In this paper, the synthesis of nickel ferrite and its use for voltammetric determination of uric acid (UA) are presented. Nickel ferrite was synthesized via a hydrothermal process using spherical carbons as hard template followed by calcination at 500 °C. It was found that iron and nickel compositions in nickel ferrite can be controlled by the initial Fe/Ni molar ratio. The stoichiometric nickel ferrite (NiFe2O4) with hollow spherical morphology was obtained from a reaction mixture with Fe/Ni molar ratio of around 1.2–1.5. Glassy carbon electrode modified with nickel ferrite was employed to quantitatively determine UA by different pulse voltammetric method. Under the optimum conditions, the anodic peak current was linearly proportional to UA concentration in the range of 0.398 to 6.761 μM. The detection limit (3σ) was found to be 0.15 μM. The proposed method has been employed to determine UA in human urine samples with acceptable recoveries of 95.15–104.8%. On the other hand, the results obtained from this method were also compared with that from standard HPLC method, showing no statistical difference.