In the article, the authors analyze the effect of a fuel mixture (iso-octane, butanol and ethanol) on the generation of engine vibrations. The paper presents the results in the form of frequency response (using the Fast Fourier Transform – FFT) for three mixtures of different proportions. The measurements were made with the use of accelerometers and data acquisition cards, conditioning the received signal. The vibration component, in the form of acceleration, will be subjected to a FFT and presented in graphical form (periodogram). The authors put a special emphasis on a comparative analysis, indicating changes in harmonics, which may be a potential cause of engine degradation.