Use of a bioreactor consisting of sequentially aligned L-glutamate dehydrogenase and L-glutamate oxidase for the determination of ammonia by chemiluminescence.
A chemiluminometric method for the automated flow injection analysis of ammonia is described. The essence of the invention is the use of a bioreactor consisting of both immobilized L-glutamate dehydrogenase (GLDH) and L-glutamate oxidase (GLXD), which are sequentially aligned in this order in a minicolumn measuring 2.0 X 20 mm. The unidirectional constant flow of liquid through the column reactor minimizes the reversed diffusion of the solutes so that the following sequence of reactions is ensured. Thus, ammonia to be determined is first transformed by GLDH into L-glutamate, which then produces hydrogen peroxide by GLXD. Hydrogen peroxide in the effluent from the column is then determined by its chemiluminescence upon admixing with luminol and potassium ferricyanide. The present method gives linearity of the standard curve for ammonia up to 1.0 mM. It is at least 100 times more sensitive than the conventional method for ammonia assay using ultraviolet absorption measurement.
Murachi
T
T
Department of Clinical Science and Laboratory Medicine, Faculty of Medicine, Kyoto University, Japan.
Tabata
M
M
D000594:Amino Acid Oxidoreductases / Q000378:metabolism*; D000641:Ammonia / Q000032:analysis*; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005969:Glutamate Dehydrogenase / Q000378:metabolism*; D007700:Kinetics; D008163:Luminescent Measurements; D011511:Proteus / Q000201:enzymology; D013302:Streptomyces / Q000201:enzymology
D004800:Enzymes, Immobilized; D000641:Ammonia; D000594:Amino Acid Oxidoreductases; D005969:Glutamate Dehydrogenase; C023061:L-glutamate oxidase