Persistence and Fate of Antibiotics in Sewage
Prof. Dr. Klaus Kü[email protected]
Pharmaceutical Sewage During Influenza Pandemic
Somerville College, University of Oxford, 3 March 2009
Department of Environmental Health Sciences
Some DataConsumption of Antibiotics
Medicine Veterinary Growth Promoters
65 %
6 %29 %USA 23 000 t ?
Other countries ?
World ?
EU: total 13 216 t (1999)
* FEDESA 2001
** Estimation based on data of the Union of Concerned Scientists (2001)
(only until 2005 in the EU)
Some Data
• 250 different active compounds authorized in Germany(antibiotics, antimycotics)
• Consumption: Germany 3-6 g per capita and year (regional differences)
•
• Data Europe:
Alexy et al., Acta Hydrochim. Hydrobiol. 34, 587-592, 2006
http://www.esac.ua.ac.be/
0
20
40
60
80
100
120
Penic
illin
s
Cephal
osp.
Sulfo
namid
es
Amin
oglyko
s.
Tetra
cycl
ines
Mac
rolid
es
Glyco
peptid
es
Quinolo
nes
Other
s
t/y
r
Prescriptions by Practitioners
Hospitals
Emissions of Antibiotics Into Municipal SewageUsage 1998: 412 t (Germany)
Emission 1998: 277 t (Germany)Thereof hospitals 92 t (Germany)
Kümmerer und Henninger, Clinical Microbiology and Infection, 9, 1203-1211, 2003
What Happens in the Sewage Treatment Plant?
© Abwasserzweckverband Breisgauer Bucht, http://www.azv-breisgau.de/
• Antibiotics: 0 -100%
• Cytotoxics 0 -100%
• Hormones: 20 - 80%
• Statins (Lipid lowering) 0 - 50%
• Pain killers 0 - 50%
• Anti depressivs 0 - 50%
• Disinfectants 0 -100%
• Diagnostics 0 - 20%
Elimination of Pharmaceuticalsin Municipal STPs
…., Golet et al. 2002, Färber et al. 2002, Kolpin et al. 2002, Sacher et al. 2001, Kümmerer et al. 2000, Hamscher et al. 2002,
Zuccato et al. 2001, Hirsch et al. 1999, Ternes 1998ff, Heberer 1996ff, Richardson und Bowron 1985, ….
Elimination in the environment is the sum of:
• Biotransformation
• Photolysis
• Hydrolysis
• Oxidation/Reduction
• Sorption
Key Processes for Elimination
Key Properties for Elimination
• Water solubility
• Partition coefficients (log Pow, log Koc)
• Redox potential
• Degree of saturation
• Types of functional groups
• Electrical charges
• Acid/base constants (pKas)
Roxithromycin
0
20
40
60
80
100
Zulauf Ablauf Belebt-schlamm
Überschuss-schlamm
Kon
zent
ratio
n [n
g/l]
oder
[µ
g/kg
TS
]
Structure Matters: Impact of Structure on Elimination in a STP
Clarithromycin
0
20
40
60
80
100
Zulauf Ablauf Belebt-schlamm
Überschuss-schlamm
Ko
nze
ntr
atio
n [
ng
/l] o
der
[µ
g/k
g T
S]
Con
cent
ratio
n(n
g/L;
µg
/kg
d.m
.)
Con
cent
ratio
n(n
g/L;
µg
/kg
d.m
.)
influent influentsludge effluent sludgeeffluent surplus sludge
surplussludge
Alexy, Sacher, Wenz, Kümmerer, Acta hydrochim. Hydrobiol., 2006
Roxithromycin
0
20
40
60
80
100
Zulauf Ablauf Belebt-schlamm
Überschuss-schlamm
Kon
zent
ratio
n [n
g/l]
oder
[µ
g/kg
TS
]
Structure Matters: Impact of Structure on Elimination in a STP
Clarithromycin
0
20
40
60
80
100
Zulauf Ablauf Belebt-schlamm
Überschuss-schlamm
Ko
nze
ntr
atio
n [
ng
/l] o
der
[µ
g/k
g T
S]
Con
cent
ratio
n(n
g/L;
µg
/kg
d.m
.)
Con
cent
ratio
n(n
g/L;
µg
/kg
d.m
.)
influent influentsludge effluent sludgeeffluent surplus sludge
surplussludge
NOCH2OC2H5OCH3
Alexy, Sacher, Wenz, Kümmerer, Acta hydrochim. Hydrobiol., 2006
Pharmaceuticals Are Different!Ciprofloxacin
OH
N
OH
O
N
HN
O
F
Ciprofloxacin
Acidic groups
Basic groups
Optimised for biological activity and stability.Several different functional groups within on and the same molecule.
Physical Chemical Instability of Antibiotics
0,00
0,03
0,06
0,09
0,12
0,15
0,18
k [
1/d
]
4°C Dunkelheit 15°C Dunkelheit 20°C Dunkelheit 20°C Lichtdarknessdarkness darkness light
Kümmerer and co-workers, unpublished
Stability Profile of Ceftriaxone(Fitted According to Arrhenius-Equation)
0
1
2
3
4681012141618205 10 15 20 25
Ko
nze
ntr
ati
on
[m
g/l
]
0 [mg/l]1 [mg/l]2 [mg/l]3 [mg/l]C
on
cen
tra
tio
n
DAY Temperature
Kümmerer and co-workers, unpublished
Degradation of Structurally Closely Related Penicillins
Piperacillin Amoxicillin
N
S
OOH
H
HN
O
O
Benzylpenicillin
SNH
O
H2N
OH
O
N
HOO
N
S
OOH
H
HN
O
O
HNN
O
N
O
O
0
20
40
60
80
100
0 5 10 15 20 25 30day
Elim
inat
ion
in %
HPLC-UVVis: biol. non biol.
DOC: biol. non biol.
Piperacillin
Day
Elimination and BiodegradationZahn Wellens Test (OECD 302 B)
Kümmerer et al., Chemosphere, in press
m/z 518 (+mode)Piperacillin
m/z 536 (+ mode)hydrolysedPiperacillin
N
S
O
O H
H
H N
O
O
HNN
O
N
O
O
NH
S
O
O H
H
H N
O
HNN
O
N
O
O
O HO
(n. biotic) hydrolysis
Degradation of Pieracillin (LC-MS/MS ion trap, -UV-Vis, DOC)
+H2O
Kümmerer et al., Chemosphere, in press
0
20
40
60
80
100
0 5 10 15 20 25 30day
Elim
inat
ion
[%
]
0
20
40
60
80
100
0 5 10 15 20 25 30day
Elim
inat
ion
in %
HPLC-UVVis: biol. non biol.
DOC: biol. non biol.
Piperacillin Amoxicillin
Day
Elimination and BiodegradationZahn Wellens Test (OECD 302 B)
Kümmerer et al., Chemosphere, in press
SNH
O
H2N
OH
O
N
HO
O
SNH
O
H2N
O
HN
HO
OOH
OHS
NH
O
H2N
OH
O
HN
HO
OOH
+ H2O -C6H5
m/z 366 (+ mode) m/z 307 (+ mode)m/z 388 (+ mode)
Amoxicillin hydrolysiertes Amoxicillin hydrolysiertes Amoxicillinohne Benzenring
non biotic biotic
???
?
Amoxicillin hydrolysed Amoxicillin hydrolysed Amoxicillinwithout phenol ring
Bio-degradation
Bio-mineralization
(n. biotic) Hydrolysis
-C6H6O
Degradation of Amoxicillin (LC-MS/MS ion trap, -UV-Vis, DOC)
Kümmerer et al., Chemosphere, in press
t1/2 Benzylpenicillin ≈ 2 * t1/2 Amoxicillin
Photo-degradation of Ciprofloxacin
Vasconcelos T. G., Henriques D.M., König, A., Martins A.F., Kümmerer K., Chemosphere, accepted
0 2 4 6 8 10 12 Time [min]0
20
40
60
80
100
Intens.mAU
0 2 4 6 8 10 12 Time [min]0
20
40
60
80
100
Intens.mAU
0 2 4 6 8 10 12 Time [min]0
20
40
60
80
100
Intens.mAU
Ciprofloxacin
Photo-transformation products 4 min Photo-transformation products 2 min
Vasconcelos T. G., Henriques D.M., König, A., Martins A.F., Kümmerer K., Chemosphere, accepted
N
F
N
O O
OH
HN
NN
O O
OH
HN CH3
O
N
F
NH
O O
OH
NH2
NNH
O O
OH
NH2
N
HO
N
O O
OH
HN
Product 1
Product 4
Product 3
Product 2
CIPRO
N
F
N
O
HN
Product 5
Photo-degradation of Ciprofloxacin
0
0,5
1
1,5
2
2,5
3
3,5
4
4,5
5
0 1 7 14 28days
Cip
ro C
on
ce
ntr
ati
on 0 min
2 min4 min
Vasconcelos T. G., Henriques D.M., König, A., Martins A.F., Kümmerer K., Chemosphere, accepted
0 min: Ciprofloxacin
Biodegradability of Photo Products of Ciprofloxacin
• Advanced filtration technologies (reversed osmosis, membrane filtration, nanofiltration), and (photo)oxidation technologies are emerging.
• Efficiency may depend strongly on the type of compound. Reaction products?
• Resistance and bio-membrane (enrichment, residence time)
• Costs?
• Storm water?
• Infiltration of the ground before STP?
• Not compatible with green chemistry.
Advanced Effluent Treatment (1)
• Working for new compounds?
• Not possible in less developed countries
Advanced Effluent Treatment (2)
Summary - Questions
• Which compound (pH, metabolites, transformation products)?
• Which bacterial consortium?• Significance of resistance?• Temperature?• Concentration?• Primary degradation vs. mineralization?• Transformation products still active?• Properties?
EC50
5-Fluorouracil (5-FU): 260 µg/l
Amoxicillin: 2,000 µg/l
5-FU + Amoxicillin: 6 µg/l
Cefazolin: 120,800 µg/l
5-FU + Cefazolin: 8 µg/l
Mixture: 5-FU – ß-Lactams Growth Inhibition Test with Bacteria (EN ISO 10712 :
1995)
Factor: 4*102 (5-FU) – 1,5 *104 (Cefazolin)
Alexy and Kümmerer, in preparation