This study compared the acute effects of 5 ppm hydrogen sulfide (H2S) inhalation (50 % of its occupational exposure limit) on the biochemical properties of skeletal muscle in exercising men and women. Twenty-five healthy volunteers, 13 men and 12 women, completed two 30-minute submaximal tests at 50% of their predetermined maximal aerobic power (VO2max) while breathing 0 ppm (control) or 5 ppm H2S from a specially designed flow system in a single-blind manner. Immediately after exercise, biopsies were obtained from the vastus lateralis muscle under local anaesthesia. They were subsequently analyzed for concentrations of the following markers of anaerobic and aerobic metabolism: lactate (La), lactate dehydrogenase (LDH), citrate synthase (CS), and cytochrome oxidase (CytOx). Repeated measures analysis of variance indicated that in men, the CS concentration decreased significantly (p = 0.006) as a result of H2S exposure. There was also a tendency for their La and LDH concentrations to increase and CytOx concentration to decrease in the presence of H2S, but these changes were not significant (p > 0.05). In women no significant changes were observed in any of these biochemical properties. These results suggest that (1) exposure to H2S at 50% of its OEL might inhibit aerobic metabolism during exercise in healthy men, thereby increasing their dependency on anaerobic metabolism; and (2) there could be a significant gender difference in the acute response to sub-OEL exposures of H2S.