The interaction between Bovine Hemoglobin (BHb) and Minodiaq was investigated at different temperatures by fluorescence spectroscopy, as well as the effect of some metal ions (Mg2+, Zn2+, Cu2+, Mn2+, Ni2+, V5+, Mo6+, Cr6+) on the BHb-Minodiaq system. Results showed that Minodiaq could quench the intrinsic fluorescence of BHb, and the quenching mechanism was a dynamic quenching process. The hydrophobic force played an important role on the conjugation reaction between BHb and Minodiaq. Synchronous spectra and CD spectra revealed that the microenvironment and the conformation of BHb were changed during the binding reaction. Studies on the interaction between BHb and drug will facilitate interpretation of the drugÂs metabolism and transporting process in the blood, and will help to explain the relationship between structures and functions of BHb.