Neotropical poison frogs possess alkaloid-based antipredator defenses which they sequester from a diet of arthropods such as oribatid mites and myrmicine ants. Alkaloid sequestration is still poorly understood and although several studies have examined its uptake, most experiments directly feed alkaloids to the frogs. Here, we examined the alkaloid uptake system in the poison frog species Dendrobates auratus by feeding it an alkaloid-containing prey item, the red imported fire ant Solenopsis invicta (Formicidae, Myrmicinae). Captive bred frogs were either fed live ants or fruit flies dusted with powdered ants for 4 months. Using GC–MS, we confirm that S. invicta contain previously described piperidine alkaloids known as solenopsins; however, none of these piperidine alkaloids was detected in the skin of D. auratus, suggesting the frogs are incapable of sequestering solenopsins from S. invicta. It is possible that D. auratus are unable to sequester fire ant piperidines due to their long hydrocarbon side chains, a feature that makes them structurally different than most known alkaloids in poison frogs.