The ball screw is a critical device for precision linear motion control that has widespread applications in industrial robots, computer numerical control (CNC) machines, and high-precision leveling systems, among others. Because high-precision positioning is ensured by the addition of a preload to the ball screw system, it is crucial to detect and monitor the loss of preload at the earliest possible stage of degradation. The degradation process of the ball screw can be characterized in two stages: the initial reduction of preload without backlash, followed by a loss of preload with an emergence of backlash. To explore the change of the ball screw dynamics caused by degradation, a novel fixed cycle feature test (FCFT) is implemented in combination with multi-level mass experiments and a run-to-failure (RTF) test. The relationships of the ball screw dynamics with preload, worktable mass, and axis position are investigated, with a focus on the axial natural frequency as an indicator of preload loss. Experimental results validate the axial natural frequency’s role as a reliable early detector of preload loss for interventive use in a prognostic system.