Abstract: Objective: To optimize the extraction process of hemin in the enzymatic hydrolysis blood residue.Method:Taking the enzymatic hydrolysis blood residue of preparation hemoglobin peptide as raw material, the hydrochloric acid consumption A, solvent consumption B, mixing time C were optimized by the L9 ( 34) orthogonal design.Results: The optimum extration condition were as follows: hydrochloric acid to acetone ratio = 4 ∶ 100, 5 folds of Acidic acetone, stirring 20min. The purity of extracted hemin could reach more than 97%, the iron conten could reach more than 7.8%.Conclusion: This method was easy to operate, had better reproducible and could be used for industrial production.