Insectivorous bats regularly experience dramatic and sometimes rapid changes in nutrient stores, yet our ability to study these changes has been limited by available techniques. Plasma metabolite analysis has proven effective for studying individual rates of mass change in birds but has not been validated for other taxa. We tested the effectiveness of plasma metabolite analysis by conducting a study with captive big brown bats (Eptesicus fuscus) and little brown bats (Myotis lucifugus) in the field. In the lab, we varied food availability to induce various rates of mass change. As predicted, individual rate of mass change was positively correlated with plasma triglyceride concentration, but there was no relationship with plasma β-hydroxybutyrate concentration, whereas such a relationship has been found in birds. In the field, we collected blood samples from postlactating females as they emerged in the evening (fasted) and when they returned from feeding in the morning. Plasma triglyceride concentration was greater in fed bats than fasted bats, and the increase was less when rain limited foraging. Contrary to predictions, β-hydroxybutyrate concentration was also greater in fed bats than fasted bats. Analysis of plasma triglyceride concentration provides a technique for assessing individual feeding state and rate of mass change of bats and will facilitate further study of bat nutritional ecology and energetics.