There is mounting scientific evidence from both animal, and in vitro studies that suggest deleterious effects on human health from inhaled uranium. Serum albumin plays a significant role in carrying, buffering and nutrition. In order to study the interaction between uranium and proteins, as well as the in vivo mechanism of uranium metabolic pathway and biological effect, the orthogonal design experiments in four levels and the three factors were carried out in this research. The results showed that the best reaction conditions between uranyl ions and BSA (Bovine Serum Albumin) was pH3.0, when uranyl ions concentration was 0.60 mg/mL, BSA concentrations was 0.45 mg/mL. In this research the FTIR (Fourier-transform infrared) spectra was also used to analysis the interaction between the uranium and BSA. The FTIR spectra indicated that the -O-H, -C=O of BSA contributed to the major binding groups and the uranyl ions ccould change the BSA conformation of BSA when interaction each other under different pH conditions.