OBJECTIVE: To describe and analyze interpretations of measurements performed on tympanometry in neonates with probe tone of 226 and 1000 Hz. MATERIAL AND METHODS: We studied 73 newborns, 38 females and 35 males, aged between 4 and 29 days old, otoacoustic emissions in both ears and no risk factors for hearing loss. The research was conducted with tone test of 226 and 1000 Hz in the Department of Speech Pathology, Nossa Senhora da Conceição Hospital of Porto Alegre, using the immittance Interacoustics Audiotest 425h. In statistical analysis we applied the chi-square McNemar, Student's t test for paired samples and ANOVA. RESULTS: Statistically significant differences (p <0.05) in the comparison between the genders in the extent of compliance with a tone of 226Hz (p = 0.011) in the left ear. Comparing the probe tones of 226 and 1000 Hz, as complacency, we observed a statistically significant difference between the ears, OD (p = 0.003) and OS (p = 0.001). Comparing the probe tones and ears statistical significance was found on the following: compliance at 226Hz (p = 0.002), pressure peak at 226 Hz (p <0.001); complacency at 1000Hz (p = 0.004), peak pressure 1000Hz (p = 0.045). We observed a higher incidence of single peak with 1000 Hz probe tone and double peak with a tone of 226Hz in both ears (p <0.001). CONCLUSION: The analysis of measurements obtained in tympanometries found a statistically significant difference between the probe tones of 226 and 1000 Hz and ear (right and left) in neonates.