In this study, five kinds of iron ores, limonite, hematite, manganese ore, magnetite and lava rock, were used as the in situ desulfurizers in the anaerobic digestion reactors to investigate their effects on controlling H2S in biogas. The results show that the addition of the five iron ores could significantly control the content of H2S in biogas, with the best performance for limonite. As limonite dosages increase (10-60 g/L), the contents of H2S in biogas were evidently decreased in the digesters with different initial sulfate concentrations (0-1000 mg/L). After the anaerobic digestion, the removed sulfur was mostly deposited on the surface of limonite. A possible mechanism of H2S control in biogas by limonite was proposed preliminarily, including adsorption, FeS precipitation, and Fe (III) oxidation. The results demonstrated that limonite was a promising in situ desulfurizer for controlling H2S in biogas with low cost and high efficiency.