Tl ϩ reduction on gramicidin-modified dioleoylphosphatidylcholine-coated mercury electrodes has been studied by the impedance method in different experimental conditions. On one hand, experiments at twelve gramicidin concentrations in 0.1 M KCl solutions have been performed in order to clarify whether radial diffusion plays some role. On the other hand, KCl, KNO 3 , KClO 4 , MgCl 2 , and CaCl 2 solutions have been used as supporting electrolyte to study the influence of ion nature on both the mass transfer and the electron transfer of the process. In all the cases the frequency dependence of the admittance data fits well to a Randles circuit and the Warburg coefficient, , and the irreversibility coefficient, pЈ, can be obtained at every dc potential. However, the potential dependence of can only be explained if a mechanism is assumed, including two chemical steps, one preceding and one following the electron transfer ͑CEC mechanism͒ even at very high gramicidin concentrations. The impedance equations for reactions on partially blocked electrodes have been deduced, but the results do not conform completely to this model, so other explanations are proposed for the origin of the two chemical steps. Moreover, the standard rate constant, k s , and the charge transfer coefficient, ␣, for the electron transfer were obtained from pЈ. These parameters are not sensitive to gramicidin concentration or to the nature of the supporting electrolyte. On the contrary, the mass transfer parameters are strongly dependent on these variables. The behavior can be understood at the light of the mechanisms for ion movement in biomembranes.Several approaches have been proposed for application of the electrochemical methods to the understanding of ion transport and electron transfer through biological membranes. 1 Among them, the use of phospholipid-coated mercury electrodes was initially proposed by Miller 1 and later extensively developed by Nelson. [2][3][4] In this case, the electrode metal is coated with a film consisting in half a bilayer of highly organized phospholipids with the polar heads directed toward the electrolyte solution, 2 contrary to what was observed on gold electrodes. 5 The charge density on these films under different experimental conditions and the electron and proton transfer processes that biologically relevant compounds undergo when they are incorporated in these self-assembled monolayers have been investigated by Moncelli et al. [6][7][8][9][10][11] Incorporation of gramicidin in the phospholipid film allows studying ion transport in an environment similar to the situation of the transport through gramicidin channels in biological membranes. In this respect, the reduction of aqueous Tl ϩ ions on gramicidinmodified phospholipid-coated mercury electrodes is interesting because it involves the transfer of only one electron and the ion has the size and charge adequate to pass through the gramicidin channels. Nelson et al. have extensively studied this system by voltammetry and chronoamperometry under different exp...