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1 Supporting Information for A model to estimate the population contributing to the wastewater using samples collected on census day Jake W. O Brien* a, Phong K. Thai a, Geoff Eaglesham a, Christoph Ort b,c, Andreas Scheidegger c, Steve Carter d, Foon Yin Lai a, Jochen F. Mueller a a The University of Queensland, The National Research Centre for Environmental Toxicology (Entox), 39 Kessels Road, Coopers Plains, QLD 4108, Australia b The University of Queensland, Advanced Water Management Centre (AWMC), St. Lucia, QLD 4072, Australia c Eawag, Swiss Federal Institute of Aquatic Science and Technology, CH 8600 Dübendorf, Switzerland d Queensland Health Forensic and Scientific Services, 39 Kessels Road, Coopers Plains, QLD 4108, Australia *j.obrien2@uq.edu.au * Tel. +61 (0) , Fax +61 (0) Number of pages: 4 Number of figures: 2 Number of tables: 2 0
2 An example copy of the R code and calibration data to run the model can be downloaded at: SI 1. Total daily wastewater volume versus population As no rainfall occurred during the sampling period at any of the STPs, the relationship between the total daily volume of wastewater and the census population was assessed with a linear regression (Figure 1). This indicated a strong correlation with an R 2 = 0.99 and that each person within a catchment contributes 250 ± 10 L of wastewater per day. However, this method for population estimation is fraught with uncertainties such as the daily variability of water consumption, infiltration/exfiltration of water into/out of the sewer, seasonal variations in water consumption, commercial inputs of wastewater and so on 2, 3. Discussion with STP operators, however, identified that most of their population estimates are based solely on wastewater volume where a certain volume of water is assigned as a Population Equivalent. Our continuous flow monitoring data from three STPs indicate that wastewater entering the plant during rain events may increase more than 100% and thus this method for assessing population size is not applicable during such events. Figure SI 1. Population versus daily wastewater volume for 10 different STPs. The black line is the best fit. Dotted lines represent the 95% confidence interval while continuous grey lines represent the 95% prediction interval. 1
3 2
4 SI 2. Chemical mass loads for each wastewater catchment Table SI 1. Chemical mass loads [kg/day] as measured in the wastewater samples for each site STP ID Date Sampled (D/M/Y) ABS Census Count STP Estimat e acesulfame atenolol caffeine codeine frusemide gabapentin Chemical Mass Load [kg/day] carbamazaepine hydrochlorothiazide ibuprofen iopromide naproxen norfloxacin paracetamol salicylic acid 1 10/08/2011 3,682 3, >LOQ /08/2011 8,995 10, /08/ ,698 28, >LOQ /08/ ,005 26, /08/ ,513 40, /08/ , , /08/ , , /08/ ,104 96, /08/ ,104 96, /08/ , , /08/ , , /08/ , , /08/ , , /08/ , , * * measurement considered to be an outlier LOQ = limit of quantification 2
5 SI 3. Population versus daily mass load of iopromide outlier Linear regressions were plotted for the measured mass loads of all potential chemical population size biomarkers against the population size; it appears that iopromide contains an outlier. Including this outlier changed the slope of the regression from y = 5.95 e-7 to y = 2.22e-6 (Figure 2). We therefore decided to exclude this value from our calibration data. Figure SI 2. Population versus daily mass load of iopromide with and without the outlying value. The black line is the line of best fit. The grey lines indicate the 90% prediction interval. SI 4. Instrumental parameters for analysis of pharmaceutical and personal care products Table SI 2. LCMS/MS parameters using an AB/Sciex API 5500Q mass spectrometer (AB/Sciex, Concord, Ontario, Canada) with an electrospray ionization (ESI) interface coupled to a Shimadzu Nexera HPLC system (Shimadzu Corp., Kyoto, Japan). Separation was achieved on a Luna C-18 (2) (3 µm 100 Å LC Column 150 mm x 3 mm, Phenomenex) column using a mobile phase gradient of 1 to 95 % acetonitrile with 0.1% formic acid. 3
6 Source Compound Retention Time (RT) Quantitation Ion/Transition Confirmation Ion/Transition MRM (Q1 > Q3) DP EP CE CXP MRM (Q1 > Q3) DP EP CE CXP ESI + 3,4-Dichloroaniline > > Sigma-Aldrich Australia Acetyl-Sulfamethoxazole-d > > Dr Ehrenstorfer Supplier Acetyl-Sulfamethoxazole-d > > Toronto Research Chemicals Ametryn > > Restek Atenolol > > Cerilliant Atenolol-d > > Toronto Research Chemicals Atorvastatin > > Cerilliant Atrazine > > Restek Atrazine-d > > Dr Ehrenstorfer Bromacil > > Restek Caffeine > > Cerilliant Caffeine-d > > CDN Isotopes Carbamazepine > > Cerilliant Carbamazepine-d > > CDN Isotopes Carbaryl > > Restek Cephalexin > > Dr Ehrenstorfer Chlorpyriphos > > Restek Ciprofloxacin > > Sigma-Aldrich Australia Ciprofloxacin-d > > CDN Isotopes Citalopram > > Cerilliant Codeine > > Cerilliant Cyclophosphamide > > Dr Ehrenstorfer Dapsone > > Dr Ehrenstorfer DEET > > Restek Desethyl-Atrazine > > Restek Desisopropyl-Atrazine > > Restek Desmethyl-Citalopram > > Cerilliant Desmethyl-Diazepam > > Cerilliant Diazepam > > Cerilliant Diazinon > > Restek Diclofenac > > Cerilliant Diclofenac-d > > Dr Ehrenstorfer Diuron > > Restek Doxylamine > > United States Pharmacopeial Convention Erythromycin > > Cambridge Isotope Laboratories Erythromycin-Hydrate > > Sigma-Aldrich Australia Flumeturon > > Restek Fluoxetine > > Cerilliant Fluoxetine-d > > Isotec Stable Isotopes Gabapentin > > Cerilliant Haloxyfop-Etoyl > > Dr Ehrenstorfer Haloxyfop-Methly > > Accustandard Hexazinone > > Restek Ifosfamide > > Dr Ehrenstorfer Indomethacin > > Dr Ehrenstorfer Iopromide > > Dr Ehrenstorfer Lincomycin > > Dr Ehrenstorfer Metolachlor > > Restek Metoprolol > > Cerilliant Naproxen +ve > > Cerilliant Norfloxacin > > Sigma-Aldrich Australia Norfloxacin-d > > Sigma-Aldrich Australia Oxazepam > > Cerilliant Oxycodone > > Cerilliant Paracetamol > > Cerilliant Phenytoin > > Cerilliant Praziquantel > > Dr Ehrenstorfer Primidone > > Cerilliant Prometryn > > Restek Propoxur > > Restek Propranolol > > Cerilliant Ranitidine > > Cerilliant Ranitidine-d > > Toronto Research Chemicals Roxithromycin > > Dr Ehrenstorfer Sertraline > > Cerilliant Simazine > > Restek Simazine-d > > Dr Ehrenstorfer Sulfsalazine > > Sigma-Aldrich Australia Sulphadiazine > > Dr Ehrenstorfer Sulphamethoxazole > > Cambridge Isotope Laboratories Sulphathiazole > > Dr Ehrenstorfer Tebuthiuron > > Restek Temazepam > > Cerilliant Temazepam-d > > Cerilliant Terbutryn > > Restek Tramadol > > Cerilliant Trimethoprim > > Dr Ehrenstorfer Tylosin > > Dr Ehrenstorfer Venlafaxine > > Cerilliant ESI - Amoxicilin -ve > > Sigma-Aldrich Australia 2,4,5-T > > Restek 2,4,5-TP > > Restek 2,4-D > > Restek 2,4-D 13C > > Restek 2,4-DB > > Restek 2,4-DP > > Restek Acesulfame > > Dr Ehrenstorfer Acesulfame-d > > Toronto Research Chemicals Acetylsalicylic Acid > > Dr Ehrenstorfer Atorvastatin > > Cerilliant Chloramphenicol > > Cambridge Isotope Laboratories Clopyralid 4 190> > Sigma-Aldrich Australia Dalapon > > Restek DCPA (internal standard) > > ChemServices Dicamba > > Restek Diclofenac > > Cerilliant Diclofenac-d > > CDN Isotopes Fluroxypyr > > Restek Fluvastatin > > United States Pharmacopeial Convention Furosemide > > Cerilliant Gemfibrozol > > Cerilliant Haloxyfop > > Accustandard Hydrochlorothiazide > > Cerilliant Hydrochlorothiazide-C13d > > Toronto Research Chemicals Ibuprofen > > Sigma-Aldrich Australia Iopromide > > Dr Ehrenstorfer MCPA > > Dr Ehrenstorfer MCPB > > Restek Mecoprop > > Restek Naproxen > > Cerilliant Picloram > > Restek Salicylic Acid > > Cerilliant Triclopyr > > Restek Triclosan > > Sigma-Aldrich Australia Warfarin > > Cerilliant Notes: MRM = Multiple Reaction Monitoring, DP = Declustering Potential, EP = Entrance Potential, CE = Collision Cell Energy, CXP = Collision Cell Exit Potential ESI = Electrospray Ionisation 4
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