TY - JOUR
T1 - Inhibitors of sterol synthesis. Chromatography of acetate derivatives of oxygenated sterols
AU - Kudo, K.
AU - Emmons, G. T.
AU - Casserly, E. W.
AU - Via, D. P.
AU - Smith, L. C.
AU - St. Pyrek, J.
AU - Schroepfer, G. J.
N1 - Copyright:
Copyright 2004 Elsevier B.V., All rights reserved.
PY - 1989
Y1 - 1989
N2 - The separation of the acetate derivatives of a number of oxygenated sterols was achieved by medium pressure liquid chromatography on silica gel columns and by normal and reversed phase high performance liquid chromatography. We have explored the application of these chromatographic systems for the analysis of oxygenated sterols of plasma samples from two normal human subjects. The addition of highly purified [14C]cholesterol to plasma permitted the detection and quantitation of oxygenated sterols formed by autoxidation of cholesterol during processing of the samples. Special attempts to suppress autoxidation of cholesterol included the use of an all-glass closed system for saponification and extraction under argon followed by rapid removal of cholesterol from the polar sterols by reversed phase medium pressure liquid chromatography. Chromatographic analyses of the [3H]acetate derivatives of the polar sterols provided a sensitive approach for the detection and quantitation of the individual oxygenated sterols. Oxygenated sterols detected in plasma included cholest-5-ene-3β,26-diol, (24S)-cholest-5-ene-3β,24-diol, and cholest-5-ene-3β,7α-diol. After correction for their formation by autoxidation of cholesterol during processing of the samples, very little or none of the following sterols were observed: cholest-5-ene-3β,7β-diol, 5α,6α-epoxy-cholestan-3β-ol, 5β,6β-epoxy-cholestan-3β-ol, and cholestane- 3β, 5α,6β-triol, and the 25-hydroxy, 22R-hydroxy, 21-hydroxy, 20α-hydroxy, and 19-hydroxy derivatives of cholesterol.
AB - The separation of the acetate derivatives of a number of oxygenated sterols was achieved by medium pressure liquid chromatography on silica gel columns and by normal and reversed phase high performance liquid chromatography. We have explored the application of these chromatographic systems for the analysis of oxygenated sterols of plasma samples from two normal human subjects. The addition of highly purified [14C]cholesterol to plasma permitted the detection and quantitation of oxygenated sterols formed by autoxidation of cholesterol during processing of the samples. Special attempts to suppress autoxidation of cholesterol included the use of an all-glass closed system for saponification and extraction under argon followed by rapid removal of cholesterol from the polar sterols by reversed phase medium pressure liquid chromatography. Chromatographic analyses of the [3H]acetate derivatives of the polar sterols provided a sensitive approach for the detection and quantitation of the individual oxygenated sterols. Oxygenated sterols detected in plasma included cholest-5-ene-3β,26-diol, (24S)-cholest-5-ene-3β,24-diol, and cholest-5-ene-3β,7α-diol. After correction for their formation by autoxidation of cholesterol during processing of the samples, very little or none of the following sterols were observed: cholest-5-ene-3β,7β-diol, 5α,6α-epoxy-cholestan-3β-ol, 5β,6β-epoxy-cholestan-3β-ol, and cholestane- 3β, 5α,6β-triol, and the 25-hydroxy, 22R-hydroxy, 21-hydroxy, 20α-hydroxy, and 19-hydroxy derivatives of cholesterol.
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M3 - Article
C2 - 2794792
AN - SCOPUS:0024324848
SN - 0022-2275
VL - 30
SP - 1097
EP - 1111
JO - Journal of Lipid Research
JF - Journal of Lipid Research
IS - 7
ER -