A reversible surface-confined system for potentiometric coulometry studies was fabricated by the deposition of a liquid film on an electrode surface, and the system was used to detect charge accumulation due to the peroxidase (POD)-catalyzed reduction of H2O2. The components of the liquid film were 1-ethyl-3-methylimidazolium bis(nonafluorobutanesulfonyl)imide as the electrolyte, dibutyl phthalate as the medium, and ferrocene as the redox-active mediator. When the thickness of the liquid film was less than that of the diffusion layer in the minimum potential step of the apparatus, a symmetric voltammogram, according to the Nernst equation, was recorded. The reduction of H2O2 by POD was carried out on the liquid-film-modified electrode and studied by potentiometric coulometry. The addition of H2O2 shifts the electrode potential positively, due to the oxidation of ferrocene. The change in the potential is in agreement with the Nernst equation. Thus, potentiometric coulometry, carried out on the liquid-film-modified electrode, enables the determination of the substrate without the requirement of a calibration curve. (C) 2016 Elsevier B.V. All rights reserved.