An Improved Method for the Separation and Quantification of Major Phospholipid Classes by LC-ELSD
An improved HPLC procedure for the separation of phospholipids is described. The method described utilizes a solvent mixture of acetonitrile-methanol–water-trifluoroacetic acid (100:25:1.7:2.5, v/v) as the mobile phase, which is more compatible with the pump than mobile phases containing inorganic acids. Separation was by isocratic elution on a Hypersil silica column coupled to an evaporative light scattering detector. Complete separation of phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidylcholine (PC) and sphingomyelin (SM) was achieved in less than 20 min. The detection limits for PS, PE, PC and SM were 50, 50, 80 and 150 ng (S/N = 3), respectively. Human, bovine and porcine erythrocyte ghost membranes and animal tissues have been successfully analyzed for their phospholipid contents.
KeywordsColumn liquid chromatography Evaporative light scattering detector Phospholipids Trifluoroacetic acid
The study was supported by grants from the National 863 Program of China (Grants No. 2006AA02A143) and “Special Research Foundation” (Grants No. 09JK784 and 08JK469) by Education Department of Shaanxi Province.
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