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A precise and simple method for measuring catalase activity in biological samples

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Abstract

The current study describes a simple, precise, and accurate protocol for measuring the activity of catalase, which controls the rate-limiting step of the dissociation of hydrogen peroxide reactions. The current protocol assesses catalase activity by incubating catalase samples with suitable concentrations of hydrogen peroxide dissolved in a phosphate buffer (pH 7.4). After the incubation period, a working solution that contained vanadate (V) and pyridine-2,6-dicarboxylic acid was added to stop the enzymatic reaction. The reaction between undissociated hydrogen peroxide and the added reagent forms a stable orange-colored chelate complex known as oxo-peroxo-pyridine-2,6-dicarboxylato-vanadate (OPDV) that demonstrates maximum absorbance at 435 nm. To optimize the formation of the method (the OPDV-CAT assay), we applied the Box–Behnken design (BBD) by utilizing the response surface methodology (RSM) as an index of precision of the assay. This novel method was validated against a Bland–Altman plot analysis of catalase activity using the carbonato-cobaltate method in matched samples. The comparison between the two methods resulted in a correlation coefficient equal to 0.9968, demonstrating that the new method is just as effective as the reference method.

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Acknowledgements

We thank all of our colleagues, especially Dr. Muhannad Musa Karim and Dr. Jassim Al-Shammari for their continuous encouragement and helpful scientific comments.

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Authors and Affiliations

  1. Chemistry Department., College of Science, University of Babylon, p.o. 51002, Hilla City, Babylon Governorate, Iraq

    Mohammed A. Kadhum & Mahmoud H. Hadwan

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  1. Mohammed A. Kadhum
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  2. Mahmoud H. Hadwan
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Correspondence to Mahmoud H. Hadwan.

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Kadhum, M.A., Hadwan, M.H. A precise and simple method for measuring catalase activity in biological samples. Chem. Pap. 75, 1669–1678 (2021). https://doi.org/10.1007/s11696-020-01401-0

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