Dietary intake of PCDD/Fs and dioxin-like PCBs of the Belgian population.

The World Health Organization recommends assessing human exposure to contaminants on a regular basis. In order to assess the current dietary exposure of the Belgian adult population to PCDD/Fs and dioxin-like PCBs and to update exposure estimates of 2000-2001, a total diet study was designed. The mean dietary intake of PCDD/Fs and dioxin-like PCBs in the Belgian adult population in 2008 was estimated to be 0.72pgTEQkgbw(-1)d(-1) (middle bound concentrations, TEF of 1998) based on occurrence data of 2008 and national food consumption data of 2004. This value is clearly below the Tolerable Weekly Intake (TWI) of 14pgTEQkgbw(-1)week(-1) set by the Scientific Committee on Food of the European Commission and below the provisional tolerable monthly intake of 70pgTEQkgbw(-1)month(-1) set by the Joint FAO/WHO Expert Committee on Food Additives. Considering the cumulative distribution, the intake was less than 1pgTEQkgbw(-1)d(-1) for more than 80% of the population, and less than 2pgTEQkgbw(-1)d(-1) for the entire population. When using the 2005 TEF instead of the 1998 TEF, the mean dietary intake in the Belgian adult population was estimated to be 0.61pgTEQkgbw(-1)d(-1).

PCB and organochlorine pesticides in home-produced eggs in Belgium.

The level of polychlorinated biphenyls (PCB) and persistent organochlorinated pesticides (OC) in home-produced eggs was investigated in Belgium. The concentration of dichlorodiphenytrichloroethane (DDT) is above the norm for 17% of the eggs collected during the spring on 58 different locations. For PCB, aldrin, dieldrin, and chlordane, 3-5% of the samples are above the norm too. These levels are surprisingly high for compounds banned for about 30 years. Higher concentrations in home-produced eggs are expected compared to battery eggs because of contact with the environment and especially the soil. For ten selected locations, the concentration in soils, excreta and feed was measured, but no simple correlation between egg and feed or soil level could be established. Hexachlorohexane, endosulfan, endrin, methoxychlor and nitrofen were not detected in any sample.

Assessment of the chemical contamination in home-produced eggs in Belgium: general overview of the CONTEGG study.

This overview paper describes a study conducted for the Belgian Federal Public Service of Health, Food Chain Safety and Environment during 2006-2007. Home-produced eggs from Belgian private owners of hens were included in a large study aiming to determine concentration levels of various environmental contaminants. By means of the analyses of soil samples and of kitchen waste samples, obtained from the same locations, an investigation towards the possible sources of contaminants was possible. Eggs, soils, faeces and kitchen waste samples were checked for the presence of dioxins, PCBs (including dioxin-like PCBs), organochlorine pesticides, trace elements, PAHs, brominated flame retardants and mycotoxins. The study design, sampling methodology and primary conclusions of the study are given. It was found that in some cases dioxin-like compounds were present at levels that are of concern for the health of the egg consumers. Therefore, measures to limit their contamination in eggs, produced by hens of private owners, were proposed and deserve further attention.

Validation of the analysis of the 15+1 European-priority polycyclic aromatic hydrocarbons by donnor-acceptor complex chromatography and high-performance liquid chromatography-ultraviolet/fluorescence detection.

The European Commission recommends to monitor the 16 polycyclic aromatic hydrocarbons (PAHs) possessing both genotoxic and carcinogenic properties. Since robust analytical methods specific for this set of European PAH are lacking, a new method for their analysis in food is proposed. The donnor-acceptor complex chromatography (DACC) is used as clean-up step and high-performance liquid chromatography-ultraviolet/fluorescence detection (HPLC-UV/FLD) is used for detection and quantification. The method has been validated for analysis of PAH in oil and in dried plants and bears very good results for all compounds.

PCBs and PCDD/FS in fish and fish products and their impact on the human body burden in Belgium.

The concentrations of marker PCBs (28, 52, 101, 118, 138, 153, 180) in fish have been assessed with GC-MS: an average concentration of 540 ng-PCB g(-1) fat (5.02 ng-PCB g(-1) wet weight) was observed. The average concentration of PCDD/Fs, assessed with the CALUX bioassay, amounted to 64 pg-CALUX-TEQ g(-1) fat (0.58 pg-CALUX-TEQ g(-1) wet weight) and that of PCDD/Fs + dioxin-like PCBs amounted to 131 pg-CALUX-TEQ g(-1) fat (1.18 pg-CALUX-TEQ g(-1) wet weight). Results of the PCB congeners analyses show that PCB-153 is the most abundant congener in almost all samples, with also main contributions of the 138- and 180-congeners. For some species such as the sand sole and lemon sole, a fairly constant PCB content, independent of the fat percentage, was observed. For a second group of species such as whelks, cod, and whiting, a positive correlation was observed between their PCB concentration (ng g(-1) fat) and their % of fat. The relationship between marker PCBs and PCDD/Fs concentrations, when plotted on a log scale, fits a straight line (correlation coefficient r = 0.83). With our results on fish and literature data for other food products, intake of marker PCBs and PCDD/Fs could be calculated for the adult population in Belgium (19-60 years old). The Total Daily Intake (TDI) of marker PCBs (ng-PCB day(-1)) ranges between 1690 and 2210. The TDI of PCDD/Fs (pg-CALUX day(-1)) ranges between 80.5 and 122, that of PCDD/Fs + dioxin-like PCBs amounts to 151. When PCDD/Fs in fish are assessed with GC-HRMS, the TDI can be lower. The relative importance of fish regarding marker PCB intake amounts to 15-19%, while regarding PCDD/Fs intake it amounts to 34-51%. Using TDI, the body burden evolution of marker PCBs and PCDD/Fs, with age has been calculated.

Validation of the CALUX bioassay for PCDD/F analyses in human blood plasma and comparison with GC-HRMS.

Following the dioxin crisis of 1999, several studies were conducted to assess the impact of this crisis on the dioxin body burden in the Belgian population. The Scientific Institute of Public Health identified a population from whom plasma samples were available and from whom, during the follow up survey, plasma samples were obtained in 2000. In total, 496 samples were collected for GC-HRMS and CALUX analyses to verify statistical assessment conclusions. This study was seen as an opportunity to validate the CALUX bioassay for biological sample analysis and to compare toxic equivalency (TEQ) values obtained by the reference GC-HRMS technique and by the screening method. This article focuses on the validation results of the CALUX bioassay for the analyses of the dioxin fractions of blood plasma. The sample preparation is based on a liquid-liquid extraction, followed by an acid silica in series with an activated carbon clean-up. A good recovery (82%) and reproducibility (coefficient of variation less than 25%) were found for this method. Based on 341 plasma samples, a significant correlation was established between the bioassay and chemical method (R = 0.64). However, a proportional systematic error was observed when the results obtained with the CALUX bioassay were regressed with the results from the GC-HRMS analyses. The limit of quantification (LOQ) used to calculate TEQ values from the GC-HRMS determinations, the use of the relative potency values instead of the toxic equivalent factor and the potential of CALUX bioassay to measure all compounds with affinity for the AhR may partly explain this proportional systematic error. Nevertheless, the present results suggest that the CALUX bioassay could be a promising valid screening method for human blood plasma analyses.

Importance of clean-up for comparison of TEQ-values obtained by CALUX and chemo-analysis.

This paper presents Chemically Activated LUciferine gene eXpression (CALUX) TEQ-values obtained for nine plasma samples following two different purification procedures, one of them involving fractionation. CALUX results obtained for the dioxin (DX) and dioxin + PCB (DX + PCB) fractions were then compared to the GC-HRMS TEQ-values calculated for the 17 polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (17 PCDD/F) and 17 PCDD/F + 4 cPCB congeners, respectively. The overestimation of the CALUX (DX fraction) TEQ-values in comparison with the chemo-analyses of the 17 PCDD/F is mainly explained by the presence of other AhR agonists, like brominated compounds. Otherwise, the constancy of the CALUX (DX + PCB fraction) TEQ-value which compares to increasing the GC-HRMS (17 PCDD/F + 4 cPCB) TEQ results raises questions concerning (1) the significance of CALUX results obtained without fractionation as well as (2) the toxicological effect of a cocktail of contaminants on the human health.

TEQ-value determinations of animal feed; emphasis on the CALUX bioassay validation.

Polyhalogenated aromatic hydrocarbons, such as polychlorinated dibenzo-p-dioxins are a large and diverse group of environmental pollutants. Their tendency to accumulate in the food chain and their toxicity make monitoring necessary. The reference analysis method is laborious and very expensive, therefore cheap and rapid bioassays have been developed. The chemical-activated luciferase bioassay (CALUX) bioassay uses a recombinant cell line, which responds to dioxins and dioxin-like molecules with Ah receptor (AhR)-dependent induction of firefly luciferase in a dose related response. The CALUX was tested for its use in the screening of feed. Aliquots of 20g of enriched feed were extracted with a toluene:methanol mixture (20:4 v/v) and extracts were defatted on 33% H(2)SO(4) silica columns and purified on carbon columns. Only the dioxin and furan fraction was analysed, the PCB fraction was discarded. The precision of the method is acceptable and in compliance with an R.S.D. <30% as suggested for cell-based bioassays in the Commission Directive 2002/70/EC of July 2002. The results evidence good agreement between TEQ-values obtained by either CALUX or GC-HRMS. The method is now routinely in use for a feed screening programme designed by the Federal Agency for the Safety of the Food chain. Approximately, 25 samples are analysed weekly. From the obtained results approximately 10% was confirmed by GC-HRMS. The false positive ratio is 1% and no false negatives were found, making the use of the CALUX technology advantageous.

Supercritical fluid extraction and accelerated solvent extraction of dioxins from high- and low-carbon fly ash

This study investigates the replacement of Soxhlet extraction by supercritical fluid extraction (SFE) or accelerated solvent extraction (ASE) for the removal of dioxins from municipal waste incinerator fly ash. SFE is very matrix dependent; higher percent recoveries versus Soxhlet extraction can be obtained for low-carbon-level fly ash, but only a few percent of dioxins can be extracted from high-carbon-level fly ash. The addition of large quantities of toluene in the extraction cell prior to extraction of high-carbon fly ash improves the recovery of the lowest chlorinated dioxins (approximately 90%), but a maximum of 20% of the octachlorodibenzo-p-dioxins can be extracted. Since large quantities of toluene are needed to improve the recoveries, ASE with toluene was tested. Recoveries similar to Soxhlet extraction can be obtained in 2 h at 80 degrees C. Increasing the temperature to 150 degrees C increases the extraction rate and yields recoveries of approximately 110-160% compared to 48-h Soxhlet extraction for all congeners for both low- and high-carbon fly ashes. These results question the choice of Soxhlet extraction as a reference method for dioxin determination.