Cited by 377Redox biology and oxidative stressCancer and biochemical researchAntioxidant Activity and Oxidative Stress

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Simple spectrophotometric assay for measuring catalase activity in biological tissues

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Hadwan, Mahmoud Hussein·2018·BMC Biochemistry

DOI 10.1186/s12858-018-0097-5PMC PMC6091033

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Abstract

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 3 ) 3 ]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.

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Fig. 1 The spectrophotometric properties of the carbonato-cobaltate(III) complex that correlates with the activity of catalase enzyme. a A carbonato-cobaltate(III) complex. b Decreased color intensity in association with increased catalase activity. c Absorption spectra obtained for the colorimetric products of the present assayCC BY · PubMed Central

Fig. 2 The absorbance of carbonato-cobaltate(III) complex ([Co(CO3)3]Co) as a function of the incubation time of H2O2 alone (●) or H2O2 mixed with catalase enzyme (0.86 U mL−1) (▲)CC BY · PubMed Central

Fig. 3 The catalase activities measured by the carbonato-cobaltate(III) complex method versus those expected for red blood cell homogenate at different dilutionsCC BY · PubMed Central

Fig. 4 The catalase activity of liver and kidney tissues homogenates measured with the carbonato-cobaltate(III) complex method in comparison to that with the dichromate methodCC BY · PubMed Central

Fig. 5 The catalase activity of Escherichia coli and Staphylococcus aureus measured with the carbonato-cobaltate(III) complex method in comparison to that with the Iwase et al. [26]CC BY · PubMed Central

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Authors: Hadwan, M. H.
Affiliations: University of Babylon (IQ)
Corresponding author: Hadwan, Mahmoud Hussein
ORCID: Hadwan, Mahmoud Hussein
Venue: BMC Biochemistry
Publisher: Springer Science and Business Media LLC
Published: 2018-08-03
Volume: 19
Issue: 1
Pages: 7-7
ISSN: 1471-2091 (ISSN-L)
DOI: 10.1186/s12858-018-0097-5
PMC: PMC6091033
OpenAlex: W2885891351
Cited by: 377
References: 26
Citations by year: 2026: 43 · 2025: 77 · 2024: 60 · 2023: 63 · 2022: 51 · 2021: 43 · 2020: 30 · 2019: 9
FWCI: 7.29
Type: journal article
Language: en
Open access: hybrid
doi.org
Licence: cc-by
creativecommons.org/licenses/by/4.0/ (tdm)
Locations: BMC Biochemistry (publishedVersion, cc-by)
Europe PMC (PubMed Central) (publishedVersion, other-oa)
PubMed Central (publishedVersion, cc-by)
PubMed (publishedVersion)
DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cancer and biochemical research
Health Sciences › Medicine › Biochemistry (medical) › Antioxidant Activity and Oxidative Stress
Keywords: catalase activity, spectrophotometric assay, hydrogen peroxide decomposition, erythrocyte lysate, tissue homogenates, erythrocytes, liver tissue, kidney tissue, bacterial catalase
MeSH terms: Bacteria, Catalase, Cobalt, Erythrocytes, Humans, Hydrogen Peroxide, Kidney, Liver, Spectrophotometry, Ultraviolet, Reproducibility of Results, Artifacts, Enzyme Assays

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Simple spectrophotometric assay for measuring catalase activity in biological tissues

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