One of the main components in ballasted railway track systems is the rail pad. It is installed between the rail and the sleeper to attenuate wheel/rail interaction loads, preventing the underlying railway sleepers from excessive stress waves. Generally, the dynamic design of tracks relies on the available data, which are mostly focused on the structural condition at a specific toe load. Recent findings show that track irregularities could significantly amplify the loads on railway tracks. This phenomenon gives rise to a concern that the rail pads may experience higher effective preloading than anticipated in the past. On this ground, this paper highlights the significance of accounting for effects of preloading on dynamic properties of polymeric rail pads. An innovative test rig for controlling preloads on rail pads has been devised. A non-destructive methodology for evaluating and monitoring the dynamic properties of the rail pads has been developed based on an instrumented hammer impact technique and an equivalent single degree-of-freedom system approximation. Based on the impact-excitation responses, some of the selected rail pads have been tested to determine such modal parameters as dynamic stiffness and damping constants in the laboratory. The influence of large preloads on dynamic properties of both new and worn rail pads is demonstrated in this paper. Additionally, the design criteria, which has been used to take into account the influence of the level of preload on dynamic properties of generic rail pads, are discussed.