In many operational scenarios, hypohydration can be corrected with oral rehydration following the work interval. Although rare, there are potential situations that require extended intervals of uncompensable heat stress exposure while working in personal protective equipment (PPE). Under these conditions, retention of body water may be valuable to preserve work capacity and reduce cardiovascular strain. We conducted a pilot study comparing intramuscular atropine sulfate versus saline placebo to establish the safety profile of the protocol and to provide pilot data for future investigations. Five, healthy, heat-acclimated subjects completed this crossover design laboratory study. Each subject performed up to one hour of exertion in a hot environment while wearing a chemical resistant coverall. Atropine sulfate (0.02 mg/kg) or an equivalent volume of sterile saline was administered by intramuscular injection. Core temperature, heart rate, perceptual measures, and changes in body mass were measured. All five subjects completed the acclimation period and both protocols. No adverse events occurred, and no pharmacologically induced delirium was identified. Change in body mass was less following exercise influenced by atropine sulfate (p = 0.002). Exertion time tended to be longer in the atropine sulfate arm (p = 0.08). Other measures appeared similar between groups. Intramuscular atropine sulfate reduced sweating and tended to increase the work interval under uncompensable heat stress when compared to saline placebo. Heart rate and temperature changes during exertion were similar in both conditions suggesting that the influence of an anticholinergic agent on thermoregulation may be minimal during uncompensable heat stress. BACKGROUND thermoregulation; cholinolytic; anticholinergic; reaction time.