Abstract
A new method for the simultaneous determination of cortisol and cortisone in serum by high-performance liquid chromatography with fluorimetric detection has been developed. The two steroids were derivatized by treatment with 9-anthroyl nitrile in triethylamine-acetonitrile to give the fluorescent esters through the 21-hydroxyl group. These derivatives were efficiently separated on a straight-phase Cosmosil 5SL column with hexane-ethyl acetate (6:5) as eluent. The detection limit was 20 fmol, using the 9-anthroyl derivative of prednisolone as an internal standard. The use of Sep-Pak C18 and Clin-Elut cartridges proved to be efficient for the clean-up of cortisol and cortisone in serum samples.
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