Materials used for biosensor development normally include silicon, glass, and synthetic polymers, however, paper is a practical and cheap option for the reduction of manufacturing costs with a wide range of applications. Paper-based biosensors have been widely produced, yet poorly characterized on the interaction of different type of molecules with its intricate microstructure. In this work, five ink solutions were prepared as model samples to examine their diffusion and imbibition behavior on grade 3MM chromatography paper. Different mathematical models, previously reported for porous matrices, were fitted and results revealed that upward wicking (r2 ≥ 0.90) equations described the experimental data during the initial stage (< 5 s) and yielded similar permeability values to those calculated from the matrix structural properties. The diffusion coefficient was determined up to attaining equilibrium using the diffusion equation in a cylinder element (r2 ≥ 0.90). This study enabled the characterization of the performance from 3MM chromatography paper, by using ink as a surrogate model of small molecules (e.g. mycotoxins) or small colloidal particles.