Dietary supplements have been increasingly used by gym users and are often consumed without the guidance of a health professional. Moreover, the indiscriminate supplements use can have adverse health effects, such as changes in liver and kidney function. The aim of this study was to verify the association between dietary supplements intake with alterations in the liver and kidney function among gym users. A cross-sectional study was conducted with 594 gym users (mean age 37 (sd 14) years, 55·2 % women) from a city in southern Brazil. A questionnaire was used to evaluate the use of dietary supplements. The markers of the liver (alanine aminotransferase, aspartate aminotransferase (AST), alkaline phosphatase, γ-glutamyltransferase) and renal (creatinine and urea) function were also evaluated on a subsample of the study population. Data were analysed by binary logistic regression, adjusted for sex, age and education. The prevalence of dietary supplement intake was 36·0 %. Individuals who intake dietary supplements showed a higher prevalence to present slight alterations in the AST enzyme and in the urea after adjustments for potential confounders. In conclusion, the use of dietary supplement was associated with slight alterations in AST enzyme and in the urea among gym users. These findings show the importance of using supplements correctly, especially with guidance from professionals trained to avoid possible risks to health.