Details zur Publikation

Kategorie Textpublikation
Referenztyp Zeitschriften
DOI 10.1002/etc.5490
Lizenz creative commons licence
Titel (primär) Validation of an SH-SY5Y cell-based acetylcholinesterase inhibition assay for water quality assessment
Autor Lee, J.; Huchthausen, J.; Schlichting, R.; Scholz, S.; Henneberger, L.; Escher, B.I.
Journal / Serie Environmental Toxicology and Chemistry
Erscheinungsjahr 2022
Band/Volume 41
Heft 12
Seite von 3046
Seite bis 3057
Sprache englisch
Topic T9 Healthy Planet
Keywords Acetylcholinesterase inhibitors; dissolved organic matter; effects-based monitoring; neurotoxicity; SH-SY5Y cells; volatility
Abstract The acetylcholinesterase (AChE) inhibition assay has been frequently applied for environmental monitoring to capture insecticides such as organo(thio)phosphates (O(T)P) and carbamates. However, natural organic matter such as dissolved organic carbon (DOC) co-extracted with solid-phase extraction from environmental samples can produce false-negative AChE inhibition in free enzyme-based AChE assays. We aimed to evaluate whether disturbance by DOC can be alleviated in a cell-based AChE assay using differentiated human neuroblastoma SH-SY5Y cells. The exposure duration was set at an optimum of 3 h considering the effects of O(T)Ps and carbamates. Since loss to the airspace was expected for the more volatile OTPs (chlorpyrifos, diazinon, and parathion), the chemical loss in this bioassay set-up was investigated using a solid-phase microextraction followed by chemical analysis. The three OTPs were relatively well retained (loss < 34%) during 3 h of exposure in the 384-well plate but higher losses occurred upon prolonged exposure, accompanied by slight cross-contamination of adjacent wells. AChE inhibition by paraoxon-ethyl was not altered in the presence of up to 68 mgc/L Aldrich humic acid used as surrogate for DOC. Binary mixtures of paraoxon-ethyl and water extracts showed concentration-additive effects. These experiments confirmed that the matrix in water extracts does not disturb the assay unlike purified enzyme-based AChE assays. The cell-based AChE assay proved to be suitable for testing water samples with effect concentrations causing 50% inhibition of AChE at relative enrichments of 0.5 to 10 in river water samples, which were distinctly lower than corresponding cytotoxicity, confirming the high sensitivity of the cell-based AChE inhibition assay and its relevance for water quality monitoring.
dauerhafte UFZ-Verlinkung
Lee, J., Huchthausen, J., Schlichting, R., Scholz, S., Henneberger, L., Escher, B.I. (2022):
Validation of an SH-SY5Y cell-based acetylcholinesterase inhibition assay for water quality assessment
Environ. Toxicol. Chem. 41 (12), 3046 - 3057