Dissolved H(2) and CO(2) were measured by an improved manual headspace-gas chromatographic method during fermentative H(2) production with N(2) sparging. Sparging increased the yield from 1.3 to 1.8 mol H(2)/mol glucose converted, although H(2) and CO(2) were still supersaturated regardless of sparging. The common assumption that sparging increases the H(2) yield because of lower dissolved H(2) concentrations may be incorrect, because H(2) was not lowered into the range necessary to affect the relevant enzymes. More likely, N(2) sparging decreased the rate of H(2) consumption via lower substrate concentrations.