High performance reversed phase chromatography of cholesterol and cholesteryl esters of human plasma lipoproteins

Cholesterol and cholesteryl esters were separated according to their carbon number and number of double bonds by high performance reversed-phase chromatography (HPRC) using acetonitrile/chloroform/methanol (1:1:1, v/v) as a mobile phase. It was found that within the same equivalent carbon number (ECN) category, cholesteryl esters with the highest number of double bonds eluted ahead of those with a lower number of double bonds, and with the cis isomers eluting ahead of their trans partners. Thus, cholesteryl oleate (C27-18:1c) elutes ahead of cholesteryl palmitate (C27-16:0) and ahead of cholesteryl elaidate (C27-18:1t). Human lipoprotein, as well as rat liver cholesteryl esters, were separated using this technique.