OxyFile #221

Spectrophotometric determination of hydrogen peroxide: 
catalase activity and rates of hydrogen peroxide removal 
by erythrocytes.


Author:   Masuoka N; Wakimoto M; Ubuka T; Nakano T;

Source:   Clin Chim Acta 1996 Oct 29; 254(2):101-12

Abstract:

A new method of hydrogen peroxide determination for the 
measurement of catalase activity and rates of hydrogen 
peroxide removal by erythrocytes was described. Hydrogen 
peroxide was determined by converting it to the indamine 
dye with a water-soluble ironporphyrin and measuring the 
absorbance at 590 nm. This method was applied to the assay 
of catalase in hemolysates from human, rat and mouse blood. 
The activities obtained were in agreement with those obtained 
by other methods including UV method. The present method 
was also applied to the determination of rates of hydrogen 
peroxide removal by intact erythrocytes from human subjects, 
rats and mice. Data suggested that normal erythrocytes 
have substantial capacity to remove extracellular hydrogen 
peroxide. From the measurement of catalase activity in 
erythrocytes treated with 3-amino-1,2,4-triazole and rates 
of hydrogen peroxide removal by the erythrocytes, it is 
deduced that rate constants related to the hemoglobin content 
(k/g Hb) for hydrogen peroxide removal by catalase in normal 
and acatalasemic erythrocytes are 42.0 +/- 6.0 and 8.0 
+/- 3.0, respectively.