It is unknown if females and males use jaw muscles similarly during exercise. This pilot study assessed jaw elevator muscle duty factors (DFs = time of muscle activity/total recording time) at repeated sessions to test if DFs are reliable and different between sexes during exercises in two environments. Ten female and seven male subjects recruited from university soccer teams provided informed consent. Surface electromyography was recorded from masseter and temporalis muscles during biting and leg-extension laboratory exercises. Average activities to produce 20 N bite-forces for each muscle and subject determined thresholds (5-80%·T20 N) for subject-specific DF calculations during exercises performed in laboratory and natural environments. Subjects self-recorded via portable electromyography equipment during in-field leg-extension and weight-lifting exercises. Effects of variables on DFs were assessed via ANOVA (α = 0.05) and simple effects testing (Bonferroni-adjusted p ≤ 0.012). All subjects used jaw muscles during exercises in both environments. DFs between laboratory sessions were reliable (R = 0.84). During laboratory exercises, male temporalis DFs were significantly higher than female DFs from both muscles (p ≤ 0.001). During in-field exercises females had higher DFs during weight-lifting while males had higher DFs during leg-extensions. In-field sex differences were significant at most thresholds and showed larger effect sizes for leg-extension compared to weight-lifting exercises.