This study developed a method to perform the simultaneous concentration and selective separation of66(ultra) trace persistent toxic substances in Antarctic waters. The substances included 30 polychlorinated biphenyls, 17 organochlorine pesticides, 16 polycyclic aromatic hydrocarbons, 3hexabromocyclododecanes. Solid phase extraction was performed using a C18 membrane and silica gel column. Gradient elution was conducted using organic solvents with different polarities; as a result, the efficiency of the C18 film is improved and the interferences from impurities and target compounds are eliminated. Extracts were subsequently analyzed through gas chromatography or liquid and gas chromatography coupled to tandem mass spectrometry. Method validation yielded the following values:recoveries of all target analytes in the Antarctic water ranged from 87.3% to 117.6% and reproducibility as percent relative standard deviation was lower than 5%. Quantification limits ranged from 0.004 mg L 1[2_TD$IF]to0.030 mg L 1. The established method improved the recoveries and reduced the limits of detection.Results indicated the method exhibited good performance in the simultaneous concentration and selective separation of 66(ultra) trace organic pollutants; Therefore, the proposed sample pretreatment can potentially eliminate the effects of various classes of impurities to some extent. This study developed a method to perform the simultaneous concentration and selective separation of 66 (ultra) trace persistent toxic substances in Antarctic waters. The substances included 30 polychlorinated biphenyls, 17 organochlorine pesticides, 16 polycyclic aromatic hydrocarbons, 3hexabromocyclododecanes. Solid phase extraction was performed using a C18 membrane and silica gel column. Gradient elution was conducted using organic solvents with different polarities; as a result, the efficiency of the C18 film is improved and the interferences from impurity and target compounds are eliminated. and gas chromatography coupled to tandem mass spectrometry. Method validation yielded the following values: recoveries of all target analytes in the Antarctic water ranged from 87.3% to 117.6% and reproducibility as percent relative standard deviation was lower than 5%. Quantification limits ranged from 0.004 mg L 1 [2_TD $ I F] to 0.030 mg L 1. The established method improved the recoveries and reduced the limits of detection. Results indicate the method exhibiting good performance in the simultaneous concentration and selective separation of 66 (ultra) trace organic pollutants; Therefore, the proposed sample pretreatment can potentially eliminate the effects of various classes of impurities to some extent.