Limited studies have investigated the effects of serum carotenoids on the risk of non-Hodgkin lymphoma (NHL), and the findings have been inconclusive. This study aims to assess the association between serum total or specific carotenoid levels and NHL risk. This 1:1 matched, hospital-based case–control study enrolled 512 newly diagnosed (within 1 month) NHL patients and 512 healthy controls who were matched by age (±5 years) and sex in Urumqi, China. Serum carotenoid levels were measured by HPLC. Conditional logistic regression showed that higher serum total carotenoid levels and their subtypes (e.g. α-carotene, β-carotene, β-cryptoxanthin and lycopene) were dose-dependently associated with decreased NHL risk. The multivariable-adjusted OR and their 95 % CI for NHL risk for quartile 4 (v. quartile 1) were 0·31 (95 % CI 0·22, 0·48; Pfor trend < 0·001) for total carotenoids, 0·52 (95 % CI 0·33, 0·79; Pfor trend: 0·003) for α-carotene, 0·63 (95 % CI 0·42, 0·94; Pfor trend: 0·031) for β-carotene, 0·73 (95 % CI 0·49, 1·05; Pfor trend: 0·034) for β-cryptoxanthin and 0·51 (95 % CI 0·34, 0·75; Pfor trend: 0·001) for lycopene. A null association was observed between serum lutein + zeaxanthin and NHL risk (OR 0·89, 95 % CI 0·57, 1·38; Pfor trend: 0·556). Significant interactions were observed after stratifying according to smoking status, and inverse associations were more evident among current smokers than past or never smokers for total carotenoids, α-carotene and lycopene (Pfor heterogeneity: 0·047, 0·042 and 0·046). This study indicates that higher serum carotenoid levels might be inversely associated with NHL risk, especially among current smokers.