Pharmaceuticals and Transformation Products in Hospital Wastewater - PowerPoint PPT Presentation

Pharmaceuticals and Transformation Products in Hospital Wastewater and a Rural Conventional Wastewater Treatment Plant Lydia Niemi Mark Taggart 1 , Kenneth Boyd 1 Zulin Zhang 2 , Stuart Gibb 1 1 Environmental Research Institute, University of the Highlands and Islands, Castle Street, Thurso KW14 7JD UK 2 The James Hutton Institute, Craigiebuckler Aberdeen AB15 8QH UK

Pharma: ‘Emerging’ Environmental Contaminants • Pharma extensively used: >102 mil prescriptions in Scotland (2016/17) • Enter environment mainly with WWTP effluent Point source Wastewater treatment Surface water Environmental Removal? Effects? Cl/H 2 O 2 ? h ⱱ Degradation? bio/h ⱱ ? 2 2 Comber et al. 2018; Information Services Division, 2018; Poirier-Larabie et al. 2016

Research Obje jectives 1. Monitor hospital impact on pharma in municipal wastewater 2. Determine pharma change within conventional WWTP 3. Characterise transformation product presence Study location: Wick, Caithness County, Scottish Highlands 3

Study Sites & Sampling Wick WWTP Sampling frequency: 1x per week, 7 weeks May – July 2018 • • Only 24h A&E ‘major’ injuries unit 13500 PE in region (>6800 km 2 ) • 171 L/sec max. flow • 50 medical/surgical beds • Conventional AS • North Sea discharge WWTP WWTP WWTP WWTP Caithness Combined Primary Secondary Final General Influent Sample Sample Effluent Hospital 2 3 4 5 1 4

Target pharmaceuticals Molecular Prescription Water Solubility Prioritised Class Molecular Structure Weight pK a LogP Items, (mg/L) Compound (g/mol) Scotland Paracetamol Analgesic 151.1 9.4 0.4 14000 2680000 No Yes, UK and Diclofenac NSAID 296 4.1 4.5 50000 283150 EU Ibuprofen NSAID 206 4.4 3.9 21 325281 Yes, UK Macrolide Yes, UK and Clarithromycin 748 8.9 3.1 0.33 254270 Antibiotic EU Trimethoprim Antibiotic 290 7.1 0.9 400 481168 Yes, UK Carbamazepine Anticonvulsant 236 13.9 2.4 17 216405 Yes, UK Fluoxetine Antidepressant 309.3 10.1 4.1 14000 844744 Yes, UK 17a-ethynyl Synthetic Yes, UK and 296 10.3 3.6 11 444944 Hormone EU estradiol

Sample Processing & Analysis Bruker Triple • 1L 0.7µm filtration, 1 surrogate spike Quadrupole QQQ HPLC-ESI-MS/MS • 1L SPE Oasis Prime • ESI +/- 2 HLB, elution 12mL 1:1(v:v) Ace:EtOAc • 0.5mL reconstitute Thermo Exactive 3 1:1(v:v) MilliQ:MeOH Orbitrap Q1 UPLC-HESI-MS • Instrumental 4 • HESI +/- analysis 6

HPLC-MS/MS Detection 0102 3 YUAN hospital project 0674 Paracetamol (PAR) 1: MRM of 4 Channels ES+ 3.13 152 > 110 100 1.75e6 3.13 min, 152 > 110 m/z Detection frequency (%) of ESI + mode pharma in % wastewater samples. Limit of quantification (LOQ, 0 YUAN hospital project 0638 2: MRM of 4 Channels ES+ ng/L). 11.11 Trimethoprim (TRI) 291 > 230 100 6.46e5 11.11 min, 291 > 230 m/z PAR TRI CBZ CLAR FLX % Hospital 0 discharge 85 85 100 57 14 YUAN hospital project 0641 3: MRM of 4 Channels ES+ 17.19 237 > 194 (n=7,%) 100 Carbamazepine (CBZ) 2.21e5 WWTP 17.19 min, 237 > 194 m/z Influent 100 100 100 71 14 % (n=7,%) WWTP 0 Primary 100 100 100 66 n.d. YUAN hospital project 0641 4: MRM of 8 Channels ES+ 17.97 749 > 158 100 Clarithromycin (CLA) (n=6,%) 1.20e7 WWTP 17.97 min, 749 > 158 m/z Secondary 50 100 100 100 100 % (n=6,%) WWTP 0 YUAN hospital project 0641 4: MRM of 8 Channels ES+ Effluent 85 100 100 100 100 18.40 310 > 44 Fluoxetine (FLX) 100 2.13e4 (n=7,%) 18.40 min, 310 > 44 m/z LOQ 0.78 0.78 0.81 0.81 3.60 % (ng/L) 19.51 22.17 24.98 0 Time -0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 26.00 28.00 30.00 32.00 HPLC separation ESI + mode pharma in a hospital discharge sample. 7

HPLC-MS/MS Detection 0207 3 Detection frequency (%) of ESI - mode pharma in YUAN hospital project 0838 1: MRM of 1 Channel ES- 2.06 205 > 161 Ibuprofen, IBU 100 1.70e6 wastewater samples. Limit of quantification (LOQ, 2.06 min, 205 > 161m/z ng/L). 17a-ethynyl estradiol (EE2) not detected (n.d.). % IBU DCF EE2 2.63 0 Hospital YUAN hospital project 0855 2: MRM of 4 Channels ES- 100 57 n.d. discharge (n=7,%) 3.29 294 > 250 Diclofenac, DCF 100 5.78e6 WWTP 3.29 min, 294 > 250 m/z 85 57 n.d. Influent (n=7,%) WWTP 83 83 n.d. % Primary (n=6,%) WWTP secondary 100 66 n.d. (n=6,%) WWTP 0 Time 100 57 n.d. -0.00 2.00 4.00 6.00 8.00 10.00 12.00 14.00 16.00 18.00 20.00 22.00 24.00 effluent (n=7,%) HPLC separation ESI - mode pharma in a hospital discharge sample. LOQ (ng/L) 0.78 0.77 4.01 8

Pharma Concentrations pK a = 9.4 LogP= 0.4 Hospital Discharge (n=7) pK a = 10.1 WWTP Influent (n=7) LogP= 4.1 WWTP Primary sample (n=6) WWTP Secondary sample (n=6) WWTP Effluent (n=7) 9

Pharma Concentrations & Hospital Im Impact Hospital Discharge (n=7) * WWTP Influent (n=7) WWTP Primary sample (n=6) WWTP Secondary sample (n=6) WWTP Effluent (n=7) * indicates significant difference (p<0.05) between hospital discharge and WWTP influent, Welch two sample t-test 10

Pharma Removal in WWTP 11

Transformation Products (T (TP) T: FTMS {1,1} + p ESI Full ms [100.00-2000.00] TP1 RT: 0.00 - 32.02 PARENT COMPOUND 253.0970 Carbamazepine (CBZ) 100 253.0970 m/z ( Δ 0 ppm) RT: 13.51 AA: 22031274 90 17.51 min 237.1020 m/z ( Δ 1 ppm) 100 80 70 50 60 RT: 11.99 RT: 10.89 AA: 658122 210.0913 AA: 67737 50 0 RT: 10.21 271.1075 TP1 Carbamazepine-10,11-epoxide 40 * * Acridine ( Δ 1 ppm) AA: 15212112 180.0808 100 30 10.21 min 20 RT: 11.91 50 10 293.0893 5 134.0711 RT: 8.99 AA: 4379074 356.1422 RT: 13.60 425.1308 491.2063 0 AA: 1996168 AA: 207358 100 150 200 250 300 350 400 450 500 0 m/z RT: 11.11 TP2 Dihydroxycarbamazepine AA: 1626879 T: FTMS {1,1} + p ESI Full ms [100.00-2000.00] TP2 100 269.0922 100 11.11 min 269.0922 m/z ( Δ 0 ppm) 90 50 80 70 0 60 0 2 4 6 8 10 12 14 16 18 20 22 24 279.1591 50 Time (min) 40 Carbamazepine and 2 transformation products in secondary 30 149.0233 244.0970 310.1185 sample, week 2. 198.0916 370.0831 20 207.1381 136.0215 405.2457 182.9809 339.2376 • ID after direct/indirect photolysis and biological degradation 10 414.2638 475.2084 509.4 0 • 1 potential carcinogenic compound - acridine * 100 150 200 250 300 350 400 450 500 m/z 12 Mathon et al. 2016; Lekkerkerker-Teunissan et al. 2012; Donner et al. 2013

Transformation Products (T (TP) PARENT COMPOUND TP RT: 0.00 - 32.01 0 RT: 3.19 RT: 4.66 Paracetamol (PAR) Methoxy-paracetamol (M-PAR) AA: 58082215 AA: 3307590 100 100 3.19 min, 152.0701 m/z ( Δ 1 ppm) 4.66 min, 182.0813 m/z ( Δ 2 ppm) 80 80 60 60 40 40 RT: 9.15 20 20 RT: 1.19 RT: 2.08 RT: 4.03 RT: 6.93 RT: 10.65 RT: 12.32 RT: 16.48 RT: 8.41 RT: 10.60 RT: 13.29 AA: 1744131 AA: 10096 AA: 238132 AA: 214673 AA: 590705 AA: 107399 AA: 19839 AA: 61201 AA: 12026 AA: 18746 AA: 16229 0 0 0 0 2 4 6 8 10 12 14 16 18 20 0 2 4 6 8 10 12 14 16 18 20 Time (min) Time (min) UHPLC chromatograms PAR (primary sample), M- PAR (secondary sample), week 1. PAR PAR n.d. M-PAR n.d. M-PAR Influent Primary Secondary Effluent • PAR effective biological degradation (avg 96% WWTP WWTP WWTP WWTP Influent Primary Secondary Effluent removal) 56307 122806 33 1890 PAR • Methoxy-PAR formed during biological treatment 7616-127437 15596-273859 n.d.-61 360-4248 (ng/L) (100%) (100%) (50%) (100%) Ibáñez et al. 2017

Conclusions & Significance 1 st study of pharma behaviour in rural Scottish • Highlands WWTP • Pharma observable in hospital discharge, but other significant sources impacting municipal wastewater • Wick WWTP ineffective for complete pharma removal • <50% avg removal TRI, CBZ; <0% avg removal DCF, FLX • Transformation product formation and persistence observed • Wick harbour and rural environment potential impact from pharma pollution • Tidal zones and estuaries are sink for pharma/organic pollutants (Letsinger et al. 2019; Alygizakis et al. 2016) 14

Acknowledgements Supervisors : Stuart Gibb, Zulin Zhang, Mark Taggart, Kenny Boyd Project contributors : Sylvain Massière (Université de Montpellier, France), Scottish Water Research funders : The Scottish Government’s Hydro Nation Scholars Programme Thank you! lydia.niemi@uhi.ac.uk @LydiaNiemi 15