Positively-charged hierarchical PEDOT interface with enhanced electrode kinetics for NADH-based biosensors

Poly(ethylenedioxythiophene) (PEDOT) has attracted considerable attention as an advanced electrode material for electrochemical sensors and biosensors, due to its unique electrical and physicochemical properties. Here, we demonstrate the facile preparation of a positively-charged and hierarchical micro-structured PEDOT electrochemical interface with enhanced electrode kinetics for the electrooxidation of NADH. Processable PEDOT colloidal microparticles (PEDOT CMs) were synthesised by template-assisted polymerisation and were then utilised as building blocks for the fabrication of hierarchically-structured electrodes with a larger accessible electroactive surface (2.8 times larger than that of the benchmark PEDOT:PSS) and inter-particle space, thus improving electrode kinetics. The intrinsic positive charge of the PEDOT CMs further facilitated the detection of negatively-charged molecules by electrostatic accumulation. Due to the synergistic effect, these hierarchically structured PEDOT CMs electrodes exhibited improved NADH electrooxidation at lower potentials and enhanced electrocatalytic activity compared to the compact structure of conventional PEDOT:PSS electrodes. The PEDOT CMs electrodes detected NADH over the range of 20-240 mu M, with a sensitivity of 0.0156 mu A/mu M and a limit of detection of 5.3 mu M. Moreover, the PEDOT CMs electrode exhibited a larger peak separation from the interferent ascorbic acid, and improved stability. This enhanced analytical performance for NADH provides a sound basis for further work coupling to a range of NAD-dependent dehydrogenases for applications in biosensing, bio-fuel cells and biocatalysis.