We studied the association between intake of non-prescription analgesics and semen quality and male reproductive hormone levels in a cross-sectional study among 1493 men. The men provided one semen (n = 1493) and blood sample (n = 1056) and filled in questionnaires on use of non-prescription analgesics (paracetamol, NSAIDs and combination drugs (yes/no)). Adjusting for age, study and other covariates, we observed no association between intake of non-prescription analgesics and markers of semen quality. Adjusting for age and time of day of blood sampling, users of non-prescription analgesics had a 10.4% (95% confidence interval (CI) 4.0–17.1%) higher testosterone level than non-users. When we stratified by medication type, the association between analgesics and higher testosterone was observed between users of non-steroidal anti-inflammatory drugs (NSAIDs) and combination drugs but not paracetamol. This study suggests that use of non-prescription analgesics is associated with slightly higher serum testosterone levels than non-use.