Document gaojeZNRkLyL8BQ6YRJ0KmppN

Supporting Information: Quantitation of Total PFAS including Trifluoroacetic Acid with Fluorine Nuclear Magnetic Resonance (19F-NMR) Authors: Dino CamdziclI, Rebecca A. DickmanIT, Abigail S. Joyce3, Joshua S. Wallace1'2, P. Lee Ferguson3, Diana S. Aga1,2* 1. Department of Chemistry, University at Buffalo SUNY, Buffalo, NY 14260 2. RENEW Institute, University at Buffalo SUNY, Buffalo, NY 14260 3. Depaitment of Civil and Environmental Engineering, Duke University, Durham, NC 27708 *Corresponding author -- @buffalo.edu TThese authors contributed to this work equally. Supporting Information includes: 13 pages, 5 tables, 1 figure, and 1 equation Si Table of Contents: Materials and Methods................................................................................................................S3 Experimental Materials......................................................................................................S3 Sample Preparation and Analysis.......................................................................................S3 Table S1. PFAS analytes (n=32) and their respective abbreviations, target surrogate, chemical shift, and supplier for the analytes targeted..............................................................S5 Table S2. Extrapolation of limit of detection (LOD) based on scans performed on PFOS solutions........................................................................................................................................S7 Equation S1. Quantification of PFAS analytes via Internal Standard in 19F-NMR..............S7 Table S3. LC-HRMS Quantification of WWa............................................................................S8 Table S4. LC-HRMS Quantification of WWb............................................................................S9 Table S5. PFAS detected with HRMS suspect screening........................................................S10 Figure S1. F-NMR spectra of the landfill leachate extract.....................................................S12 References...................................................................................................................................S13 S2 Materials and Methods Experimental materials Perfluoropropionic acid (PFPrA) reference standard and chromium (III) acetylacetonate (Cracac4) were purchased from Acros Organics (Pittsburgh, PA). Perfluoro(2ethoxyethane)sulfonic acid (PFEESA) and Perfluoro-3-methoxypropanoic acid (PFMPA) were purchased from Synquest Laboratories (Alachua, FL). Nonafluoro-3,6-dioxaheptanoic acid (NFDHA) was purchased from Apollo Scientific (Stockport, UK). Reference standards for Trifluoroacetic acid (TFA) and hexafluorobenzene (HFB) were purchased from Sigma Aldrich (Milwaukee, WI) and Trifluoro sulfonic acid (TFMS) was purchased from Alfa Aesar (Tewksbury, MA). The remaining 29 PFAS standards, 13C-labelled PFOA (MPFOA) standard, and a mixture of isotopically labelled PFAS standards (MPFAC - 24 ES) (Table S1) were purchased from Wellington Laboratories, LLC (Overland Park, KS). Deuterated methanol was purchased from Cambridge Isotope Laboratories (Andover, MA). Liquid chromatography-mass spectrometry (LCMS) grade acetonitrile and methanol for instrumental analysis was obtained from Omnisolv through Millipore Sigma (Saint Louis, MO). American Chemical Society (ACS) grade glacial acetic acid and ammonium acetate were obtained from J. T. Baker (Philipsburg, NJ). Waters BEH S3 X-BridgeTM (3.5 m particle size, 2.1 mm internal diameter, 150 mm length) analytical column was obtained from Waters (Milford, MA). Sample Preparation and Analysis Two wastewater samples were analyzed for the comparison of TOP and LC-HRMS to 19F- NMR. The TOP wastewater (WWa) (10.0 mL) was prepared using a TOP assay and analyzed as discussed in Martin et al. 2019. After this, WWa was lyophilized (-40C) and reconstituted in D4MeOH (500 L) with Cracac4 (4 mg/mL) and analyzed with 19F-NMR analysis. A second wastewater (WWb) sample (2.0L) was lyophilized, diluted in D4-MeOH (1.4 mL), and split for individual 19F-NMR and LC-HRMS analysis. The 19F-NMR WWb aliquot was reduced 20%, followed by the addition of Cracac4 (4 mg/mL) and hexafluorobenzene, an internal standard for quantification in all 19F-NMR analyses. The LC-HRMS WWb aliquot (200 L) was dried under N2 and resuspended in 25:75 (v/v) 5mM ammonium acetate (pH 3.8): MeOH (700 L) then analyzed by LC-HRMS analysis for targeted quantitation (n= 27) and PFAS suspect screening. Fluoromatch FlowTM 2.2 was used for suspect screening, matching theoretical exact masses to the EPA PFAS master list and known standards. Data dependent acquisition was performed for LCHRMS samples, with any detections being manually inspected to ensure validity according to peak shape and fragmentation spectra. Detections were then graded according to the Schymanski1 scale. Reference standards were used to confirm level 1 detections, 2a confirmed using reference spectra from Massbank of North America to compare to sample spectra, and 2b confirmed using a precursor match to the EPA PFAS Master list, -0.25 to 0.1 amu mass defect, and at least one common PFAS fragment. If an unknown PFAS had multiple potential isomers and fragmentation S4 is not sufficient to assist in the identification of the specific isomer, the identification as labeled as level 3. The WW samples both underwent sample preconcentration through lyophilization to improve the method LOD. Lyophilization removes water from frozen samples at the sublimation point, so unlike SPE, lyophilization is inherently less biased because the highly polar low molecular weight PFAS are not lost during pre-concentration. S5 Table S1. PFAS analytes (n=32) and their respective abbreviations, target surrogate, chemical shift, and supplier for the analytes targeted in the F-NMR and LCMS analysis, organized by class. The isotopically labelled surrogates identified are used for isotope dilution quantification in LC-MS analysis. NA denotes not applicable for analytes that are not amenable to the analysis by the LC-MS method used in this study. The reported chemical shift is the observed shift for the terminal -CF3 group of each PFAS. Analyte Carboxylates Trifluoroacetic acida Perfluoropropionic acid Perfluorobutanoic acid Perfluoropentanoic acid Perfluorohexanoic acid Perfluoroheptanoic acid Perfluorooctanoic acid Perfluorononanoic acid Perfluorodecanoic acid Perfluoroundecanoic acid Perfluorododecanoic acid Perfluorotridecanoic acid Perfluorotetradecanoic acid Sulfonic acid Trifluoromethane sulfonic acid Perfluorobutanesulfonic acid Perfluoropentanesulfonic acid Perfluorohexanesulfonic acid Abbreviation Isotopically labelled surrogate F-NMR -CF3 chemical shift (ppm) TFA PFPrA PFBA PFPeA PFHxA PFHpA PFOA PFNA PFDA PFUdA PFDoA PFTrDA PFTeDA NA NA 13C4-PFBA 13C5-PFPeA 13C5-PFHxA 13C4-PFHpA 13C8-PFOA 13C9-PFNA 13C6-PFDA 13C7-PFUDA 13C2-PFDoA 13C2-PFDoA 13C2-PFTeDA -77.0 -84.9 -82.5 -82.6 -82.5 -82.4 -82.4 -82.4 -82.4 -82.4 -82.4 -82.4 -82.4 TFMS PFBS PFPeS PFHxS NA 13C3-PFBS 13C3-PFHxS 13C3-PFHxS -80.2 -82.3 -82.5 -82.4 Supplier Sigma Aldrich Acros Organics Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Alfa Aesar Wellington Laboratories Wellington Laboratories Wellington Laboratories S6 Perfluoroheptanesulfonic acid Perfluorooctanesulfonic acid Perfluorononanesulfonic acid Perfluorodecanesulfonic acid Aminated Perfluorooctanesulfonamide 2-(N-Methylperfluorooctanesulfonamido) acetic acid 2-(N-Ethylperfluorooctanesulfonamido) acetic acid Fluorotelomers Fluorotelomer sulfonic acid 4:2 Fluorotelomer sulfonic acid 6:2 Fluorotelomer sulfonic acid 8:2 PFHpS PFOS PFNS PFDS FOSA NMeFOSAA NEtFOSAA 4:2 FTS 6:2 FTS 8:2 FTS 13C8-PFOS 13C8-PFOS 13C8-PFOS 13C8-PFOS 13C8-FOSA d3-N-MEFOSAA d5-EtFOSAA 13C2-4:2 FTS 13C2-6:2 FTS 13C2-8:2 FTS -82.4 -82.4 -82.4 -82.4 -82.4 -82.4 -82.4 -82.7 -82.4 -82.4 Carboxylate Ethers Hexafluoropropylene oxide dimer acid HFPO-DA NA -82.9 Perfluoro(2-ethoxyethane)sulfonic acid PFEESA NA -88.3 Perfluoro-4-methoxybutanoic acid PFMBA NA -56.8 Perfluoro-3-methoxypropanoic acid PFMPA NA -56.9 Nonafluoro-3,6-dioxaheptanoic acid NFDHA NA -57.0 aTFA was not included in the initial training set for method development, but was added later for detection confirmation Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Wellington Laboratories Synquest Sigma Aldrich Synquest Apollo Scientific S7 S8 Table S2. Extrapolation of limit of detection (LOD) based on scans performed on PFOS solutions in D4-MeOH with Cracac4. The S/N ratios for the -CF3 shift was used to extrapolate the methods LOD, which was concluded to be 50 g/L. Concentration (g/L): Transients (nt): S/N: 2,000 100 12 2,000 1800 48 500 1800 12 500 16000 36 50 16000 3.6 Equation S1. Quantification of PFAS analytes with internal standard, hexafluorobenzene (HFB). # () C = # ( 3) Where: . C: ( 3) : () Ex. TOP total PFAS S9 C = 1210..1789 63 4.53 10 5 C:195.04 Table S3. Total oxidizable precursor quantitation using liquid chromatography with mass spectrometry. Results are expressed as M for each carboxylate and sulfonate end product of the oxidation reaction. A total of 6.66 M of PFAS were detected in the WWa sample. Analyte PFBS PFPeS PFHxS PFBA PFPeA PFHxA PFHpA PFOA PFNA PFDA Total PFAS Concentration (Mole / L) 1.57E-05 4.00E-06 3.81E-06 1.92E+00 3.45E+00 1.14E+00 1.59E-01 7.26E-05 2.85E-05 3.89E-06 6.66E+00 Concentration (g/L) 4.71E-03 1.40E-03 1.52E-03 4.11E+02 9.11E+02 3.58E+02 5.79E+01 3.01E-02 1.32E-02 2.00E-03 1.74E+03 S10 Table S4. LC-HRMS quantitative targeted analysis results for individual PFAS detected in WWb (M) and the calculated total PFAS within the sample. The total PFAS detected was 6.926 M. Analyte 6:2 FTS 8:2 FTS N-Me-FOSAA N-Et-FOSAA PFHxS PFOA PFOS PFBA PFBS PFDA PFHpA PFHxA PFNA PFPeA PFPeS PFUdA Total PFAS Concentration (Mole / L) 0.738 0.009 0.124 0.033 0.065 1.365 0.096 0.738 0.034 0.136 0.625 2.065 0.047 0.844 0.006 0.002 6.926 Concentration (g/L) 316 4.75 70.8 19.3 26.0 565 48.0 158 10.2 69.9 228 649 21.8 223 2.10 1.13 2412 S11 Table S5. PFAS detected with high resolution mass spectrometry suspect screening. Detections were supported by an exact mass match (+5 ppm mass error), at least 2 structurally relevant fragments, and isotopic pattern matches. Homologous series are considered detections separated by a single -CF2 unit and are denoted by *,,. Name 1-(ethoxymethyl)-3(2,2,3,3,4,4,5,5,6,6,7,7,8,8,8- pentadecafluorooctyl)urea 2,2,3-trifluoro-3- ((1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8hexadecafluoro-8- (trifluoromethoxy) octyl)oxy) propanoic acid Retention Chemical m/z time Formula (min) 499.0533 11.95 C12H11F15N2O2 626.9533 14.09 C12H2F22O4 Fragments C4F9 C2F5, C5F11 Detection Confidence 3c Structure O F2 F2 F2 O N N C C C C C C CF3 H H F2 F2 F2 F2 F2 F2 F2 F H O 3c F3C O C C C C C C C C O C C OH F2 F2 F2 F2 F2 O OH N-(Perfalcuiodro1b-eutthaynloeyslt)egrlutamic 370.0542 14.86 C11H12F7NO5 CC190HH71F1F6N7NOO3,3 3a 3,4,4,5,5,6,6,7,7,8,8,8dodecafluorooct-2-enoic acid* 356.9794 16.46 C8H2F12O2 C7F11 1b N-Ethyl-N- C4F9, [(nonafluorobutyl)sulfonyl]glycine 383.9960 17.87 C8H8F9NO4S C4F9SO2, 2b C6H7F9NSO2 Unde3c,4af,4lu,5o,r5o,o6c,6t-,27-,7e,n8o,8ic- acid 338.9885 17.89 C8H3F11O2 CC52FF151, 3c F2 O F3C C C N F2 H O O F2 F2 F F2HC C C C C C F2 F2 O OH F2 O N F3C C C C S F2 F2 O F2 F2 F F2HC C C C C C F2 F2 O OH O OH S12 6:2 Fluorotelomer carboxylic acid 376.9855 18.11 C8H3F13O2 Name Retention Chemical m/z time Formula (min) 3h,e4x,4a,d5e,c5a,6fl,u6o,7r,o7d,8ec,8-,29-,e9n,1o0ic,1a0c,i1d0*- 456.9728 19.60 C10H2F16O2 5,5,6,6,7,7,8,8,9,9,10,10,10Tridecafluoro-2-hydroxydecanoic acid 421.0118 20.32 C10H7F13O3 5:3 Fluorotelomer carboxylic acid 341.0042 20.70 C8H5F11O2 7:2 Fluorotelomer carboxylic acid 476.9791 21.31 C8H2F12O2 6:3 Fluorotelomer carboxylic acid 391.0012 22.43 C9H5F13O2 7:3 Fluorotelomer carboxylic acid 440.9979 24.06 C10H5F15O2 CO2F, 1b C7F11 Fragments Detection Confidence C9F15 2c C7F9, 3c C7F7 C7F7, 1b C7HF8 CO2F, 2c C9F15 C7F7, C8F9, 2c C8HF10 C8F9, C9F11, 1b C9HF12 F2 F2 F2 F3C C C C C C F2 F2 O OH Structure F2 F2 F2 F F3C C C C C C C C F2 F2 F2 O OH F2 F2 F2 F3C C C C C C F2 F2 O OH OH F2 F2 F3C C C C C F2 F2 O OH F2 F2 F2 F2 F3C C C C C C C C F2 F2 F2 O OH F2 F2 F3C C C C C C F2 F2 F2 O OH F2 F2 F2 F3C C C C C C C F2 F2 F2 O OH S13 * unsaturated polyfluorocarboxylic acid homologous series n:2 Fluorotelomer carboxylic acid homologous series n:3 Fluorotelomer carboxylic acid homologous series Figure S1. F-NMR spectra of the landfill leachate extract, with a 100 g/L PFOS standard addition to elucidate the triplet character of the -CF3 peak. Peaks are labeled at (A) trifluoroacetic acid (TFA), (B) -CF3 of per- and polyfluoroalkyl substances (PFAS), and (C) hexafluorobenezene. In this example, (B) shows the clear triplet peak of the -CF3 shift from the PFOS standard addition. This supports the environmental detection of -CF3 proximal to the LOD. A -76.5 -77.0 B -82.0 -82.5 -83.0 Shift (ppm) C -164.5 -165.0 -165.5 4000000 3800000 3600000 3400000 3200000 3000000 2800000 2600000 2400000 2200000 2000000 1800000 1600000 1400000 1200000 1000000 800000 600000 400000 200000 0 -200000 Intensity S14 References: 1. Schymanski, E. L.; Jeon, J.; Gulde, R.; Fenner, K.; Ruff, M.; Singer, H. P.; Hollender, J., Identifying Small Molecules via High Resolution Mass Spectrometry: Communicating Confidence. Environ. Sci. Tech. 2014, 48 (4), 2097-2098. S15