New Spectrophotometric Method for the Assessment of Catalase Enzyme Activity in Biological Tissues

Abstract

Background: Catalase is a vital antioxidant enzyme that dismutates H2O2 into water and molecular oxygen. Many protocols have been developed to measure catalase enzyme activity. Spectrophotometric methods are the most common assays that used to assess catalase enzyme activity. Methods: Because the rate-limiting step during catalase enzyme activity depends upon the dissociation of hydrogen peroxide, the developed assay measures the reaction between a hydroquinone/anilinium sulfate/ammonium molybdate reagent and Unreacted Hydrogen Peroxide, which results in the production of a purple, disubstituted quinone compound with a maximum absorbance value at 550 nm. Results: To clarify the precision of the developed method, the coefficients of variation were determined to be 2.6% and 4.7% for within run measurements and between run measurements, respectively. This method returned results that correlated well (r = 0.9982) with the results returned using the peroxovanadate method to assess catalase enzyme activity. Additionally, we examined the use of the newly developed hydroquinone assay to measure catalase enzyme activity in liver and bacterial homogenate samples. Conclusion: These results demonstrated that this assay can be used for scientific research and routine health applications because it is inexpensive, simple, accurate, and rapid. This method is suitable for use in clinical pathology laboratories because it is simple and produces precise and reproducible results.

Activity of antioxidant enzymes in hepatocytes of mice with lymphoma under the action of thiazole derivative in complex with polymeric nanocarrier

“…Сatalase (CAT) activity was measured by the method of Hamza et al [7] with an absorption wavelength of 410 nm and was expressed in nmoles of H 2 O 2 /min×mg of protein. Superoxide dismutase (SOD) activity was measured by the method of Paoletti et al [17] and was expressed as unit active/min×mg protein.…”

Protective effects of Leuzea uniflora ( Rhaponticum uniflorum ) on the brain mitochondrial function in white rats at hypoxia/reoxygenation

“…The energetic state of the rats' brain was estimated taking into account the content of adenosine triphosphate (ATP), lactic and pyruvic acids contents and their ratio (Patergnani et al 2014). The state of pro-and antioxidant systems was evaluated according to the concentration of malonic dialdehyde (MDA) (Kikugawa et al 1992), activity of superoxide dismutase (SOD) (Boriskin et al 2019), catalase (Hamza and Hadwan 2020), pyruvate kinase (PK) (Osterman at al. 1973), glutathione peroxidase (GP) and glutathione reductase (GR) (Pinto and Bartley 1969), as well as according to the content of reduced glutathione (GSH) (Shaik and Mehvar 2006) in the brain homogenate.…”