A general tenet in stress physiology is that the hypothalamic-pituitary-adrenal (HPA) axis predominantly produces one glucocorticoid (GC) in response to stressors. Researchers typically measure only the GC believed to be dominant in their species of interest; however, the two main glucocorticoids produced by vertebrates - cortisol and corticosterone - show variation across species in absolute levels, relative proportions, and stress responsivity. Therefore, generalizing assumptions and focusing on a single GC may not tell the whole story. In this study, we measured both cortisol and corticosterone in water samples collected from five species of Dendrobatid poison frogs. Understanding the full picture of the physiological stress response in poison frogs is of special interest due to their susceptibility to environmental stressors and conservation status. Counter to the dogma that corticosterone is dominant and more abundant in amphibians, we found cortisol was more abundant than corticosterone in four of the five species. Dendrobates tinctorius was unique in releasing equal amounts of cortisol and corticosterone. When we stimulated the HPA axis in D. tinctorius, we found corticosterone was responsive to adrenocorticotropic hormone (ACTH), while cortisol did not change. Comparing plasma and water samples, we found water-borne cortisol was positively correlated with cortisol in plasma and the relationship between plasma and water-borne corticosterone was positive but not statistically significant. These results challenge the broadly accepted assumption that corticosterone is 'dominant' in amphibians and add to the growing literature highlighting the importance of measuring both GCs to understand (stress) physiology.