Abstract
In order to increase hydrogen yield and shorten the lag time, low frequency ultrasonic (LFU) pretreatment was applied to stimulate the bio-activity of the seed bacteria, using waste activated sludge as the inoculum. A pretreated, sterilized sludge sample was seeded as the substrate for bio-hydrogen production. A Unisense microsensor multimeter with a hydrogen microelectrode was employed to examine the hydrogenase activities at different ultrasonic densities and exposure times. The optimum conditions were achieved at an ultrasonic density of 125 W L−1 and a reaction time of 10 s. A maximum hydrogen yield of 13.03 mL H2/g TS was obtained by LFU pretreatment under these optimal conditions, which was 18% higher than that obtained when seeding the raw sludge sample. To gain a better understanding of the end product distribution, three-dimensional excitation–emission matrix (3D-EEM) fluorescence spectroscopy was performed to characterize the dissolved organic matter (DOM) and extracellular polymeric substances (EPS) in the sludge samples during the anaerobic fermentation process. Hydrogen production from NADH/NAD+ conversion was also proved by EEM.
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