance liquid chromatography is described. Baseline resolution of phosphatidylinositol, phosphatidylserine, phosphatidylethanolamine, phosphatidylcholine, lysophosphatidylcholine, and sphingomyelin is achieved in less than 40 min on a silica column. Lipids are injected in 10 microliter of chloroform-diethyl ether 1:2 (v/v) and eluted with a solvent mixture of acetonitrile-methanol-sulfuric acid 100:3:0.05 (v/v/v) at a flow rate of 1 ml/min. Neutral lipids and cardiolipin elute with the solvent front. Chromatography of a radioactive cell lipid extract indicates a recovery of better than 97%. The procedure is sensitive enough to permit the analysis of the main phospholipids present in a monolayer culture containing about 100 micrograms of cell protein.
D000818:Animals; D001011:Aorta / Q000032:analysis; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004727:Endothelium / Q000032:analysis; D005227:Fatty Acids / Q000032:analysis; D009131:Muscle, Smooth, Vascular / Q000032:analysis; D010713:Phosphatidylcholines / Q000302:isolation & purification; D010743:Phospholipids / Q000302:isolation & purification*; D010944:Plants / Q000032:analysis; D013025:Glycine max
D005227:Fatty Acids; D010713:Phosphatidylcholines; D010743:Phospholipids