USE OF THE ORTHO-PHENYLENEDIAMINE FLUORESCENCE SYSTEM IN THE ENZYMATIC ASSAY OF SERUM URIC-ACID

A manual enzymatic method is described for sensitive fluorometric determination of uric acid in human serum. This method is based on an enzymatic reaction with uricase to form hydrogen peroxide from uric acid and the following oxidation of o-phenylenediamine with peroxidase and hydrogen peroxide for the production of a fluorescence compound. The specificity and the selectivity in the method are due to the uricase reaction and the fluorometry, respectively. The formed fluorescence in the reaction mixture is measured at 410 nm (an excitation) and 550 nm (an emission). This enzymatic method can determine uric acid at 30-1000-mu-M, with a between-assay relative standard deviation of 4.35% or less. A good correlation is obtained between the present method and the colorimetric kit method.