The invention discloses a method for extracting black wolfberry seed oil by virtue of subcritical fluid. The method comprises the steps: (1) placing dry black wolfberry seeds and an extracting agent into an extraction kettle of a subcritical fluid extraction apparatus according to a mass-volume ratio of 1:1 to 1: 20, continuously extracting for 1 to 20 times to obtain extract, wherein the extraction time is 10 to 15 minutes per time; (2) separating the extract in the step (1) by utilizing a pressure reduction heating method under the condition that the separation pressure is 0.10 to 1.95 MPa and the separation temperature is 10 to 90 DEG C, to obtain an extraction solution; (3) standing the extraction solution for 10 to 90 minutes under the room temperature, to obtain the black wolfberry seed oil. By adopting the method, not only can the production cost be effectively reduced, the extraction and separation time is shortened, and the consumption of solvent and energy consumption in the process flow can be reduced; moreover, the method is simple in process flow, high in operability, high in extraction efficiency, environment-friendly, pollution-free, good in safety and repeatability and capable of realizing the industrialized continuous mass production.