Spotted hyenas (Crocuta crocuta) exhibit an array of behavioral and morphological characteristics that set them apart from other mammals: females are heavier and more aggressive than males, and females have external genitalia that closely resemble those of the male. Because androgenic hormones might mediate the expression of these traits, androgens are of great interest in this species. Past work on circulating androgens in wild hyenas has been limited, in part because of small sample sizes. In this study we validated a non-invasive method of monitoring variation in androgens by measuring total androgen metabolites in the feces of wild and captive spotted hyenas with an enzyme immunoassay. HPLC analysis revealed multiple immunoreactive androgen metabolites in fecal extracts from both males and females. LHRH challenge in three male and two female hyenas in captivity caused an increase in fecal androgens one to three days after LHRH injection. Furthermore, presence of bone in the diet did not affect fecal androgen concentrations in captive female hyenas. In wild spotted hyenas, time of day of fecal deposition, time elapsed between deposition and freezing of the sample, and time elapsed between freezing and extraction did not systematically affect fecal androgen concentrations. Finally, in wild hyenas, fecal androgen patterns mirrored plasma testosterone patterns in that adult immigrant males had higher concentrations than adult natal males, and pregnant females had higher concentrations than lactating females. These methods can therefore be used in future studies addressing relationships among fecal androgens, social status, reproductive state, and behavior in spotted hyenas.