Renewable Reagentless Hydrogen Peroxide Sensor Based on Direct Electron Transfer of Horseradish Peroxidase Immobilized on Colloidal Gold-modified Electrode

A novel renewable reagentless hydrogen peroxide (H(2)O(2)) sensor based on the direct electron transfer of horseradish peroxidase (HRP) is proposed. The direct electrochemistry of HRP immobilized on a colloidal gold-modified carbon paste electrode (Au-CPE) was investigated using electrochemical methods. The immobilized HRP displayed a pair of redox peaks in 0.1M phosphate buffer (PB), pH 7.0, with a formal potential of -0.346 V. The response showed a surface-controlled electrode process with an electron transfer rate constant of 6.04+/-0.18s(-1) determined in the scan rate range from 120 to 500 mV/s. The biosensor displayed an excellent electrocatalytic response to the reduction of H(2)O(2) without the aid of an electron mediator. The sensor surface could be renewed quickly and reproducibly by a simple polish step. The calibration range of H(2)O(2) was 0-0.3mM with linear relation from 0.48 to 50 microM and a detection limit of 0.21 microM at 3 sigma. The response showed Michaelis-Menten behavior at higher H(2)O(2) concentrations. The K(app)(M) value of HRP at HRP-Au-CPE was determined to be 3.69+/-0.71 mM.