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Human metabolites and transformation products of cyclophosphamide and ifosfamide: analysis, occurrence and formation during abiotic treatments

Abstract

This study describes a gas chromatography-mass spectrometry analytical method for the analysis of cytostatic cyclophosphamide (CP), ifosfamide (IF) and their selected metabolites/transformation products (TPs): carboxy-cyclophosphamide (carboxy-CP), keto-cyclophosphamide (keto-CP) and 3-dechloroethyl-ifosfamide/N-dechloroethyl-cyclophosphamide (N-decl-CP) in wastewater (WW). Keto-cyclophosphamide, CP and IF were extracted with Oasis HLB and N-decl-CP and carboxy-CP with Isolute ENV+ cartridges. Analyte derivatization was performed by silylation (metabolites/TPs) and acetylation (CP and IF). The recoveries and LOQs of the developed method were 58, 87 and 103 % and 77.7, 43.7 and 6.7 ng L−1 for carboxy-CP, keto-CP and N-decl-CP, respectively. After validation, the analytical method was applied to hospital WW and influent and effluent samples of a receiving WW treatment plant. In hospital WW, levels up to 2690, 47.0, 13,200, 2100 and 178 ng L−1 were detected for CP, IF, carboxy-CP, N-decl-CP and keto-CP, respectively, while in influent and effluent samples concentrations were below LOQs. The formation of TPs during abiotic treatments was also studied. Liquid chromatography-high-resolution mass spectrometry was used to identify CP and IF TPs in ultrapure water, treated with UV and UV/H2O2. UV treatment produced four CP TPs and four IF TPs, while UV/H2O2 resulted in five CPs and four IF TPs. Besides already known TPs, three novel TPs (CP-TP138a, imino-ifosfamide and IF-TP138) have been tentatively identified. In hospital WW treated by UV/O3/H2O2, none of the target metabolites/TPs resulted above LOQs.

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Acknowledgments

This work was financially supported by the EU through the EU FP7 project CytoThreat (fate and effects of cytostatic pharmaceuticals in the environment and the identification of biomarkers for an improved risk assessment on environmental exposure (Grant Agreement No.: 265264), by the Slovenian Research Agency (Program Groups P1-0143, Project L1-5457, Project J1-6744 and Young Researcher Grant to M. Č.) and the Slovene Human Resources Development and Scholarship Fund (Ad futura). The authors would like to thank the staff at the hospital and the WWTP involved in the study for their collaboration and help during sampling campaign.

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Correspondence to Ester Heath.

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Česen, M., Kosjek, T., Busetti, F. et al. Human metabolites and transformation products of cyclophosphamide and ifosfamide: analysis, occurrence and formation during abiotic treatments. Environ Sci Pollut Res 23, 11209–11223 (2016). https://doi.org/10.1007/s11356-016-6321-1

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Keywords

  • Cyclophosphamide
  • Ifosfamide
  • Cytostatic
  • Metabolites
  • Transformation products
  • Occurrence
  • Wastewater
  • Abiotic treatment