Previous studies concerning the relationship of serum testosterone levels to aging in normal men have yielded apparently inconsistent results. Studies performing blood sampling in the morning have often shown an age-related decrease in testosterone levels, while those using afternoon samples have failed to show such a decrease. These results suggested to us the possibility that the circadian rhythm in serum testosterone levels might be altered with normal aging in men. Hourly blood samples were obtained for 24 h from 1 young (mean age, 52.2 yr) and 12 old (mean age, 17 yr) healthy men. Total testosterone levels were measured by RIA. The circadian rhythm in serum testosterone levels found in normal young men was markedly attenuated or absent in healthy elderly men; the early morning rise in testosterone levels characteristic of young men was not present in old age. Mean testosterone levels for the entire 24-h day were lower in healthy old men than in young men. These results demonstrate a clear decrease in serum testosterone levels in healthy old men compared to those in young men and provide an explanation for the inability to demonstrate an age-related decline in testosterone levels in earlier studies using serum samples obtained in the afternoon.