Measurement of human- dispensed pharmaceuticals in complex

matrices, like digestates and blackwater

Tobias Wätzel, Katrin Krause & Tobias Schnabel

EWA 18th International Symposium

IFAT 2016, Munich, Germany 31.05.2016 – 01.06.2016

Blackwater and Digestates in innovative sanitation systems

Background

characterization

pharmaceutical pollution

Description of matrix, challenges for process controlling

Expectancy values

Normative substances for standardization

Distracting, complex molecules

Developed procedure for process validation using GC-MS instead of LC-MS/MS

Procedure for sample preparation

Advantages of GC- MS comparing to LC-MS/MS

Lessons learned and future applications of the new method

Cost reduced, pharmaceutical measurement in mostly ALL liquid matrices

Scientific usage and economic relevance

Continuous development for reliable results in sludge and solids

Contents

The background of the project:

- big networking project with different scientific and practical members (called

KREIS)

- the main aim: “Linking sustainable energy generation to urban

wastewater engineering”

- closed loop for wastewater with combined energy recovery

Main focus and working task of the Chair in the whole project:

- separation of municipal wastewater in different technical parts for optimizing

wastewater treatment (BW, GW, RW, PW)

- specified degradation of pharmaceuticals and pharmaceutical residues

in wastewater

1

Backround

blackwater greywater rainwater

strategy: separation of the different technical compounds for a specified

treatment application

The HAMBURG WATER Cycle ®

Separation of municiple wastewater in the different

technical compounds…

http://www.cghauling.com/wp-content/uploads/2013/02/green-waste.jpg

Greenery waste

Reasons for this experimental investigation

biowaste

worms-fresh-compost-scraps.jpg

• 4 targets of blackwater treatment including recovery of energy from

organic residues

• digester gas recovery

• pharmaceutical degradation

• Aprroving treatment and biogas production stability of anaerobic

treatment plants with a decentralized focus

• Organic to liquid doseing, pumping, homogenise

• Are they combinable?

• Are they conflicted?

• 2 challenges involved

• increasing of treatment efficiency by increasing digester gas production

• degradation of pharmaceuticals for decreasing of environmental pollution

using anaerobic treatment conditions (CSTR + UASB)

• Are this targets disputed?

Providing a comprehensive study using the CSTR and UASB- technique

with a variation of different reactor loadings by hydrolysis- substrate

feeding

Main aims of the research activities

Organic C to CH4

Reducing

pharmaceutical load

Stable treatment performance

Resource efficiency (C,N,P)

- human- related intruding of pharmaceuticals and pharmaceutical residues in different

environmental compartments (water and soil)

- antidromic effect; increasing of analytical quality and increasing of pharmaceutical

consumption detection of pharmaceuticals and residues in every

environmental area (groundwater, surface water, soil etc.)

- insufficient long- time- and big- scale- studies about human- and environmental- toxic

effects and result of re- formation and interaction between different substances

(„World´s experiment“) Sattelberger, 1999

- human- depending loop of pharmaceuticals and residues direct and indirect

intruding into soil and water vs. direct and indirect recovery of freshwater, producing

of animals and food plants

- Results: antibiotic resistance, infertility, hormonal balance

The problem with pharmaceuticals in wastewater and the environment

Constructed pilot plant- input material and parameters/blackwater

Diclofenac Ibuprofen Metformin Metoprolol Amoxicillin Carba-

mazepine

85%-Percentile

[µg/l] 28,45 411,5 1765 47,75 1015 141,5

Value Number [n] 20 20 20 20 20 20

Sampling

Duration [d] 370 370 370 370 370 370

acetic

acid

propionic

acid TP DOC DIC TOC

TN

dissolved COD

mg/l mg/l mg/l g/l g/l g/l g/l g/l

85%-

Percentile 690 295,4 169,9 1,4 1 3,6 1,4 7,4

Value

Number [n] 11 10 8 11 11 7 7 11

Sampling and analytical procedure: - 1m3 of blackwater was sampled from Lübeck- Flintenbreite once a month - stored at 4°C in climate chambers - directly homogenized before feed to the different duplicates - weekly analyzing of dry and organic matter - physic- chemical analyzing twice a month - pharmaceutical analyzing twice a month - hygienic analyzing once a month

same procedure for the resulting digestate

- projection period: 6 months

- construction period: - CST- reactors: 3 months - UASB- reactors: 6 months

- treatment capacity for blackwater: - low- load conditions ca. 20l/d - high- load conditions ca. 76l/d

- used co- substrates for variation of reactor loading:

- biowaste, hydrolyzed organic materials (greenery, kitchen waste, biowaste), fat separator deposits

Main data of the pilot plant for the anaerobic treatment of separated blackwater

(in extracts) chemical structure of:

Amoxicillin Metformin Diclofenac

pharmaceutical substance effect in human body

average of daily

dose

[1][2]

[ddd]

pbt-index (pbt) [1]

degradation performance[%]

aerobic anaerobic with co-

substrate [11] [12] [13]

Diclofenac anti- rheumatic agent 10.015.675 4 (301) <10 [3] 93

Ibuprofen anti- rheumatic agent 13.363.692 2 (002) 90 [4] 29

Metoprolol selective- β receptor

blocker

16.377.224 4 (301) 60[4][5] 36

Metformin oral anti- diabetic agent 8.214.307 5 (302) 50 [6] 95

Amoxicillin antibiotic

agent

1.187.403 6 (303) 30 [7] 75

Carbamazepine anti- convulsing agent 1.166.333 4 (301) <10 [8] 44

17- alpha-

Ethynilestradiol sexual hormone 21.477.195 9 (333) 96 [9] 94

Sulfamethoxazol antibiotic

agent

198.734 6 (303) 28 [10] 82

Metabolism, daily doses, pbt- index & degradation performances of selected pharmaceuticals [1] SLL (2009), [2] Baumgartner & Lutters (2011), [3] Kümmerer et al. (2011), [4] Jekel & Dott (2013), [5] Rosal et al. (2010), [6] Scheuerer et al. (2009 & 2012), [7] Valvo et al. (1997), [8] LANUV-NRW (2007), [9] Tennhard (2004), [10] UBA (2005), [11] Wätzel et al. (2013 & 2014), [12] Wätzel & Kraft (2014)

Is that the end of it??

No!- Diclofenac as one example!

Balancing of pharmaceuticals during the anaerobic treatment

!!!!! !!!!!

!!!!?? !!!!??

!?!?!?

State of knowledge

good

moderate

poor

http://www.vfa.de/static/generated/12817-ffdl2011-s25.jpg

Human produced metabolites

Raw substance

Unknown products

Human produced metabolites

Raw substance

Anaerobic metabolites

Unknown products

CSTR

Balancing of pharmaceuticals during the anaerobic treatment

State of knowledge

good

moderate

poor

Human produced metabolites

Raw substance

Unknown products

Human produced metabolites

Raw substance

Anaerobic metabolites

Unknown products

CSTR

Balancing of pharmaceuticals during the anaerobic treatment- how to

solve that problems?

State of knowledge

good

moderate

poor

Identified problems and future works in our field: -Balancing of treatment efficiency not proofed using raw substances -Unknown products of both sites of the system; efficiency rates -Ratio of the pharmaceutical inactivation during the process? -„look at that what you want to see!“ –> problems with the target analytics -Possible options for solving some of that problems –> Non- targets, new measurement methods

Balancing of pharmaceuticals during the anaerobic treatment- heavy

matrices

Balancing of pharmaceuticals during the anaerobic treatment- new

options

Advantages/Disadvantaged for pharmaceutical measurement

GC- MS LC-MS/MS

- Reduction of matrix clean up of impurities

- Clean up vs. transformation of origin sample

- Increased stability of measurement by ionisation using EI- electron impact/ Diclorprop as standard substance

- Decreased stability of measurement by using ESI- electron spray injection Ionsuppression depending on salt, fats, humic acid etc.

- Diclorprop as internal standard for all measurements

- Internal, deuterated C13- standard substances are needed for each molecule

- Possibilities for non- target screening using stable molecule mass library

- At this juncture: method only useable for acetous pharmaceuticals which have to be derivate

- Valid method can be used for all “well- known” raw pharmaceutical substances

Balancing of pharmaceuticals during the anaerobic treatment- significant

GC- MS measurements

Lessons learned and future applications of the new

method

- Cost reduced, pharmaceutical measurement in mostly ALL liquid matrices

- Equipment/Invest of GC- MS new: 60000€

- Equipment/Invest of LC- MS/MS new: 150000€ – 200000€

- Reducing the analytical prize/sample around 2/3

- Scientific usage and economic relevance

- Fast and approved measurements for validation of experimental results in lab- as well as pilot

scale

- Potential and useful solution for process controlling and validation in municipal and innovative

wastewater treatment systems

- Continuous development for reliable results in sludge and solids

- Increasing the number of pharmaceutical substances for GC- MS detection

- Development of non- abrasive and non- distracting methods for sample clean

up and preparation

- Development of non- abrasive and non- distracting methods for solid

analyzing (activated sludge, granular sludge, soil and concrete)

BAUHAUS- UNIVERSITÄT WEIMAR (2013): Bewertung der Stoffwechselvorgänge zum Abbau von Arzneimitteln. Master- Thesis FACHAGENTUR NACHWACHSENDE ROHSTOFFE (2010): Leitfaden Biogas – von der Gewinnung zur Nutzung; 5.Auflage FAHLENKAMP, H.; NÖTHE, T.; NOWOTNY, N.; RIES, T.; HANNICH, C. B.; PEULEN, C.; KUHN-JOERESSEN, S.; VON SONNTAG, C.; PEHL, C.; POPPE, A.; KRAFT, A.; OBENAUS, F.; JAGEMANN, P.; REUPERT, R.; SIELEX, K.; STOCK, H.-D.; SPITELLER, M.; ZÜHLKE, S.; LIGON, A.; DÜPUTELL, D.; GROTEHANS, J., JANTZEN, E. (2006): Untersuchungen zum Eintrag und zur Elimination von gefährlichen Stoffen in kommunalen Kläranlagen, Teil 2.- MUNLV – NRW 1.16.100.1. ISOE (2008): Pharmaceuticals for Human Use: Options of Action for Reducing the Contamination of Water Bodies; Institute for Social-Ecological Research (ISOE) GmbH; Practical Guideline MOGENSEN C.E. (2003): Microalbuminuria and hypertension with focus on type 1 and type 2 diabetes; Journal of Intern Medicine; p. 45- 66. PALMQUIST, H.; JÖNSSON, H. (2003): Urine, faeces, greywater and biodegradable solid waste as potential fertilisers, ecosan- closing the loop; proceedings of the 2nd international symposium on ecological sanitation; 7th – 11th April 2003; Lübeck, Germany ROENNEFAHRT, I. (2005): Verbrauchsmengen in der Bewertung des Umweltrisikos von Humanarzneimitteln; In: Umweltbundesamt (Hrsg); Arzneimittel in der Umwelt – Zu Risiken und Nebenwirkungen fragen Sie das Umweltbundesamt; Dessau, Germany; UBA-text publishing 29/05 RWTH Aachen (2009): Eliminierung von Spurenstoffen aus Krankenhausabwässern mit Membrantechnik und weitergehenden Behandlungsverfahren - Pilotprojekt Kreiskrankenhaus Waldbröl; Institut für Siedlungswasserwirtschaft SCHARF, S.; GANS, O.; SATTELBERGER, R. (2002): Arzneimittelwirkstoffe im Zu- und Ablauf von Kläranlagen; Federal Environment Agency of Austria. SIEGRIST, H., JOSS, A., BOEHLER, M., BUETZER, S., HOLZAPFEL, M., MOOSER, H., (2007): Treatment of toilet wastewater for reuse in a membrane bioreactor; Wat. Sci. Technol.; 56(5); p. 63-70 Tennhardt, L. (2004): Potenzial technischer Abwasser- und Klärschlammbehandlungs-verfahren zur Elimination endokrin aktiver Substanzen; Beiträge zu Abfallwirtschaft und Altlasten, Schriftenreihe des Institutes für Abfallwirtschaft und Altlasten der Technischen Universität Dresden; Vol. 33 UBA (2002): Arzneimittelwirkstoffe im Zu- und Ablauf von Kläranlagen; Berichte des österreichischen Umweltbundesamt UBA (2011): Zusammenstellung von Monitoringdaten zu Umweltkonzentrationen von Arzneimitteln Universität Dortmund (2003): Untersuchungen zum Eintrag und zur Elimination von gefährlichen Stoffen in kommunalen Kläranlagen Teil 1; Fachbereich Chemietechnik, Lehrstuhl Umwelttechnik URBAN, I. (2009): Anaerobe Kommunalabwasserbehandlung – Einsatz und Bemessung von UASB-Reaktoren; Gottfried Wilhelm Leibnitz-University Hannover; Publication of Institute for Urban Sanitation und Waste Management ZWIENER, C (2007): Occurrence and analysis of pharmaceuticals and their transformation products in drinking water treatment; Anal. Bioanal. Chem. 387; p. 1159-1163

References

Thank you for your attention!

Special thanks goes to the BMBF for funding and also to the involved

colleges from the b.is and from the other participated partners in the project

M.Sc. Tobias Wätzel Bauhaus- Universität Weimar bauhaus institute for infrastructure solutions (b.is) Chair of Biotechnology in Resources Management tobias.waetzel@uni-weimar.de