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

Author(s):  
Mahmoud H. Hadwan ◽  
AbdulRazzaq S. Alsalman ◽  
Lamia A. Almashhedy ◽  
Abdulsamie H. Altaee ◽  
Asad M. Hadwan
2021 ◽  
Author(s):  
mahmoud hussein hadwan ◽  
Abdul Razzaq S. Alsalman ◽  
Lamia A. Almashhedy ◽  
Abdulsamie H. Alta'ee ◽  
Asad M. Hadwan

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 is used to optimize the new method (RSM). 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 SHO 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 SHO activity in biological tissues. The present method's analytical recovery yielded high specific findings.


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