Bound Iron and Unsaturated Iron-binding Capacity of Serum; Rapid and Reliable Quantitative Determination

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Abstract
Siderophilin-bound serum or plasma iron is determined by adjustment of serum sample with concentrated phosphate buffer to a pH value at which constituent proteins remain in solution but siderophilin-bound iron is wholly dissociated and available to combine with a suitable chromogenic agent, (1,1[image]-dipyridine, 2,2[image],2[image]-terpyridine, 4,7-diphenyl-1,10-phenanthroline). Ascorbic acid is used as the iron reductant. The method for determination of iron-free siderophilin involves addition to serum of a known amount of iron in excess of that capable of being bound, followed by direct analysis of serum sample at pH about 8.0 with the chromogenic agent, without acidification or removal of constituent proteins. The difference between the quantity of iron added to serum and that found to be in excess gives the unsaturated iron-binding capacity of the serum.