A new kinetic, automated assay to determine the ferroxidase activity of ceruloplasmin

Abstract

A new kinetic and automated assay was developed to determine ceruloplasmin ferroxidase activity. Ferrous ions are turned into ferric ions via catalytic activity of the ferroxidase enzyme. Acetohydroxamic acid, a chromogen, forms a colored complex with ferric ions. This reaction was measured kinetically. Significant and strong correlations were obtained between the new acetohydroxamic method and the p-phenylenediamine oxidase (r = 0.988, p < 0.001), o-dianisidine oxidase (r = 0.981, p < 0.001), norfloxacine oxidase (r = 0.989, p < 0.001) and nephelometric methods (r = 0.861, p < 0.001). This reliable, applicable, user-friendly, and low-priced method can be performed fully automatically or with manual spectrophotometry, and can be used to measure the ferroxidase activity of ceruloplasmin.