The exploration of novel nanomaterials employed as substrate to construct glucose biosensors is still of significance in the field of clinical diagnosis. In this work, NiO/Mn2O3 nanoflake arrays were synthesized by hydrothermal approach in combination with calcination process. As-prepared NiO/Mn2O3 nanoflake arrays were utilized to construct electrochemical biosensors for glucose detection. NiO/Mn2O3 nanoflake arrays were investigated systematically by scanning electron microscopy (SEM), X-ray diffractionmeter (XRD), transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS) and energy dispersive X-ray spectroscopy, the formation mechanism of NiO/Mn2O3 nanoflake arrays was proposed. As-prepared glucose biosensors based on NiO/Mn2O3 nanoflake arrays were characterized by cyclic voltammgrams and chronoamperometry. The results indicated that glucose biosensors based on optimized NiO/Mn2O3 nanoflake arrays exhibited a high sensitivity of 167.0 mu A mM(-1) Cm-2 and good anti-interference ability, suggesting the NiO/Mn2O3 nanoflake arrays are an attractive substrate for the construction of oxidase-based biosensors. (C) 2017 Elsevier B.V. All rights reserved.