Abstract: Superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and glutathione s-transferase (GST) in gills and liver of Boleophthalmus pectinirostris were selected as biomarkers to explore the acute biotoxic effects of different concentrations of crude oil in intertidal sediment on Boleophthalmus pectinirostris after 48 hours. The results showed that with the increase of pollution concentration, the activities of SOD, GSH-Px and GST increased firstly and then dropped down, indicating that the enzyme activities of gills and liver of Boleophthalmus pectinirostris were activated at low concentration while inhibited at high concentration. The maximum induction times of SOD, CAT and GST activities in gills of Boleophthalmus pectinirostris were higher than those in liver, which showed that the gills of Boleophthalmus pectinirostris were more sensitive to crude oil. Combined with the significance of antioxidant enzyme activities in different tissues, it was found that the SOD and GST in gills were more sensitive to crude oil pollution and more suitable as effective biomarkers for monitoring marine crude oil pollution.