Water Consulting Services

Pharmaceuticals in watersNiina Vieno

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Over 3000 chemicals are used in human and veterinary medicine

• Sources in the environment:

• Wastewater treatment plants• Via human urine and feces (sewage)

• Via disposal of old pharmaceuticals (in Finland 29 000 kg)

• Hospitals

• Pharmaceutical industry

• Solid waste• For example, in Finland 32 000 kg of

pharmaceuticals are incorrectly disposed via solid waste and end up in landfills

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Harmful effects in the environment have been proven

1 μg/l of diclofenac and propranolol induced:

• Reduced growth• Reduced ability to attach to the underlying substrate• Bioconcentration into the mussel

Source: Ericson et al. 2010, Aquat Toxicol. 99(2):223-31

Baltic Sea blue mussel

Photo: www.blueoceans.org

Unpublished data ofBlue Mussels in the Gulfof Finland:

• Low concentrations(< 10 ng/l) ofpharmaceuticals inducestress in blue mussel.

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Zebrafish embryos:

30 ng/l concentration of diclofenac at 90-minute exposure has inducedoxidative stress

reduced ability to detoxify the reactive intermediates or to repair the resulting damage

Photos: ehsc.oregonstate.edu & www.landesbioscience.com

Source: Feito et al. 2012 Ecotoxicology. 21(1):289-96

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Wild European perch:

Antidepressant oxazepam has been noted to alter behavior and feeding rate of at concentrations measured in effluent-influenced surface waters.

Source: Brodin et al. 2013, Science 339, 814-815

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Pharmaceuticals bioconcentrate in fish

• Fish were caught from a Finnish lake that receives treated municipal wastewater via a small river.

• Diclofenac, ibuprofen and naproxen were detected in bile samples of nearly all the fish

• Concentrations in the lake were up to 300 ng/l (diclofenac)

Source: Brozinski et al. 2013, ES&T47, 342-348

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17α-ethinylestradiol (EE2) is an endocrine disruptor: there are effects already at a concentration of 0.1 ng/l

Fathead minnow:

• LOEC= 1 ng/l for:• plasma vitellogenin induction (measures

estrogenic sensitivity) in both sexes• ultrastructural changes in the testes and

livers

• LOEC= 0.1 ng/l for:• significant increase in the mean number of

eggs spawned per pair

Source: Pawlowski et al. 2004. Ecotoxicol Environ Saf. 2004, (3):330-45.

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EU’s environmental legislation will consider pharmaceuticals for the first

time ever in the revised Water Framework Directive

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LIST OFPRIORITY

SUBSTANCESAND

PRIORITY HAZARDOUS SUBSTANCES

Use of these compounds should be reduced or banned.

Water Framework Directive (2000/60/EC)

WATCH LIST OF EMERGING

POLLUTANTS

CURRENT SITUATIONPROPOSAL (alongside

the list of priority substances

These compounds should be monitored-> can be classified as priority substance

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Pharmaceuticals and hormones proposed to be added in the watch

list

Inclusion isconfirmed!

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Pharmaceuticals in the “watch list” means that member states shall:

• monitore them in surface waters and report to European Commission (mandatory)

• monitore them in wastewater effluents (voluntary)

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Classification of pharmaceuticals as HELCOM Core indicators

• Diclofenac and EE2 has been suggested

• Discussion also about addition of natural estrogens (E1, E2 and E3) and possibly complemented with an in vitro yeast assay.

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Occurrence of pharmaceuticals in wastewater depends on the

consumption

Spai

n

Austri

a

Austra

lia

Japa

n

Germ

any

USA

Finl

and

0100200300400500600700800900

1000

Diclofenac

Con

sum

pti

on

mg

/in

h/y

ear

Spai

n

Austri

a

Austra

lia

Japa

n

Germ

any

USA

Finl

and

02000400060008000

1000012000140001600018000200002200024000

Ibuprofen

Con

sum

pti

on

mg

/in

h/y

ear

Dnf Dnf

Dnf= data not found

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…next important thing is human metabolism

Carba

maz

epin

e

Ibup

rofe

n

Diclof

enac

Met

opro

lol

Keto

prof

en

Napro

xen

Sulfa

met

hoxa

zole

Norflo

xacin

Cipro

floxa

cin

Acebu

tolo

l

Bezafi

brat

e

Sota

lol

Ofloxa

cin

Ateno

lol

0%10%20%30%40%50%60%70%80%90%

100%

Excreted as parent compound

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Diclofenac-1--o-acyl glucuronide (DCF-G)

4´-hydroxy-diclofenac (4´-OH-DCF)

5-hydroxy-diclofenac (5-OH-DCF)

Diclofenac (DCF)

3´-hydroxy-diclofenac(3´-OH-DCF)

4´,5-dihydroxy-diclofenac(4´,5-OH-DCF)

R, R´, R´´= not definitely identified ligands, presumably one per molecule

About 1 % is excreted unchanged

About 11 % is excreted conjugated

36 % 10%

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Sewage concentrations follow consumption pattern

0100200300400500600700800900

1000

Diclofenac con-sumption

Con

sum

pti

on m

g/i

nh

/year

Spai

n

Austri

a

Austra

lia

Japa

n

Germ

any

USA

Finl

and

0.0

1.0

2.0

3.0

4.0

5.0

6.0

7.0

8.0

Diclofenac, max influent concentration

Co

nce

ntr

ati

on

, g

/l

Dnf Dnf Dnf Dnf

Dnf= data not found

Niina Vieno

Spai

n

Austri

a

Austra

lia

Japa

n

Germ

any

USA

Finl

and

02000400060008000

1000012000140001600018000200002200024000

Ibuprofen

Con

sum

pti

on

mg

/in

h/y

ear

Spain

Austri

a

Austra

lia

Japa

n

Germ

any

USA

Finland

0.0

5.0

10.0

15.0

20.0

25.0

30.0

35.0

Ibuprofen, influent concentration

Conce

ntr

ati

on,

g

/l

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Sewage concentrations can easily be estimated by calculations

A= consumption of a pharmaceutical (as grams per inhabitant per year)P= number of inhabitants serviced by the sewage treatment plante%= the percentage of pharmaceuticals excreted unmetabolizedQ= influent flow rate of the sewage treatment plant (m3/d)

Concentration of a pharmaceutical in influent ofa sewage treatment plant in g/l:

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Carba

maz

e...

Diclof

enac

Met

opro

lol

Keto

prof

en

Napro

xen

Norflo

xacin

Cipro

floxa

cin

Acebu

tolo

l

Bezafi

brat

e

Sota

lol

Ofloxa

cin

Ateno

lol

0.0

1.0

2.0

3.0

4.0

5.0

6.0 Calculated

Measured

Se

wa

ge

co

nce

ntr

ati

on

, g

/l

Calculated values are often lower because deconjugation is not considered in the equation.

10 % excreted unchanged60 % excreted as glucuronides

10 % excreted unchanged70 % excreted as glucuronides

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Concentrations in Finnish sewage (n= 13-21)

Ofloxa

cin

Norflo

xacin

Acebu

tolo

l

Carba

maz

epin

e

Diclof

enac

Cipro

floxa

cin

Ateno

lol

Sota

lol

Bezafi

brat

e

Met

opro

lol

Keto

prof

en0

0.5

1

1.5

2

2.5

Co

nce

ntr

ati

on

, g

/l

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Concentrations in Finnish sewage

Naproxen Ibuprofen0

2

4

6

8

10

12

14

16

18C

on

ce

ntr

ati

on

, g

/l

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In comparison: Black wastewater

PAR TRI TC MET DOX PRO CBZ CET IBP DCF

g/L

0

50

100

150

200

250

300BW UASB PN AMX

>1000 g/L

DOX PRO CBZ CET

g/L

0

1

2

3

4

5

6

7BW= Black waterUASB= Upflow Anaerobic Sludge

BlanketPN= Partial nitritationAMX= Anammox

PAR= paracetamolTRI= trimethoprimTC= tetracyclineMET= metoprololDOX= doxycyclinePRO= propranololCBZ= carbamazepineCET= cetirizineIBP= ibuprofenDCF= diclofenac

Source: De Graaff et al. 2011. Water Research, 45(1), 375-383.

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Elimination in wastewater treatment

• In conventional treatment plants, elimination of pharmaceuticals from the water phase occurs:

• Via sorption to sludge

• Via biodegradation/biotransformation

• Compounds do not evaporate

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Sorption= Absorption + Adsorption

SLUDGE FLOC

CELL

Hydrophobic compound Hydrophilic, ionic, compound

Absorption Adsorption- desorption

SLUDGE FLOC

CELL

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Sorption can be estimated by Kd value

• Sorption of a compound to sludge can be considered negligible when

• Kd value is < 500 L/kgSS (i.e. logKd < 2.7)

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logKd (primary sludge) logKd (activated sludge)

Ciprofloxacin 4.3

Norfloxacin 4.2

Sulfamethoxazole 2.05 – 2.6

Carbamazepine < 1.3 0.09

Diclofenac 2.3 – 2.7 1.2 – 2.1

Ibuprofen <1.3 0.85

Naproxen 2.34

Atenolol 1.6

Metoprolol 0.5

Sotalol 1.6

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Biodegradation: direct- or cometabolism

• Direct metabolisation= bacteria use the

compound as their primary carbon source

• Cometabolisation= bacteria break down or

partially convert the compound but do not

use it as the primary carbon source

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Pharmaceuticals are primarily degraded cometabolically

• Bacteria secrete enzymes to break down

large organic molecules into monomers

that are small enough to be ingested.

• The enzymes have the potential to degrade

or to transform small organic pollutant

molecules present in wastewater.

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• E Coli secrete -glucuronidase enzyme

• It is capable to deconjugate the -glucuronated pharmaceuticals excreted by the human body.

• Ammonia monooxygenase (AMO) is secreted by autotrophic ammonia oxidizing bacteria during the oxidation of ammonium to hydroxylamine in aerobic nitrification.

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Deconjugation of diclofenac b-O-acyl glucuronide can occur in wastewater

treatment plant

• In microbiological degradation tests:

• DCF-G has been noted to deconjugate to form equimolar DCF within 7 d of incubation.

• Reaction rate constant were 0.28 d-1 in the inoculum and 0.12 d-1 under sterile conditions.

• This suggests that abiotic hydrolysis occurs but the reaction is enhanced by micro-organisms.

Source: Lee et al. 2012, Chemosphere, 87, 969-974

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Biodegradation varies greatly between pharmaceuticals

• Ibuprofen is highly biodegradable at any process configurations

• Diclofenac and carbamazepine are hardly biodegradable

• Suggestions that process parameters such as increase of solids retention time or applying membrane bioreactor could enhance biodegradation.

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Example for diclofenac

0%

10%

20%

30%

40%

50%

60%

70%

80%

90%

100%

Conventional activated sludge Activated sludge with biological nutrient removal Membrane bioreactor

35 %

29%

54%

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Pharmaceuticals degrade to metabolites and complete

mineralization is rare

DICLOFENAC

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Removal rates are often calculated from influent and effluent

concentrations

• In Finnish sewage treatment plants

Compound Removal rate (%)

Ciprofloxacin 84 %

Carbamazepine -121 %

Diclofenac 17 %

Ibuprofen 95 %

Ketoprofen 81 %

Naproxen 91 %

Atenolol 60 %

Metoprolol 17 %

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Example of mass flow analysis of diclofenac in Finland

PRETREATMENT ACTIVATEDSLUDGE

324 kg 305 kg

19 kg (6 %)

270 kg (83 %)

4 kg (1 %)

31 kg (10 %)

Sludge Sludge

Biodegradation or-transformation

ENVIRONMENTYearlydischarge tosewer ca. 60 mg/inh

Evaporation

0 %

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Elimination can be enhanced by tertiary treatment

• Oxidation techniques• Ozonation, O3/UVA/TiO2

• Fenton treatment

• UV based technologies

• Catalytical techniques• Nanocatalysts• Photoelectrocatalysis

• Adsorption and filtration techniques• Sand filtration• Ultra and nanofiltration• Reverse and forward osmosis• Innovative sorbents

• Other techniques• Electrochemical incineration• Sonolysis and ultrasound• Irradiation techniques• Liquid-core microcapsules

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Formation of oxidation products needs to be considered

DICLOFENAC DICLOFENAC-2,5-IMINOQUINONE

Rate constant with O3 is about 5 % of that of diclofenac

Reaction with O3

Source: Sein et al. 2008, ES&T, 42, 6656-6662

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Removal from aqueous phase is usually incomplete and pharmaceutical residues

remain in the effluentsTreated wastewater from Finnish WWTPs (n= 13-21)

Ofloxa

cin

Norflo

xacin

Acebu

tolo

l

Carba

maz

epin

e

Diclof

enac

Cipro

floxa

cin

Ateno

lol

Sota

lol

Bezafi

brat

e

Met

opro

lol

Keto

prof

en

Napro

xen

Ibup

rofe

n0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

Co

nce

ntr

ati

on

, g

/l

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In comparison: Antibiotic ciprofloxacin in treated wastewater

in India

Finland: Max 0.13 g/l (i.e. 0.00013 mg/l)

Patancheru/ India: 31 000 g/l (i.e. 31 mg/l)

Defined daily dose for ciprofloxacin is 500–1000 mg.

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Rivers carry a significant load of the

pharmaceutical residues

River Aura empties into the Baltic Sea at Turku, Finland.

Example: River Aura

Distance from Aura WWTP (km)

c (ng/l)

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Concentrations in Finnish rivers (n= 64)

Ofloxa

cin

Norflo

xacin

Acebu

tolo

l

Carba

maz

epin

e

Diclof

enac

Cipro

floxa

cin

Ateno

lol

Sota

lol

Bezafi

brat

e

Met

opro

lol

Keto

prof

en

Napro

xen

Ibup

rofe

n0

10

20

30

40

50

60

Co

nce

ntr

ati

on

, n

g/l

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Marine environment is

the final sink for pharmaceuticals

Photo: www.helcom.fi

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Source: E-PRTR (European Pollutant and Transfer Register)

-> Waste and waste water management

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Measurement data in the Baltic Sea is scarce

Compound Therapeutic use

Max concentrations as ng/l

Germany

Stockholm

Helsinki

Bezafibrate Lipid regulator 0.3

Carbamazepine Antiepileptic 6.8 Detected

Clofibric acid Metabolite of fibrates (lipid regulators)

0.4

Diclofenac Anti-inflammatory

0.4 Detected

Gemfibrozil Lipid regulator 0.1

Ibuprofen Anti-inflammatory

0.1 Detected

Naproxen Anti-inflammatory

0.5 Detected

Oxazepam Antidepressive 20

Propyphenazone Analgesic 0.2

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EE2 (i.e. 17α-ethinylestradiol) is the most potent out of estrogens

Source: Hazardous Substances in the Baltic Sea. Baltic Sea Environment Proceedings No. 120 B. Available at: www.helcom.fi

EE2: 2.1– 17.9 ng/l

EE2: Up to 14.1 ng/l

EE2: Up to 3.9 ng/l

EE2: 1.6–3.0 ng/l

EE2: 1.7–3.2 ng/l

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Most problematic are those pharmaceuticals that are not eliminated in the WWTP or in the

environmentIBUPROFEN

Breaks down easilyby biotransformation

Only a small fraction entersthe environment

CARBAMAZEPINE

Is persistent in wastewater treatment

Is persistent in the environmentand is detected also in ground w.aters

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Whose responsibility it is

to reduce the environmental

load of pharmaceuticals?

PHARMACEU-TICAL

INDUSTRY

WASTEWATER

TREATMENT PLANTS

EVERYONETOGETHER!

AUTHORITIES

DOCTORS &CONSUMERS

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Contact information

Niina Vienopuh. 050 5448431niina.vieno@envieno.com

www.vesiuutisvuoto.fi