The invention relates to the technical field of biologic molecule purification and separation, and in particular relates to a liquid moving gun inserting device for biologic molecule purification and separation. The liquid moving gun inserting device comprises a mounting frame, a hollow tube, a baffle and an elastic buffer piece, wherein the hollow tube penetrates through the lower frame side of the mounting frame; the hollow tube is positioned in the frame by virtue of the baffle mounted on the hollow tube; the elastic buffer piece is arranged between the upper frame side of the mounting frame and the baffle; in the process of inserting a to-be-mounted gun, under the action of a sliding block mechanism, the mounting frame moves downwards, when the lower end of the hollow tube is contacted with a gun head, the hollow tube and the mounting frame perform certain relative movement, and because of the compression force generated when the elastic buffer piece is compressed, the lower end of the hollow tube is abutted with the gun head in a buffered manner, so that the gun head can be stably and reliably inserted and taken out from the lower end of the hollow tube, furthermore the inserted gun head is prevented from tilting, and moreover good sealing of the lower end of the hollow tube and the gun head is achieved.