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

Mohammed A. Kadhum and Mahmoud H. Hadwan

Chemistry Department., College of Science, University of Babylon, Hilla City, Iraq

 

E-mail: mahmoudhadwan@gmail.com

Received: 16 July 2020  Accepted: 20 October 2020

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.

Graphic abstract

Keywords: Box–Behnken design; Catalase activity; Pyridine-2,6-dicarboxylic acid; Response surface methodology; Spectrophotometry; Vanadate (v)

Full paper is available at www.springerlink.com.

DOI: 10.1007/s11696-020-01401-0

 

Chemical Papers 75 (4) 1669–1678 (2021)

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