The interaction between Holo-Transferin (HTF) and Colchicine (COL) was investigated in the present of Pb+2 ions under physiological conditions by using synchronous fluorescence spectra. The synchronous fluorescence spectra show a slight change of tryptophan residue micro-environment. Synchronous fluorescence spectra show that the structure of the tyrosine residue environment was altered by interaction of the COL and Pb+2 ions with HTF. The fluorescence intensity of HTF decreased regularly beside a small blue shift with increasing concentrations of COL and Pb+2 ions. The intrinsic fluorescence of HTF was quenched in the presence of drug and ion. Interaction of drugs with HTF and HTF-Pb+2 can elucidate the properties of drug-protein and ion- protein complex, as it may provide useful information about the structural feature that determines the therapeutic effectiveness of ion and drugs. Therefore, it has become a significant research field in life science, chemistry, biotechnology and clinical medicine.