Abstract
Pyrene-1-carbonyl fluoride (PCF) was synthesized as a precolumn fluorescent labeling reagent for alcohols and amines for use in high performance liquid chromatography (HPLC). PCF reacted with primary and phenolic hydroxyl groups in dichloromethane at 100 degrees C for 30 min in the presence of 4-dimethylaminopyridine (DMAP) to give the corresponding fluorescent pyrene esters and also reacted with primary amines in acetonitrile at room temperature for 2 min in the presence of DMAP to give the amides. The PCF esters of corticosteroids were separated by normal-phase chromatography on a Cosmosil 5 SL column with hexane-ethyl acetate (6:5, v/v) and the PCF amides of primary amines were separated by reversed-phase chromatography on a TSK gel ODS-80 TM column with methanol-water (10:3, v/v). The detection limits (S/N = 3) of cortisone and 2-phenylethylamine were 600 fmol and 800 fmol for an injection volume of 10 microliters, respectively.
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