Abstract: Aqueous enzymatic extraction( AEE) of liver oil from Ommastrephes bartrami was performed,the enzyme type,ratio of material to solvent,reaction temperature,extraction time and enzyme amount were investigated,meanwhile the fatty acid compositions of squid liver oil were analyzed by GC- MS in this paper.The single factor test and orthogonal experiment showed the optimal conditions for the aqueous enzymatic extraction of squid liver oil were as follows: neutral protease was the optimal hydrolytic enzyme,ratio of material to solvent was1∶ 0( g / m L),reaction temperature was 55 ℃,extraction time was 6 h,enzyme amount was 1800 U / g and the maximum oil extraction yield( 79.21%) was obtained under the optimal conditions.The main fatty acids of squid liver oil were C16∶ 0( 16.27%),C18 ∶ 1n- 9( 11.68%),C20 ∶ 1n- 11( 12.94%),EPA( 9.81%) and DHA( 23.61%),and the total amount of n-3 polyunsaturated fatty acids was up to 35.97%,and the ratio of ∑n-3 PUFA to ∑n-6 PUFA was7.99,indicating that the liver oil from Ommastrephes bartrami had high nutritional values and development prospects,which could be used as the important dietary source of n-3 polyunsaturated fatty acids,such as EPA and DHA especially.