The invention relates to a detection method for the content of lipid-soluble vitamins. The detection method comprises the steps that a classical pretreatment mode of a saponification reaction is adopted, but traditional liquid-liquid extraction is replaced with ultrasonic extraction, and a to-be-detected solution and a standard solution are obtained through pretreatment; the standard solution and the to-be-detected solution are detected through liquid chromatography to obtain the retention time and the peak area of all components in a lipid-soluble vitamin standard substance and all components in to-be-detected feed, and a corresponding curve of the concentration and the peak area of the lipid-soluble vitamin standard substance is drawn; the retention time and the peak area of all the components of the to-be-detected feed are compared with a standard curve, and the content of the lipid-soluble vitamins in the to-be-detected feed is calculated. Compared with the prior art, the detection method has the advantages that the pretreatment process is simple, the usage amount of poisonous and harmful petroleum ether is decreased, and the method is more environmentally friendly and safer; in addition, a detection result is high in precision, the effect is good, and simultaneous separation and detection on the vitamin A, the vitamin D3 and the vitamin E are achieved in a short time.