Objectives To describe the prevalence of hearing loss among 13 year old adolescents, and to examine the change in prevalence between ages 9 and 13 years. Methods This study was embedded within Generation R, a population-based prospective cohort study from fetal life onwards in the Netherlands. Pure-tone thresholds were obtained at 0.5 to 8 kHz, and tympanometry was performed. Sensorineural hearing loss (SNHL) was defined as a low-frequency and/or high-frequency pure-tone average of more than 15 dB HL in one of both ears. Audiometric signs suggestive of noise-induced hearing loss (NIHL) included the presence of a notch and/or high-frequency hearing loss. The study was conducted from April 2012 to October 2015, and April 2016 to September 2019. Results A total of 4572 adolescents with a mean age of 13 years and 7 months (SD, 5 months) were included, of whom 2334 (51.0%) were girls. Within the cohort, 6.4% (95% CI, 5.7%-7.2%) were estimated to have SNHL, and 12.4% (95% CI, 11.5%-13.4%) met the criteria of NIHL. In total, 3675 participants were included in the longitudinal analysis. The prevalence of SNHL decreased from 8.0% to 5.3% between ages 9 and 13 years (P < .001). The prevalence of NIHL increased from 9.8% to 11.7% (P = .004), due to an increase in number of participants with a notch. Conclusions The prevalence of SNHL significantly decreased by 2.7% (95% CI, 1.6%-3.9%) between ages 9 and 13 years, probably due to a change in alertness during assessment at the age of 13 years. Other possible explanations include the presence of selection bias or a decline in prevalence of conductive hearing loss. The number of participants with audiometric signs suggestive of NIHL increased by 1.9% (95% CI, 0.5%-3.3%). Level of Evidence Level 3.