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For the quantitative determination of iron, total iron-binding capacity in human serum. The iron content of the human body may be divided into three classes: iron in storage, iron in use, and iron in transport. It is of fundamental importance to note that a specimen should be analyzed for both iron and iron binding capacity because of the need for both values in the differential diagnosis of various types of anemia and liver diseases. The iron in serum is dissociated from its Fe (III) - transferrin complex by the addition of an acidic buffer containing hydroxylamine. This addition reduces the Fe (III) to Fe (II). The chromogenic agent, Ferene, forms a highly colored Fe (II) - complex that is measured photometrically at 560 nm. The unsaturated iron binding capacity (UIBC) is determined by adding Fe (II) iron to serum so that they bind to the unsaturated iron binding sites on transferrin. The excess Fe (II) ions are reacted with Ferrozine to form the color complex, which measured photometrically. The difference between the amount of Fe (II) added and the amount of Fe (II) measured represents the unsaturated iron binding. The total iron binding capacity (TIBC) is determined by adding the serum iron value to the UIBC value.
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