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Evaluation of the recombinant cell-based bioassay as a screening method for PCDD/Fs monitoring in fish tissue

Stypula-Trebas S., Warenik M., Gembal M., Malagocki P., Piskorska-Pliszczynska J.

Environmental Biotechnology

2009

Vol. 5, No. 1

32-41

Among 210 congeners only 17 highly toxic, 2,3,7,8-chlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) are of toxicological concern. They exhibit high binding affinity to an intracellular aryl hydrocarbon receptor (AhR), causing harmful effects at exposure levels of thousands of times lower than most environmental toxicants. The Chemically Activated LUciferase gene expression bioassay (CALUX) utilizes recombinant cells that were transfected with a luciferase reporter gene, which responds to dioxin-like compounds with the induction of luciferase in a time-, dose-, AhR-dependent and chemical-specific manner. The bioassay evaluation concerning the European Community (EC) requirements for the PCDD/Fs determination for the official control of foodstuffs was performed on salmon tissue. In order to evaluate the bioassay performance characteristics, recovery range, limit of detection (LOD), limit of quantification (LOQ) and precision were determined. The results revealed that combining a dichloromethane: hexane extraction, an acid silica plus activated carbon clean-up provides reliable, reproducible (CV = 9-20%) measurements with acceptable recovery (78%) and sensitivity at the required ppt range. Due to the low cost and high throughput characteristics of the CALUX assay, food monitoring for PCDD/Fs may benefit from use of this bioassay as a prescreening tool to select and prioritize samples for subsequent analysis by high resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS). Although the bioassay may not be able to specify identity of the reactive substances, it may serve as a very useful tool for the evaluation of contamination sources.

Biotest – nowe narzędzie do kompleksowej oceny stanu środowiska

Kursa-Mikołajczak M., Kuczyńska A., Wolska L.

Analityka : nauka i praktyka

2005

nr 2

13-15