Metabolite profiles of live or dead carp (Cyprinus carpio) exposed to endosulfan sulfate using a targeted GC–MS analysis

Abstract

Endosulfan sulfate is a major oxidized metabolite of endosulfan, which is a broad-spectrum chlorinated cyclodiene insecticide. In this study, GC–MS-based metabolic profiles of dead or live carp (Cyprinus carpio) exposed to endosulfan sulfate were investigated to elucidate the molecular toxicological effects of endosulfan sulfate on carp. Three different extraction methods were compared, and a 50% methanol solution was chosen as an efficient extraction method. Carp was exposed to endosulfan sulfate at a concentration of 8ppb for 2days. After exposure, the whole body of the fish was homogenized with liquid N2, extracted with the 50% methanol solution and dried before TMS derivatization for GC–MS analyses of the dead and live carp. A SIM (selected ion monitoring)-library of 373 metabolites was applied after GC–MS analysis to detect 146 metabolites in carp. Based on the one-way ANOVA results (P < 0.001) and fold changes of metabolites in dead carp versus control (fold change > 1.5 or < 0.667), 30 metabolites were identified as biomarkers that were significantly different in the metabolic profiles among the control, dead and live carp. A metabolic pathway analysis using MetaboAnalyst 4.0 revealed that those biomarkers were important for the living or death response to endosulfan sulfate. The pathways indicated by the metabolic pathway analysis included starch and sucrose metabolism, galactose metabolism, glycerolipid metabolism, the citrate cycle and linoleic acid metabolism. These results suggest that these pathways underwent significant perturbations over the exposure period.