Application of the AhR Reporter Gene Assay for the Determination of PCDD/Fs and DL-PCBs in Feed Samples.

Abstract

IntroductionPolychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (DL-PCBs) belong to a well-known group of pollutants. Present in feedstuffs, they bioaccumulate in tissues of food-producing animals. Food is the source of over 90% of human PCDD/Fs and DL-PCBs intake. Thus, feed control is one of the pillars of the EU strategy and a mean of reducing human exposure. The article presents AhR based reporter gene bioassay method for PCDD/Fs and DL-PCBs analysis in feed and its validation results.Material and MethodsAnalytes were extracted from samples with fat. Subsequently, fat and other interferences were removed from extract using sulphuric acid modified silica. Extract was further cleaned and PCDD/Fs separated from DL-PCBs using carbon column. Contaminants detection was performed using H1L6.1c3 cell line, which produces luciferase in response to AhR ligands present in extract.ResultsPerformance characteristics (repeatability, reproducibility, and apparent recovery) fulfil the requirements of Regulation 2017/771/EU. The positive correlation between bioassay and reference HRGC-HRMS method was confirmed. Moreover, the role of screening method used in connection with the confirmatory HRGC-HRMS method in providing feed and food safety has been discussed.ConclusionBioassay is a useful method for dioxin and DL-PCBs analysis, allowing cost reduction of monitoring programmes with minimal risk of false negative results.