PURPOSE: This study aimed to evaluate the effects of complete external ear canal occlusion on hearing thresholds with aging. The goal was to decide which tuning fork is more appropriate to use for the contralateral occlusion test (COT), in individuals of different ages. METHODS: Forty-two normal hearing subjects between 21 and 67 years were divided into three age groups (20-30 years, 40-50 years, and 60-70 years). Participants underwent sound field audiometry tests with warble tones, with and without ear canal occlusion. Each ear was tested with the standard frequencies (250, 500, 1000, and 2000 Hz). The contralateral ear was suppressed by masking. RESULTS: Hearing thresholds showed an increase as the frequency increased from 20.85 dB (250 Hz, 20-30 years group) to 48 dB (2000 Hz, 60-70 years group). The threshold differences between occlusion and no occlusion conditions were statistically significant and increased ranging from 11.1 dB (250 Hz, 20-30 years group) to 32 dB (2000 Hz, 20-30 years group). We found statistically significant differences for the three age groups and for all evaluations except to 500 Hz difference and average difference. The mean hearing loss produced by occlusion at 500 Hz was approximately 19 dB. We found no statistically significant differences between right and left ears and gender for all measurements. CONCLUSION: We conclude that the use of the 512 Hz tuning fork is the most suitable for COT, and its use may allow clinicians to distinguish mild from moderate unilateral conductive hearing loss.