degradation of pharmaceuticals in the aquatic environment. · 1. monitoring the presence of...

Degradation of pharmaceuticals in theaquatic environment.

Investigation of transformation products

10th November 2016Clara Boix

1. General Introductiona) Water quality

b) Emerging contaminants

c) Analytical methodology: UHPLC-MS

2. Degradation of organic contaminants in the aquatic environment

3. Degradation of omeprazole in water:a) Hydrolysis

b) Photo-degradation

c) Chlorination

4. Biodegradation of five pharmaceuticals in water

5. Conclusions

2

3







c) Chlorination


5. Conclusions

- Human and veterinary pharmaceuticals

- Drugs of abuse

- Metabolites

- Transformation Products (TPs)

Water quality

The importance of monitoring contaminants!

4

Emerging contaminant

MetabolismPharmaceuticals

+Metabolites

Environment

WWTP

Pharmaceuticals+

Metabolites+

TPs

Pharmaceuticals

5

Some TPs/metabolites are as hazardous as, or even more,than the parent compound

Pharmaceutical transformation:

6

LiquidChromatography

Quadrupole Time-Of-Flight(Q)TOF

ElectroSpray

Analytical methodology:

MassSpectrometry

ESI MSLC

7

UHPLC-(Q)TOF MS (Q Time-Of-Flight)







c) Chlorination


5. Conclusions

8

9







c) Chlorination


5. Conclusions

Degradation of organic compounds

WWTP

1. Hydrolysis2. Biodegradation (Surface water)3. Photo-degradation (Sun light)

1. Biodegradation (Active sludge)2. Photo-degradation (UV)3. Chlorination

Environment

10

11

Degradation methodology:

Degradation experiments

Hydrolysis Water MilliQ / darkness

Photo-degradation

Biodegradation

Chlorination NaClO 1% w/v

Controlsample

Spikedsample

SunTest (Sun light)

Mercury lamp (UV)

Surface Water

Active Sludge

LC-QTOF MS

-20

oC

50µL

0.5-1.0mg/L







c) Chlorination


5. Conclusions

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13







c) Chlorination


5. Conclusions

C17H19N3O3S[M+H]+: 346.1225

Omeprazole

Scarcely found inwater samples

DEGRADATIONSTUDY

Highly consumed

Whyomeprazole isnot present inwaters?

-Proton pump inhibitor-Long-lasting reduction ofgastric acid production

14

Sampling 2-mL aliquotsat different time

intervalsLC-(Q)TOF MS

Objective: Detailed study of omeprazole degradation focused on the formation ofits possible transformation products (TPs) in waters

15

DEGRADATION

PROCEDURECONDITIONS

DEGRADATIONTIME

HYDROLYSISDarkness

Room temperature0-17 days

CHLORINATIONNaClO (1%)Darkness

Room temperature0-4 days

UV

PHOTO-DEGRADATION

Mercury lampUV radiation (254nm)

Glass vessels0-60 hours

SUNLIGHT

PHOTO-DEGRADATION

Irradiance: 500 W/m2

Day light filter (Suntest XL)Quartz vessels

0-90 hours

Lab

ora

tory

-co

ntr

olle

dd

egr

adat

ion

exp

eri

me

nts

:

Data are acquired byLC-QTOF MS (MSE)

Data processed byMetaboLynx XSApplication manager

Elemental compositionproposal

(parent ion information)Structure proposal

(fragment ions information)

DATA PROCESSING

MSE

LE

HE

Detection of 17chromatographic peaks.

Possible TPs!

Elucidation

How a TP is identifiedand elucidated?

1 2

3

CompoundRetention

Time(min)

Elementalcomposition

Experimentalmass(m/z)

MassError

(mDa)

Omeprazole 5.98 C17H20N3O3S+ 346.1211 -1.4

Hydrolysis

OTP 1 1.93 C9H14NO2+ 168.1030 0.5

OTP 2 2.37 C9H12NO3+ 182.0822 0.5

OTP 3 3.86 C17H18N3O4+ 328.1313 1.6

OTP 4 4.09 C17H20N3O2+ 298.1556 0.0

OTP 5 5.68 C17H20N3O2S+ 330.1271 -0.5

Photo-degradation

OTP 6 1.71 C9H12NO4+ 198.0773 0.7

OTP 7 -103.45 - 3.554.82 - 5.03

C8H8N3O3+ 194.0565 -0.1

Chlorination

OTP 11 - 12 7.21 - 7.52 C17H17N3O3Cl+ 346.0942 -1.6

OTP 13 7.70 C17H18N3O4SCl2+ 430.0405 1.0

OTP 14 8.23 C17H16N3O3Cl2+ 380.0556 -1.3

OTP 15 – 16 8.34 - 8.46 C17H17N3O4SCl3+ 464.0010 0.5

OTP 17 9.15 C16H14N3O4SCl2+ 414.0081 -0.1

Total 17omeprazole

TPs

5 TPs hydrolysis

5 sunlight photo-degradation TPs

7 Chlorination TPs

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WWTP

Primarytreatment

Primarytreatment

Secondarytreatment

Primarytreatment

Secondarytreatment

Tertiarytreatment

Hydrolysis TPs

HydrolysisBiodegradation TPs

HydrolysisBiodegradation

ChlorinationUV TPs

Environment

OmeprazoleHydrolysis

BiodegradationChlorination

UV and SunlightTPs

IWW EWW

SW

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OMEPRAZOLE

Searching of 17omeprazole TPs in

IWW, EWW and SW

% positive findings

Compound name IWW EWW SW

(n=15) (n=10) (n=27)

Omeprazole - - -

OTP 1* - - 11

OTP 2* - - -

OTP 3* - - -

OTP 4* - 10 7

OTP 5 - 90 26

OTP 6* - - -

OTP 7-10* - - -

OTP 11-12* - - -

OTP 13* 7 50 11

OTP 14* - - -

OTP 15-16* - - -

OTP 17* - - -

Positive findings in water samples

19

*N

ot

rep

ort

ed

inlit

era

ture

yet







c) Chlorination


5. Conclusions

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21







c) Chlorination


5. Conclusions

Compound Therapeutic group StructureElemental

composition

Ofloxacin Antibiotic (Quinolone) C18H20FN3O4

Venlafaxine Antidepressant C17H27NO2

Irbesartan Antihypertensive C25H28N6O

Ibuprofen Analgesic/anti-inflammatory C13H18O2

Gemfibrozil Lipid Regulator C15H22O3

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Objective: Biodegradation study of 5 pharmaceuticals using surface water(SW) and active sludge (AS)

Biodegradation

Surface water (SW)

Active sludge (AS)

65 days

35 days

TSS 100mg/L

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UHPLC-(Q)TOF MS

24

(b)

(a)0.00

0.20

0.40

0.60

0.80

1.00

1.20

0 5 10 15 20 25 30 35 40

No

rma

lise

dA

rea

t (days)

Sewage sludgeexperiments

IRBESARTAN

VENLAFAXINE

OFLOXACIN

IBUPROFEN

GEMFIBROZIL

0.00

0.20

0.40

0.60

0.80

1.00

1.20

0 10 20 30 40 50 60 70N

orm

alis

ed

Are

a

t (days)

Surface waterexperiments

IRBESARTAN

VENLAFAXINE

OFLOXACIN

IBUPROFEN

GEMFIBROZIL

Surface Water(SW)

Active Sludge(AS)

25

0,0

0,5

1,0

1,5

2,0

2,5

3,0

3,5

4,0

4,5

5,0

0 10 20 30 40 50 60 70Lo

gar

ea

Degradation time (d)

GHB1 Hydrolysis

GHB1 Biodegradation0,0

0,5

1,0

1,5

2,0

2,5

3,0

3,5

4,0

0 10 20 30 40 50 60

log

are

a


IbH1

IbH2a

IbB3

0,0

0,5

1,0

1,5

2,0

2,5

3,0

0 5 10 15 20 25 30 35

Log

area


OB1

0,0

0,5

1,0

1,5

2,0

2,5

3,0

3,5

4,0

4,5

5,0

0 5 10 15 20 25 30 35

Log

area


VB1aVB1bVB2VB4

0,0

0,5

1,0

1,5

2,0

2,5

3,0

3,5

4,0

4,5

5,0

0 5 10 15 20 25 30 35

Log

are

a


IH1bIB3aIB3bIB4IB5

IrbesartanAS

VenlafaxineAS

OfloxacinAS Ibuprofen

SW + AS

GemfibrozilSW + AS

ISW1b

IbSW1

IbSW2a

GSWB1 SW

GSWB1 ASBio

de

grad

atio

nTP

so

f5

ph

arm

ace

uti

cals

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Biodegradation Detected TPs

PharmaceuticalSurfaceWater

ActiveSludge

Irbesartan 3 5

Venlafaxine - 6

Ofloxacin - 1

Ibuprofen 4 2

Gemfibrozil* 1 1

* For gemfibrozil, the TP detected in SW and AS was the same

Search of pharmaceuticals + TPs + metabolites, previouslyinvestigated, in SW and EWW samples

Pharmaceuticals

TPs biodegradation (SW)

TPs biodegradation (AS)

Metabolites “common fragment”

5

8

14

3

30 Compounds

Search in:38 EWW and 18 SW

27

Positive findings (%)

EWW (n=38) SW (n=18)

Irbesartan 92 39ISW1b 87 6IB3a* 84 22IB3b* 89 22IB4* 32 11IB5* 79a 22a

Valsartan 79 33Venlafaxine 87 22

VB1a 92 17VB1b 92a 17a

V1* 58 6V2* 87 11

Ofloxacin 82 17Ibuprofen 11 6IbSW2a 16 11IbSW2b 8a 0IbB4* 34 50

Ib1 21 6Gemfibrozil 24 22

GSWB1* 71 33 a Only the [M+H]+/[M-H]- was observed

Up to 20positivefindings

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Importance ofthese studies in

theenvironmental

field

* Not reported in literature yet







c) Chlorination


5. Conclusions

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30







c) Chlorination


5. Conclusions

1. Monitoring the presence of omeprazole in the aquatic environmentshould be focused on omeprazole TPs instead of the parent compound.

2. From the best of our knowledge, a large number of these TPs had notbeen previously reported in scientific literature, for this, in a future, anevaluation of their toxicity/dangerousness in the aquatic environment isrequired.

3. Biodegradation TPs of pharmaceuticals should be included in monitoringprograms, to have a more realistic overview of the impact ofpharmaceuticals on water quality.

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