Fluorimetry as a Simple and Sensitive Method for Determination of Catalase Activity

Saeed Naazeri; Mosayeb Rostamian; Bahram Yaghmaei; Mehdi  Hedayati

Zahedan Journal of Research in Medical Sciences

The Official Journal of Zahedan University of Medical Sciences

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Fluorimetry as a Simple and Sensitive Method for Determination of Catalase Activity

Authors:

Saeed Naazeri¹,

Mosayeb Rostamian²,

Bahram Yaghmaei³,

Mehdi Hedayati^4,*

1Department of Clinical Biochemistry, Faculty of Medicine, Shahid Beheshti Univer sity of Medical Sciences, Tehran, Iran

2Biotechnology and Biochemistry Departments, Pasteur institute of Iran , Tehran, Iran

3Department of Clinical Biochemistry, Faculty of Medicine, Shahid Beheshti Univer sity of Medical Sciences , Tehran, Iran

4Cellular & Molecular Endocrine Research Center Research Institute for Endocrine Sciences Shahid Beheshti University of Medical Science, Tehran, Iran

Zahedan Journal of Research in Medical Sciences:Vol. 16, issue 2; 64-67

Published online:Jun 29, 2013

Article type:Research Article

Received:Jul 12, 2012

Accepted:Oct 10, 2012

How to Cite:Saeed NaazeriMosayeb Rostamian Bahram YaghmaeiMehdi HedayatiFluorimetry as a Simple and Sensitive Method for Determination of Catalase Activity.16(2):64-67.

Abstract

Background: Catalase enzyme plays an important role in the anti-oxidation defense of body so it is important to measure its activity. Nowadays catalase activity measurement is performed by expensive imported kits in various scientific fields. The purpose of this study was to design a sensitive fluorimetry method for measuring catalase activity with improved sensitivity, accuracy and speed.

Materials and Methods: In this study, the reaction of hydrogen peroxide with peroxidase (as a reaction accelerator) was used in fluorimetry for catalase activity measuring in serum samples in order to increase the sensitivity of the assay. The sensitivity and intra- and inter-assay accuracy, verification test, recovery and parallelism tests, comparison method and correlation and coherence investigation methods were also performed. In order to increase the accuracy and speed of reading, the assay was performed in microplates and reading was done in fluorimetry plates.

Results: The percentage of intra- and inter-assay variation coefficients were measured 3.8-6.6 % and 4.1-7.3%, respectively. Comparison of the results of mentioned method for 50 serum samples with common colorimetric method showed a good correlation (0.917). In assessing the accuracy, the recovery percent was obtained 91% to 107%. The test sensitivity was measured 0.02 IU/ml.

Conclusion: The fluorimetry method by microplate reading has a sufficient precision, accuracy and efficiency for catalase activity measuring as well as speed of measurement. Thus it can be an alternative method to conventional imported colorimetric methods.

Keywords

Fluorimetry

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