In this work, the diffusion of thiocholine ion into an enzyme-loaded polypyrrole film was evaluated by different methods, and the results were compared to identify the most suitable method. The enzyme-loaded polypyrrole film was coated with a thin layer of gelatin and gluteraldehyde so as to prevent enzyme leaching. Diffusion coefficients under normal and prepolarized conditions were calculated by five different methods, namely, the Cottrell method, the method of Peerce and Bard, the theoretical impedance model, the electrochemically stimulated conformational relaxation (ESCR) method, and the direct impedance measurement method.
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