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Fig. 1 Workflow to evaluate metabolic changes due to a transient exposure of BPF during the developmental phase of life. (A) Zebrafish larvae (7 dpf) were transiently exposed to BPF (1 mg/L, 48 h) and then maintained in standard tank water without BPF for 5 months. (B) Brain, eye, liver, fin, heart, and muscle tissue as well as serum were subjected to two sequential extractions. The first extraction was performed with 2:2:1 (acetonitrile:methanol:water), followed by a second extraction with isopropanol (100%). (C) Polar metabolites were profiled from the 2:2:1 extract by using hydrophilic interaction liquid chromatography (HILIC) coupled to high-resolution mass spectrometry. Lipids were profiled from a 1:1 mixture of the 2:2:1 and isopropanol extracts by using reversed-phase (RP) liquid chromatography coupled to triple quadrupole mass spectrometry. Created with BioRender.com.