Abstract
A capillary electrophoretic (HPCE) method that can be used to quantitatively determine trace amounts of iron has been developed and applied to determine the iron level in human serum. After precipitation of serum proteins, Fe(III) in the serum is reduced to Fe(II) with hydroxylamine hydrochloride, and a stable Fe(II)-1,10-phenanthroline complex is formed by adding 1,10-phenanthroline to the supernatant containing 2.5 m
M ammonium acetate-acetic acid at pH 5.0. The Fe(II)-1,10-phenanthroline complex, [Fe(C
12H
8N
2)
3]
2+, has a very strong absorbance at 270 nm (with a molar absorptivity of 9.2 · 10
4. By measuring the absorbance of [Fe(C
12H
8N
2)
3]
2+ at 270 nm, the iron level in human serum can be precisely quantified. The interference from copper, a major interference in serum, can be totally eliminated due to the complete separation of [Fe(C
12H
8N
2)
3]
2+ and the Cu(II)-1,10-phenanthroline complex. In addition, other problems that usually occurred with conventional spectrophotometric methods, such as co-precipitation and occlusion of iron during sample pretreatment, are significantly minimized due to the ability to wash the precipitate and the higher detection sensitivity. With this method, a single drop (10 μl) of serum would be sufficient to determine the serum iron concentration. The method is reliable, sensitive, rapid and reproducible. Thus it is highly suitable for use in the clinical laboratory.