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3-(7'-Methoxycoumarin-3'-carbonyl)-benzoxazoline-2-thione (MCBT) was synthesized as a precolumn fluorescent labeling reagent for amantadine for use in high-performance liquid chromatography (HPLC). Amantadine was derivatized quantitatively into a fluorescent compound through the amino group by treatment with MCBT at room temperature for 5 min in aqueous 95% acetonitrile solution in the presence of 4-dimethylaminopyridine. The derivative was subjected to HPLC on TSK gel ODS-80TM (250 x 4.6 mm i.d.) with methanol-water (10:1) as the mobile phase and monitored with an excitation wavelength of 355 nm and an emission wavelength of 405 nm. The limit of detection was 2.0 micrograms/ml in the urine. This method was satisfactory with respect to simplicity and precision to quantify amantadine spiked in the urine.

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.

A fluorescence high-performance liquid chromatographic method for the determination of corticosteroids is described. Corticosteroids were labeled quantitatively by treatment with 1-(difluoro-1,3,5-triazinyl)-2-methylisoindole in an alkaline medium at room temperature for 10 s and the fluorescent adducts were eluted isocratically on a reversed-phase column by a mobile phase of aqueous methanol. The detection limit of cortisone was 3 pmol for an injection volume of 10 microliter.

Anthracene-1-carbonyl azide was found to be a sensitive fluorescent derivatization reagent for primary and secondary alcohols for high-performance liquid chromatography. Reaction conditions were investigated for cholestanol. The reagent reacts with alcohols in benzene in the presence of 4-dimethylaminopyridine to give the corresponding fluorescent urethanes at 100 degrees C for 50 min. The detection limit for cholestanol was 250 fmol for an injection volume of 10 microliters. Tertiary alcohols and phenols do not produce fluorescent derivatives under these conditions.