Agilent infinity 2

Water samples were collected at 0 h (immediately after dosing) and 24 h (before treatment renewal) of exposure from each treatment and stored at −20 °C in a dark environment until analysis. All samples were analyzed using an Agilent Infinity II (Agilent, Santa Clara, CA, USA) liquid chromatography instrument attached to a SCIEX Triple Quad 5500+ LC-MS/MS (SCIEX, Framingham, MA, USA). A Zorbax Diol (4.6 mm ID, 12.5 mm, 6 μm particle size) attached to an Agilent InfinityLab Poroshell 120 EC-C18 column (4.6 mm ID, 100 mm, 2.7 μm particle size) was used for separation of PFOS and each sample run with a ramping LC solvent gradient with methanol and nanopure water, each containing 20 mmol/L ammonium acetate. Two multiple reaction monitoring (MRM) transitions were employed for each PFOS, one for quantitation and the other for confirmation of the PFOS. Measured PFOS treatment concentrations are available in the Supplementary Materials (Tables S2–S5).

Water samples were taken at 0 and 24 h of exposure from each treatment and stored at −20 °C in a dark environment until analysis. Due to the lengthy chromatographic method and large volume of solvent used per run, water samples were pooled from each replicate which resulted in one measurement per concentration. All samples were directly injected (10 µL) into an Agilent Infinity II (Agilent, Santa Clara, CA, USA) liquid chromatography instrument attached to a SCIEX Triple Quad 5500+ LC-MS/MS (SCIEX, Framingham, MA, USA). A Zorbax Diol (4.6 mm ID, 12.5 mm, 6 μm particle size) attached to an Agilent InfinityLab Poroshell 120 EC-C18 column (4.6 mm ID, 100 mm, 2.7 μm particle size) was used for separation of PFAS and each sample run with a ramping LC solvent gradient with methanol and nanopure water, each containing 10 mmol/L ammonium acetate. A flow rate of 0.4 mL/min was used, and the column was maintained at room temperature. Two multiple reaction monitoring (MRM) transitions were employed for each PFAS, one for quantitation and the other for confirmation of the PFAS (Table S2). The PFAS water concentrations are available in the Supplementary Materials (Table S3).

Central carbon metabolism metabolites were analyzed using an Agilent 6470 liquid chromatography triple quadrupole mass spectrometer (LC-QqQ-MS) coupled with an Agilent Infinity II ultra-high-performance liquid chromatography (UHPLC) system (Agilent Technologies, Santa Clara, CA, USA) as per Beale et al. [31 (link)], and after Gyawali et al. [32 (link)].