Elimination of the "chromogen oxidase" activity of bilirubin oxidase added to obviate bilirubin interference in hydrogen peroxide/peroxidase detecting systems

• Published in: Clinical chemistry (Baltimore, Md.) Vol. 31; no. 12; pp. 2007 - 2008
• Main Author: Maguire, GA
• Format: Journal Article
• Language: English
• Published: Washington, DC Am Assoc Clin Chem 01.12.1985; American Association for Clinical Chemistry
• Subjects: 5'-Nucleotidase; Analytical, structural and metabolic biochemistry; Bilirubin; Biological and medical sciences; Catalysis; Chromogenic Compounds - metabolism; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Hydrogen Peroxide - analysis; Indicators and Reagents; Nucleotidases; Oxidation-Reduction; Oxidoreductases; Oxidoreductases - metabolism; Oxidoreductases Acting on CH-CH Group Donors; Peroxidases - analysis; Assay; Technical progress; Hydrogene peroxyde; 5'-Nucleotidase; Enzyme; Clinical biology; Interference; Peroxidase; Application
• ISSN: 0009-9147; 1530-8561
• DOI: 10.1093/clinchem/31.12.2007

The use of bilirubin oxidase to remove interference by bilirubin in hydrogen peroxide/peroxidase detecting systems is hampered by its inherent "chromogen oxidase" activity (its ability to oxidize the chromogens used in the systems). This unwanted activity is greater than 99% inhibited by 0.5 mmol/L cyanide, 97% inhibited by 20 mmol/L azide. At these same concentrations, they inhibit bilirubin oxidase activity by 95% and 73%, respectively. Sequential addition of reagents allows the use of bilirubin oxidase without interference by the chromogen oxidase activity.