▎ 摘　　要

The graphene nanosheets and carbon nanospheres mixture (GNS-CNS) was prepared by electrolyzing graphite rob in KNO(3) solution under constant current, which was characterized by TEM, AFM, SEM, FT-IR, XRD, XPS, TGA and UV-vis. The nano-mixture can keep stable in water for more than one month. Based on this kind of mixture material, a novel electrochemical biosensing platform for glucose determination was developed. Cyclic voltammetry of glucose oxidase (GOD) immobilized on GNS-CNS/GCE exhibited a pair of well-defined quasi-reversible redox peaks at -0.488 V (E(pa)) and -0.509V (E(pc)) by direct electron transfer between the protein and the electrode. The charge-transfer coefficient (alpha) was 0.51, the electron transfer rate constant was 2.64 s(-1) and the surface coverage of HRP was 3.18 x 10(-10) mol cm(-2). The immobilized GOD could retain its bioactivity and catalyze the reduction of dissolved oxygen. The glucose biosensor has a linear range from 0.4 to 20 mM with detection limit of 0.1 mM. Moreover, the biosensor exhibits acceptable reproducibility and storage stability. The fabricated biosensor was further used to determine glucose in human plasma sample with the recoveries from 96.83% to 105.52%. Therefore, GOD/GNS-CNS/GCE could be promisingly applied to determine blood sugar concentration in the practical clinical analysis. (C) 2011 Elsevier B.V. All rights reserved.