Validation and use of the CALUX-bioassay for the determination of dioxins and PCBs in bovine milk.

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.     

Dioxins,furans, and non-ortho PCBs in Canadian Arctic seabirds

This is the first account of PCDDs, PCDFs, and non-ortho PCBs in Canadian Arctic seabirds. Livers and eggs of thick-billed murres, northern fulmars, and black-legged kittiwakes were collected in 1975 and 1993 from Prince Leopold Island in Lancaster Sound, Canada. Detectable concentrations of PCDDs, PCDFs, and non-ortho PCBs were found in all the Arctic seabird samples analyzed. Of the PCDD congeners assayed, only 2,3,7,8-substituted PCDDs were detected in the samples, whereas non-2,3,7,8-substituted PCDFs were found in addition to 2,3,7,8-substituted PCDFs in some of the samples. The predominant PCDD/F congener found in the livers of all three species was 2,3,4,7,8-PnCDF, both in 1975 and 1993. Concentrations of most dioxins and furans decreased in the fulmars and kittiwakes between 1975 and 1993 but increased in the murres. Of the non-ortho PCBs measured, PCB-126 occurred in the highest concentrations and contributed the majority of the non-ortho PCB-TEQ in all three species in both years. The highest concentrations of dioxins and furans as well as the highest TEQ values were found in the northern fulmar livers in both 1975 and 1993. Concentrations of some of the PCDDs and PCDFs are among the highest reported for Canadian Arctic biota.     

Evidence for a specific pattern of polychlorinated dibenzo-p-dioxins and dibenzofurans in bivalves

The occurrence of the polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDDs/PCDFs) was studied in four different bivalve species (mussels, oysters, carpetshells, and cupped oysters) collected in three different sampling stations at the Spanish Coast from 1994 onward. The analysis revealed an unexpected isomer distribution of chlorosubstituted dioxins and furans characterized by a complex mixture of TCDF and TCDD, including both toxic and nontoxic isomers and highly chlorinated congeners, mainly OCDD. Principal component analysis revealed similarities between identical species collected from the different sites. A dioxin fingerprint could be easily distinguished between bivalve species collected in the same sampling point. The dioxin fingerprint found in filter-feeders was also compared to other marine species showing remarkable differences. Due to the habits of the bivalve species, sediment samples were also analyzed within this study. No similarities in the pattern were found for both matrices, the sediments and the bivalves. In the absence of a potential source of these toxicants affecting these species the hypothesis of a specific metabolism of the bivalves is suggested. No values higher than 4 pg WHO-TEQ/g whole weight were determined, and all samples met the current limit proposed by European Regulations. Independently of the bivalve species considered, the major contribution to the total WHO-TEQ comes from 2,3,7,8-TCDF and 2,3,4,7,8-PeCDF, at about the same levels of the 2,3,7,8-TCDD and 1,2,3,7,8-PCDD.     

Severe PCDD/F and PBDD/F pollution in air around an electronic waste dismantling area in China

The presence of chlorinated and brominated compounds in electronic waste (EW) results in the formation of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and polybrominated dibenzo-p-dioxins and dibenzofurans (PBDD/Fs) during the EW dismantling process. In this study, we investigated the dioxins present in ambient air around the EW dismantling area Guiyu in Guangdong, China. Atmospheric PCDD/F (tetra to octa) abundances and toxic equivalent (TEQ) values were 64.9-2365 pg/m3 and 0.909-48.9 pg of W-TEQ/m3, respectively; these are the highest documented values of these compounds found in ambient air in the world. PBDD/Fs (eight 2,3,7,8-substituted congeners) were also found at high pollution levels (concentrations of 8.124-61 pg/m3 and 1.6-2104 pg of I-TEQ/m3). Profiles of the 2,3,7,8-PCDD/F homologues in the air of Guiyu differed from typical urban air patterns reported in the literature, and the concentration of homologues increased with the chlorination degree of 2,3,7,8-PCDD/Fs except for OCDF. The severe dioxin pollution present in Guiyu substantially influences the adjacent area of Chendian, where atmospheric PCDD/F and 2,3,7,8-PBDD/F levels are higher than those of common urban areas in the world. Our tentative inhalation risk assessment showed that residents in Guiyu are at a high risk of exposure to dioxins. The total PCDD/F intake doses far exceed the WHO 1998 tolerable daily intake limit of 1-4 pg of W-TEQ kg(-1) day(-1).     

In vitro transactivation potencies of black-footed albatross (Phoebastria nigripes) AHR1 and AHR2 by dioxins to predict CYP1A expression in the wild population

Our previous studies have detected high levels of dioxins and related compounds (DRCs) including polychlorinated dibenzo-p-dioxins (PCDDs), furans (PCDFs), and coplanar PCBs (Co-PCBs) in the black-footed albatross (BFA), Phoebastria nigripes, from the North Pacific region. We have also cloned two aryl hydrocarbon receptors, AHR1 and AHR2, of the BFA. To evaluate the sensitivity to DRCs in the BFA and to assess the status of cytochrome P450 1A (CYP1A) induction in the wild population, this study investigated the mRNA expression levels of BFA AHR1 and AHR2 and also the transactivation potencies of each AHR by 15 selected DRC congeners. Quantitative real-time PCR of BFA AHR mRNAs showed that hepatic AHR1 is more highly expressed than AHR2. Transactivation by graded concentrations of individual DRCs was measured in COS-7 cells, where BFA AHR1 or AHR2 was transiently transfected. For congeners that exhibited AHR-mediated dose-dependent activities, 50% effective concentration (EC(50)) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) relative potencies (REPs) were estimated. Based on the estimates of the REPs, TCDD induction equivalency factors (IEFs) were determined. For BFA AHR1, PeCDF was equipotent to TCDD, but other congeners exhibited lower IEFs. For BFA AHR2, PCDD/F congeners except OCDD/F showed IEFs ≥ 1.0. Using BFA AHR1- or AHR2-IEFs and hepatic concentrations of DRCs in North Pacific BFAs, TCDD induction equivalents (IEQs) were calculated. We further constructed nonlinear regression models on the relationships between BFA AHR1- or AHR2-IEF derived total IEQ or WHO-TEF derived total TEQ and ethoxyresorufin-O-deethylase activity (EROD) in the liver of wild BFAs. The results indicated that the relationships of BFA AHR1- and AHR2-based IEQs and EROD were predictable from BFA AHR1- and AHR2-mediated transactivation by TCDD, respectively. Collectively, these results suggest that the in vitro assay incorporating the AHR of species of concern would be a useful tool to predict the sensitivity to DRCs in the species and CYP1A induction in the wild population.     

Comparison of five in vitro digestion models to study the bioaccessibility of soil contaminants

Soil ingestion can be a major exposure route for humans to many immobile soil contaminants. Exposure to soil contaminants can be overestimated if oral bioavailability is not taken into account. Several in vitro digestion models simulating the human gastrointestinal tract have been developed to assess mobilization of contaminants from soil during digestion, i.e., bioaccessibility. Bioaccessibility is a crucial step in controlling the oral bioavailability for soil contaminants. To what extent in vitro determination of bioaccessibility is method dependent has, until now, not been studied. This paper describes a multi-laboratory comparison and evaluation of five in vitro digestion models. Their experimental design and the results of a round robin evaluation of three soils, each contaminated with arsenic, cadmium, and lead, are presented and discussed. A wide range of bioaccessibility values were found for the three soils: for As 6-95%, 1-19%, and 10-59%; for Cd 7-92%, 5-92%, and 6-99%; and for Pb 4-91%, 1-56%, and 3-90%. Bioaccessibility in many cases is less than 50%, indicating that a reduction of bioavailability can have implications for health risk assessment. Although the experimental designs of the different digestion systems are distinct, the main differences in test results of bioaccessibility can be explained on the basis of the applied gastric pH. High values are typically observed for a simple gastric method, which measures bioaccessibility in the gastric compartment at low pHs of 1.5. Other methods that also apply a low gastric pH, and include intestinal conditions, produce lower bioaccessibility values. The lowest bioaccessibility values are observed for a gastrointestinal method which employs a high gastric pH of 4.0.     

Polychlorinated naphthalenes, -biphenyls, -dibenzo-p-dioxins, and -dibenzofurans in double-crested cormorants and herring gulls from Michigan waters of the Great Lakes

Concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), naphthalenes (PCNs), and biphenyls (PCBs) were measured in eggs of double-crested cormorants and herring gulls collected from Michigan waters of the Great Lakes. Concentrations of PCNs in eggs of double-crested cormorants and herring gulls were in the ranges of 380-2400 and 83-1300 pg/g, wet wt, respectively. Concentrations of 2,3,7,8-substituted PCDDs and PCDFs were 10-200 times less than those of PCNs in eggs whereas those of total PCBs (380-7900 ng/g, wet wt) were 3-4 orders of magnitude greater. While the profile of PCB isomers and congeners between double-crested cormorants and herring gulls was similar, the PCN isomer profile differed markedly between these two species. PCN congeners 66/67 (1,2,3,4,6,7/1,2,3,5,6,7) accounted for greater than 90% of the total PCN concentrations in herring gulls, whereas their contribution to total PCN concentrations in double-crested cormorants ranged from 18 to 40% (mean, 31%). The ratios of concentrations of PCDDs to PCDFs were greater in herring gulls than in double-crested cormorants collected from the same locations, suggesting the ability of the former to metabolize PCDF congeners relatively rapidly. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) equivalents (TEQs) contributed by PCNs in double-crested cormorant and herring gull eggs were 2-3% of the sum TEQs of PCBs, PCDDs, PCDFs, and PCNs. PCB congener 126 (3,3',4,4',5-PeCB) accounted for 57-72% of the total TEQs in double-crested cormorant and herring gull eggs.     

Characterization and inventory of PCDD/F emissions from the ceramic industry in China

The ceramic industry is considered to be a potential source of dioxins (polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), considering the widespread distribution of dioxins in kaolinitic clays. Nevertheless, studies on the emission of dioxins from the ceramic industry are still very scarce. In this study, raw clays and stack gases from six typical ceramic plants in China were collected and analyzed to estimate the emission of dioxins from the ceramic industry. Dioxin profiles in raw clays were characterized by the domination of the congener octachlorodibenzo-p-dioxin (OCDD), and the contents of other congeners declined with the decreasing degree of chlorination. During the ceramic firing process, a considerable amount (16.5-25.1 wt % of the initial quantity in raw clays) of the dioxins was not destroyed and was released to the atmosphere. Dechlorination of OCDD generated a broad distribution within the PCDD congeners including a variety of non-2,3,7,8-substituted ones with the mass abundance of 0.4-3.6%. Based on the mean concentrations measured in this study, the inventory of PCDD/Fs from the manufacturing of ceramics on the Chinese scale was estimated to be 7.94 kg/year; the corresponding value on the I-TEQ basis is 133.6 g I-TEQ/year. This accounts for about 1.34% (I-TEQ basis) of the total emission of dioxins to the environment in China. The results suggest that the ceramic industry is a significant source of dioxins in the environment.     

Analytical methods for the determination of polychlorinated dibenzodioxins and dibenzofurans in human milk

A method is described for the determination of polychlorinated dibenzodioxins and dibenzofurans in breast milk. After addition of potassium oxalate, the compounds are extracted, along with fat and other lipophilic compounds, using ethanol, ethyl ether and pentane. An aliquot of the fat is fortified with seven 13C-labelled dioxins and furans. Gel permeation chromatography on Bio-Beads S-X3 is used for removal of fat followed by column chromatography on florisil, charcoal and acid alumina. Except for 2,3,7,8-tetrachlorodibenzodioxin (2,3,7,8-TCDD), all other congeners are determined by combined capillary gas chromatography/mass spectrometry by use of negative chemical ionization employing methane as reagent gas. MS analysis is performed in the SIM mode (selected ion monitoring). For each isomer group two ions are monitored. Octachloronaphthalene is used as a syringe spike for the quantification based on a calibration run of standards. The calibration mixture, which contains the spike as well as all 2,3,7,8-chlorine substituted dioxins and furans and the 13C-labelled surrogates at a concentration level of 3-5 pg/microliters, is injected twice every day. For the determination of 2,3,7,8-TCDD a mass selective detector is used operating in electron impact mode. The MS analysis is also performed in SIM mode monitoring two ions for TCDDs and one ion for 13C-2,3,7,8-TCDD. Dependent on the level of each congener in breast milk, the coefficient of variation of the method varies between 3% for 2,3,4,7,8-pentachlorodibenzofuran and 40.9% for octachlorodibenzofuran. More than 200 human milk samples, mostly from nursing mothers living in North Rhine-Westphalia, have been analysed for residues of polychlorinated dibenzodioxins and polychlorodibenzofurans.(ABSTRACT TRUNCATED AT 250 WORDS)     

Polychlorinated dibenzo-p-dioxin and dibenzofuran concentration profiles in sediments and flood-plain soils of the Tittabawassee River,Michigan

Concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in sediments and flood-plain soils collected along the Tittabawassee River in Michigan ranged from 102 to 53,600 pg/g, dry wt. Mean PCDD/PCDF concentrations in downstream sediment and soil were from 10- to 20-fold greater than those found at locations upstream of Midland, Michigan. Concentrations of PCDD/PCDF in sediments and flood-plain soils from the Tittabawassee watershed were comparable to those found in industrialized areas such as the Housatonic and lower Passaic Rivers in the U.S. Concentrations of PCDDs/PCDFs in soil and sediment were not correlated with total organic carbon (TOC) in sediments or soils. OCDD and 2,3,7,8-TeCDF were the predominant congeners in sediment/soil collected from locations downstream of Midland, Michigan. Principal component analysis of the PCDD/PCDF congener profile suggested the presence of sources originating from a mixture of chlorophenol and other chlorinated compound production. Mass balance analysis of TCDD equivalents (TCDD-EQs) derived from H4IIE-luc bioassay of sediment extracts and 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TEQs) estimated from instrumental analysis suggested that PCDDs/PCDFs were the major dioxin-like compounds present in sediments. A significant correlation existed between bioassay-derived TCDD-EQs and instrumentally measured TEQs (r2 = 0.94).     

Characterization of trophic transfer for polychlorinated dibenzo-p-dioxins, dibenzofurans, non- and mono-ortho polychlorinated biphenyls in the marine food web of Bohai Bay, North China

Many investigations have highlighted the bioaccumulation of dioxins in animals, but little is known about the trophodynamics of dioxins in the food web. In this study, the trophic transfer of nine dibenzo-p-dioxin (PCDD) congeners, eleven dibenzofuran (PCDF) congeners, and twelve non-, mono-ortho polychlorinated biphenyl (non- and mono-ortho PCBs) congeners in a marine food web were determined. The concentrations of PCDDs, PCDFs, non- and mono-ortho PCBs were analyzed in phytoplankton/ seston, zooplankton, three invertebrate species, six fish species, and one seabirds species collected from Bohai Bay, representing approximately 4 trophic levels based on stable nitrogen isotope values. Positive relationships were found between trophic levels and lipid equivalent concentrations of non- and mono-ortho PCBs except for PCB-77, PCB-81, PCB-126, PCB-156, and PCB-167, indicating bioaccumulation of these compounds in this food web. But lipid equivalent concentrations of low chlorinated 2,3,7,8-substituted-PCDD/Fs did not exhibit statistically significant trends with trophic levels. And lipid equivalent concentrations of high chlorinated 2,3,7,8-substituted-PCDD/Fs and three non-2,3,7,8-substituted-PCDD/Fs declined significantly with increasing trophic levels providing that these isomers undergo trophic dilution. The similarity in log Kow values for non-, mono-ortho PCBs, non-2,3,7,8-substituted-PCDD/Fs, and some 2,3,7,8-substituted-PCDD/Fs suggests that the difference of trophic transfer is mainly due to their different metabolic transformation rates.     

Predicting toxicity equivalents in fish tissue

Toxicity equivalents is a measure of "dioxin-like" toxicity contributed by polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (dioxins), and polychlorinated dibenzofurans (furans). Calculation of toxicity equivalents require the use of analytical procedures that are expensive and slow, making them impractical for routine analysis. U.S. Environmental Protection Agency National Fish Tissue Study (2003) data were used to determine the most significant predictors of toxicity equivalents with multiple regression analysis. The strongest predictive model (P < 0.0001, R2 = 0.97) included five compounds (PCB-118; PCB-126; 2,3,7,8-TCDD; 1,2,3,6,7,8-HxCDD; 2,3,4,7,8-PeCDF). However, the required lower limit of detection for an analytical method measuring these congeners is 0.1 ppt and would not provide much benefit over the current analytical method. An alternative model (P < 0.0001, R2 = 0.68) that included three PCBs (PCB-138, PCB-153, PCB-118) would require a limit of detection of 1,000 ppt and be more practical. This research demonstrates that the measurement of selected compounds can be used to estimate toxicity equivalents and consequently serve as the impetus for the development of lower cost, rapid analytical methods for analysis of fish.     

Levels and trends of PCDD/Fs and DL-PCBs in Polish animal feeds, 2004–2017

Feed control is essential for the safety of animal-origin food. It is estimated that more than 80% of human exposure to dioxins and related compounds comes from the consumption of animal-origin food, and farm animals are exposed to dioxins mainly through the feed. A monitoring programme for dioxins (polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs)) and dioxin-like polychlorinated biphenyls (DL-PCBs) was conducted in the Polish feed market between 2004 and 2017. Using two complementary ISO 17025-accredited analytical methods (bioassay and high-resolution gas chromatography/high-resolution mass spectrometry (HRGC/HRMS)), 2919 samples of plant, animal and minerals origin were analysed. The total number of samples exceeding the maximum limit (ML) was 52, which is 1.8% of all tested samples. They all contained PCDD/Fs in concentrations over the ML, and in 16 samples the limit for the sum of PCDD/F/DL-PCBs was also exceeded. The highest dioxins level was found in feedstuffs derived from Baltic fish; the fish oil was followed by fish meal. Low-chlorinated furans (2.3,7,8 TCDF, 2,3,4,7,8-PCDF and 1,2,3,7,8-PeCDF) were dominant and these congeners accounted for the toxicity expressed in WHO toxic equivalents (WHO-TEQ). In this category, 155 out of the 498 samples (32%) exceeded the action threshold (AT) for PCDD/Fs and in 10 samples (2.0%) for DL-PCBs. Non-compliant fish meals were found in 34 (5.6%) of the tested samples, in 0.7% of compound feeds, and in 1.5% of plant-origin materials. Seven dried plant-derived materials contained PCDD/F level above ML, including alfalfa, maize, apple, basil and beet pulp pellets containing molasses. The dry vegetable materials were mostly contaminated by the 2,3,7,8-furans, the congeners arising from a faulty drying process. No significant time trend can be seen. From the obtained results, it can be concluded that concentrations of PCDD/Fs and DL-PCBs in Polish feed materials were sufficiently low to ensure low concentrations in animal-derived products.     

Distribution, characteristics, and worldwide inventory of dioxins in kaolin ball clays

Distribution, characteristics, and global inventory of dioxins (polychlorinated dibenzo-p-dioxins [PCDDs] and dibenzofurans [PCDFs] and dioxin like polychlorinated biphenyls) in kaolin clays collected from 10 countries were investigated. Dioxins were found in all kaolin clay samples analyzed, at total concentrations ranging from 1.2 pg/g (Brazil) to 520,000 pg/g (USA). Dioxin concentrations in kaolin clays from a few countries (e.g., Brazil and UK) were lower than those reported for background soils in Japan. Dioxin profiles in kaolin clays were characterized by the domination of the congener octachlorodibenzo-p-dioxin (OCDD), and the concentrations of other congeners decreased in the order of reduction in the levels of chlorination. Furthermore, specific distribution of congeners, with predominant proportions of 1,4,6,9-substituted PCDDs within each homologue group, was found in most clay samples. The ratios of concentrations of PCDD to PCDF and 1,2,3,7,8,9-HxCDD to 1,2,3,6,7,8-HxCDD indicated differences in the profiles found for anthropogenic sources (including pentachlorophenol) and kaolin clays. Concentrations of PCDD/Fs in kaolin clays, except for American ball clays, did not exceed the environmental criteria set by the Law Concerning Special Measures against Dioxins in Japan. Based on the average concentrations measured in our study, inventories of PCDD/Fs from the production/usage of ball clays on a global scale were estimated to be 650 kg/yr; the corresponding value on a TEQ basis is 2400 g-TEQ/yr. More than 480 kg of OCDD is estimated to be released annually from the production of kaolin clays worldwide, suggesting that kaolin clays can be a major contributor for additional source of dioxins, especially OCDD, in the environment.     

Open dumping site in Asian developing countries: a potential source of polychlorinated dibenz-p-dioxins and polychlorinated dibenzofurans

Open landfill dumping areas for municipal wastes in Asian developing countries have recently received particular attention with regard to environmental pollution problems. Because of the uncontrolled burning of solid wastes, elevated contamination by various toxic chemicals including dioxins and related compounds in these dumping sites has been anticipated. In this study, concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and coplanar polychlorinated biphenyls (PCBs) were determined in soils from dumping sites in the Philippines, Cambodia, India, and Vietnam. Residue concentrations of PCDD/Fs and coplanar PCBs in dumping site soils were apparently greater than those in soils collected in agricultural or urban areas far from dumping sites, suggesting that dumping sites are potential sources of PCDD/Fs and related compounds. Observed PCDD/F concentrations in soils from dumping sites in the Philippines and Cambodia were comparable or higher than those reported for dioxin-contaminated locations in the world (e.g., near the municipal waste incinerators and open landfill dumping sites). Homologue profiles of PCDD/Fs in dumping site soils from the Philippines and, to a lesser extent, from Cambodia and India reflected patterns of samples representing typical emissions, while profiles of agricultural or urban soils were similar to those of typical environmental sinks. This result suggests recent formation of PCDD/Fs in dumping site areas and that open dumping sites are a potential source of dioxins in Asian developing countries. Uncontrolled combustions of solid wastes by waste pickers, generation of methane gas, and low-temperature burning can be major factors for the formation of dioxins in dumping sites. Elevated fluxes of PCDD/Fs to soils in dumping sites were encountered in the Philippines, Cambodia, India, and Vietnam-Hanoi, and these levels were higher than those reported for other countries. Considerable loading rates of PCDD/Fs in the dumping sites of these countries were observed, ranging from 20 to 3900 mg/yr (0.12-35 mg TEQ/yr). PCDD/F concentrations in some soil samples from the Philippines, Cambodia, India, and Vietnam-Hanoi exceeded environmental guideline values, suggesting potential health effects on humans and wildlife living near these dumping sites. The estimated intakes of dioxins via soil ingestion and dermal exposure for children were higher than those for adults, suggesting greater risk of dioxin exposure for children in dumping sites. To our knowledge, this is the first comprehensive study on PCDD/Fs contamination in open dumping sites of Asian developing countries. On the basis of the result of this study, we have addressed a new environmental issue that open dumping sites are potential sources of PCDD/Fs and related compounds, and dioxin contamination in dumping sites may become a key environmental problem in developing countries.     

Determination of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in 2,4,5-trichlorophenoxyacetic acid, 2,4,5-trichlorophenoxyacetic acid-alkylesters and in the corresponding herbicide formulations (author's transl)]

A method is described for the determination of 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) in 2,4,5-Trichlorophenoxyacetic Acid (2,4,5-T), 2,4,5-T-Akylesters and commercial herbicide formulations containing these substances. TCDD is extracted from alkaline solution with n-hexane and purified by column chromatography on silica gel and Florisil. TCDD is determined by gas-liquid chromatography with EC-detection. For TCDD-additions of 20 micron/kg to herbicide formulations recoveries of dioxin are 96--104%. The detection limit is about 5 microgram/kg 2,4,5-T.     

Dioxin and furan formation in FBC boilers

Fluidized bed combustion (FBC) is increasingly being used for municipal solid waste incineration and for various applications in which dioxin and furan (PCDD/F) formation are concerns. Despite the fact that fluidized bed combustion is a relatively low temperature system, current evidence shows that they perform relatively well. One hypothesis is that sulfur, which is contained in some of the fuels used in FBC (e.g. coal in cofiring situations), helps to reduce dioxin production. This paper endeavors to examine this speculation and finds that such benefits depend on the CI/S ratio and that at higher sulfur addition levels dioxins, furans, and polycyclic aromatic hydrocarbon (PAH) production may actually increase. This and the reasons for it are discussed.     

Food risk characterization from exposure to persistent organic pollutants and metals contaminating eels from an Italian lake

Monitoring environmental contaminants in edible animal species is important for the assessment of environmental quality and for food safety as well. In this study, the occurrence of 2,3,7,8 dioxins, and furans, dioxin-like polychlorinated biphenyls, six non-dioxin-like polychlorinated biphenyl indicators (ndl-PCBs), seven polybrominated diphenylethers (PBDEs), four polycyclic aromatic hydrocarbons (PAHs) and four metals (As, Hg, Ni, Pb) was evaluated in 90 European eels (Anguilla anguilla) from Lake Garda. The results on eel samples showed a frequent contamination of dl-PCBs, found at concentrations higher than the maximum limit (ML) provided by European Union in 41 % of the eels. Two PBDE congeners (BDE 47 and BDE 99) were detected in almost all the samples. The contamination by PAHs was found negligible; only one sample had a high concentration of benzo[a]pyrene. Concerning metals, Pb showed concentrations higher than the maximum level (ML) in 77% of the samples, while Ni content could be a cause of acute toxicity due to dermatitis. Extreme consumers (95th percentile) are subjected to an actual risk attributable to dioxins, furans, and mainly dl-PCBs and PBDEs. The contamination of Lake Garda eels is therefore a matter of concern and a constant monitoring of the pollution levels of its freshwater is mandatory.     

Determination of brominated flame retardants and brominated dioxins in fish collected from three regions of Japan

The concentrations of brominated dioxins which are polybrominated dibenzo-p-dioxins/polybrominated dibenzofurans (PBDD/DFs) and mono-bromo polychlorinated dibenzo-p-dioxins/dibenzofurans, polybrominated diphenyl ethers (PBDEs) and tetrabromobisphenol A (TBBPA) were investigated in a total of 45 fish samples collected from three regions in Japan. In the brominated dioxins, 1,2,3,4,6,7,8-heptabromodibenzofuran (HpBDF) was the most abundant congener, and it was found in seven fish samples at 0.10-25.6 pg/g wet weight (ww). The highest concentration of 1,2,3,4,6,7,8-HpBDF was found in the pike eel. Regarding other congeners, 2,3,7,8-tetrabromodibenzo-p-dioxin was detected in the sea bream at 0.02 pg/g ww, and 2,3,7,8-tetrabromodibenzofuran was detected in the conger eel at 0.03 pg/g ww. 3-Bromo-2,7,8-trichlorodibenzofuran was detected in the Sardinella zunasi and the conger eel at 0.01 pg/g ww and 0.02 pg/g ww, respectively. Using toxic equivalency factors of chlorinated dioxins, we calculated the PBDD/DFs concentrations of these fish samples at 0.001-0.256 pg TEQ/g ww. PBDEs were detected in all of the fish samples. The concentrations of total PBDEs were 0.01-2.88 ng/g ww. The seerfish and the yellowtail containd PBDEs in high concentrations. The most dominant congener in most of the fish was 2,2',4,4'-tetrabromo diphenyl ether. TBBPA was detected in 29 fish samples at 0.01-0.11 ng/g ww. The mean level of TBBPA was about one-tenth or less of the total level of PBDEs. A good correlation was obtained between total PBDEs and fat content. On the other hand, no correlation was obtained between TBBPA and fat content. The daily intakes from fish were estimated to be 0.58 ng/kg body weight (bw)/day for total PBDEs, 0.03 ng/kg bw/day for TBBPA, and 0.01 pg TEQ/kg bw/day for brominated dioxins in the case assuming that the average bw of a Japanese adult person is 50 kg and that the average fish consumption is 82 g/day. For PBDEs, the provisionally calculated value was much less than the lowest observed adverse effect level value (1 mg/kg bw/day). For brominated dioxins, the daily intake was at a very low level compared with the Japanese daily intake of polychlorinated dioxins from fish. Even if the value of PBDD/DFs is added to the amount of chlorinated dioxin exposure, it was estimated that it is less than the tolerable daily intake (4 pg TEQ/kg bw/day) in Japan.     

Dioxin concentrations in breast milk of Vietnamese nursing mothers: a survey four decades after the herbicide spraying

In an operation by United States Armed Forces during 1961 to 1971, large quantities of herbicides were sprayed in South Vietnam. These herbicides contained 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-tetraCDD), the most toxic congener of dioxins. Several decades after the herbicide spraying ceased, dioxin concentrations in the environment and human remained elevated in the sprayed areas. Breast milk samples from 520 nursing mothers residing in areas including the hot spots as well as the sprayed and unsprayed areas were collected to quantify the levels of dioxins. The total toxic equivalents of 2,3,7,8-substitued PCDDs/PCDFs in breast milk of mothers living in the hot spots, and the sprayed and unsprayed areas were 14.10 pg/g lipid, 10.89 pg/g lipid, and 4.09 pg/g lipid for primiparae and 11.48 pg/g lipid, 7.56 pg/g lipid, and 2.84 pg/g lipid for multiparae, respectively, with significant differences in the values among the three areas. In the hot spots, dioxin levels were highly correlated with the residency of mothers after adjustment for their age and parity.