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Screening for Endocrine Activity in Water Using Commercially-available In Vitro Transactivation Bioassays

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In Vitro Bioassay Evidence for Chemical Mixture Propagation from the Environment to Humans.

Beate Escher^1,2,3, Martin Scholze⁴, Maria Margalef⁵

¹Department of Cell Toxicology, Helmholtz Centre for Environmental Research - UFZ, Leipzig 04318, Germany.

Environmental Science & Technology

|June 2, 2026

Summary

This summary is machine-generated.

Complex chemical mixtures in the environment and food impact neurodevelopment and reproductive health pathways. While current contamination levels pose no immediate risk, further toxicological study is needed due to unannotated molecular features contributing to bioactivity.

Keywords:

adverse outcome pathway concentration addition iceberg modeling in vitro bioassay mixture effects new approach methodologies

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Area of Science:

Environmental toxicology
In vitro bioassay development
Human health risk assessment

Background:

Environmental and food samples contain complex chemical mixtures.
These mixtures can affect critical biological pathways, including neurodevelopment and reproduction.
Understanding mixture toxicity is crucial for public health.

Purpose of the Study:

To assess the in vitro bioactivity of complex chemical mixtures from environmental and food samples.
To identify specific biological pathways affected by these mixtures.
To evaluate potential health risks associated with average contamination levels.

Main Methods:

Extraction of organic chemicals from diverse matrices (water, food, blood).
Testing extracts in 22 in vitro bioassays targeting neurodevelopmental and reproductive pathways.
Application of concentration addition models for mixture effects.
Nontarget analysis to detect unannotated molecular features.

Main Results:

Most bioassay endpoints were responsive, with stronger effects in fish and human samples.
Mixture effects in key bioassays (e.g., neurite outgrowth inhibition, androgen receptor antagonism) followed concentration addition.
Designed mixtures explained only a small fraction of total bioactivity, highlighting the role of unannotated features.
Preliminary effect-based trigger values suggest no immediate health risks at average contamination levels.

Conclusions:

Complex environmental and food chemical mixtures exhibit significant bioactivity on neurodevelopmental and reproductive pathways.
Unannotated molecular features contribute substantially to the overall mixture effects.
While current average contamination levels may not pose immediate risks, the complexity and multivalent bioactivity warrant further toxicological investigation.