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 comp lexes. It involves successively a gel filtration technique to separate and distinguish the complexed from the free metallic ions, and a spec trometric technique, inductively coupled plasma atomic emission spectr ometry (ICP-AES), which allows us to calculate accurately the concentr ation of ionic metallic species in solution. In the chromatographic st ep, we applied a large-zone Hummel and Dreyer method. Thus, fractions can be collected throughout the chromatographic experiment and their m etal concentration measured by ICP-AES, at constant and known protein concentration. This method has been tested on the copper complex of bo vine serum albumin. Results of our study are in good agreement with pr evious studies on this complex. (C) 1997 Elsevier Science B.V.