Abstract: Fresh black-bone chicken blood as the main raw material to study the enzymatic hydrolysis process conditions, the study showed that intermittent ultrasonic broken processing under the conditions had a better haemolytic effect than mechanical crushing.By comparing the hydrolysis ability of AS1.398 neutral protease, pepsin, papain, alkaline protease on black-bone chicken blood, AS1.398 neutral protease was selected for enzymatic hydrolysis; then on the basis of single factor and orthogonal experiments to determine the optimal enzymatic conditions for AS1.398 neutral protease were:the enzymolysis liquid initial pH7.5, enzyme dosage 8000U /g, the hydrolysis temperature 45℃, substrate concentration 8% , the enzyme solution time 6h. which the degree of hydrolysis reached 22.22% , hemoglobin content was 0.62 mg /mL.