Simple method for the measurement of the hydrogenotrophic methanogenic activity of anaerobic sludges

The specific hydrogenotrophic activity of anaerobic sludges is usually assayed by gas chromatographic analysis for methane in the headspace of sealed test vials. Gas is sampled with a pressure lock syringe which allows quantification independent of the pressure prevailing in the vials. An alternative method was developed using pressure transducer monitoring of the decrease in headspace gas pressure as the H2/CO2 substrate is converted to CH4. Application of a simple formula related the decrease at each sample point to millilitres of CH4 produced and gave values for the specific hydrogenotrophic activity of granular anaerobic sludge which were in good agreement with the values obtained by the more labor-intensive gas chromatographic method. The simplicity of the method facilitates multiple replicate analyses and allows more accurate determination of initial rates than is achievable by the gas chromatographic method which is prone to analytical error at the very low concentrations of CH4 present in the headspace during the early stages of the assay. Mass transfer of H2 from headspace to liquid was found to be rate-limiting and to result in significant under-estimation of the specific hydrogenotrophic activity of the granular sludge. A test protocol, which used a vial volatile suspended solids concentration between 1.7 and 8 g l-1; a 1:5 ratio between liquid and headspace; incubation of the vials horizontally with vigorous shaking (180 rev./min) and an initial H2/CO2 (80/20) gas pressure of 100-150 kPa was found to give reproducible and maximal values for the specific hydrogenotrophic activity of the test sludge.