The single factor experiments and response surface optimization experiments were used to study the extraction processing of oil from crayfish waste.The fatty acid composition and oxidation stability of the oil were also analyzed.Experimental results showed that optimal processing conditions were: 78.88% alcohol,rate of liquid to material 6.65:1,temperature 40℃,extraction time 1h,2 times.The optimal model was: Extraction rate(%)=10.73+0.12A+0.65B-0.072C-0.43A2-0.56B2+0.027C2-0.31AB+0.012AC-0.033BC,prediction value of extraction rate was 11.04%,the actual extraction rate was 0.96%.By fatty acid analysis of crayfish refining oil,the results were: 14 kinds of fatty acid were determined,47.98% of the total fatty acids were unsaturated fatty acids,saturated fatty acids was 52.02%,essential fatty acids was at 17.26%,EPA和DHA was 7.35% of polyunsaturated fatty acids and cis-fatty acids was to 95.65% of unsaturated fatty acid.This indicated that crayfish oil had good nutritional value.Oxidation stability studies showed that oxidation resistance time of crayfish oil was 0.86h.This study provided a theoretical basis of oil extraction and storage.