Presence of emerging per- and polyfluoroalkyl substances (PFASs) in river and drinking water near a fluorochemical production plant in the

Wouter A. Gebbink †,* , Laura van Asseldonk †, Stefan P.J. van Leeuwen †

† RIKILT, Wageningen University & Research, 6700 AE Wageningen, the Netherlands.

* Corresponding: RIKILT, Wageningen University and Research, P.O. Box 230, NL 6700 AE

Wageningen, the Netherlands. Email: [email protected] ; Tel: +31 (0)317481453

SUPPORTING INFORMATION

16 Pages; 7 Tables; 8 Figures

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Page Table S1. Target compounds and selected instrumental parameters for quantification of S3 each compound by UPLC/ESI MS/MS. Table S2. Sampling location and information of the river and drinking water samples. S4 Table S3. Mobile phase gradient program LCMSMS analysis. S5 Table S4. Recoveries of native PFASs spiked to MilliQ water at 3 concentrations and S5 MQLs. Table S5. Internal standard recovery in river and drinking water samples (n=18). S6 Table S 6. Mobile phase gradient program LCQExactive analysis. S6 Figure S1. Pattern of detected PFCAs and PFSAs in river water samples (R118) and S7 drinking water samples (D16). Figure S 2. Concentrations of ∑PFCA+PFSA (ng/L; top figure) and ∑PFCA+PFSA pattern S8 (bottom figure) reported in the literature at sampling locations nearby the sampling locations of the present study.

Figure S3. Chromatograms of detected C 2n H2n F2n O2 homologues with n = 3, 4, 5, 6, 7, and 8 S9

in river water sample R13 (Top Figure), and MS/MS spectrum of C 6H6F6O2 (233.0199 m/z) as an example spectrum (Bottom Figure). Figure S4. Adjusted mass defect plot for detected homologue series in river water samples S10 collected downstream from a fluorochemical production plant in the Netherlands. Table S7. Formula and possible structures of detected emerging PFASs in river water S11 and/or drinking water collected in 2016 in the Netherlands.

Figure S5. Relative abundance (based on instrumental area count) of detected C 2n H2n F2n O2 S12 homologues in river water samples (R118).

Figure S6. Chromatograms of detected C2n H2n+2 F2n SO 4 homologues with n = 2 and 3 in S13

river water sample R13 (Top Figure), and MS/MS spectrum of C 4H6F4SO 4 (224.9850 m/z) as an example spectrum (Bottom Figure).

Figure S7. Chromatograms of detected C 2n+1 H2n F2n+4 SO 4 homologues with n = 2, 3 and 4 in S14

river water sample R13 (Top Figure), and MS/MS spectrum of C 9H8F12 SO 4 (438.9879 m/z) as an example spectrum (Bottom Figure).

Figure S8. Chromatograms of detected C 2n H2F4n SO 3 homologues with n = 2 and 3 in river S15

water sample R13 (Top Figure), and MS/MS spectrum of C 4H2F8SO 3 (280.9523 m/z) as an example spectrum (Bottom Figure). References S16

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Table S1. Target compounds and selected instrumental parameters for quantification of each compound by UPLC/ESIMS/MS. Compound Chemical name Precursor Product Collision Retention Internal standard ion (m/z) ion (m/z) energy (eV) time (min) used 13 PFBS Perfluorobutane sulfonate 299 80 75 2.87 C3PFBS 299 99 40 18 PFHxS Perfluorohexane sulfonate 399 80 104 3.98 O2PFHxS 399 99 42 18 PFHpS Perfluoroheptane sulfonate 449 80 102 4.29 O2PFHxS 449 99 102 13 PFOS Perfluorooctane sulfonate 499 80 100 4.57 C4 PFOS 499 99 94 13 PFBA Perfluorobutanoic acid 213 169 10 0.99 C4PFBA 13 PFPA Perfluoropentanoic acid 263 219 15 1.62 C3PFPA 13 PFHxA Perfluorohexanoic acid 313 269 14 2.89 C2PFHxA 313 119 24 13 PFHpA Perfluoroheptanoic acid 363 319 12 3.53 C4PFHpA 363 169 24 13 PFOA Perfluorooctanoic acid 413 369 14 3.89 C4PFOA 413 169 24 13 PFNA Perfluorononanoic acid 463 419 16 4.18 C5PFNA 463 169 26 13 PFDA Perfluorodecanoic acid 513 469 16 4.45 C2PFDA 513 219 26 13 GenX Tetrafluoro2(heptafluoropropoxy)propanoic acid 329 169 18 3.16 C3GenX (HFPO-DA, 329 285 6 PFPrOPrA )* 13 ADONA Dodecafluoro3H4,8dioxnonanoic acid 377 251 16 3.67 C4PFHpA 377 85 36 13 6:2 Cl PFESA 9chlorohexadecafluoro 3oxanonane 1sulfonate 531 351 36 4.75 C4 PFOS (F-53B, 531 83 32 9Cl-PF3ONS )* Note: product ions in bold were used for quantification, the other product ions were used for confirmation. * Alternative acronyms used the in literature.

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Table S2. Sampling location and information of the river and drinking water samples. Sampling Waterbody City Sample type Lat °N Long °E Sampling number 1 date River water R1 Breeddiep Hoek van Holland Downstream 51.979580 4.116420 25.10.2016 R2 Downstream 51.915056 4.249758 25.10.2016 R3 Het Scheur Downstream 51.899634 4.348026 25.10.2016 R4 Hoogvliet Downstream 51.861766 4.341127 25.10.2016 R5 Ridderkerk Downstream 51.899898 4.581301 28.10.2016 R6 Kinderdijk Downstream 51.888315 4.628266 28.10.2016 R7 Oude Maas Zwijndrecht Downstream 51.807939 4.593047 25.10.2016 R8 Alblasserdam Downstream 51.861980 4.651010 28.10.2016 R9 Downstream 51.769818 4.629435 25.10.2016 R10 Oude Maas Dordrecht Downstream 51.814985 4.658521 25.10.2016 R11 Noord Papendrecht Downstream 51.834023 4.673907 28.10.2016 R12 Beneden Papendrecht Downstream 51.823413 4.687687 28.10.2016 R13 Dordrecht Downstream 51.821784 4.711317 25.10.2016 R14 Beneden Merwede Sliedrecht Upstream 51.823728 4.745576 28.10.2016 R15 BovenHardinxveld Upstream 51.821290 4.888000 28.10.2016 R16 Haaften Upstream 51.812423 5.214936 25.10.2016 R17 AmsterdamRijnkanaal Utrecht Control site 52.088738 5.079698 28.10.2016 R18 Wageningen Control site 51.961233 5.689544 28.10.2016

Drinking water D1 Drinkingwater Zwijndrecht 25.10.2016 D2 Drinkingwater Dordrecht 25.10.2016 D3 Drinkingwater Papendrecht 28.10.2016 D4 Drinkingwater Sliedrecht 28.10.2016 D5 Drinkingwater Utrecht 28.10.2016 D6 Drinkingwater Wageningen 28.10.2016 1 see Figure 1 map for sampling locations

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Table S3. Mobile phase gradient program LCMSMS analysis. Time (min) Mobile phase A (%) 1 Mobile phase B (%) 2 Flow (mL/min) 0.0 85 15 0.3 0.1 85 15 0.3 0.5 80 20 0.3 3.0 30 70 0.3 6.0 0 100 0.3 8.0 0 100 0.3 9.0 15 85 0.3 12.0 15 85 0.3 1 Mobile phase A = 2 mM ammoniumacetate in water 2 Mobile phase B = Acetonitrile

Table S4. Recoveries of native PFASs spiked to MilliQ water at 3 concentrations and MQLs. PFASs Spike at 5 Spike at 10 Spike at 25 MQL (ng/L) ng/L ng/L ng/L PFBA 87% 102% 102% 2 PFPA 95% 90% 86% 4 PFHxA 115% 110% 113% 0.1 PFHpA 103% 99% 106% 0.05 PFOA 105% 94% 100% 0.3 PFNA 114% 109% 119% 0.03 PFDA 109% 94% 100% 0.03 PFBS 91% 101% 105% 0.04 PFHxS 102% 90% 101% 0.02 PFHpS 101% 107% 93% 0.02 PFOS 103% 97% 105% 0.03 GenX 84% 81% 95% 0.2 ADONA 91% 87% 89% 0.01 6:2 ClPFESA 107% 111% 105% 0.02

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Table S5. Internal standard recovery in river and drinking water samples (n=18). Internal Standard Recovery (%; ± SD) 13 C4PFBA 46 ± 19% 13 C3PFPA 88 ± 8% 13 C2PFHxA 90 ±6% 13 C4PFHpA 95 ± 8% 13 C4PFOA 93 ± 11% 13 C5PFNA 96 ± 12% 13 C2PFDA 83 ± 15% 13 C3PFBS 101 ± 8% 18 O2PFHxS 108 ± 12% 13 C4PFOS 96 ± 15% 13 C3GenX 66 ± 15%

Table S6. Mobile phase gradient program LCQExactive analysis. Time (min) Mobile phase A (%) 1 Mobile phase B (%) 2 Flow (mL/min) 0.0 100 0 0.3 0.1 100 0 0.3 2.0 55 45 0.3 8.0 0 100 0.3 14.5 0 100 0.3 15 .0 100 0 0.3 20 .0 100 0 0.3 1 Mobile phase A = 2 mM ammonium formate + 0.002% formaic acid in water 2 Mobile phase B = 2 mM ammonium formate + 0.002% formaic acid in methanol/water (95/5)

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Figure S1. Pattern of detected PFCAs and PFSAs in river water samples (R118) and drinking water samples (D16).

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Figure S2. Concentrations of ∑PFCA+PFSA (ng/L; top figure) and ∑PFCA+PFSA pattern (bottom figure) reported in the literature at sampling locations nearby the sampling locations of the present study.

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Figure S3. Chromatograms of detected C2n H2n F2n O2 homologues with n = 3, 4, 5, 6, 7, and 8 in river water sample R13 (Top Figure), and MS/MS spectrum of C 6H6F6O2 (233.0199 m/z) as an example spectrum (Bottom Figure). In the chromatograms the mass deviation (in ppm) of the experimental mass compared to the theoretical mass is indicated for each peak.

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Figure S4. Adjusted mass defect plot for detected homologue series in river water samples collected downstream from a fluorochemical production plant in the Netherlands. PFCAs, PFSA, and C2n H2F4n SO 3 were CF 2 normalized, while C 2n H2n F2n O2, C2n H2n+2 F2n SO 4, and C2n+1 H2n F2n+4 SO 4 were CH 2CF 2 normalized.

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Table S7. Formula and possible structures of detected emerging PFASs in river water and/or drinking water collected in 2016 in the Netherlands. Homologue Formula Possible structures

C2n H2n F2n O2 C6H6F6O2

C8H8F8O2

C10 H10 F10 O2

C12 H12 F12 O2

C14 H14 F14 O2

C16 H16 F16 O2

C2n H2n+2 F2n SO 4 C4H6F4SO 4 or

C6H8F6SO 4 or

C2n H2F4nSO 3 C4H2F8SO 3

C6H2F12 SO 3

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Figure S5. Relative abundance (based on instrumental area count) of detected C 2n H2n F2n O2 homologues in river water samples (R118). At sampling location R9 and R1518 no C2n H2n F2n O2 homologues were detected.

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Figure S6. Chromatograms of detected C2n H2n+2 F2n SO 4 homologues with n = 2 and 3 in river water sample R13 (Top Figure), and MS/MS spectrum of C 4H6F4SO 4 (224.9850 m/z) as an example spectrum (Bottom Figure). In the chromatograms the mass deviation (in ppm) of the experimental mass compared to the theoretical mass is indicated for each peak.

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Figure S7. Chromatograms of detected C2n+1 H2n F2n+4 SO 4 homologues with n = 2, 3 and 4 in river water sample R13 (Top Figure), and MS/MS spectrum of C 9H8F12 SO 4 (438.9879 m/z) as an example spectrum (Bottom Figure). In the chromatograms the mass deviation (in ppm) of the experimental mass compared to the theoretical mass is indicated for each peak.

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Figure S8. Chromatograms of detected C2n H2F4n SO 3 homologues with n = 2 and 3 in river water sample R13 (Top Figure), and MS/MS spectrum of C 4H2F8SO 3 (280.9523 m/z) as an example spectrum (Bottom Figure). In the chromatograms the mass deviation (in ppm) of the experimental mass compared to the theoretical mass is indicated for each peak.

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References

1. Moller, A.; Ahrens, L.; Surm, R.; Westerveld, J.; van der Wielen, F.; Ebinghaus, R.; de Voogt, P., Distribution and sources of polyfluoroalkyl substances (PFAS) in the River watershed. Environ. Pollut. 2010, 158, (10), 32433250.

2. Heydebreck, F.; Tang, J. H.; Xie, Z. Y.; Ebinghaus, R., Alternative and Legacy Perfluoroalkyl Substances: Differences between European and Chinese River/Estuary Systems (vol 49, pg 8386, 2015). Environ. Sci. Technol. 2015, 49, (24), 1474214743.

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