Titanium dioxide (TiO(2)) nanoparticles (NPs) are widely used as an important kind of biomaterials due to their large surface area, enhanced chemical reactivity and easy penetration into cells. Nano TiO(2) with pure anatase phase was successfully prepared by solvothermal method. Its particle size was about 21 nm, while the larger specific surface area of TiO(2) was 77.43 m(2)/g. The interaction of colloidal TiO(2) with human serum albumin was studied by using absorption spectra and fluorescence spectra. The apparent binding constants (K) were 345.780 × 10(5), 4.376 × 10(5), 0.035 × 10(5) at 298, 303 and 308 K, respectively. In addition, the number of binding sites (n) was gradually decreased with the increase of temperature, which indicated that the quenching mechanism of albumin by colloidal TiO(2) was static fluorescence quenching process. Based on fluorescence resonance energy transfer, the energy transfer efficiency (E) and critical transfer distance (r(0)) between donor (human serum albumin, HSA) and acceptor (colloidal TiO(2)) were calculated to be 0.862 and 6.244 nm, which suggested that non-radiative energy transfer occurred between TiO(2) and HSA. Furthermore, the conformational changes of HSA were shown by synchronous fluorescence.