The invention belongs to the field of Chinese herbal medicine quality control, and particularly relates to an ephedrae decoction standard substance and a preparation method and a quality detection method thereof. According to the preparation method of the ephedra decoction standard substance, freeze-dried powder preparation is changed into standard decoction preparation, operations such as concentration and drying are not needed, and volatilization of some important components (such as cinnamaldehyde) is avoided. Based on the ephedra decoction standard substance prepared by the method, the original taste and flavor of the 'one bowl' of ancient prescription ephedra decoction can be restored. The method can be used for guiding actual industrial production. According to the ephedra decoction standard substance quality detection method, quantitative determination of ephedrine hydrochloride, pseudoephedrine hydrochloride and amygdalin and qualitative analysis of liquiritin, cinnamic acid, cinnamaldehyde and glycyrrhizic acid are fused, meanwhile, new chromatographic conditions are developed, the separation degree is better, the stability is better, the detection time is short, and the efficiency is high.