What’s in Your Sewage? - Trace Organic / Liquid Extraction Salt out with NaCl Acidify pH

What’s in Your Sewage?Dave Humphries, Jeff Charrois, Kristina Stefanizyn

Alberta Innovates – Technology FuturesVegreville, Alberta

Why do we care?(courtesy of J. Lavengood, U of Illinois)

B. Wilson, University of KansasEnvir. Sci. Technology 2003

■ “our results suggest that these 3 PPCPs{triclosan, ciprofloxacin, tergitol} may potentially influence both structure and function of algal communities in streams receiving WWTP effluents”

Where does sewage come from?

■ Direct Deposit■ Down the Drain Disposal

■ Unused medicine■ Bathing & Showering

■ Creams, lotions, skin patches, sweat glands■ Mostly Human

Some History

■ 2002 Trent University – Metcalfe et al■ 9 Acidic drugs in sewage effluents {Canada}■ Includes: Ibuprofen, Naproxen, Gemfibrozil

■ 2004 University of Hamburg – Weigel et al■ Selected Pharmaceuticals and caffeine in

sewage {Norway}■ DEET, Caffeine, Ibuprofen, Triclosan

Some More History

■ 2006 Abo Akademi Univ – Vieno et al■ Neural and basic pharmaceuticals in STPs

and recipient rivers { Finland}■ Carbamazepine, 4 fluoroquinoline antibiotics

■ 2010 Touro University – Ruhoy et al■ Bathing and Showering: Underappreciated

sources of water pollution from medicines {USA}

■ Topically applied creams, lotions, ointments, DEET

What Did We Look For?■ Antibiotics + Sulfa Drugs■ Antibacterials■ Hormones■ Artificial Sweeteners■ Nonylphenol Ethoxylates■ Analgesics (Ibuprofen, etc)■ Caffeine, DEET, Musks■ Illegal Drugs■ Metabolites■ 71 compounds in all

Many scans

■ DRUGA - GC■ DRUGN - LC■ Hormones -LC■ NPE - LC■ Sweeteners - LC■ Musks - GC

Compounds DrugA Scan19 Compounds & Metabolites

■ ASA 0.01 µg/l■ Clofibric Acid 0.01 µg/l ■ Ibuprofen 0.005 µg/l■ Gemfibrizol 0.005 µg/l■ Fenoprofen 0.005 µg/l■ Naproxen 0.005 µg/l■ Ketoprofen 0.01 µg/l■ Diclofenac 0.01 µg/l■ Bezafibrate 0.1 µg/l

Compounds DrugA Scan (cont’d.)

■ Indomethacin 0.05 µg/l■ Carbamazapine 0.01 µg/l■ Tolfenamic Acid 0.005 µg/l■ Caffeine 0.02 µg/l■ Triclosan 0.025 µg/l■ Methyl Triclosan 0.01 µg/l■ 3,3,4-Trichlorocarbanilide 0.025 µg/l■ Meclofenamic Acid 0.01 µg/l■ DEET 0.025 µg/l

DrugA Scan(Method)

■ Surrogates■ Ibuprofen-d3■ Gemfibrizol-d6■ Caffeine-d9■ Carbamazepine-d10■ Methyl Triclosan 13C12■ Triclosan-13C12■ DEET-d7

DrugA Scan(Method)

■ Liquid / Liquid Extraction■ Salt out with NaCl■ Acidify pH <2■ Dry with acidified sodium sulphate

■ Diazomethane Derivatization■ GC/Iontrap-MS

■ DB5 column

Compounds DrugN Scan

■ Cotinine {Metabolite of Nicotine}■ Cocaine & Benzoylecgonine■ 9 Sulfa Drugs■ 11 Antibiotics■ Codeine■ Methamphetamine■ Prozac & Metabolite■ Acetaminophen

DrugN Scan(Method)

■ SPE – Oasis HLB■ Waters LC/MS/MS

■ C18 water/methanol/formic acid■ Deuterated Surrogates

■ Cotinine-d4■ Norfluoxetine-d5■ Methamphetamine-d9■ Benzoylecgonine-d3■ Chloramphetamine-d5■ Acetaminophen-13C2

Compounds Hormone Scan

■ Estrone 0.05 µg/l■ B-Estradiol 0.50 µg/l■ 17a-Ethynylestradiol 1.0 µg/l■ Estriol 0.5 µg/l■ Progesterone 0.002 µg/l■ Testosterone 0.002 µg/l

Hormone Scan(Method)

■ 500 mL Sample Size pH 2■ SPE – Oasis HLB■ Waters LC/MS/MS

■ Water/acetonitrile/ammonium acetate■ C8 column

■ Surrogates■ B-Estradiol-d2■ Progesterone-d9

Compounds – Musk Scan

■ Cashmeran 0.02 µg/l■ Celestolide 0.02 µg/l■ Phantolide 0.02 µg/l■ Musk Ambrette, Ketone 0.02 µg/l■ Galaxolide 0.02 µg/l■ Traesolide 0.02 µg/l■ Musk Xylene 0.02 µg/l■ Tonalide 0.02 µg/l■ Musk Moskene, Tibetene 0.02 µg/l

Musk Scan(Method)

■ Liquid / Liquid■ Saltout with NaCl■ pH adjust to 6.0■ GC/Iontrap-MS

■ DB5 column■ Surrogates

■ Tonalide-d3■ Musk Xylene-d15

Compounds – NPE Scan

■ Nonylphenol Isomers 0.1 µg/l■ Nonylphenol Monoethoxylate 1.0 µg/l■ Nonylphenol Diethoxylate 0.2 µg/l■ Nonylphenol Triethoxylate 0.2 µg/l■ Nonylphenol Tetraethoxylate 0.2 µg/l■ Nonylphenol Pentaethoxylate 0.2 µg/l■ Bisphenol A 0.1 µg/l

MethodsNonylphenol Ethoxylate Scan

■ SPE – Oasis HLB■ 500 ml sample size■ pH 7-8

■ Waters LC/MS/MS■ C18 water/acetonitrile/ammonium acetate

■ Surrogate■ BisphenolA-13C12

Compounds – Artificial Sweeteners

■ Cyclamate 0.5 µg/l■ Sucralose 2.0 µg/l■ Acesulfame 0.5 µg/l■ Saccharin 0.5 µg/l■ Aspartame 0.5 µg/l■ Neotame 0.1 µg/l

MethodsArtificial Sweeteners Scan

■ SPE – Oasis HLB■500 ml sample■pH 2

■ Waters LC/MS/MS■C18■Water/acetonitrile/acetic acid

Did We Find Anything?

Caffeine

0.0570.04526.2

µg/Lµg/Lµg/L

After Membrane FilterFinal Effluent

Partial Processed Effluent

Headache

Pain Relievers

7.8817.413.8Codeine

ndnd107.3Acetaminophen0.2190.0869.65Ibuprofen

2.11.610.998Diclofenac

ndnd47.8Salicylic acid

µg/Lµg/Lµg/L

After Membrane

FilterFinal Effluent

Partial Processed

Effluent

Artificial Sweeteners

ndndndSucralose

ndndndCyclamate

ndnd0.2Saccharin0.30.30.1Acesulfame

ndndndSucralose

µg/Lµg/Lµg/L

After Membrane

FilterFinal Effluent

Partial Processed

Effluent

Antibacterials

0.0480.0210.469Methyl Triclosan

0.1400.1882.63Triclosanµg/Lµg/Lµg/L

After Membrane

FilterFinal Effluent

Partial Processed

Effluent

Nonylphenol Ethoxylates

1.822.174.95NP isomers

ndnd1800NPE17700079000126000NPE2

137000125000172000NPE3

10000800011000NPE4

140001000030000NPE5

µg/Lµg/Lµg/L

After Membrane

FilterFinal Effluent

Partial Processed

Effluent

MISC

0.2550.2272.62Bisphenol A

0.0500.0560.296Methamphetamine0.0470.0490.250DEET

0.0440.0452.69Benzoylcgonine

nd0.0261.58Cocaine

nd0.4528.4Cotinine

µg/Lµg/Lµg/L

After Membrane

FilterFinal Effluent

Partial Processed

Effluent

Questions ?