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High-Throughput Miniaturized Biotransformation Testing Using Activated Sludge Enables Rapid Chemical Persistence

Sarah B Partanen¹, Nicolas Mueller¹, Kathrin Fenner^1,2

¹Swiss Federal Institute of Aquatic Science and Technology, Eawag, 8600 Dubendorf, Switzerland.

Environmental Science & Technology Letters

|November 17, 2025

Summary

Developing new chemicals requires rapid environmental persistence assessments. A new miniaturized biotransformation assay provides accurate data within 48 hours, supporting safe-by-design chemical development.

Keywords:

SSbD (Safe and Sustainable by Design)biodegradation biotransformation chemical screening environmental fate persistence assessment well plates

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Area of Science:

Environmental Chemistry
Chemical Engineering
Microbiology

Background:

Chemical development poses risks to human and ecosystem health.
Safe-by-design (SSbD) approaches and regulatory frameworks necessitate efficient chemical assessment tools.
Existing persistence tests are time-consuming and resource-intensive, hindering early-stage development.

Purpose of the Study:

To develop and validate a miniaturized, high-throughput biotransformation assay for environmental persistence assessment.
To support the principles of Safe and Sustainable by Design (SSbD) in chemical development.

Main Methods:

A miniaturized biotransformation assay was developed using municipal activated sludge.
The assay was validated using 33 pesticides and pharmaceuticals.
Biotransformation rate constants were correlated with large-volume control tests.

Main Results:

The assay demonstrated strong correlation with large-volume controls (R² > 0.84).
Consistent biotransformation rankings were observed across different conditions (Spearman correlations > 0.8).
The test requires minimal sample volume (2 mL) and provides data within 48 hours.

Conclusions:

The miniaturized assay is a rapid and reliable method for assessing chemical persistence.
This tool enables early-stage persistence assessment, directly supporting SSbD principles.
The assay's design is suitable for automation and integration into chemical design workflows.