This paper describes lost motion analysis for a novel 6-DOF ultra-precision positioning stage. In the case of flexure hinge based precision positioning stage, lost motion is generated when the displacement of actuator is not delivered completely to the end-effector because of the elasticity of flexure hinge. Consequently, it is need to compute amount of lost motion to compensate the motion or to decide appropriate control method for precision positioning. Lost motion analysis for the vertical actuation unit is presented. The analysis results are presented in two ways: analytic and numerical analyses. It is found that they closely coincide with each other by 1% error. In finite element analysis result, the amount of lost motion is turned out to be about 3%. Although, the amount is not so large, it is necessary procedure to check the lost motion to establish the control method.