The complexation of charged compounds by humic acids represents the process of exceptional environmental importance. Nevertheless, traditional methods utilized in the complexation studies do not address the way, how these interactions affect the transport of ions in humic-rich environments. To overcome this dilemma, the diffusion cells technique is proposed as an innovative reactivity mapping technique. Using this method, the diffusion of methylene blue was studied in aqueous solutions and in agarose gels with and without the addition of humic acids. Experimental results clearly illustrate the immobilizing effects of humic acids on the transport of methylene blue in gels. The partitioning of methylene blue at the solution-gel interface and the specific interactions between methylene blue and humic acids is discussed on the basis of experimental data. Effective structural parameters of hydrogels (effective porosity, tortuosity factor) were calculated, as well as some standard diffusion and interaction parameters (diffusion and partition coefficients and apparent equilibrium constants). (C) 2013 Elsevier Ltd. All rights reserved.