Objective To obtain high-quality peony seed oil.Methods Using peony seeds as materials, this study investigated the effects of moisture content, microwave power, microwave time, and kernel-shell mass ratio on peony seed oil yield. Response surface methodology was then used to optimize the process conditions for peony seed oil preparation via microwave pretreatment-pressing methods. The quality of peony seed oil obtained by microwave pretreatment-pressing methods was compared with that extracted by supercritical CO₂ extraction and solvent extraction.Results The optimal process conditions for microwave pretreatment-pressing methods to prepare peony seed oil were listed below: moisture content of 8.6%, microwave power of 1 000 W, microwave time of 5 min, and kernel-shell mass ratio of 15∶1. Under these conditions, the oil yield from peony seeds was (26.74±0.58)%. Although there were differences in the physicochemical indicators of peony seed oil produced by the three methods, all met the standards for edible vegetable oil. The extraction methods were non-selective for fatty acid extraction, with no isomerization or oxidation of fatty acids. However, the peony seed oil prepared by microwave pretreatment-pressing methods contained higher levels of trace components such as squalene, tocopherols, and total phenols, which gave it better antioxidant activity.Conclusion Microwave pretreatment-pressing methods are effective processing methods for obtaining high-quality peony seed oil.