Analysis of Amino Acids by High Performance Liquid Chromatography with Pre-column Derivatization of 2-(9-Carbazol)-ethyl-chloroformate

A new chloroformate fluorescence reagent, 2-(9-carbazol)-ethyl -chloroformate(CEOC), is used for the analysis of amino acids. In a one-step procedure, CEOC reacts with amino acids to form stable derivatives. Maximal yields of derivatization are observed with the amount of the derivant reagent being equal to 4 times of amount of amino acids. The method in conjunction with a gradient elution offers a baseline separation on a reversed_phase Hypersil BDS C18 column. The fluorescent derivatives are detected with a fluorescence monitor at λex of 293 nm and λem of 365 nm. The average detection limits reaches 22.8 fmol.