Stable isotopes are commonly used to examine various aspects of animal ecology. The use of stable isotopes generally proceeds under the implicit assumption that resource use is the only factor driving variation in stable isotope levels; however, a wealth of studies demonstrate that a range of common ecological factors can affect the behavior of stable isotopes in animal tissues and potentially confound inferences. For example, studies of some invertebrates and endothermic vertebrates show that animals fasted for ecologically realistic time periods have higher nitrogen (δ¹⁵N) or lower carbon (δ¹³C). We examined whether realistic fasting would influence the stable isotope composition of one of the most metabolically efficient ectothermic vertebrates, the Eastern Red-backed Salamander, Plethodon cinereus. We fasted salamanders for 7, 14, 21, 28, or 35-day intervals and examined whether δ¹⁵N or δ¹³C levels of tissues changed between fasted and fed animals. We investigated whether body condition (body mass to length and C:N [an index of lipid levels]) declined in fasted animals and whether there was a relationship between C:N and δ¹⁵N or δ¹³C. Body mass to length index and C:N, δ¹³C, and δ¹⁵N of tail and liver tissues did not differ between fasted and fed animals between 7 and 35 days. Because of their extreme metabolic efficiency, vertebrate ectotherms such as lungless salamanders (Plethodontidae) may not show the effects of fasting on stable isotopes observed in endothermic vertebrates and some invertebrates. This difference should lead to simpler interpretation of stable isotope results from field studies of these animals.