Objective:This study aimed to compare the effects of different extraction methods including cold pressing extraction, soxhlet extraction and subcritical butane extraction on the yield, physicochemical properties, fatty acid composition and in vitro antioxidant activity of elderberry seed oil.Methods:Elderberry seed oils were extracted from elderberry seed by cold pressing extraction, soxhlet extraction and subcritical butane extraction. The physicochemical parameters and in vitro antioxidant activity of oil were determined with reference to national standards, and the fatty acid composition was analyzed by GC-MS.Results:Among the three extraction methods, the highest oil yield was obtained by subcritical butane extraction (28.8%), followed by soxhlet extraction (27.6%) and cold pressing extraction (18.8%). The acid value and peroxide value of elderberry seed oil obtained by subcritical butane extraction were the lowest, and its iodine value was the highest, compared to other methods. The fatty acid composition of elderberry seed oils obtained from the three extraction methods were similar, mainly including linolenic acid, linoleic acid and palmitic acid, whereas the contents were different, with the highest linolenic acid content (46.94%) in the elderberry seed oil obtained by subcritical butane extraction. The elderberry oils obtained by three extraction methods had good antioxidant activity, and the elderberry oils obtained by subcritical butane extraction had the strongest scavenging ability of DPPH free radicals and ABTS free radicals.Conclusion:Different extraction methods had certain influence on the quality of elderberry seed oil, and subcritical butane extraction had good effect on extraction rate and quality of elderberry seed oil, which was an ideal method for the extraction of elderberry seed oil.