A simple, sensitive, and mild method for the determination of amino compounds based on a condensationreaction with fluorescence detection has been developed.9-(2-Hydroxyethyl)acridone reacts with coupling agent
N,
N'-carbonyldiimidazole at ambient temperature to formactivated amide intermediate 9-(2-acridone)oxyethylcarbonylimidazole (AOCD). The amide intermediate (AOCD)preferably reacts with amino compounds under mildreactions in the presence of 4-(dimethylamino)pyridine(base catalyst) in acetonitrile to give the correspondingsensitively fluorescent derivatives with an excitation maximum at
ex 404 nm and an emission maximum at
em 440nm. The labeled derivatives exhibit high stability underreversed-phase conditions. The fluorescence intensitiesof derivatives in various solvents or at different temperatures were investigated. The method, in conjunction witha gradient elution, offers a baseline resolution of thecommon amine derivatives on a reversed-phase C
18column. The LC separation for the derivatized aminesshows good reproducibility with acetonitrile-water including 2.5% DMF as mobile phase. The relative standarddeviations (
n = 6) for each amine derivative are <4.5%.The detection limits (at a signal-to-noise ratio of 3) perinjection were 0.16-12.8 ng/mL. Further research forthe field of application, based on the AOCD amideintermediate as derivatization reagent, for the determination of free amines in real water samples is achieved.