Polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins and dibenzo-p-furans (PCDD/Fs), organochlorine pesticides (OCPs), polybrominated diphenyl ethers (PBDEs), hexabromocyclododecane diastereomers (HBCDs), and perfluorinated compounds (PFCs) were analyzed in popular farmed fish such as salmon, trout, tilapia, and pangasius and in farmed shrimp. The samples originated from southeast Asia, Europe, and South America. Results show the following: (i) Carnivorous species contained higher contaminant concentrations than omnivorous species. (ii) Contaminant concentrations generally decreased per species in the following order of salmon > trout >> tilapia approximately equal to pangasius approximately equal to shrimp. (iii) Most contaminant concentrations decreased in the following order of PCBs approximately equal to dichloro-diphenyl-trichloroethanes (DDTs) >> hexachlorobenzene approximately equal to pentachlorobenzene approximately equal to dieldrin approximately equal to PBDEs approximately equal to alpha-HBCD approximately equal to perfluorooctane sulfonate (PFOS) >> World Health Organization toxic equivalents (WHO-TEQ) [PCDD/Fs and dioxin-like (dl)-PCBs]. (iv) Contaminant concentrations were very low (mostly <1 ng/g wet weight) and far below the European and Dutch legislative limits. (v) Contaminant concentrations in farmed shrimp, pangasius, and tilapia were lower than those in wild fish, whereas contaminant concentrations in farmed salmon and trout were higher than those in lean wild marine fish. From the five species investigated, salmon is predominantly responsible (97%) for human exposure to the sum of the investigated contaminants. The contribution of trout, tilapia, pangasius, and shrimp is small (3%) because contaminant concentrations and consumption volumes were much lower.
There is a strong need for the development of relatively cheap and rapid bioassays for the determination of dioxins and related compounds in food. A newly developed CALUX (Chemical-Activated LUciferase gene eXpression) bioassay was tested for its possible use to determine low levels of dioxins in bovine milk. Data show that this mammalian cell-based test is very sensitive for 2,3,7,8-substituted dioxins and related PCBs, thereby reflecting the relative potencies of these compounds in comparison to TCDD (TEF-values). The limit of detection was about 50 fg of TCDD. Furthermore, the response obtained with a mixture of dioxins was additive, in accordance with the TEF-principle. Milk fat was isolated by centrifugation followed by clean-up of the fat with n-pentane, removal of the fat on a 33% H2SO4 silica column, and determination of Ah receptor agonist activity with the CALUX-bioassay. An equivalent of 67 mg fat was tested per experimental unit, resulting in a limit of quantification around 1 pg i-TEQ/g fat. To investigate the performance of the method, butter fat was cleaned and spiked with a mixture of 17 different 2,3,7,8-substituted PCDD and PCDF congeners at 1, 3, 6, 9, 12 and 15 pg TEQ/g fat, as confirmed by GC/MS. In this concentration range, the method showed a recovery of TEQs around 67% (58-87%). The reproducibility, determined in three independent series showed a CV varying between 4% and 54%, with the exception of the sample spiked at 1 pg i-TEQ (CV 97%). The repeatability determined with the sample spiked at 6 pg i-TEQ/g showed a CV of 10%. Testing of 22 bovine milk samples, taken at different sites in The Netherlands, in the CALUX-assay showed combined dioxin and dioxin-like PCB levels equivalent to 1.6 pg TCDD/g fat (range 0.2-4.6). GC/MS analysis of these samples revealed an average level of 1.7 pg i-TEQ/g fat, varying between 0.5 and 4.7 pg i-TEQ/g fat. All five samples showing a GC/MS determined dioxin content of more than 2 pg i-TEQ/g fat gave a response in the CALUX-assay corresponding to more than 2 pg TCDD/g fat. These data clearly show that the CALUX-bioassay is a promising method for the rapid and low cost screening of dioxins in bovine milk.
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