Peripheral androgen action largely determines the occurrence of hirsutism in women. Although serum 5 alpha-androstane-3 alpha, 17 beta-diol (3 alpha-diol) glucuronide signifies skin 5 alpha-reductase activity and has been used as a marker of hirsutism and peripheral androgen metabolism, other C19 androgen conjugates have recently been measured and may also be useful markers of hirsutism in women. In addition to normal controls we studied both hirsute and nonhirsute patients with polycystic ovarian syndrome who had similar levels of circulating androgen precursors. In these three groups we measured various C19 sulfates and glucuronides including serum 3 alpha-diol glucuronide. Serum androgen precursors were elevated, but were equal in the hirsute and nonhirsute patients. Serum androsterone sulfate and glucuronide, and 3 alpha-diol sulfate and glucuronide clearly differentiated the hirsute from the nonhirsute group. Among the conjugates, androsterone glucuronide was most reflective of the difference between the two groups (100.3 +/- 28.0 versus 42.9 +/- 4.0 ng/ml, p less than 0.05). In hirsute compared with nonhirsute patients with polycystic ovarian syndrome, serum 3 alpha-diol glucuronide was increased by the smallest amount (32%), followed by androsterone sulfate (38%), 3 alpha-diol sulfate (59%), and androsterone glucuronide with the largest increase (134%). Serum androsterone glucuronide and 3 alpha-diol glucuronide both correlated with androstenedione and dehydroepiandrosterone sulfate in hirsute women but not in nonhirsute women. These data suggest that besides serum 3 alpha-diol glucuronide, other C19 sulfate and glucuronide conjugates may reflect peripheral androgen action.