Ammonia was used as catalyst to synthesize spherical silica particles by Stober process. More details about the effect of ammonia concentration on the silica powders were investigated. With increase of ammonia concentration from 0.05 to 1.75 mol/L, it was found that particle size increased from 0.068 to 0.91 μm and number density of silica particles decreased rapidly from 9242.40 x 10(10) to 4.62 x 10(10)/mL. Besides, the ratio of standard deviation and the particle size decreased with the increase of ammonia concentration. These results were well consistent with prediction of aggregation model. It was proved that ammonia resulted in persistently high pH values of solutions, which were vital to form large silica spheres. In the formation process of silica spheres, solution hydroxyl concentration was reduced, which might be attributed to transfer of negative charge in hydroxyl groups to silica spheres.