Abstract
A reversed phase high performance liquid chromatographic (HPLC) method was developed for direct profiling and determination of adrenal cholesteryl ester composition. Cholesteryl adrenate and cholesteryl cervonate, which are not commercially available, were synthesized as markers. Lipid extracts of rat adrenal homogenates or lipid droplets were individually applied to a conditioned silica gel-60 column which separated cholesteryl esters from other native lipids. The eluted cholesteryl ester fraction was then analyzed by HPLC. With cholesteryl heptadecanoate as internal standard, seven adrenal cholesteryl esters were detected and quantified: cholesteryl cervonate, cholesteryl arachidonate, cholesteryl adrenate, cholesteryl myristate, cholesteryl oleate, cholesteryl palmitate, and cholesteryl stearate. Among them, cholesteryl adrenate appeared to be the major sterol ester stored in the rat adrenal.
Highlights
A reversed phase high performance liquid chromatographic (HPLC) method was developed for direct profiling and determination of adrenal cholesteryl ester composition
Because cholesteryl adrenate (CE 22:4) and cholesteryl cervonate (CE 225) were not commercially available, we present a protocol for the synthesis of these lipids as markers
We have found that cholesteryl linoleate (34.54 min) can be separated from cholesteryl adrenate (32.86 min) and X4 peak (35.99 min), but the adrenal cholesteryl linoleate content is too low to be measured by the present method
Summary
A reversed phase high performance liquid chromatographic (HPLC) method was developed for direct profiling and determination of adrenal cholesteryl ester composition. Cholesteryl adrenate and cholesteryl cervonate, which are not commercially available, were synthesized as markers. Lipid extracts of rat adrenal homogenates or lipid droplets were individually applied to a conditioned silica gel-60 column which separated cholesteryl esters from other native lipids. The eluted cholesteryl ester fraction was analyzed by HPLC. With cholesteryl heptadecanoate as internal standard, seven adrenal cholesteryl esters were detected and quantified: cholesteryl cervonate, cholesteryl arachidonate, cholesteryl adrenate, cholesteryl myristate, cholesteryl oleate, cholesteryl palmitate, and cholesteryl stearate. Cholesteryl adrenate appeared to be the major sterol ester stored in the rat adrena.Cheng, B., and J. Analysis of adrenal cholesteryl esters by reversed phase high performance liquid chromatography.J Lipid Res. 1994. Analysis of adrenal cholesteryl esters by reversed phase high performance liquid chromatography.J Lipid Res. 1994. 35: 1115-1121
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