The Bilirubin Interferences in the Peroxidase-Coupled Spectrophotometric Determination of Hydrogen Peroxide

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In determining hydrogen perokide in any reagent system in which peroxidase is present, it is noted that presence of bilirubin jeopardizes the results in two ways, firstly in chemical reaction and secondary in physical manifestation. The mechanisms of bilirubin interferences in above procedure are as follows: (1): Bilirubin as a hydrogen donor of peroxidase interferes with the peroxidase reaction. (2): Overlapping of the spectra, namely, for bilirubin and for chromophore in the peroxidase reaction is commonly obserbed in determining hydrogen peroxide. To minimize the bilirubin interferences as in (1) and (2) in the peroxidase reaction syst ems with 4-aminoantipyrine (4AA)-phenol or 4AA-diethylaniline (DEA) as the chromophore, investigations were carried out. In the peroxidase reaction system with AA4-phenol, the chemical interferences descri bed in (1) can be minimized by following procedures;(a) decreasing the pH value of pho sphate buffer,(b) decreasing the concentration of phosphate buffer,(c) decreasing the pero xidase activity,(d) decreasing the concentration of 4AA,(e) increasing the concentration of phenol. In the peroxidase reaction system with 4AA-DEA, the chemical interferences described in (1) can be minimized by decreasing the pH value and/or the concentration of phosphate buffer. The spectral interferences described in (2) can be averted by decreasing the pH value of phosphate buffer and using longer than 500nm wavelength in measurement.
Japan Society of Clinical Chemistryの論文

The Bilirubin Interferences in the Peroxidase-Coupled Spectrophotometric Determination of Hydrogen Peroxide

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