Silver ion high pressure liquid chromatography provides unprecedented separation of sterols: Application to the enzymatic formation of cholesta-5,8-dien-3 beta-ol

We report that silver ion HPLC provides remarkable separations of C-27 sterols differing only in the number or location of olefinic double bonds. This technique has been extended to LC-MS, analysis of purified components by GC, CC-MS, and H-1 NMR, and to its use on a semipreparative scale. The application of this methodology for the demonstration of the catalysis, by rat liver microsomes, of the conversion of 7-dehydrocholesterol to cholesta-5,8-dien-3 beta-ol is also presented.