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

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.