The invention provides a method for separating beta-elemene from zedoary turmeric oil. The method comprises the steps of fully mixing normal hexane, methyl alcohol and water according to a volume ratio of 2 : (0.3 to 1.7) : (1.7 to 0.3), and standing, so that liquid is layered to obtain an upper phase as a flow phase and a lower phase; taking the lower phase, and adding silver nitrate to form a fixed phase of high-speed countercurrent chromatography; weighing the zedoary turmeric oil, dissolving the zedoary turmeric oil through mixed solution of the upper phase and the lower phase with the volume ratio of 1 to 1, feeding a sample, separating through high-speed countercurrent chromatography, detecting through an ultraviolet detector, collecting outflow liquid containing the beta-elemene, evaporating to remove a solvent and drying to obtain a beta-elemene product. The separating method disclosed by the invention is simple and convenient to operate, high efficiency and quickness are achieved, and solvent consumption is small; the silver nitrate has a pi coordination effect on the beta-elemene, the beta-elemene can be selectively separated from the zedoary turmeric oil, and the separation effect is excellent.