The experimental design methodology was applied for modeling and optimizing the operation parameters on photocatalytic degradation of chloramphenicol (CAP) using TiO2 as photocatalyst in a photoreactor. Three experimental parameters (including pH, TiO2 concentration and CAP initial concentration) were adopted to obtain the preliminary information. The multivariate experimental design was employed to establish a quadratic model as a functional relationship between the degradation rate of CAP and three experimental parameters. The interaction effects and optimal parameters were obtained by using Design Expert software. The optimal values of the operation parameters under the related constraint conditions were found at pH 6.4, TiO2 concentration of 0.94 g/L and CAP initial concentration of 19.97 mg/L, respectively. The degradation rate of CAP approached 85.97% under optimal conditions. The regression analysis with R2 value of 0.9519 had a good agreement between the experimental results and the predictive values. In addition, pH and TiO2 concentration had a significant influence on the degradation rate of CAP.