Abstract: OBJECTIVE To determine the optimal parameters in extraction of seed oil from Annona squamosa. METHODS The extraction efficiency of four extraction methods， including percolation extraction， soxhlet extraction， ultrasonic extraction and SFE-CO₂ extraction were compared. The optimal parameters of the best extraction method were determined， and the chemical composition of the oils was analyzed by GC-MS. RESULTS The yields of the oils extracted by the soxhlet extraction method were highest (22.28±0.21%)， and the optimal parameters were obtained as follows: Petroleum ether were used as extraction solvent， the extraction temperature was 80℃， and the extraction time was 90 min. GC-MS analysis results showed that the main chemical composition was palmitic acid (9.92%)， linoleic acid (20.49%)， oleic acid (56.50%) and stearic acid (9.14%). CONCLUSION The optimum extraction method was stable. Annona squamosa seeds are rich in unsaturated fatty acids and have a certain economic value.