Amperometric Hydrogen Peroxide Biosensor Based on Immobilization of Hemoglobin on a Glassy Carbon Electrode Modified with Fe3O4/Chitosan Core-Shell Microspheres

Abstract

Novel magnetic Fe3O4/chitosan (CS) microspheres were prepared using magnetic Fe3O4 nanoparticles and the natural macromolecule chitosan. Then, using an easy and effective hemoglobin (Hb) immobilization method, an innovative biosensor with a Fe3O4/CS-Hb-Fe3O4/CS “sandwich” configuration was constructed. This biosensor had a fast (less than 10 s) response to H2O2 and excellent linear relationships were obtained in the concentration range of 5.0 × 10-5 to 1.8 × 10-3 M and 1.8 × 10-3 to 6.8 × 10-3 M with a detection limit of 4.0 × 10-6 M (s/n = 3) under the optimum conditions. The apparent Michaelis-Menten constant Km was 0.29 mM and it showed the excellent biological activity of the fixed Hb. Moreover, the biosensor had long-time stability and good reproducibility. The method was used to determine H2O2 concentration in real samples.