Liquid chromatographic separation and stereoselective detection of L- 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 dichlorophenosulphonyl 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.
Domínguez
E
E
Department of Applied Microbiology, University of Lund, Sweden.
Marko-Varga
G
G
Carlsson
M
M
Gorton
L
L
D000594:Amino Acid Oxidoreductases; D000596:Amino Acids / Q000302:isolation & purification*; D001391:Azo Compounds / Q000032:analysis; D002384:Catalysis; D002853:Chromatography, Liquid; D004800:Enzymes, Immobilized; D005976:Glutaral / Q000032:analysis; D007202:Indicators and Reagents; D010783:Photometry; D013237:Stereoisomerism
D000596:Amino Acids; D001391:Azo Compounds; D004800:Enzymes, Immobilized; D007202:Indicators and Reagents; D000594:Amino Acid Oxidoreductases; D005976:Glutaral