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Fluorometric Assessment of Sulfhydryl Oxidase Activity: Optimization by Response Surface Methodology

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Abstract

Sulfhydryl oxidase was studied using a spectrofluorometric assay. The current protocol operates by using a combination of hemoglobin (HB) and hematin (HT) as a peroxidase mimic to catalyze the H2O2-dependent oxidation of thiamine. The response surface methodology (RSM) is used to optimize the new method. The current method is very accurate, sensitive, and linear up to 200 IU. When compared to the colorimetric method, the method produced a satisfactory correlation. The novel protocol is being used to evaluate asthenospermic patients' and fertile men's seminal sulfhydryl oxidase activity. The current protocol was used to determine reference values for seminal sulfhydryl oxidase activity. Due to the fact the newly developed spectrofluorometric method is more sensitive and precise than other colorimetric methods, and because thiamine is less expensive than other types of probes used in colorimetric and spectrofluorometric methods, it is likely to find widespread use among scientists studying sulfhydryl oxidase activity in biological tissues. The present method's analytical recovery yielded high specific findings.

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Acknowledgements

We thank all of our colleagues for their continuous encouragement and helpful scientific comments.

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Authors and Affiliations

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

    Mahmoud H. Hadwan & Lamia A. Almashhedy

  2. College of Medicine, University of Babylon, Hilla, Iraq

    AbdulRazzaq S. Alsalman & Abdulsamie H. Altaee

  3. Chemistry Department, College of Education, University of Al-Qadisiyah, Ad Diwaniyah, Iraq

    Asad M. Hadwan

Authors
  1. Mahmoud H. Hadwan
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  2. AbdulRazzaq S. Alsalman
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  3. Lamia A. Almashhedy
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  4. Abdulsamie H. Altaee
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  5. Asad M. Hadwan
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Contributions

The authors confirm contribution to the paper as follows: study conception and design: Mahmoud Hadwan; data collection: Abdul Razzaq S. Alsalman, Lamia A. Almashhedy, Abdulsamie H. Alta'ee, Mahmoud H. Hadwan, Asad M. Hadwan; analysis and interpretation of results: Abdul Razzaq S. Alsalman, Lamia A. Almashhedy, Abdulsamie H. Alta'ee, Mahmoud H. Hadwan; draft manuscript preparation: Abdul Razzaq S. Alsalman, Lamia A. Almashhedy, Abdulsamie H. Alta'ee, Mahmoud H. Hadwan, Asad M. Hadwan. All authors reviewed the results and approved the final version of the manuscript.

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Correspondence to Mahmoud H. Hadwan.

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Ethics Committee (University of Babylon/ College of Science/ Iraq), Ref. no.: 3155 Date: 11/9/ 2019.

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Highlights

• A novel technique for detecting the activity of the sulfhydryl oxidase enzyme in biological samples was investigated

• The spectrofluorometric method was applied

• The validation procedure demonstrated that the diagnostic technique works with biological material

• The protocol may be used as a diagnostic tool and is appropriate for research purposes

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Hadwan, M.H., Alsalman, A.S., Almashhedy, L.A. et al. Fluorometric Assessment of Sulfhydryl Oxidase Activity: Optimization by Response Surface Methodology. J Fluoresc 32, 381–388 (2022). https://doi.org/10.1007/s10895-021-02861-7

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  • DOI: https://doi.org/10.1007/s10895-021-02861-7

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