The present study contains an experimental analysis of the vibratory response in a low-cylinder engine motorcycle at varying suspension preloads. Three different speed bumps of varying heights were used to subject the motorcycle to different vibrations. The analysis was carried out in three domains: time, frequency, and time-frequency. A triaxial accelerometer was used to measure the vibrations at the seat of the vehicle. The results indicated that the suspension system became more differentiated as the height of the bumps increased. However, for lower bumps, the action of the three spring preloads studied was quite similar. Quantitatively, only the higher bump showed a significant difference between the set preloads. The spectral distribution revealed that the frequency of interest was below 20 Hz for all the studied cases, which is in the same range of human body natural frequencies. The findings of this research can be utilized to enhance the design of low-cost motorcycles, thereby improving the safety and comfort of their drivers and passengers. This study constitutes a significant step towards developing an affordable system capable of gathering sufficient data to support the creation of evidence-based public health policies and propose new transport industry standards based on field measurements.