Simple spectrophotometric assay for measuring catalase activity in biological tissues

• Published in: BMC biochemistry Vol. 19; no. 1; p. 7
• Main Author: Hadwan, Mahmoud Hussein
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 03.08.2018; BioMed Central
• Subjects: Antioxidants; Bacteria; Biochemical assays; Biological activity; Biological research; Catalase; Chemical properties; Cobalt; Electrodes; Enzymes; Hydrogen peroxide; Methodology; Methods; Oxidation; Peroxidase; Spectrophotometry; Tissues; Tissues (Anatomy); Carbonato-cobaltate (III) complex; Catalase activity; Bicarbonate; Hydrogen peroxide; Cobalt
• ISSN: 1471-2091; 1471-2237; 1471-2091
• DOI: 10.1186/s12858-018-0097-5

The details of a precise, accurate, and sensitive spectrophotometric method for measuring catalase activity are presented here. The assay was established for biological samples and depends on the rapid formation of a stable and colored carbonato-cobaltate (III) complex. Samples exhibiting catalase activity are incubated with hydrogen peroxide solution for 2 min prior to rapid mixing of the incubation enzymatic reaction mixture with cobalt-bicarbonate reagent, which assesses non-reacting hydrogen peroxide. Catalase activity is always directly proportional to the rate of dissociation of hydrogen peroxide. Hydrogen peroxide acts to oxidize cobalt (II) to cobalt (III) in the presence of bicarbonate ions; this process ends with the production of a carbonato-cobaltate (III) complex ([Co (CO ) ]Co). The formed end product has two maximum absorbance peaks: 440 nm and 640 nm. The 440-nm peak has been utilized for assessing catalase activity. The catalase activity results of the current method for erythrocyte lysate homogenates were computationally identical to those of the dichromate method (r = 0.9950). The coefficient of variation was calculated to determine the imprecision of the current assay. The within-run and between-run results were 2.96 and 3.83%, respectively. This method is appropriate for analyzing bacteria, red blood cells and liver and kidney tissue homogenates.