Amidst tightening regulations, the proliferation of next-generation per- and polyfluoroalkyl substances (PFAS) necessitates a deeper understanding of their environmental fate and potential risks. Here, we conducted a comprehensive assessment of PFAS in the water and sediment of Taihu Lake, incorporating both nontarget and target screening, seasonal and geographical variation analysis, and risk prioritization. A total of 58 PFAS from 13 classes were identified, revealing complex PFAS contamination. In addition to short-chain perfluoroalkyl carboxylates (PFCAs) and sulfonates (PFSAs), bis(trifluoromethanesulfonyl)imide (Ntf2) and perfluoro-2,5-dimethyl-3,6-dioxo-heptanoic acid (C7 HFPO-TA) exhibited relatively high concentrations in water, with median values of 21.7 and 5.72 ng/L, respectively. Seasonal and geographical variation analysis revealed elevated levels of C7 HFPO-TA, Ntf2, and perfluorohexanoic acid (PFHxA) in the northeastern areas, suggesting transport via water diversion project. Multicriteria risk prioritization identified four high priority PFAS (Ntf2, C7 HFPO-TA, PFHxA, and perfluorooctanoic acid (PFOA)) in water and two high priority PFAS (hexafluoropropylene oxide dimer acid (HFPO-DA) and PFHxA) in sediment. Overall, this study revealed Ntf2 and C7 HFPO-TA as priority PFAS in Taihu Lake, underscoring the urgent necessity of evaluating risks associated with these emerging PFAS.