ABSTRACT
The step in the determination of the haemoglobin absorption spectra of any animal species is to measure, with the highest attainable accuracy, the absorptivity of a single derivative at a single wavelength. The absorbance at 540 nm as haemiglobincyanide, of the stroma-free haemoglobin solution of which the haemoglobin concentration had been determined on the basis of the iron content, was measured. After colour developement, measure the absorbance of the solutions at 520 nm and use the ensuing calibration line to calculate the iron concentration of the haemoglobin stock solution. For the preparation of the haemoglobin stock solutions, however, differences in procedure were unavoidable, mainly because of differences in the solubility of haemoglobin. Since haemoglobin monomer contains only one atom of iron, the substance concentration of iron in the haemoglobin stock solution is equal to the substance concentration of total haemoglobin.
