The separation safety of wedged self-locking anti-vibration mechanism was analyzed and the checking methods were given in this paper. Through the detailed analysis on the principle of the mechanism and the biggest deformation calculated by dimension chain as so as the maximal positive pressure calculated by using finite element method, the analysis method of the separation safety of wedged self-locking anti-vibration mechanism was put forward by the calculation and analysis of the maximal positive pressure. The numerical example shows that the method is correct and with the calculated results, the applicable scope of the mechanism is given.