According to the experimental design principle of response surface methodology, four factors and three levels of response surface experiment was designed to optimize the preparation process of the tilapia scales collagen peptide iron chelate. A mathematical model was established to describe peptide iron chelate rate as a function of pH, peptide iron mass ratio, time and temperature. The results show that pH, peptide iron mass ratio and time have a significantly effect on the peptide iron chelate rate. The optimal preparation conditions is pH 5.4, temperature 40 ℃, the peptide iron mass ratio 3:1, and time 41 min. Under the optimal preparation process, the practical value of peptide iron chelate rate is 90.49%, which has no significant difference with the predicted value.