Mahmoud Hussein Hadwan, Ahed Kamil Rahi, Esraa Rafied Abass, Asad M. Hadwan, Rawa M. Mohammed, Abdulsamie Hassan Alta’ee, Abdul Razzaq Alsalman, Muntadher M. Hadwan, Zainab Abbas Al-Talebi
{"title":"A new spectrophotometric method for measuring ceruloplasmin ferroxidase activity: an innovative approach","authors":"Mahmoud Hussein Hadwan, Ahed Kamil Rahi, Esraa Rafied Abass, Asad M. Hadwan, Rawa M. Mohammed, Abdulsamie Hassan Alta’ee, Abdul Razzaq Alsalman, Muntadher M. Hadwan, Zainab Abbas Al-Talebi","doi":"10.1007/s10534-024-00635-9","DOIUrl":null,"url":null,"abstract":"<div><p>Ferroxidases are enzymes that participate in the iron metabolism of different organisms. They catalyze the oxidation of ferrous iron, Fe<sup>2</sup>⁺, into ferric iron, Fe<sup>3</sup>⁺, which is essential in iron homeostasis and physiological functioning. The present study describes a novel spectrophotometric method of serum ceruloplasmin ferroxidase activity. This method is easy to perform; it is also sensitive, specific, and rapid. In this method, ferrous ions are used as a substrate for the enzyme, with either salicylic acid or sulfosalicylic acid being taken as a chromogenic compound. These chromogens easily form a colored complex with ferric ions but are not formed with ferrous ions. In the enzymatic reaction, the ceruloplasmin ferroxidase enzyme catalyzes the oxidation of ferrous to ferric ions. The resulting increase in ferric ion concentration is then measured spectrophotometrically, following the formation of the colored complex. The complex formed has maximum absorbance at 540 nm in the case of salicylic acid and 490 nm in the case of sulfosalicylic acid. Comparatively, it was tested against the standard method to ascertain the new method’s effectuality and reliability for assaying ferroxidase activity. The determined correlation coefficient amounted to 0.99, showing a strong correlation between the results obtained by the two methods. This new spectrophotometric technique offers a simplified, sensitive, specific, and fast means of estimating ferroxidase activity. It avoids using concentrated strong acids in the procedure and correlates excellently with the standard technique. This sets up a potential alternative for accurately determining ferroxidase activity in biological samples.</p><h3>Graphical abstract</h3>\n<div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":491,"journal":{"name":"Biometals","volume":"37 6","pages":"1699 - 1712"},"PeriodicalIF":4.1000,"publicationDate":"2024-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biometals","FirstCategoryId":"99","ListUrlMain":"https://link.springer.com/article/10.1007/s10534-024-00635-9","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Ferroxidases are enzymes that participate in the iron metabolism of different organisms. They catalyze the oxidation of ferrous iron, Fe2⁺, into ferric iron, Fe3⁺, which is essential in iron homeostasis and physiological functioning. The present study describes a novel spectrophotometric method of serum ceruloplasmin ferroxidase activity. This method is easy to perform; it is also sensitive, specific, and rapid. In this method, ferrous ions are used as a substrate for the enzyme, with either salicylic acid or sulfosalicylic acid being taken as a chromogenic compound. These chromogens easily form a colored complex with ferric ions but are not formed with ferrous ions. In the enzymatic reaction, the ceruloplasmin ferroxidase enzyme catalyzes the oxidation of ferrous to ferric ions. The resulting increase in ferric ion concentration is then measured spectrophotometrically, following the formation of the colored complex. The complex formed has maximum absorbance at 540 nm in the case of salicylic acid and 490 nm in the case of sulfosalicylic acid. Comparatively, it was tested against the standard method to ascertain the new method’s effectuality and reliability for assaying ferroxidase activity. The determined correlation coefficient amounted to 0.99, showing a strong correlation between the results obtained by the two methods. This new spectrophotometric technique offers a simplified, sensitive, specific, and fast means of estimating ferroxidase activity. It avoids using concentrated strong acids in the procedure and correlates excellently with the standard technique. This sets up a potential alternative for accurately determining ferroxidase activity in biological samples.
期刊介绍:
BioMetals is the only established journal to feature the important role of metal ions in chemistry, biology, biochemistry, environmental science, and medicine. BioMetals is an international, multidisciplinary journal singularly devoted to the rapid publication of the fundamental advances of both basic and applied research in this field. BioMetals offers a forum for innovative research and clinical results on the structure and function of:
- metal ions
- metal chelates,
- siderophores,
- metal-containing proteins
- biominerals in all biosystems.
- BioMetals rapidly publishes original articles and reviews.
BioMetals is a journal for metals researchers who practice in medicine, biochemistry, pharmacology, toxicology, microbiology, cell biology, chemistry, and plant physiology who are based academic, industrial and government laboratories.