出版社：American Society for Biochemistry and Molecular Biology
摘要：A technique of column chromatography using factice, a hydrophobic polymer, as stationary phase, and aqueous acetone as moving phase is described. With this liquid-gel system, the major nonpolar lipid classes (cholesterol esters, triglycerides, cholesterol, diglycerides, and monoglycerides) are easily separated from each other. Furthermore, separations of components with different fatty acid composition within the same lipid class can also be obtained. Thus, cholesterol esters with fatty acid chain lengths from 2 to 18 carbons are eluted as individual peaks. Similar separations are possible with glycerides or with mixtures of methyl esters of fatty acids. However, with the exception of cholesterol esters, lipids within a given class containing fatty acids of equivalent polarity (2-carbon shortening equivalent to one double bond) are inseparable. Elution is performed with a single solvent mixture and detection of lipid in the effluent stream can be achieved by the use of automatic differential refractometry. In this way, the entire procedure can be automatically performed. Free fatty acids and phospholipids are poorly separated and must be removed from the lipid mixture prior to chromatography.
