The invention relates to the technical field of test tube holders and particularly provides a test tube holding device including a handle; a sliding groove is formed in the handle; a spring is fixedlyconnected to the rear end of the sliding groove; an adjustment seat is fixedly connected to the front end of the spring, extends upwardly out of an adjustment opening in the handle and is movable relative to the adjustment opening; a metal tie is fixedly connected to the front end of the adjustment seat, passes through a through hole in the front end of the handle and is fixedly connected to theend portion of an annular trough in a fixing ring; the fixing ring is fixedly connected to the handle through a connection plate; a position limiting trough is arranged at a position, corresponding tothe metal tie, on the connection plate. By adjusting the metal tie by the spring, a to-be-held test tube can be wrapped by the metal tie, so that the test tube is more convenient and safe to move.