Abstract

The generation of fast electrochemical response with high sensitivity is a significant parameter for enzymatic biosensors. However, establishing nanocomposite based modified electrode with fast electron shuttling between enzymes and the electrode surface for determination of hydrogen peroxide (H2O2) is highly challenging. Herein, the present work aims to develop second generation horseradish peroxidase (HRP) biosensors using Au nanochains dropcasted over electrochemically reduced graphene oxide-chitosan (ERGO-CHIT), a bio-nanocomposite film modified glassy carbon electrode (GCE) for reduction of H2O2 is demonstrated. The experimental conditions including effect of pH, loading of enzyme on the electrode surface and concentration of redox mediator as electrolyte were optimized. As a result, the HRP/Au/ERGO-CHIT/GCE electrode shows a larger enhancement in cathodic current signal with sharp peak response for reduction of H2O2, which can be attributed to the presence of Au nanochains on the modified electrode which improves the electron transfer between heme (Fe2+/Fe3+) center of enzymes on the electrode and the redox mediator in the electrolyte solution. The presence of HQ redox mediator in the electrolyte solution have provided stable peak response and offered more beneficiary in lowering the operating potential for the detection of H2O2 (-0.1 V), thereby reducing the influence from interference species. From the amperometric measurements, the HRP/Au/ERGO-CHIT/GCE modified electrode revealed high sensitivity (368.01 mu A mM(-1) cm(-2)) and possesses wide linear range (0.01 to 6.31 mM) which is mainly due to the high surface area and conductivity offered by Au nanochains which are located between ERGO and HRP. The resultant HRP/Au/ERGO-CHIT/GCE was found to possess good operational stability, high reproducibility, repeatability with low detection limits (4 mu M), and thus it could be applicable for sensitive and rapid responsive detection of H2O2 in biological, clinical and environmental samples.