Hydrogen production from diluted molasses by anaerobic fermentation bacteria was investigated in a three-compartment anaerobic baffled reactor (ABR) with an effective volume of 27.48 L. After being inoculated with aerobic activated sludge and operated at chemical oxygen demand (COD) of 5000 mg/L and temperature of 35 degrees C for 26 days, the ABR achieved stable ethanol-type fermentation. The liquid fermentation products, including volatile fatty acids (VFAs) and ethanol, stabilized at 1254, 2053, and 2761 mg/L in the three compartments, respectively. Effluent pH, oxidation-reduction potential (ORP), and alkalinity ranged at 4.3-4.4, -241 to -249 mV, and 306-334 mg CaCO3 /L, respectively. The hydrogen yield of the ABR was 32.51 L/d at the stable operation status, specific hydrogen production rate of anaerobic activated sludge was 0.13 L/g MLVSS d, and the substrate conversion rate was 0.13 L/g COD. Hydrogen yields, fermentation types, and acclimatization durations varied in each compartment, with the 1st compartment having lowest hydrogen yield but longest acclimatization duration and the 2nd and 3rd compartments having higher hydrogen yields but shorter acclimatization durations. The study found that the individual compartment configuration in the ABR system provided a favorable environment for different types of anaerobic bacteria. Compared with complete stirring tank reactor (CSTR), the ABR system had a better operation stability and microbial activity, which led to higher substrate conversion rate and hydrogen production ability. (C) 2007 International Association for Hydrogen Energy. Published by Elsevier Ltd. All rights reserved.