Biohydrogen production by fermentation has become a promising technology developed in the world. In this study, diluted normal molasses wastewater is used as the raw material, and biohydrogen production efficiency and operation characteristics of an anaerobic baffled reactor (ABR) are studied. The effect of hydraulic retention time (HRT) on biohydrogen production efficiency and operating characteristics of ABR is extensively discussed. Experimental results showed that methanogen residuals were still observed in the last three compartments under HRT of 24 h and COD(Chemical oxygen demand) concentration of 8000 mg/L. Meanwhile, the first three compartments presented an ethanol fermentation type. The characteristics of butyric acid fermentation in Compartment IV were also enhanced. The average removal efficiency of COD was reduced to 15.4%. The average rates of biohydrogen production and specific biohydrogen production were 12.85 and 360.22 L/kg COD, respectively. The extension of HRT was beneficial to enrich hydrogen-producing acetogens and could increase the production rate of biohydrogen by a factor of 1.65. However, with the decrease in the bioactivity of acidogenic fermentation bacteria, the biohydrogen production efficiency of ABR was significantly reduced when HRT was longer than 30 h. The specific biogas production rate decreased from 191 to 92 L/(kg MLVSS·d). The specific biohydrogen production rate also decreased from 24.34 to 2.7 L/(kg MLVSS·d).