Separation of acidic and neutral lipids by aminopropyl-bonded silica gel column chromatography.
The separation of acidic and neutral lipids by aminopropyl-bonded silica gel column chromatography is presented. Total lipid extracts from Escherichia coli and human spermatozoa were loaded onto pre-packed aminopropyl-bonded silica gel columns and the lipids separated into four fractions. Non-polar lipids including cholesterol esters, triglycerides, diglycerides, monoglycerides and cholesterol, were eluted with 4 ml of isopropanol-chloroform (1:2, v/v) (fraction 1); free fatty acids were eluted with 4 ml of 2% acetic acid in diethyl ether (fraction 2); neutral polar lipids, including phosphophatidylethanolamine, phosphatidylcholine, sphingomyelin and neutral glycolipids, were eluted with 4 ml of methanol (fraction 3); and, finally, polar acidic lipids, including phosphatidylglycerol, cardiolipin, phosphatidylinositol, phosphatidylserine, seminolipid lipid A and acidic glycosphingolipids, were eluted with 4 ml of chloroform-methanol-0.8 M sodium acetate (60:30:4.5, v/V/V) (fraction 4). The recoveries for the different lipids ranged between 89 and 98% and the intra-assay variation, expressed as the standard deviation, was less than 5%.
Alvarez
J G
JG
Department of Obstetrics and Gynecology, University of Pennsylvania, School of Medicine, Philadelphia 19104-6080.
Touchstone
J C
JC
D002853:Chromatography, Liquid / Q000379:methods*; D002855:Chromatography, Thin Layer; D004926:Escherichia coli / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008055:Lipids / Q000737:chemistry / Q000302:isolation & purification*; D008297:Male; D011437:Propylamines / Q000737:chemistry; D058428:Silica Gel; D012822:Silicon Dioxide / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013094:Spermatozoa / Q000737:chemistry
D008055:Lipids; D011437:Propylamines; D058428:Silica Gel; D012822:Silicon Dioxide