Abstract

Hydrogen peroxide (H2O2) is a highly relevant metabolite in many biological processes, including the oral microbiome. To study this metabolite, we developed a 25 μm diameter, highly sensitive, nonenzymatic H2O2 sensor with a detection limit of 250 nM and a broad linear range of 250 nM to 7 mM. The sensor used the synergistic activity of the catalytically active Pt nanoparticles on a high surface area multiwalled carbon nanotube and conducting ionic liquid matrix to achieve high sensitivity (2.4 ± 0.24 mA cm-2 mM-1) for H2O2 oxidation. The unique composite allowed us to miniaturize the sensor and couple it with a Pt electrode (25 μm diameter each) for use as a dual scanning electrochemical microscopy probe. We could detect 65 ± 10 μM H2O2 produced by Streptococcus gordonii (Sg) in a simulated biofilm at 50 μm above its surface in the presence of 1 mM glucose and artificial saliva solution (pH 7.2 at 37 °C). Because of its high stability and low detection limit, the sensor showed a promising chemical image of H2O2 produced by Sg biofilms. We were also able to detect 30 μM H2O2 at 50 μm above the biofilm in the presence of the H2O2-decomposing salivary lactoperoxidase and thiocyanate, which would not otherwise be possible using an existing H2O2 assay. Thus, this sensor can potentially find applications in the study of other important biological processes in a complex matrix where circumstances demand a low detection limit in a compact space.

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