ZnFe2O4 nanoparticles were prepared by hydrothermal reduction approach. A considerable amount of α-Fe2O3 was segregated in the as-synthesized sample, which diffused into the tetrahedral and octahedral sites of the ZnFe2O4 spinel structure with increasing the annealing temperature. The introduction of Fe3+ into the tetrahedral positions was observed by Mössbauer spectra. Magnetization measurements showed an unusual ferrimagnetic behavior of the ZnFe2O4 phase, even at room temperature, confirming the introduction of Fe3+ into the tetrahedral sites of the spinel structure. Catalytic activity measured in the oxidative dehydrogenation of 1-butene reaction increased with increasing annealing temperature, indicating that those interactions of Fe3+ in tetrahedral and octahedral positions also promotes the activity and selectivity to butadiene formation.