Method Development for Analysis of Pharmaceuti-Cals in Environmental
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Umweltforschungsplan des Bundesministeriums für Umwelt, Naturschutz, Bau und Reaktorsicherheit Forschungskennzahl 3715674130 UBA-FB-00 [trägt die UBA-Bibliothek ein] Method development for analysis of pharmaceuti- cals in environmental samples von Prof. Dr. Thomas Ternes Bundesanstalt für Gewässerkunde, Koblenz Dr. Georg Dierkes Bundesanstalt für Gewässerkunde, Koblenz Lise Boulard Bundesanstalt für Gewässerkunde, Koblenz Alexander Weizel Bundesanstalt für Gewässerkunde, Koblenz Bundesanstalt für Gewässerkunde Am Mainzer Tor 1 56068 Koblenz Im Auftrag des Umweltbundesamtes Abschlussdatum Februar 2018 UBA Texts Pharmaceuticals in environmental samples Abstract Since the last decades the increasing currency of pharmaceuticals in the aquatic environment and their risk potential for the aquatic life has become an urgent issue. The number of pharmaceuticals detecta- ble in the low µg/L range in the environment is rising each day. Against this background the aim of this project was the development, optimization, validation and comparison of analytical methods for quan- tification of selected pharmaceuticals in different environmental matrices (water, sediment, suspend- ed particular matter and biota). High sensitive LC-MS-MS methods for determination of extreme polar, polar and hormonal pharmaceuticals in water samples were developed and validated. Using these methods occurrence and distribution of the selected pharmaceuticals in water with different waste water content were determined. For analysis of pharmaceuticals in particular matter and biota sam- ples extraction and clean-up procedures were tested, but method development is still in process. Fur- thermore a monitoring campaign was set up and water samples, suspended particular matter and bio- ta (fish) samples from six different sampling sites were taken. Kurzbeschreibung Die Verbreitung von Arzneimittelrückständen in Gewässern und deren Gefährdungspotential für Was- serlebewesen hat in den letzten Jahren beunruhigende Ausmaße angenommen. Immer mehr Arznei- mittelwirkstoffe werden in Konzentrationen bis in den unteren µg/L-Bereich in der aquatischen Um- welt nachgewiesen. Vor diesem Hintergrund ist das Ziel dieses Projekts analytische Nachweisverfah- ren und Probenahmekonzepte für ausgewählte Arzneimittelwirkstoffe und verschiedene Matrizes (Wasser, Sediment, Schwebstoff, Biota) zu entwickeln, optimieren, validieren und vergleichen. Leis- tungsstarke LC-MS-MS Methoden wurden für den Nachweis von extrem polaren, polaren und hormo- nellen Wirkstoffen in Wasserproben entwickelt und validiert. Mit Hilfe dieser Methoden wurden an- schließend das Vorkommen und die Verteilung der ausgewählten Arzneimittelwirkstoffe in Wasser- proben aus Gewässern mit unterschiedlichen Abwasseranteilen ermittelt. Für die Analytik von Schwebstoff- und Biotaproben wurden Extraktions- und Aufreinigungsverfahren getestet, allerdings befinden sich diese Methoden noch in der Entwicklung. Des Weiteren wurde ein Monitoringskonzept entwickelt und Wasser-, Schwebstoff und Biotaproben aus sechs verschiedenen Gewässern entnom- men. 4 UBA Texts Pharmaceuticals in environmental samples Table of content Table of figures ................................................................................................................................................. 6 Table of tables .................................................................................................................................................. 7 Abbreviations.................................................................................................................................................... 8 1 Introduction .......................................................................................................................................... 13 2 Work package 1: Categorization and prioritization .............................................................................. 14 3 Work package 2: Method development and conception of a monitoring program ............................ 16 3.1 Method development ........................................................................................................... 16 3.1.1 Suspect-Screening ............................................................................................................ 16 3.1.1.1 Method 16 3.1.1.2 Results 17 3.1.2 Extreme polar pharmaceuticals (without hormones) ...................................................... 18 3.1.2.1 Material and Methods 19 3.1.2.2 Result and discussion 22 3.1.2.3 Conclusions 40 3.1.3 Middle polar pharmaceuticals ......................................................................................... 43 3.1.4 Hormone .......................................................................................................................... 43 3.1.4.1 Materials and Methods 45 3.1.4.2 Results and Discussion 48 3.1.4.3 Conclusions 57 3.1.5 Sediment and suspended particulate matter .................................................................. 58 3.2 Conception of a monitoring program ................................................................................... 62 4 Work package 3: Environmental monitoring ........................................................................................ 63 4.1 Rhine ..................................................................................................................................... 63 4.2 Occurrence of pharmaceuticals in surface waters ............................................................... 70 4.3 Comparison of suspect screening and target analysis .......................................................... 79 5 Summary and elaboration of recommendations (Work package 4): ................................................... 81 6 References ............................................................................................................................................ 83 7 Appendix ............................................................................................................................................... 90 5 UBA Texts Pharmaceuticals in environmental samples Table of figures Figure 1: Scheme of the data processing-steps for the used in-house Suspect-Screening .............................. 17 Figure 2: Identified suspect ............................................................................................................................ 18 Figure 3: Superposition of MRM transitions of a 500 ng/L multi-standard. .................................................... 23 Figure 4: Influence of the composition of the aqueous eluent and the column on the elution of metformin. ........................................................................................................ 24 Figure 5: Comparison of 2 injections of a 2000 ng/L standard with the HILIC Nucleodur. .............................. 25 Figure 6: Matrix effects .................................................................................................................................... 34 Figure 7: Post-column infusion of acyclovir-d4 ................................................................................................ 35 Figure 8: Post-column infusion of a multi-standard ........................................................................................ 36 Figure 9: Sodium adducts of emtricitabine ...................................................................................................... 37 Figure 10: Retention times and peak forms of a 1000 ng/L multi-standard dissolved in different diluents . ............................................................................................................. 38 Figure 11: Retention times and peak forms of a 1000 ng/L multi-standard measured with eluent B containing different ratio acetonitrile/Milli-Q .................................................... 39 Figure 12: Workflow of the developed method for the trace quantification of 60 steroid hormones by LC-MS/MS .................................................................................................... 46 Figure 13: Recoveries ....................................................................................................................................... 49 Figure 14: Chemical structures, extracted ion chromatogram of non-spiked WWTP effluent and high-resolution MS²-spectra. ....................................................................... 50 Figure 15: Boxplots of the recoveries of the extraction procedure with ultrasonic extraction and accelerated solvent extraction. ................................................................. 58 Figure 16: Boxplot of the recoveries obtained with different extraction solvents. MQ: Milli-Q, MeOH: methanol. ................................................................................................ 59 Figure 17: Boxplot of the recoveries obtained for different pH for the aqueous part of the extraction solvent methanol/water, 50/50, v/v. ......................................................... 60 Figure 18: Boxplot of the recoveries for different volumetric percentage of formic acid in the aqueous phase of the extraction solvent. .................................................................. 60 Figure 19: Boxplots of the recoveries obtained with different procedure ...................................................... 61 Figure 20: Map of the sampling locations along the Rhine .............................................................................