TiO 2 microspheres with a uniform particle size distribution were synthesized by hydrothermal method. After being sintered, the titania microspheres that are obtained have an average diameter of 5 µm, a surface area of 383.5 m 2 /g, an average pore volume of 0.25 cm 3 /g, and an average pore diameter of 35.9 nm. Normal phase chromatography was separated the mixture of benzene, nitrobenzene and nitro-anisole, three substances were separated well on the titania column. The microspheres possess enough rigidity to withstand high packing pressure and are very useful as a new kind of chromatographic packing material for high performance liquid chromatography (HPLC).