Determination of Amines by high performance liquid chromatography with a pre-column derivatization and fluorescence detection

2-(9-Carbazole) ethyl chloroformate (CEOC), a novel fluorescence derivatization reagent, has been firstly developed for the determiantion of amines. In a one-step procedure, CEOC reacts with the amines to form stable derivatives. Maximum yield is observed with a molar reagent excess of approximately 3 to 4 times for each of the amines. The method in conjunction with a gradient elution offers a baseline separation on a reversed-phase Hypersil BDS C-18 column. Mobile phase: A = acetonitrile/water (20/80, V/V); B = acetonitrile/water (95/5, V/V) flow rate = 1.0 mL/min; column temperature was set at 35 degreesC. The fluorescent derivatives are detected at lambda(ex)/lambda(em) = 293/365 nm. It was proved that CEOC-high performance liquid chromatography (HPLC) not only had excellent precision and accuracy but also high tolerance of the matrix of samples (including salt), therefore the method is suitable for the determination of biological materials, foods and feeds.