The electrochemical behavior of mixed Langmuir-Blodgett (LB) film of bilir ubin and phosphatidyl ethanolamine on glassy carbon electrode were investigated with cyclic voltammetry and differential pulse voltammetry. The results prove th at bilirubin Langmuir-Blodgett film has a cathodic peak of +420mV in 0.2mol ·L-1 HAc-NaAc-50%EtOH(pH=5.4) solution. The peak current increases wit h the layer number of bilirubin LB film in the range of 1 to 12 layers. The resu t of the electrochemical behavior of bilirubin in ordered LB film was also compa red with that in cast film and in aqueous solution at glassy carbon electrode. W e found that bilirubin has the similar electrochemical behavior in LB film to in cast film, but different from that in aqueous solution.