Persistence and Fate of Antibiotics in Sewage Prof. Dr. Klaus Kümmerer Pharmaceutical Sewage During Influenza Pandemic Somerville College, University of Oxford, 3 March 2009 Department of Environmental Health Sciences

Some Data Consumption of Antibiotics 65 % 6 % 29 % USA t ? Other countries ? World ? EU: total 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, , 2006 

Emissions of Antibiotics Into Municipal Sewage Usage 1998: 412 t (Germany) Emission 1998: 277 t (Germany) Thereof hospitals 92 t (Germany)  Kümmerer und Henninger, Clinical Microbiology and Infection, 9, , 2003

What Happens in the Sewage Treatment Plant? © Abwasserzweckverband Breisgauer Bucht,

Elimination of the compounds? In the ng-µg/L in the effluent!

Antibiotics: % Cytotoxics % Hormones: % Statins (Lipid lowering) % Pain killers % Anti depressivs % Disinfectants % Diagnostics % Elimination of Pharmaceuticals in 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 P ow, log K oc ) Redox potential Degree of saturation Types of functional groups Electrical charges Acid/base constants (pK a s)

Structure Matters: Impact of Structure on Elimination in a STP Concentration (ng/L; µg/kg d.m.) influent sludge effluentsludge effluent surplus sludge  Alexy, Sacher, Wenz, Kümmerer, Acta hydrochim. Hydrobiol., 2006

Structure Matters: Impact of Structure on Elimination in a STP Concentration (ng/L; µg/kg d.m.) influent sludge effluentsludge effluent surplus sludge NOCH 2 OC 2 H 5 OCH 3  Alexy, Sacher, Wenz, Kümmerer, Acta hydrochim. Hydrobiol., 2006

Compound specific approach needed ! Which Compound?

Pharmaceuticals Are Different! Ciprofloxacin Ciprofloxacin Acidic groups Basic groups Optimised for biological activity and stability. Several different functional groups within on and the same molecule.

Ciprofloxacin: Micro-species as a function of pH Zwitterion: Isoelectric point at pH 7

(Primary) Elimination vs. (Total) Mineralization What degree is needed?

Physical Chemical Instability of Antibiotics darkness light  Kümmerer and co-workers, unpublished

Stability Profile of Ceftriaxone (Fitted According to Arrhenius-Equation) Concentration DAY Temperature  Kümmerer and co-workers, unpublished

Mineralization of 14 C-Labelled Antibiotics in Laboratory Scale Sewage Treatment  T. Junker, R. Alexy, T. Knacker, K. Kümmerer, Environ. Sci. & Technol., 40 (2006), Share of Radioactivity [%] Sampling Mineralization Effluent STP Total Recovery Benzylpenicillin Cefriaxone Trimethoprím

Degradation of Structurally Closely Related Penicillins Piperacillin Amoxicillin Benzylpenicillin

HPLC-UVVis: biol. non biol. DOC: biol. non biol. Piperacillin Day Elimination and Biodegradation Zahn Wellens Test (OECD 302 B)  Kümmerer et al., Chemosphere, in press

m/z 518 (+mode) Piperacillin m/z 536 (+ mode) hydrolysed Piperacillin N S O OH H HN O O H N N O N O O N H S O OH H HN O H NN O N O O OH O (n. biotic) hydrolysis Degradation of Pieracillin (LC-MS/MS ion trap, -UV-Vis, DOC) +H 2 O  Kümmerer et al., Chemosphere, in press

HPLC-UVVis: biol. non biol. DOC: biol. non biol. Piperacillin Amoxicillin Day Elimination and Biodegradation Zahn Wellens Test (OECD 302 B)  Kümmerer et al., Chemosphere, in press

Amoxicillin hydrolysed Amoxicillin without phenol ring Bio- degradation Bio- mineralization (n. biotic) Hydrolysis -C 6 H 6 O Degradation of Amoxicillin (LC-MS/MS ion trap, -UV-Vis, DOC)  Kümmerer et al., Chemosphere, in press t 1/2 Benzylpenicillin ≈ 2 * t 1/2 Amoxicillin

Photo-degradation of Ciprofloxacin  Vasconcelos T. G., Henriques D.M., König, A., Martins A.F., Kümmerer K., Chemosphere, accepted 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 Photo-degradation of Ciprofloxacin

 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?

Properties of Mixtures of Pharmaceuticals: Expectation and Reality

Properties of Mixtures of Pharmaceuticals: Expectation and Reality

EC 50 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 : 1995) ‏ Factor: 4*10 2 (5-FU) – 1,5 *10 4 (Cefazolin)‏  Alexy and Kümmerer, in preparation

Looking forward to the discussion ?

Discussion Looking forward to the discussion !