In this work, NiO nanosheets have been successfully synthesized via a facial hydrothermal route and it was found to be an excellent catalyst for non-enzymatic glucose sensory application. The NiO nanosheets in fcc structure were examined by transmission electron microscopy (TEM), field emission scanning electron microscopy(FESEM), X-ray diffraction (XRD) and selected area electron diffraction (SEAD). The thickness of NiO nanosheets were found to be 40–50nm. This catalyst was then dropcasted on a glassy carbon electrode (GCE) for glucose sensory purposes. The electrochemical measurements were undertaken using both voltammetric method and amperometric method. The former offer a wide concentration range for glucose (up to 2.2mM) but with relatively low sensitivity, whereas the latter showed excellent sensitivity with 400.29μAmM−1cm−2 and 1μM detection limit. Thus the as-synthesized NiO nanosheets demonstrated potential application in non-enzymatic sensory of glucose.