Because diffusion of testosterone (T) into the salivary gland is thought to be largely limited to the free, biologically active fraction, salivary testosterone is expected to provide a better measure of testosterone bioavailability in the body than is plasma testosterone. Matched saliva and blood spot samples were collected from 218 Zimbabwean males (age 11-23) who were at different stages of puberty, as assessed by self-reported Tanner genital stage ratings. Testosterone concentrations in these matched samples were highly correlated (r = 0.83). Both salivary and plasma testosterone (converted from blood spot value) showed expected significant increases across puberty. However, plasma testosterone distinguished among subjects at different stages of genital development more effectively than did salivary testosterone, suggesting the former to be a better marker of testosterone bioavailability. Sex hormone-binding globulin (SHBG) levels were also measured in a subgroup of 93 of these subjects. After controlling for plasma T concentrations, we found a small but significant inverse correlation between blood spot SHBG levels and the proportion of plasma testosterone recovered in salvia, supporting the hypothesis that SHBG-related changes in T bioavailability are detectable in saliva. We conclude that salivary testosterone accurately reflects testicular production of testosterone, but that neither salivary testosterone nor plasma testosterone is clearly superior to the other as a measure of testosterone bioavailability.