LIQUID-CHROMATOGRAPHIC SEPARATION AND STEREOSELECTIVE DETECTION OF L-AMINO AND D-AMINO ACIDS WITH CATALYTIC REACTION DETECTION USING IMMOBILIZED ENZYMES

A post-column LC detection system is described for the stereoselective detection of L- and D-amino acids. The effluent of the LC column passes an immobilized enzyme reactor (IMER) containing either L- or D-amino acid oxidase. The hydrogen peroxide formed in this reactor by the oxidation of the amino acids is then transported to a second IMER containing horse-radish peroxidase. With the addition of 4-aminophenazone and dichlorophenolsulphonyl chloride to the carrier, the hydrogen peroxide is reacted to form a red-coloured complex which is detected in a flow-through photometric cell at 514 nm. Applications to the analysis of amino acids in bovine and human sera are described.