The annealing under different temperatures was performed on boron-doped nanocrystalline diamond films synthesized by hot filament chemical vapor deposition (HFCVD). The effects of annealing on the microstructure and electrochemical properties of films were systematically investigated. The results show that there are four peaks at 1157,1346,1470 and 1555 cm-1 in Raman spectra of the unannealed sample. When the films were annealed at temperatures above 800 ℃, there are only two peaks of D and G band, indicating that the hydrogen in grain boundaries significantly decreased. The area-integrated intensity ratio of D band to G band (ID/IG) reaches minimum value, revealing that the cluster number or cluster size of sp2 phase was reduced. The G peak position shifts to lower wave number, indicating an decrease in the ordering of graphitic component. The electrode exhibits the widest potential window and the highest oxygen evolution potential, and the quasi-reversible reaction occurs on the surface of the samples. The D peak is quite sharp and its intensity increases when the sample was annealed at 1000 ℃. The ID/IG value attains to the maximum value and the G peak position clearly shifts to higher value. The electrode exhibits the narrowest potential window and the lowest oxygen evolution potential, and the reversible electrochemical reaction occurs in the surface of the sample. The above results reveal that the cluster number or cluster size of sp2 phase, the amounts of trans-polyacetylene related to hydrogen in the grain boundaries, the disordering of graphitic components and the boron diffusion in the nanocrystalline diamond films give contributions to the complex change in electrochemical properties of the films with the annealing temperature increasing.