Nanostructured ZnO in a Metglas/ZnO/Hemoglobin Modified Electrode to Detect the Oxidation of the Hemoglobin Simultaneously by Cyclic Voltammetry and Magnetoelastic Resonance

In the present work, a nanostructured ZnO layer was synthesized onto a Metglas magnetoelastic ribbon to immobilize hemoglobin (Hb) on it and study the Hb's electrochemical behavior towards hydrogen peroxide. Hb oxidation by H2O2 was monitored simultaneously by two different techniques: Cyclic Voltammetry (CV) and Magnetoelastic Resonance (MR). The Metglas/ZnO/Hb system was simultaneously used as a working electrode for the CV scans and as a magnetoelastic sensor excited by external coils, which drive it to resonance and interrogate it. The ZnO nanoparticles for the ZnO layer were grown hydrothermally and fully characterized by X-Ray Diffraction (XRD), Scanning Electron Microscopy (SEM) and photoluminescence (PL). Additionally, the ZnO layer's elastic modulus was measured using a new method, which makes use of the Metglas substrate. For the detection experiments, the electrochemical cell was performed with a glass vial, where the three electrodes (working, counter and reference) were immersed into PBS (Phosphate Buffer Solution) solution and small H2O2 drops were added, one at a time. CV scans were taken every 30 s and 5 min after the addition of each drop and meanwhile a magnetoelastic measurement was taken by the external coils. The CV plots reveal direct electrochemical behavior of Hb and display good electrocatalytic response to the reduction of H2O2. The measured catalysis currents increase linearly with the H2O2 concentration in a wide range of 25-350 mu M with a correlation coefficient 0.99. The detection limit is 25-50 mu M. Moreover, the Metglas/ZnO/Hb electrode displays rapid response (30 s) to H2O2, and exhibits good stability and reproducibility of the measurements. On the other hand, the magnetoelastic measurements show a small linear mass increase versus the H2O2 concentration with a slope of 152 ng/mu M, which is probably due to H2O2 adsorption in ZnO during the electrochemical reaction. No such effects were detected during the control experiment when only PBS solution was present for a long time.