A high-resolution mass spectrometry screening method was developed and validated based on EU SANTE/11312/2021 guidelines for the analysis of 850 multi-class contaminants in commercial seafood samples. Samples were extracted using a novel sequential QuEChUP preparation method that combines the QuEChERS and QuPPe procedures. The screening detection limits (SDLs) and limits of identification (LOIs) were equal to or lower than 0.01 mg·kg⁻¹ for 92% and 78% of contaminants, respectively. This screening procedure was ultimately applied for a target screening analysis of 24 seafood samples. The concentrations of identified contaminants were assessed using semi-quantitative approach. Two identified contaminants, diuron and diclofenac, showed the highest estimated average concentrations: 0.076 and 0.068 mg·kg⁻¹ respectively in mussel samples. Suspect screening was also performed. Target and suspect screening led to the identification of mixtures of contaminants (pesticides, veterinary products, industrial chemicals and personal care products) and the assessment of their frequencies of appearance (FoA).