Reference ranges of 17-hydroxyprogesterone, DHEA, DHEAS, androstenedione, total and free testosterone determined by TurboFlow-LC–MS/MS and associations to health markers in 304 men

• Reference ranges for steroid metabolites using LC–MS/MS method
• Negative association between age and DHEA, DHEAS, androstenedione and free testosterone
• Negative association between high body mass index and steroid metabolites

We report reference ranges based on LC–MS/MS for testosterone (T), free testosterone (FT) and its precursors, i.e. 17-hydroxyprogesterone (17-OHP), dehydroepiandrosterone (DHEA), DHEA-sulfate (DHEAS) and androstenedione (Adione), in relation to different health markers and lifestyle factors.The study was based on 304 healthy men aged 30–61 years participating in a population-based cross-sectional study (Health2008). Examination program consisted of a clinical examination, completion of a self-administered questionnaire and blood sampling. Steroid metabolites were measured by a validated and sensitive LC–MS/MS method.Older age-groups were significantly associated with decreased concentrations of DHEA, DHEAS, Adione, and FT, while no significant associations with age were shown for 17-OHP or T. Participants with BMI ≥ 30 kg/m2 had lower age-related steroid metabolite z-scores compared to participants with BMI < 30 kg/m2, i.e. 17-OHP: − 0.51 vs. 0.08 (p < 0.001); DHEA: − 0.27 vs. 0.09 (p = 0.014); Adione: − 0.29 vs. 0.09 (p = 0.012); T: − 0.99 vs. 0.14 (p < 0.001); and FT − 0.55 vs. 0.05 (p < 0.001), respectively.In conclusion, this large study on serum steroid metabolites and concomitant assessment of health markers in healthy men provides age-related reference ranges, and furthermore evaluates the impact of lifestyle factors and metabolic syndrome on androgen metabolite levels.