Abstract: Leaching process parameters in a two-step oil extraction method were optimized using Elopichthys Bambusa viscera through single factor experiments and a L9 (33) orthogonal experimental design.Results showed that the fish oil extraction rate reached 64.17% after the viscera being boiled for 6～8min in 100℃ and leached under optimized conditions as follows:n-hexane was added to the viscera residue at a ratio of 2∶1 (m/v) , the viscera was leached for 4 times with 30min each time.The fish oil obtained was brown-yellow and cloudy, with a unique fish smell.The physical and chemical properties of the oil meet the grade-one crude fish oil standard.After refining, the oil was light yellow, slightly turbid with a very light fishlike smell.The acid value of the oil was 0.07mg KOH/g oil, iodine value 140g I2/100g oil, impurity 0.07%, which meet the grade-one refined fish oil standard.The peroxide value of the refined fish oil was 6.26mmol/kg, unsaponifiable matter was 1.74%, which reached to grade-two refined fish oil standard.