Stretching can lead to decreased muscle stiffness and has been associated with decreased force and power production. The purpose of this study was to investigate the acute effects of static stretching (SS) on running economy and endurance performance in trained female distance runners. Twelve long distance female (30 ± 9 years) runners were assessed for height (159.4 ± 7.4 cm), weight (54.8 ± 7.2 kg), % body fat (19.7 ± 2.8%), and maximal oxygen consumption (VO2max: 48.4 ± 5.1 ml·kg(-1)·min(-1)). Participants performed 2 sessions of 60-minute treadmill runs following a randomly assigned SS protocol or quiet sitting (QS). During the first 30 minutes (running economy), expired gases, heart rate (HR), and rating of perceived exertion (RPE) were recorded while the participant ran at 65% VO2max. During the final 30 minutes (endurance performance), distance covered, speed, HR, and RPE were recorded while the participant attempted to cover as much distance as possible. Repeated measures analyses of variance were performed on the data. Significance was accepted at p < 0.05. The SS measured by sit-and-reach increased flexibility (SS: 29.8 ± 8.3 vs. QS: 33.1 ± 8.1 cm) but had no effect on running economy (VO2: 33.7 ± 3.2 vs. 33.8 ± 2.3 ml·kg(-1)·min(-1)), calorie expenditure (270 ± 41 vs. 270 ± 41 kcal), HR (157 ± 10 vs. 160 ± 12 b·min(-1)), or endurance performance (5.5 ± 0.6 vs. 5.5 ± 0.7 km). These findings indicated that stretching did not have an adverse effect on endurance performance in trained women. This suggests that the performance decrements previously associated with stretching may not occur in trained women.