A new spectrophotometric method for measuring ceruloplasmin ferroxidase activity: an innovative approach

IF 4.1 3区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
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
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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.

Graphical abstract

测量脑磷脂铁氧化酶活性的新分光光度法:一种创新方法。
铁氧化酶是一种参与不同生物体铁代谢的酶。它们催化亚铁(Fe2⁺)氧化成铁素体(Fe3⁺),而铁素体是铁平衡和生理功能所必需的。本研究介绍了一种新型分光光度法测定血清脑磷脂铁氧化酶活性。该方法操作简便,灵敏、特异、快速。在该方法中,亚铁离子被用作酶的底物,水杨酸或磺基水杨酸被用作发色剂。这些致色剂很容易与铁离子形成有色复合物,但不会与亚铁离子形成有色复合物。在酶促反应中,脑磷脂铁氧化酶催化亚铁氧化成铁离子。在有色复合物形成后,用分光光度法测量铁离子浓度的增加。水杨酸形成的复合物在 540 纳米波长处有最大吸光度,磺基水杨酸则在 490 纳米波长处有最大吸光度。为了确定新方法在测定铁氧化酶活性方面的有效性和可靠性,我们将其与标准方法进行了比较试验。测定的相关系数为 0.99,表明两种方法得出的结果之间具有很强的相关性。这种新的分光光度法提供了一种简化、灵敏、特异和快速的估算铁氧化酶活性的方法。它避免了在操作过程中使用浓强酸,并与标准技术有很好的相关性。这为准确测定生物样本中的铁氧化酶活性提供了一种潜在的替代方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Biometals
Biometals 生物-生化与分子生物学
CiteScore
5.90
自引率
8.60%
发文量
111
审稿时长
3 months
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.
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