Determination of the change in the Gibb's free energy from the adsorption isotherm associated with the interaction between a biomolecule and an ion-exchange resin is often achieved by assuming that a Langmuirean model prevails. However, the adsorption of horse heart cytochrome c onto the tentacle-type cation exchanger LiChrospher 1000 SO-3 at pH 4.00 showed an isotherm of rectangular form. In this case the Langmuirean model is not applicable. In this paper, we propose an alternative way to deal with this situation, whereby the adsorption capacity of the adsorbent with a defined protein sample is studied as a function of displacing-ion concentration. The experimental conditions over defined ranges are then selected in order to relate this function to the change in the Gibb's free energy for the interaction between the protein and the ion exchanger. Additional comments about the general utility of the on-line adsorption vessel system employed to determine the adsorption isotherms are also made.