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 C27 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, GC-MS, and 1H 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.
Ruan
B
B
Department of Chemistry, Rice University, Houston, TX 77251-1892, USA.
Shey
J
J
Gerst
N
N
Wilson
W K
WK
Schroepfer
G J
GJ
D000818:Animals; D002250:Carbon Radioisotopes; D002775:Cholestadienols / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D002849:Chromatography, Gas / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003684:Dehydrocholesterols / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D008862:Microsomes, Liver / Q000378:metabolism*; D015394:Molecular Structure; D051381:Rats; D012834:Silver; D013261:Sterols / Q000302:isolation & purification*
D002250:Carbon Radioisotopes; D002775:Cholestadienols; D003684:Dehydrocholesterols; D013261:Sterols; D012834:Silver; C026359:cholesta-5,8-dien-3 beta-ol; C016705:7-dehydrocholesterol