Sensitive, specific time-resolved immunofluorometric assays were used to measure the concentrations of human choriogonadotropin (hCG), free beta-subunit (beta-hCG), and the core fragment of beta-hCG (c beta-hCG) in serum and urine of men and nonpregnant women without evidence of cancer. Concentrations of hCG and beta-hCG were measurable in 59-70% of serum samples and in 50-59% of urine samples. c beta-hCG was mostly undetectable in serum but measurable in 81% of urine samples. Concentrations were higher in women than in men, and hCG concentrations increased with age. Therefore, reference ranges based on the 97.5 percentile were calculated separately for women and men and for those < 50 and > 50 years. However, concentrations of hCG correlated much more strongly with those of follicle-stimulating hormone than with age. hCG concentrations in serum were similar to those reported before, but beta-hCG concentrations were below the detection limit of earlier assays, and the upper reference limit was one-fifth to one-tenth the cutoff concentrations used earlier. In urine, hCG and c beta-hCG were the major forms of hCG, and their concentrations were similar to those of hCG in serum.