Patients transported by ambulance are exposed to vibrations that can exacerbate their clinical condition. It is imperative to quantify the acceleration levels to which patients are subjected during ambulance transport. This study presents an analysis of the dynamic behaviour of ambulances when driving over uneven road surfaces. The present study differs from most previous research on ride comfort, which typically does not consider specific ambulance configurations, by aiming to quantify the discomfort experienced by patients during ambulance transport. To achieve this objective, two synthetic road profile signals were generated, based on ISO 8608 classifications. Furthermore, a full vehicle dynamic model of a standard ambulance was developed using CarSim software. The simulations yield acceleration time series at two anatomical locations on the patient’s body. The findings of this study offer significant insights into the patient’s biomechanical response, thereby establishing a foundation for the development of more effective vibration attenuation systems.