Hadwan Mahmoud Hussein, Rahi Ahed Kamil, Abass Esraa Rafied, Hadwan Asad M, Mohammed Rawa M, Alta'ee Abdulsamie Hassan, Alsalman Abdul Razzaq, Hadwan Muntadher M, Al-Talebi Zainab Abbas
Chemistry Department, College of Science, University of Babylon, Hillah, Babylon Governorate, 51002, Iraq.
Pharmaceutical Chemistry Department, College of Pharmacy, University of Babylon, Hillah, Babylon Governorate, Iraq.
Biometals. 2024 Dec;37(6):1699-1712. doi: 10.1007/s10534-024-00635-9. Epub 2024 Oct 14.
Ferroxidases are enzymes that participate in the iron metabolism of different organisms. They catalyze the oxidation of ferrous iron, Fe⁺, into ferric iron, Fe⁺, 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.
铁氧化酶是参与不同生物体铁代谢的酶。它们催化亚铁离子（Fe⁺）氧化为铁离子（Fe⁺），这在铁稳态和生理功能中至关重要。本研究描述了一种测定血清铜蓝蛋白铁氧化酶活性的新型分光光度法。该方法易于操作；它还具有灵敏、特异和快速的特点。在这种方法中，亚铁离子用作酶的底物，水杨酸或磺基水杨酸作为显色化合物。这些显色剂很容易与铁离子形成有色络合物，但不会与亚铁离子形成。在酶促反应中，铜蓝蛋白铁氧化酶催化亚铁离子氧化为铁离子。然后在形成有色络合物后，通过分光光度法测量铁离子浓度的增加。对于水杨酸，形成的络合物在540nm处有最大吸光度，对于磺基水杨酸，在490nm处有最大吸光度。相比之下，将其与标准方法进行测试，以确定该新方法在测定铁氧化酶活性方面的有效性和可靠性。测定的相关系数为0.99，表明两种方法获得的结果之间有很强的相关性。这种新的分光光度技术提供了一种简单、灵敏、特异和快速的估计铁氧化酶活性的方法。它避免了在该过程中使用浓强酸，并且与标准技术有很好的相关性。这为准确测定生物样品中的铁氧化酶活性提供了一种潜在的替代方法。
