The Rocky Mountain Arsenal (RMA), a Superfund site near Denver, Colorado, USA, has a history of various industrial processes that may have led to the release of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and polychlorinated biphenyls (PCBs). The PCDDs, PCDFs, and non-ortho- and mono-ortho-substituted PCBs cause a common set of toxic effects that are mediated through the aryl hydrocarbon receptor (AhR). The total AhR-mediated activity of complex mixtures in biota samples from the RMA and surrounding reference areas was determined by both instrumental and bioanalytical techniques. Mean concentrations of bioassay-derived 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) equivalents (TCDD-EQ) in carp eggs, owl livers, and kestrel eggs ranged from 0 to 8.5, 17 to 130, and 2.4 to 18 pg/g, respectively. For most samples analyzed, concentrations of TCDD-EQ and instrumentally derived TCDD equivalents (TEQs) were not significantly different. In a few tissue samples, concentrations of TEQs and TCDD-EQs were not equivalent. This can indicate the presence of AhR-active compounds that were not identified or quantified by instrumental techniques or the existence of nonadditive interactions among congeners when samples are analyzed by the bioassay. Overall, mean concentrations of TCDD-EQs in extracts of carp and kestrel eggs were not significantly different between groups of samples collected on and off the RMA, whereas concentrations of TCDD-EQs in several owl livers collected on the RMA were significantly greater than concentrations in owl livers from off the RMA.