Chemotherapeutics are pharmaceutical compounds the occurrence of which in the environment is of growing concern because of the increase in treatments against cancer diseases. They can reach the aquatic ecosystems after passing through wastewater treatment plants without complete removal. One of the most frequently used chemotherapeutics is 5-fluorouracil which exhibits a strong cytostatic effect. In this paper, an analytical methodology was developed, validated, and applied to determine 5-fluorouracil, its precursor, 5-fluorocytosine, and its major active metabolite, 5-fluorouridine, in hospital wastewater samples. Due to the expected low concentrations after dilution and interferences present in such a complex matrix, a very selective and sensitive detection method is required. Moreover, an extraction method must be implemented prior to the determination in order to purify the sample extract and preconcentrate the target analytes at micrograms per liter concentration levels. Solid-phase extraction followed by liquid chromatography with tandem mass spectrometry was the combination of choice and all included parameters were studied. Under optimized conditions for wastewater samples analysis, recoveries from 63 to 108% were obtained, while intraday and interday relative standard deviations never exceeded 20 and 25%, respectively. Limits of detection between 61 and 620 ng/L were achieved. Finally, the optimized method was applied to samples from hospital wastewater effluents.

Comenius University in Bratislava Faculty of Natural Sciences Department of Analytical Chemistry Bratislava Slovak Republic

Instituto de Estudios Ambientales y Recursos Naturales Universidad de Las Palmas de Gran Canaria Las Palmas de Gran Canaria Spain

University of South Bohemia in Ceske Budejovice Faculty of Fisheries and Protection of Waters South Bohemian Research Center of Aquaculture and Biodiversity of Hydrocenoses Vodnany Czech Republic

Zobrazit více v PubMed

https://gco.iarc.fr/today/data/factsheets/cancers/39-All-cancers-fact-sheet.pdf (last time accessed: April 21, 2020)

Mahnik, S. N., Lenz, K., Weissenbacher, N., Mader, R. M., Fuerhacker, M., Fate of 5-fluorouracil, doxorubicin, epirubicin, and daunorubicin in hospital wastewater and their elimination by activated sludge and treatment in a membrane-bio-reactor system. Chemosphere 2007, 66, 30-37.

Ferrando-Climent, L., Rodriguez-Mozaz, S., Barceló, D., Incidence of anticancer drugs in an aquatic urban system: From hospital effluents through urban wastewater to natural environment. Environ. Pollut. 2014, 193, 216-223.

Zhang, J., Chang, V. W. C., Giannis, A., Wang, J., Removal of cytostatic drugs from aquatic environment : A review. Sci. Total Environ. 2013, 445-446, 281-298.

Besse, J. P., Latour, J. F., Garric, J., Anticancer drugs in surface waters. What can we say about the occurrence and environmental significance of cytotoxic, cytostatic and endocrine therapy drugs? Environ. Int. 2012, 39, 73-86.

Santos, M. S. F., Franquet-Griell, H., Alves, A., Lacorte, S., Development of an analytical methodology for the analysis of priority cytostatics in water. Sci. Total Environ. 2018, 645, 1264-1272.

Martín, J., Camacho-Muñoz, D., Santos, J. L., Aparicio, I., Alonso, E., Simultaneous determination of a selected group of cytostatic drugs in water using high-performance liquid chromatography-triple-quadrupole mass spectrometry. J. Sep. Sci. 2011, 34, 3166-3177.

Fischer, J., Ganellin, R. C., Analogue-Based Drug Discovery. WILEY-VCH Verlang GmbH and Co. KGaA, Weinheim 2006.

U.S. National Library of Medicine, PubChem Database, 5-fluorouracil, https://pubchem.ncbi.nlm.nih.gov/compound/3385#section=Top (last time accessed: April 21, 2020).

Kosjek, T., Perko, S., Žigon, D., Heath, E., Fluorouracil in the environment : Analysis , occurrence , degradation and transformation. J. Chromatogr. A 2013, 1290, 62-72.

Kovalova, L., McArdell, C. S., Hollender, J., Challenge of high polarity and low concentrations in analysis of cytostatics and metabolites in wastewater by hydrophilic interaction chromatography/tandem mass spectrometry. J. Chromatogr. A 2009, 1216, 1100-1108.

Mullot, J. U., Karolak, S., Fontova, A., Huart, B., Levi, Y., Development and validation of a sensitive and selective method using GC/MS-MS for quantification of 5-fluorouracil in hospital wastewater. Anal. Bioanal. Chem. 2009, 394, 2203-2212.

Vermes, A., Guchelaar, H., Dankert, J., Flucytosine : a review of its pharmacology , clinical indications , pharmacokinetics , toxicity and drug interactions. J. Antimicrob. Chemother. 2000, 46, 171-179.

Wishart, D. S., Feunang, Y. D., Guo, A. C., Lo, E. J., Marcu, A., Grant, J. R., Sajed, T., Johnson, D., Li, C., Sayeeda, Z., Assempour, N., Iynkkaran, I., Liu, Y., Maciejewski, A., Gale, N., Wilson, A., Chin, L., Cummings, R., Le, D., Pon, A., Knox, C., Wilson, M., DrugBank 5.0: a major update to the DrugBank database for 2018, 5-fluorouracil, DOI: 10.1093/nar/gkx1037https://www.drugbank.ca/drugs/DB00544 (last time accessed: April 21, 2020).

Reynolds, E. F. J., Parfitt, K., Parsons, V. A., Sweetman, C. S., Martindale The Extra Pharmacopoeia. The Pharmaceutical Press, London 1993.

Miura, K., Kinouchi, M., Ishida, K., Fujibuchi, W., Naitoh, T., Ogawa, H., Ando, T., Yazaki, N., Watanabe, K., Haneda, S., Shibata, C., Sasaki, I., 5-FU metabolism in cancer and orally-administrable 5-FU drugs. Cancers (Basel). 2010, 2, 1717-1730.

Zounkova, R., Kovalova, L., Blaha, L., Dott, W., Ecotoxicity and genotoxicity assessment of cytotoxic antineoplastic drugs and their metabolites. Chemosphere 2010, 81, 253-260.

Maršálek, B., Zounková, R., Bláha, L., Odráška, P., Doležalová, L., Hilscherová, K., Ecotoxicity and Genotoxicity Assessment of Cytostatic Pharmaceuticals. Environ. Toxicol. Chem. 2007, 26, 2208.

Białk-Bielińska, A., Mulkiewicz, E., Stokowski, M., Stolte, S., Stepnowski, P., Acute aquatic toxicity assessment of six anti-cancer drugs and one metabolite using biotest battery - Biological effects and stability under test conditions. Chemosphere 2017, 189, 689-698.

Kovács, R., Csenki, Z., Bakos, K., Urbányi, B., Horváth, Á., Garaj-Vrhovac, V., Gajski, G., Gerić, M., Negreira, N., López de Alda, M., Barceló, D., Heath, E., Kosjek, T., Žegura, B., Novak, M., Zajc, I., Baebler, Š., Rotter, A., Ramšak, Ž., Filipič, M., Assessment of toxicity and genotoxicity of low doses of 5-fluorouracil in zebrafish (Danio rerio) two-generation study. Water Res. 2015, 77, 201-212.

Novak, M., Žegura, B., Modic, B., Heath, E., Filipič, M., Cytotoxicity and genotoxicity of anticancer drug residues and their mixtures in experimental model with zebrafish liver cells. Sci. Total Environ. 2017, 601-602, 293-300.

Nussbaumer, S., Bonnabry, P., Veuthey, J. L., Fleury-Souverain, S., Analysis of anticancer drugs: A review. Talanta 2011, 85, 2265-2289.

Nollet, L. M. L., Lambropoulou, D. A. (Eds.), Chromatographic Analysis of the Environment: Mass Spectrometry Based Approaches. Taylor and Francis Group, Boca Raton, FL 2017.

Zhang, Q., Yang, F., Ge, L., Hu, Y., Xia, Z., Recent applications of hydrophilic interaction liquid chromatography in pharmaceutical analysis. J. Sep. Sci. 2017, 40, 49-80.

Fabrizi, G., Fioretti, M., Mainero Rocca, L., Dispersive solid-phase extraction procedure coupled to UPLC-ESI-MS/MS analysis for the simultaneous determination of thirteen cytotoxic drugs in human urine. Biomed. Chromatogr. 2016, 30, 1297-1308.

Kosjek, T., Heath, E., Occurrence, fate and determination of cytostatic pharmaceuticals in the environment. TrAC - Trends Anal. Chem. 2011, 30, 1065-1087.

Santana-Viera, S., Montesdeoca-Esponda, S., Sosa-Ferrera, Z., Santana-Rodríguez, J. J., Cytostatic drugs in environmental samples: An update on the extraction and determination procedures. TrAC - Trends Anal. Chem. 2016, 80, 373-386.

Gros, M., Petrovic, M., Barceló, D., Multi-residue analytical methods using LC-tandem MS for the determination of pharmaceuticals in environmental and wastewater samples: a review. Anal. Bioanal. Chem. 2006, 386, 941-952.

Gómez, M. J., Petrović, M., Fernández-Alba, A. R., Barceló, D., Determination of pharmaceuticals of various therapeutic classes by solid-phase extraction and liquid chromatography-tandem mass spectrometry analysis in hospital effluent wastewaters. J. Chromatogr. A 2006, 1114, 224-233.

Matuszewski, B. K., Constanzer, M. L., Chavez-Eng, C. M., Strategies for the assessment of matrix effect in quantitative bioanalytical methods based on HPLC-MS/MS. Anal. Chem. 2003, 75, 3019-3030.

Buhrman, D. L., Price, P. I., Rudewicz, P. J., Quantitation of SR 27417 in human plasma using electrospray liquid chromatography-tandem mass spectrometry: a study of ion suppression. J. Am. Soc. Mass Spectrom. 1996, 7, 1099-1105.

Santana-Viera, S., Hernández-Arencibia, P., Sosa-Ferrera, Z., Santana-Rodríguez, J. J., Simultaneous and systematic analysis of cytostatic drugs in wastewater samples by ultra-high performance liquid chromatography tandem mass spectrometry. J. Chromatogr. B 2019, 1110-1111, 124-132.

Barker, S. A., de Zeeuw, R. A., Franke, J. P., Henry, M., Ingwersen, S. H., Jordan, L., Law, B., Martin, P., Matyska, M. T., Pesek, J. J., Poole, C. F., Poole, S. K., Rashid, B. A. A., Shahtaheri, S. J., Simpson, N. J. K., Stevenson, D., Wells, M. J. M., Wilson, I. D., Wynne, P. M., Solid-Phase Extraction; Principles, Techniques, and Applications. Marcel Dekker, Inc., New York, NY 2000.

Funk, W., Dammann, V., Donnevert, G., Quality Assurance in Analytical Chemistry: Applications in Environmental, Food, and Materials Analysis, Biotechnology, and Medical Engineering. WILEY-VCH, Weinheim 2007.

Liu, Y., Zhu, P., Huang, Z., Zhou, L., Shi, P., Simultaneous detection of 5-fluorocytosine and 5-fluorouracil in human cells carrying CD/5-FC suicide gene system by using capillary zone electrophoresis. J. Chromatogr. B 2018, 1076, 1-7.

Serve, K. M., Yáñez, J. A., Remsberg, C. M., Davies, N. M., Black, M. E., Development and validation of a rapid and sensitive HPLC method for the quantification of 5-fluorocytosine and its metabolites. Biomed. Chromatogr. 2010, 24, 556-561.

Mahnik, S. N., Rizovski, B., Fuerhacker, M., Mader, R. M., Determination of 5-fluorouracil in hospital effluents. Anal. Bioanal. Chem. 2004, 380, 31-35.

Tauxe-Wuersch, A., de Alencstro, L. F., Grandjean, D., Tarradellas, J., Trace determination of tamoxifen and 5-fluorouracil in hospital and urban wastewaters. Int. J. Environ. Anal. Chem. 2006, 86, 473-485.