In this work, we prepared spinel-type NiCoO (NCO) nanopowders as a low-cost and sensitive electrochemical sensor for nonenzymatic glucose detection. A facile and simple chemical bath method to synthesize the NCO nanopowders is demonstrated. The effect of pH and annealing temperature on the formation mechanism of NCO nanoparticles was systematically investigated. Our studies show that different pHs of the precursor solution during synthesis result in different intermediate phases and relating chemical reactions for the formation of NCO nanoparticles. Different morphologies of the NCO depending on pHs are also discussed based on the mechanism of growth. Electrochemical performance of the prepared NCO was characterized towards glucose, which reveals that sensitivity and selectivity of the NCO are significantly related with the final microstructure combined with constituent species with multiple oxidation states in the spinel structure.