Exposure to traffic noise and air pollution have both been associated with cardiovascular disease, though the mechanisms behind are not yet clear. We aimed to investigate whether the two exposures were associated with levels of cholesterol in a cross-sectional design. In 1993–1997, 39,863 participants aged 50–64year and living in the Greater Copenhagen area were enrolled in a population-based cohort study. For each participant, non-fasting total cholesterol was determined in whole blood samples on the day of enrolment. Residential addresses 5-years preceding enrolment were identified in a national register and road traffic noise (Lden) were modeled for all addresses. For air pollution, nitrogen dioxide (NO2) was modeled at all addresses using a dispersion model and PM2.5 was modeled at all enrolment addresses using a land-use regression model. Analyses were done using linear regression with adjustment for potential confounders as well as mutual adjustment for the three exposures. Baseline residential exposure to the interquartile range of road traffic noise, NO2 and PM2.5 was associated with a 0.58mg/dl (95% confidence interval: −0.09; 1.25), a 0.68mg/dl (0.22; 1.16) and a 0.78mg/dl (0.22; 1.34) higher level of total cholesterol in single pollutant models, respectively. In two pollutant models with adjustment for noise in air pollution models and vice versa, the association between air pollution and cholesterol remained for both air pollution variables (NO2: 0.72 (0.11; 1.34); PM2.5: 0.70 (0.12; 1.28) mg/dl), whereas there was no association for noise (−0.08mg/dl). In three-pollutant models (NO2, PM2.5 and road traffic noise), estimates for NO2 and PM2.5 were slightly diminished (NO2: 0.58 (−0.05; 1.22); PM2.5: 0.57 (−0.02; 1.17) mg/dl). Air pollution and possibly also road traffic noise may be associated with slightly higher levels of cholesterol, though associations for the two exposures were difficult to separate. •Traffic noise and air pollution are both associated with cardiovascular disease.•A potential mechanism includes raised levels of blood lipids.•Air pollution was associated with total cholesterol in a study of 36,039 elderly.•Noise was weakly associated with cholesterol before adjustment for air pollution.•It was difficult to separate effects of air pollution and noise in relation to cholesterol.