To minimize background interference in electrochemical enzymatic biosensors employing electron mediators, it is essential for the electrochemical oxidation of electroactive interfering species (ISs), such as ascorbic acid (AA), to proceed slowly, and for the redox reactions between electron mediators and ISs to occur at a low rate. In this study, we introduce a novel combination of a working electrode and an electron mediator that effectively mitigates interference effects. Compared to commonly used electrodes such as Au, glassy carbon, and indium tin oxide (ITO), boron-doped diamond (BDD) electrodes demonstrate significantly lower anodic current (i.e., lower background levels) in the presence of AA. Additionally, menadione (MD) exhibits notably slower reactivity with AA compared to other electron mediators such as Ru(NH3)63+, 4-amino-1-naphthol, and 1,4-naphthoquinone, primarily due to the lower formal potential of MD compared to AA. This synergistic combination of BDD electrode and MD is effectively applied in three biosensors: (i) glucose detection using electrochemical-enzymatic (EN) redox cycling, (ii) glucose detection using electrochemical-enzymatic-enzymatic (ENN) redox cycling, and (iii) lactate detection using ENN redox cycling. Our developed approach significantly outperforms the combination of ITO electrode and MD in minimizing IS interference. Glucose in artificial serum can be detected with detection limits of ~ 20 μM and ~ 3 μM in EN and ENN redox cycling, respectively. Furthermore, lactate in human serum can be detected with a detection limit of ~ 30 μM. This study demonstrates sensitive glucose and lactate detection with minimal interference, eliminating the need for (bio)chemical agents to remove interfering species.
Keywords: Ascorbic acid; Boron-doped diamond electrode; Electrochemical biosensor; Interfering species; Menadione; Redox cycling.
© 2024. The Author(s), under exclusive licence to The Japan Society for Analytical Chemistry.