Solid-phase extraction of estrogens and herbicides from environmental waters for bioassay analysis—effects of sample volume on recoveries


Ecotoxicological screening of surface waters can involve multiple analyses using multiple bioassay and chemical analytical methods that require enriched samples to reach low concentrations. Such broad screening of the same sample necessitates sufficient sample volume—typically several liters—to produce a sufficient amount of enriched sample. Often, this is achieved by performing parallel solid-phase extractions (SPE) where extracts are combined into a pool—this is a laborious process. In this study, we first validated our existing SPE method for the chemical recovery of an extended set of compounds. We spiked four estrogenic compounds and 11 herbicides to samples from independent rivers (1 L) and wastewater treatment plant effluents (0.5 L). Then, we investigated the effect of increased sample loading of the SPE cartridges on both chemical and biological recoveries by comparing the validated volumes with four times larger sample volumes (i.e., 4 L river water and 2 L effluent). Samples were analyzed by LC-MS/MS and three bioassays: an estrogen receptor transactivation assay (ERα-CALUX), the combined algae test, and a bacterial bioluminescence inhibition assay. Our existing SPE method was found to be suitable for enriching the extended set of estrogens and herbicides in river water and effluents with near to perfect chemical recoveries (~ 100%), except for the herbicide metribuzin (46 ± 19%). In the large volume river and effluent samples, the biological activities and concentrations of the spiked compounds were between 87 and 104% of those measured with the lower sample loading, which is adequate. In addition, the ratio between the large and original volume SPE method for the non-target endpoint (bacterial bioluminescence inhibition) was acceptable (on average 82 ± 9%). Results indicate that our current water extraction method can be applied to up to four times larger sample volumes, resulting in four times more extract volumes, without significant reductions in recoveries for the tested estrogens and herbicides.

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Author information




Eszter Simon, Inge Werner, and Etiënne Vermeirssen conceived the study. Eszter Simon, Andrea Schifferli, Thomas Bucher, Daniel Olbrich, and Etiënne Vermeirssen planned the experiments. Andrea Schifferli, Thomas Bucher, and Daniel Olbrich carried out the sampling, the sample preparation and the bioassay and chemical analyses. Andrea Schifferli, Thomas Bucher, Daniel Olbrich, and Eszter Simon performed the calculations. Eszter Simon took the lead in writing the manuscript. All authors contributed to the revision of the manuscript.

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Correspondence to Eszter Simon.

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Simon, E., Schifferli, A., Bucher, T.B. et al. Solid-phase extraction of estrogens and herbicides from environmental waters for bioassay analysis—effects of sample volume on recoveries. Anal Bioanal Chem 411, 2057–2069 (2019).

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  • Photosynthesis inhibition
  • Raphidocelis subcapitata
  • Bacterial bioluminescence inhibition
  • LC-MS/MS
  • Solid-phase extraction
  • LiChrolut EN/RP-18