Using human college-age subjects, the present study tested the commonly cited but previously untested hypothesis that yawning is facilitated by higher than normal levels of CO2 or lower than normal levels of O2 in the blood by comparing the effect on yawning of breathing 100% O2 and gas mixtures with higher than normal levels of CO2 (3 or 5%) with compressed air, the control condition. If yawning is a response to heightened blood CO2, the CO2 mixtures should increase yawning rate and/or duration. If low blood O2 produced yawning, breathing 100% O2 should inhibit yawning. The CO2/O2 hypothesis was rejected because breathing neither pure O2 nor gases high in CO2 had a significant effect on yawning although both increased breathing rate. A second study found that exercise sufficient to double breathing rate had no effect on yawning. The two studies suggest that yawning does not serve a primary respiratory function and that yawning and breathing are triggered by different internal states and are controlled by separate mechanisms.