Background: Measuring sex hormones is essential in diagnosing health issues such as testicular dysfunction, male infertility and feminization syndrome. However, there are no reports on reference intervals (RIs) in Chinese men. We conducted a nationwide multicenter study to establish RIs for seven sex hormones (luteinizing hormone [LH], follicle-stimulating hormone [FSH], prolactin [PRL], total testosterone [TT], free testosterone [FT], bioavailable testosterone [BAT] and estrogen [E2]), as well as sex hormone-binding globulin (SHBG). Methods: In 2013, 1043 apparently healthy adult men from five representative cities in China (Beijing, Hangzhou, Guangzhou, Dalian and Urumqi) were recruited; hormones were measured using an automated immunoassay analyzer. Multiple regression analysis (MRA) was performed to identify sources of variation (SVs) that might influence the hormone serum levels. RIs were computed using the parametric method. Results: Dalian and Hangzhou had significantly higher E2 values than other cities; age was a major source of variation for FSH, LH, PRL, SHBG, FT and BAT. FSH, LH and SHBG increased significantly with age, while PRL, FT and BAT decreased with age. TT showed no significant age-related changes. Median (RIs) derived without partition by age were as follows: FSH, 5.6 (1.9–16.3) IU/L; LH, 4.2 (1.6–10.0) IU/L; PRL, 189 (88–450) mIU/L; E2, 85 (4.7–195) pmol/L; SHBG, 29.4 (11.5–66.3) nmol/L; TT, 15.6 (7.4–24.5) nmol/L; FT, 0.31 (0.16–0.52) nmol/L; and BAT, 8.0 (3.7–13.2) nmol/L. RIs were also derived in accordance with between-city and between-age differences. Conclusions : RIs were established for sex hormones and SHBG in apparently healthy Chinese men in consideration of age.