Optimized Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) pretreatment methods for determination of six polychlorinated biphenyls (PCBs) residues in fish and aquatic invertebrates samples were investigated. Large volume injection (LVI) coupled gas chromatography tandem mass spectrometry (GC-MS/MS) with selected reaction monitoring (SRM) was used to provide a very sensitive and selective means of analyzing PCBs via internal calibration. Three analytical processes were validated and compared, and the florisil Celite ® 545 pretreatment method was selected. Performance characteristics, such as linearity, limit of detection (LOD), limit of quantization (LOQ), recovery and precision (relative standard deviation, RSD) were studied. The method was applied to 233 samples obtained from local markets, which was useful and thus suggested that the method could serve as a screening model for PCB residues analysis.