Abstract: OBJECTIVE To investigate the interaction between cisplatin and epirubicin in conjugation of human serum albumin(HSA). METHODS The interaction of cisplatin and epirubicin with HSA was studied by measuring fluorescence quenching spectra, synchronous fluorescence spectra. The binding constants were calculated according to Lineweaver-Burk equation and type of binding force was estimated by the calculation of thermodynamic parameters. RESULTS The fluorescence quenching spectra of HSA in the presence of aisplatin and epirubicin showed that both of them quenched the fluorescence of HSA. Cisplatin and epirubicin were static quenching. The binding site numbers of epirubicin was 1. The interaction between epirubicin and HSA was mainly driven by a hydrophobic force. When in the presence of cisplatin, the combining constants (K_A) increased. CONCLUSION Cisplatin can increase the combining constants.