The　binding　of　chloroamphenicol　(CPC)　to　bovine　serum　albumin　(BSA)　at　296　K,　303　K,　and　310　K　by　fluorescence　and　UV-visible　absorption　spectroscopy　were　investigated　under　imitated　physiological　conditions.　The　experimental　results　showed　that　the　fluorescence　quenching　mechanism　between　CPC　and　BSA　was　combined　quenching　(dynamic　and　static　quenching)　procedure　at　low　CPC　concentration,　or　a　dynamic　quenching　procedure　at　high　concentrations.　The　binding　constants,　binding　sites　and　the　corresponding　thermodynamic　parameters　of　the　interaction　system　were　calculated.　According　to　Förster　non-radiation　energy　transfer　theory,　the　binding　distance　between　CPC　and　BSA　was　calculated　to　be　3.02　nm.　Both　synchronous　fluorescence　and　FT-IR　spectra　confirmed　the　interaction,　and　indicated　the　conformational　changes　of　BSA.　The　effects　of　some　common　metal　ions　Ca2+,　Ni2+,　Mg　2+,　Fe2+,　and　Cu2+　on　the　binding　constant　between　CPC　and　BSA　were　examined.　Furthermore,　we　investigated　the　possible　sub-domains　on　BSA　that　bind　CPC　by　displacement　experiments.　©　2012　Elsevier　B.V.　All　rights　reserved.