Determination of iron in heme compounds: II. Hemoglobin and myoglobin

A rapid, sensitive assay for hemoprotein iron is described. The procedure is carried out entirely in one vessel, digestion of the protein being effected by perchloric acid-hydrogen peroxide followed by spectrophotometric determination of iron as the ferrous- o-phenanthroline complex. The complex is stabilized by carrying out the assay in 10% pyridine. The method requires minimal quantities of sample, either solid or in aqueous solution, and reproducibility is within ±1%.