Chromatographic method involving inductively coupled plasma atomic emission spectrometric detection for the study of metal–protein complexes

A chromatographic method has been used to study metal ion–protein complexes. It involves successively a gel filtration technique to separate and distinguish the complexed from the free metallic ions, and a spectrometric technique, inductively coupled plasma atomic emission spectrometry (ICP-AES), which allows us to calculate accurately the concentration of ionic metallic species in solution. In the chromatographic step, we applied a large-zone Hummel and Dreyer method. Thus, fractions can be collected throughout the chromatographic experiment and their metal concentration measured by ICP-AES, at constant and known protein concentration. This method has been tested on the copper complex of bovine serum albumin. Results of our study are in good agreement with previous studies on this complex.