In this study diclofenac sodium (DFS), an analgesic drug has been employed as an effective template for the synthesis of NiO nanostructures. The NiO nanostructures were synthesised using low temperature hydrothermal growth method, both in the presence and absence of the DFS drug. The synthesised nanostructures were studied for their structural, compositional and electrochemical properties using scanning electron microscopy, X-ray diffraction and cyclic voltammetry. The synthesised nanostructures were then utilised for the modification of glassy carbon electrode which were then utilised for the electro-catalytic enzyme free glucose sensing in alkaline media. The competitive experiments suggested that although, both nanostructures possess excellent capability of glucose sensing, the NiO nanoflakes modified electrode was found to be twice as much as sensitive (2584 A mu A mM(-1) cm(-2)) as nanoflowers based electrode (1154 A mu A mM(-1) cm(-2)). The NiO nanoflakes based sensor further demonstrated excellent anti-interference potential in the presence of common interferents like uric acid, ascorbic acid and dopamine. In addition, the successful application NiO nanoflakes based sensor to determine real blood glucose concentration further suggest its feasibility for real sample analysis.