Analytical Chemistry

BODIPY-labelled estrogens for fluorescence analysis of environmental microbial degradation

Authors

Abstract

Biodegradation of estrogen hormone micropollutants is a well-established approach towards their remediation. Fluorescently labelled substrates are used extensively for rapid, near real-time analysis of biological processes and are a potential tool for studying biodegradation processes faster and more efficiently than conventional approaches. However, it is important to understand how the fluorescently tagged surrogates compare with the natural substrate in terms of chemical analysis and the intended application. We derivatized three natural estrogens with BODIPY fluorophores by azide-alkyne cycloaddition click reaction and developed an analytical workflow based on simple liquid-liquid extraction and HPLC-PDA analysis. The developed methods allow for concurrent analysis of both fluorescent and natural estrogens with comparable recovery, accuracy, and precision. We then evaluated the use of BODIPY-labelled estrogens as surrogate substrates for studying biodegradation using a model bacterium for estrogen metabolism. The developed analytical methods were successfully employed to compare the biological transformation of 17β-estradiol (E2), with and without BODIPY fluorescent tag. Through measuring the complete degradation of E2 and the transformation of BODIPY-estradiol to BODIPY-estrone in the presence of a co-substrate, we found that BODIPY-labelled estrogens are biologically viable surrogates for investigating biodegradation in environmental bacteria.

Version notes

The introduction, results and discussion, and conclusion sections were rewritten for clarity following peer-review. The results presented are the same.

Content

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Supplementary material

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Supporting Information: BODIPY-labelled estrogens for fluorescence analysis of environmental microbial degradation
PDF document containing details of the synthesis methods (including verification results for each intermediate), the statistical analyses, specific results which are summarized in the main text, and NMR and HRMS spectra.