New ultraviolet (340 nm) method for assay of uric acid in serum or plasma.
We propose a novel enzymatic method for assay of uric acid at 340 nm, which eliminates several disadvantages of both the colorimetric and enzymatic methods now in common use. Here, uric acid is catalytically oxidized to allantoin and hydrogen peroxide. The peroxide is reacted with ethanol in the presence of catalase to form acetaldehyde and water, and the acetaldehyde is reduced by NADH in the presence of alcohol dehydrogenase to ethanol. The decrease in absorbance at 340 nm caused by oxidation of NADH is directly proportional to the concentration of uric acid in the sample. Measurement of the change in absorbance between 20 and 200 s eliminates the need for a serum blank measurement. Absorbance and concentration are linearly related to 120 mg of uric acid per liter. The new method was compared with the uricase method in which decomposition of uric acid at 293 nm is directly measured. The results for the 47 patients' sera so examined can be expressed by the linear equation y340 = 1.0078x293 + 0.122 (r = 0.9984).
D000429:Alcohol Oxidoreductases; D002374:Catalase; D005069:Evaluation Studies as Topic; D006801:Humans; D008722:Methods; D010949:Plasma / Q000032:analysis; D013056:Spectrophotometry, Ultraviolet; D014503:Urate Oxidase; D014527:Uric Acid / Q000097:blood*
D014527:Uric Acid; D000429:Alcohol Oxidoreductases; D002374:Catalase; D014503:Urate Oxidase