Previous studies have reported the relationship between effect of caffeine and many diseases. However, studies to evaluate the association between caffeine and hearing loss are contradictory. To examine the relationship of urinary caffeine metabolites with the hearing threshold in US adults, a total of 849 adults aged 20-69 years who participated in the National Health and Nutrition Examination Survey (NHANES, 2011-2012) were enrolled in this study. Urinary caffeine and its 14 metabolites were applied as biomarkers to assess caffeine exposure. Hearing loss was defined as mean pure tone averages > 25 dB HL at 500, 1000, and 2000 Hz in both ears (low frequency); and 3000, 4000, and 6000 Hz in both ears (high frequency). Univariate and multivariate linear regression analyses were conducted to examine the associations of urinary caffeine metabolites with low- and high-frequency hearing thresholds, respectively. Low-frequency hearing loss were 5.08% and 6.10% in male and female participants, respectively; and high-frequency hearing loss were 31.81% and 15.14% in male and female participants, respectively. In the unadjusted model, the P value for trend shows that urinary caffeine metabolites 137X and AAMU were significantly associated with low-frequency PTA, and that 17X, 137X, AAMU were significantly associated with high-frequency PTA, but when the model was adjusted for sex, age, education level, firearm noise exposure, occupational noise exposure, recreational noise exposure, serum cotinine, body mass index, diabetes, hypertension, these were no longer statistically significant. In conclusion, urinary caffeine metabolites were not associated with the hearing threshold shifts in US adults.