Objective: To optimize the conditions of supercritical CO2 extraction and ethanol extraction processes of yolk oil. Method: HPLC-ELSD was employed to determine the content of phosphatidylcholine with mobile phase A of methanol-water-glacial acetic acid-triethylamine (85: 15: 0.45: 0.05) and mobile phase B of n-hexane-isopropanol-mobile phase A (20: 48: 32) for gradient elution. Based on single factor tests

taking yield of yolk oil and the content of phosphatidylcholine as indexes

orthogonal tests were adopted to optimize supercritical CO2 extraction and ethanol extraction processes of yolk oil. Result: Optimum extraction conditions were as follows

moisture content of yolk oil was ≤5%

supercritical CO2 extraction temperature was 50℃

pressure was 35 MPa

flow was 50 L·h-1

extraction time was 2 h

yield of yolk oil was 42.65%

residual powder of supercritical CO2 extraction was extracted four times with four times the amount of 90% ethanol for 1.5 h of each time

the content of phosphatidylcholine was up to 17.24%. Conclusion: These optimized technologies are stable and feasible for development and application of yolk oil.