Since correct assessment of testicular function and androgenic status in humans requires multiple sampling, a sensitive and accurate radioimmunoassay (RIA) of testosterone (T) was established for male and female saliva samples. This easily collected biological fluid, which contains nonprotein-bound T, may represent an attractive alternative or a complement to total plasma T assays. In saliva samples from 5 normal males, a clear circadian rhythm was observed, and morning concentrations (135 ± 31 pg/ml) were significantly higher (p < 0.02) than evening samples (85 ± 23 pg/ml). In 11 normal females, morning saliva levels were 12.8 ± 1.8 pg/ml. The levels of T in male saliva, in response to both exogenous T administration (100 mg i.m.) and HCG stimulation (2 × 2,000 IU i.m.), accurately reflected the changes observed in plasma T, and the magnitude of increase in T levels was clearly greater in saliva than in plasma samples during the intramuscular administration of the long-acting T preparation. In males, significant correlations were observed between salivary and plasma T concentrations in morning samples (r = 0.61, p < 0.01), following HCG stimulation (r = 0.89, p < 0.05) and during T administration (r = 0.87, p < 0.05). In women, the correlation at 8 a.m. was also significant (r = 0.82, p < 0.05).

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