The Micro Estimation of Serum Iron and Iron-binding Capacity in Normals and in Disease

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Abstract
Only 0.2 ml. of serum is required. HC1 liberates the serum Fe which is allowed to react with o-phenanthroline in the presence of Na hydrosulfite after removal of protein with trichloroacetic acid. For estimation of the total Fe-binding capacity, a sufficient amt. of Fe is added to saturate the Fe-binding beta-globulin component and the excess is allowed to react with phenanthroline in the presence of Na hydrosulfite. Addition of HC1 liberates the Fe from Fe-globulin compound but not from the Fe-phenanthroline complex, which is precipitated,together with all the proteins, by trichloroacetic acid. The liberated Fe represents the total Fe-binding capacity of the serum and is estimated as for serum Fe. Free hemoglobin up to 140 mg./100 ml. of serum, if converted to carboxy-hemoglobin, will not affect the estimation. The mean, serum Fe level and percentage saturation were higher in normal adult males than in females, but they were diminished in hypochromic anemia. The percentage saturation is nearly 100% in hemochromatosis.