The invention belongs to the technical field of vaccine cultivation, in particular to a demulsification separation device for vaccine processing. Aiming at the problem of poor demulsification efficiency, the following scheme is proposed, comprising a demulsification separation box, the top outer wall of the demulsification separation box is equipped with motor, and the output shaft of the motor is connected with a rotating rod through a coupling, the bottom inner wall of the demulsification separation box is provided with a slide rail, and the inner wall of the slide rail is slidably connected with a hydraulic rod, and the top outer wall of the hydraulic rod is provided with crossbar. The present invention is equipped with a slide rail, a hydraulic rod, a sliding sleeve, a first wave rod, a second wave rod, a second stirring rod, a demulsification needle, a stirring plate, and a second demulsification hole, and the demulsification needles have different angles and lengths With different structures, the second demulsification hole is also a structure with different sizes and specifications, which can demulsify vaccines located in different positions. The second demulsification hole and demulsification needle can break the emulsified oil particles in the vaccine solution, It can comprehensively improve the stirring effect of the stirring mechanism and improve the efficiency of demulsification.