First WHO/UNEP survey of the current concentrations of persistent organic pollutants in human milk in Morocco.

The presence of persistent organic pollutants (POPs) in human milk is an indicator of the level of these substances in the environment and enables evaluation of the potential exposure of breastfed children. POPs concentrations in a Moroccan human milk pooled sample (59 donors) were determined for the first time by the WHO reference laboratory to provide a baseline and allow monitoring of the future trends for those persistent organohalogen compounds. Eighty-one different POPs, including organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), and dioxins and furans (PCDD/Fs) were identified and quantified by using gas chromatography (GC) with an electron capture detector (ECD) and confirmed by gas chromatography-mass spectrometry (GC/LRMS). The obtained results showed that the sum of DDT (sum of o,p'-DDD, p,p'-DDD, o,p'-DDE, p,p'-DDE, o,p'-DDT, and p,p'-DDT) was 237.9 ng/g, representing 94.0% of all detected OCPs levels, with a dominance of p,p'-DDE, and p,p'-DDT. Cis-heptachlor epoxide, HCB, and HCH were found at much lower levels. PCB indicator level was 60.7 ng/g of lipid, with the dominance of PCB 138, PCB 153, and PCB 180, presenting 98.3% of the total. Among the 25 PBDE congeners quantified, BDE-47, BDE-153, BDE-197, and BDE-207 were the dominant congeners account for 15.4% of the total concentration of PBDE (1.3 pg/g lipid). The lipid-adjusted level of PCDD/Fs and dl-PCBs expressed in Toxic Equivalents (TEQs: 6.65 pg/g) were lower in comparison to those found in the other countries. In summary, detected POPs levels in Moroccan human milk were much lower compared to the other countries, reflecting the effectiveness of the compliance with Stockholm Convention requirements on eliminating or reducing emissions of selected POPs. Continued monitoring is needed for these compounds, for which this is the first data available, provided by the WHO/UNEP survey in Morocco.

Chlorinated Paraffin Levels in Relation to Other Persistent Organic Pollutants Found in Pooled Human Milk Samples from Primiparous Mothers in 53 Countries.

BACKGROUND: The current production and use of chlorinated paraffins (CPs) at >1 million tons/y likely exceeds the lifetime production of polychlorinated biphenyls (PCBs). These persistent organic pollutants (POPs) are a concern to human health. OBJECTIVES: The United Nations Environment Programme conducts global surveys of human milk samples from individual countries as a noninvasive method of investigating levels and trends in human exposures to POPs such as CPs. We measured CP concentrations and assessed their relation to other POPs in pooled samples collected during 2012-2019. METHODS: We analyzed 57 official nationwide pooled milk samples from 53 countries on five continents (Africa, Central/South America, Asia, Europe, and Australia/Oceania). CP concentrations were further characterized by subgroups and compared with concentrations of 19 other POPs, including PCBs and a variety of pesticides. RESULTS: CPs were detected in pooled samples from all 53 countries, with concentrations of 23-700 ng/g lipid. CPs accounted for 18-46% of the total summed POPs in human milk, second only to dichlorodiphenyltrichloroethane (DDT). CP concentrations exceeded PCB concentrations in pooled samples from most countries. DISCUSSION: The presence of CPs in all samples, including samples from isolated locations (e.g., Pacific Island countries), emphasizes the ubiquitous presence of these compounds, whereas differences in subgroup ratios indicate a delay in the shift toward nonregulated medium-chain CPs (MCCPs) for these regions. The predominance of MCCPs in samples from many countries suggests a need for regulation and research on health effects. https://doi.org/10.1289/EHP7696.

Congener patterns of polychlorinated dibenzo-p-dioxins, dibenzofurans and biphenyls as a useful aid to source identification during a contamination incident in the food chain.

Polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and biphenyls (PCBs) are still considered among the most important groups of contaminants in the food chain. Self-control by food producers and official control by authorities are important activities that allow contaminant sources to be traced and promote further reduction in food and feed levels. Strict but feasible maximum levels were set by the EU Commission for food and feed to support this strategy, as well as action levels and thresholds. When products exceed these levels, it is important to trace the source of contamination and take measures to remove it. Congener patterns of PCDD/Fs and PCBs differ between sources and are important tools for source identification. Therefore, patterns associated with different sources and incidents relating to various feed matrices and certain agricultural chemicals were collated from published scientific papers, with additional ones available from some laboratories. The collection was evaluated for completeness by presentations at workshops and conferences. Primary sources appear to derive from 5 categories, i) by-products from production of organochlorine chemicals (e.g. PCBs, chlorophenols, chlorinated pesticides, polyvinyl chloride (PVC)), ii) the result of combustion of certain materials and accidental fires, iii) the use of inorganic chlorine, iv) recycling/production of certain minerals, and v) certain naturally occurring clays (ball clay, kaolinite). A decision tree was developed to assist in the identification of the source.

Chlorinated paraffin analysis by gas chromatography Orbitrap high-resolution mass spectrometry: Method performance, investigation of possible interferences and analysis of fish samples.

For decades, high quantities of short-chain chlorinated paraffins (SCCP) and medium-chain chlorinated paraffins (MCCP) have been widely used, for instance as plasticizers or flame retardants, leading to global pollution due to unintentional emissions from products or waste. Due to the high complexity of chlorinated paraffins with several thousand congeners there is no consensus on an analytical procedure for SCCPs and MCCPs in food samples. Amongst the multitude of methods currently in use, high-resolution mass spectrometry is particularly valuable for in-depth studies of homologue patterns. Here we analyse SCCPs and MCCPs with gas chromatography coupled to high-resolution Orbitrap mass spectrometry (GC-Orbitrap-HRMS) operated in full-scan acquisition in electron capture negative ion (ECNI) mode at 60,000 and 120,000 resolution (FWHM, m/z 200, equals roughly 30,000 and 60,000 at 5% peak height). Linear dynamic range, selectivity and sensitivity tests confirmed an excellent linearity in a concentration range of 25-15,000 pg/µL with very low limits of detection (LODs) in the low pg/µL range. Spiking experiments with high levels of native mono- and di-ortho-polychlorinated biphenyls (PCBs) and mixtures of MCCP and SCCP standards did not have a negative impact on isotope ratios of the examined homologues. Besides the [M-Cl]- fragment ions used for quantification, the mass spectra of homologues also featured [M-HCl]- ions whose abundance increased with decreasing chlorination degree. In addition, [M-HCl-Cl]- ions were detected with a relative abundance of 5-10%. Three salmon (Salmo salar) samples farmed in Norway showed a consistent CP homologue pattern which differed both from the CP pattern in a sample from Scottish aquaculture and a wild salmon sample. These measurements produce evidence that discretely different CP patterns may exist in different areas of origin. Our results demonstrate that GC/ECNI-Orbitrap-HRMS is well-suited for the analysis of CPs by overcoming a range of mass interference problems and due to its thus far unmatched sensitivity.

Do PCDD/PCDF standard solutions used in dioxin analysis pose a risk as potentially acutely toxic to lab personnel?

Laboratory safety requires protecting personnel from chemical exposures. Working with stock solutions of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/PCDFs) in routine analysis of feed and food with bioanalytical or physicochemical methods raises some concerns. Since PCDD/PCDFs are considered as possibly acutely toxic, the potential risks were evaluated to determine whether supervision of their use is necessary. Based on LD50-data for oral or dermal intake, hazard classification of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) as a substance (category 1) and in commercially available TCDD standard solutions (category 4) is different. As worst case exposure scenario during routine laboratory work it was assumed that a dose of 100 ng TCDD gets onto the skin and is absorbed. This would result in the total body burden of a 70 kg person with 15 kg fat increasing from 10 (upper range of current background levels) to ∼17 pg of toxic equivalents (TEQs) of PCDD/PCDFs per g lipid, a level commonly observed over past decades. Chloracne, the main acute effect occurring weeks after exposure, is observed at much higher blood concentrations than estimated from accidental laboratory exposure. Immunotoxicity, developmental effects and other toxic effects may occur at lower blood levels, but require longer periods to develop. Since acute toxic symptoms don't occur within an "8 h acute time window", no supervision is necessary when working with standard solutions in routine analysis. Nevertheless, precautionary measures are needed regarding long-term adverse health effects and appropriate workplace conditions must exist to ensure that additional occupational exposure to PCDD/PCDFs by laboratory personnel is negligible.

WHO/UNEP global surveys of PCDDs, PCDFs, PCBs and DDTs in human milk and benefit-risk evaluation of breastfeeding.

Since 1987, the World Health Organization (WHO) carried out global surveys on polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and polychlorinated biphenyls (PCBs) in human milk. This study presents a review of the three most recent surveys from 2000 to 2010, including DDT. The objective was to identify global quantitative differences and provide baseline information for 52 countries or provide time-trends for countries with previous data. Individual human milk samples were collected following a WHO-designed procedure and combined to form a national pooled sample. Here, we report global levels for PCDDs, PCDFs, PCBs and the sum of o,p'-DDT, p,p'-DDT, o,p'-DDE, p,p'-DDE, o,p'-DDD and p,p'-DDD (ΣDDTs). A concise risk-benefit evaluation related to human milk contamination with these persistent organic pollutants (POPs) was also done. Large global and regional differences were observed. Levels of PCDDs and PCDFs were highest in India and some European and African countries. PCB levels were highest in East and West Europe. The highest levels of ΣDDTs were found in less industrialized countries. A temporal downward trend for PCDDs, PCDFs and PCBs is indicated. A risk-benefit assessment indicates that human milk levels of PCDDs, PCDFs and PCBs are still significantly above those considered toxicologically safe, while ΣDDTs are below or around those considered safe. With respect to potential adverse health effects, a more dominant role of in utero exposure versus lactational exposure is indicated. If potential adverse effects are balanced against positive health aspects for (breastfed) infants, the advantages of breastfeeding far outweigh the possible disadvantages. Our observations provide a strong argument to plea for further global source-directed measures to reduce human exposure further to dioxin-like compounds.

Atmospheric-pressure chemical ionization tandem mass spectrometry (APGC/MS/MS) an alternative to high-resolution mass spectrometry (HRGC/HRMS) for the determination of dioxins.

The use of a new atmospheric-pressure chemical ionization source for gas chromatography (APGC) coupled with a tandem quadrupole mass spectrometry (MS/MS) system, as an alternative to high-resolution mass spectrometry (HRMS), for the determination of PCDDs/PCDFs is described. The potential of using atmospheric-pressure chemical ionization (APCI) coupled to a tandem quadrupole analyzer has been validated for the identification and quantification of dioxins and furans in different complex matrices. The main advantage of using the APCI source is the soft ionization at atmospheric pressure, which results in very limited fragmentation. APCI mass spectra are dominated by the molecular ion cluster, in contrast with the high energy ionization process under electron ionization (EI). The use of the molecular ion as the precursor ion in MS/MS enhances selectivity and, consequently, sensitivity by increasing the signal-to-noise ratios (S/N). For standard solutions of 2,3,7,8-TCDD, injections of 10 fg in the splitless mode on 30- or 60-m-length, 0.25 mm inner diameter (id), and 25 µm film thickness low-polarity capillary columns (DB5MS type), signal-to-noise (S/N) ratios of >10:1 were routinely obtained. Linearity was achieved in the region (correlation coefficient of r(2) > 0.998) for calibration curves ranging from 100 fg/µL to 1000 pg/µL. The results from a wide variety of complex samples, including certified and standard reference materials and samples from several QA/QC studies, which were previously analyzed by EI HRGC/HRMS, were compared with the results from the APGC/MS/MS system. Results between instruments showed good agreement both in individual congeners and toxic equivalence factors (TEQs). The data show that the use of APGC in combination with MS/MS for the analysis of dioxins has the same potential, in terms of sensitivity and selectivity, as the traditional HRMS instrumentation used for this analysis. However, the APCI/MS/MS system, as a benchtop system, is much easier to use.

Dioxins and PCBs in feed and food--review from European perspective.

During the 1990s, a number of adverse contamination incidents focussed the attention of the media and the general public on food safety. This led to the evaluation of safety measures with regard to dioxin intake from food. Important aspects regarding dioxins and PCBs in the food chain are reviewed here, allowing a contextual understanding of the present situation through its chronological developments. About 90-98% of the average exposure of humans to dioxins and PCBs results from dietary intake, with food of animal origin being the predominant source. Therefore, animal feed contributes considerably to the presence of these compounds in food. The detection of the "real" source of a contamination event in the food chain is a complex scientific problem and requires specific knowledge on production processes and changes of patterns during bioaccumulation. This is demonstrated by complex investigations performed in three studies on two continents to identify the source (e.g. from contamination of cow's milk in Germany, to citrus pulp pellets from Brazil as an ingredient in feed, then to contaminated lime for neutralization and finally to a landfill with residues of vinyl chloride monomer production). This example shows also the substantial economic losses resulting from incidents in the food chain and the consequences to global trade. In 2001, the EU Scientific Committee on Food established a group tolerable weekly intake (TWI) of 14 pg WHO-TEQ/kg body weight and concluded that a considerable proportion of the European population would exceed this TWI. On the global level, the Joint FAO/WHO Expert Committee on Food Additives (JECFA) provides scientific advice to the Codex Alimentarius Commission and therefore contributes to harmonized international food standards. In its evaluation of 2001, JECFA derived a provisional tolerable monthly intake (PTMI) of 70 pg TEQ/kg body weight. The sum of the median intake of PCDD/F-TEQ and PCB-TEQ exceeded the PTMI in Western European countries, was in the PTMI range in North America, but lower in Japan and New Zealand. The 90th percentile of PCDD/F-TEQ exceeded the PTMI in Western European countries and North America, the 90th percentile of coplanar PCBs in Western European countries. Therefore, in 2001 the EU Commission developed a strategy to reduce the presence of dioxins and PCBs in the environment and in the food chain. The legislative measures comprised maximum levels and action levels for feed and food, and a Rapid Alert System for detected incidents was introduced. The network of the EU Reference Laboratory and National Reference Laboratories contributes to harmonization within the EU Member States and developed analytical criteria for screening and confirmatory methods for control of feed and food. After all these efforts it is of general interest to see whether these measures had an effect. The 2012 evaluation of the European Food Safety Authority (EFSA) based on comprehensive monitoring data of 26 European countries shows a general decrease in dietary exposure of dioxins and DL-PCBs between 2002-2004 and 2008-2010, estimated to be between 16.6% and 79.3% for the different population groups. A smaller decrease was observed for NDL-PCBs. The percentage of individuals exposed above the TWI of 14 pg TEQ/kg b.w. was estimated to be between 1.0 and 52.9%. Toddlers and other children were the most exposed groups (being at the upper end of these ranges). Fish, meat and dairy products appeared to be the highest contributing food groups to dietary exposure.

Dioxins (polychlorinated dibenzo-p-dioxins and polychlorinated dibenzo-furans) in traditional clay products used during pregnancy.

Geophagy, the practice of consuming clay or soil, is encountered among pregnant women in Africa, Eastern Asia and Latin America, but also in Western societies. However, certain types of clay are known to contain high concentrations of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs). The aim of this study was to determine the PCDD/F contents of orally consumed clays purchased from Dutch and African markets. Congener patterns were compared with those of pooled human milk samples collected in eight African countries, to investigate a possible relationship with clay consumption. From the Dutch market thirteen clay products were examined, seven of African and six of Suriname origin. From seven African countries, twenty clay products were collected. All 33 clay products were screened with a cell-based bioassay and those showing a high response were analyzed by GC/HRMS. High PCDD/F concentrations were measured in three clay products from the Dutch market, ranging from 66 to 103 pg TEQ g(-1), whereas clay products from African countries were from 24 to 75 pg TEQ g(-1). Patterns and relatively high concentrations of PCDD/Fs in human milk samples from the Democratic Republic of the Congo and Côte d'Ivoire suggest a relationship with the consumption of contaminated clay. Frequent use of PCDD/F contaminated clay products during pregnancy may result in increased exposure of the mother and subsequently the developing fetus and new-born child. The use of these contaminated clays during pregnancy should be carefully considered or even discouraged.

Polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and polychlorinated biphenyls (PCBs) in breast milk of first-time Irish mothers: impact of the 2008 dioxin incident in Ireland.

The 2008 dioxin incident in Ireland resulted in elevated concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and polychlorinated biphenyls (PCBs) in Irish pork and pork products, due to the consumption of contaminated animal feed by pigs. In order to investigate any resulting impact on the Irish population, these contaminants were measured in pooled breast milk samples from 109 first-time mothers, collected in 2010. A comparison of the results with similar data from 2002 revealed generally lower concentrations of PCDD/Fs and dioxin-like PCBs in the 2010 samples, confirming the declining trend reported by many authors. Contaminant concentration levels for both 2002 and 2010 were generally slightly lower than those reported internationally, with a mean combined PCDD/F and PCB WHO-TEQ of 9.66pgg(-1)fat, for an overall pooled sample of milk from 2010. An apparent slight increase in PCDFs was observed between 2002 and 2010 (from 2.73pg WHO-TEQ g(-1)fat to 3.21pg WHO-TEQ g(-1)fat), with the main contributory congener being 2,3,4,7,8-PentaCDF. While it cannot be totally discounted that the slight increase in 2,3,4,7,8-PentaCDF and in the overall PCDF WHO-TEQ in breast milk could be attributable to consumption of Irish pork during the 2008 incident, we consider that it is more likely that this was due to other factors, including the predominantly urban/industrial sampling locations for the 2010 samples, compared to 2002.

Fourth WHO-coordinated survey of human milk for persistent organic pollutants (POPs): Belgian results.

Persistent organic pollutants (POPs) are chemicals that accumulate in the food chain and are toxic to humans and wildlife. The fourth World Health Organization (WHO) survey on POP levels in human milk (2006-2009) aims to provide baseline and trend information on human exposure to POPs. So far Belgium participated in all three previous rounds (1988, 1992, 2001). Whereas the first three rounds focused on determination of dioxins and PCBs in pooled (mixed) samples, the fourth survey comprised the analyses of individual milk samples for nine "basic POPs" (chlorinated pesticides and indicator PCBs) and of pooled milk samples for "basic POPs", "advanced POPs" (dioxins and dioxin-like PCBs) and "optional POPs" (polybrominated diphenylethers [PBDEs], polybrominated dioxins and dibenzofurans [PBrDD/F], mixed halogenated dioxins and dibenzofurans [PXDD/F] and hexabromocyclododecane [HBCD]). For the Belgian participation human milk samples were collected during the summer of 2006 from 197 women between 18 and 30 years old distributed over all Belgian provinces. The individual samples were analyzed in a Belgian Laboratory for "basic" POPs. A pooled sample was made from 178 individual samples and analyzed by the WHO Reference Laboratory for the "basic, advanced and optional" POPs. The results indicate that most organochlorinated pesticides banned 25-30 years ago were below or around detection limits in Belgian human milk samples although DDE was still found at low levels in all samples. Over the last five years the levels of marker PCBs and PCDD/Fs in Belgian human milk decreased, respectively, by 58% and 39%. For some of the other emerging or older compounds recent international data are needed to allow comparison. This shows the importance of international studies as run by WHO.

Persistent environmental contaminants in human milk: concentrations and time trends in Italy.

Breast milk monitoring studies of persistent and toxic environmental contaminants are of primary importance for carrying out an adequate risk assessment at the actual levels of human exposure and represent a major source of information on infant perinatal exposure. Milk specimens from mothers of the general population of the Venice and Rome areas were collected over the 1998-2001 period, pooled, and analyzed for selected persistent organic pollutants such as polychlorodibenzodioxins (PCDDs), polychlorodibenzofurans (PCDFs), polychlorobiphenyls (PCBs), organochlorinated pesticides (p,p'-DDE, p,p'-DDT, hexachlorobenzene), and polybromodiphenyl ethers (PBDEs), and the heavy metals Cd, Co, Cu, Hg, Mn, Pb, Sn, and Zn. The goal was to verify whether mother milk from the Venice area, whose lagoon is partly under direct industrial impact, had a contaminant load greater than that from the Rome area, primarily urban. For mothers from the Venice area, the correlation between fish and fishery product consumption and contaminant concentrations in milk was also explored, with however inconclusive results. The concentrations of PCDDs, PCDFs, dioxin-like PCBs, and organochlorinated pesticides determined in this study were compared with those available from a previous analytical work carried out on 1987 human milk pools of domestic origin: the declining trend of the aforesaid contaminants in milk is confirmed to be in agreement with what was observed in other European countries. The breast milk content of (137)Cs and (40)K radionuclides was also determined and compared with data obtained in other research programmes carried out in Italy: the health risk for breastfed infants was deemed to be not significant.

PCDD/Fs and PCBs in butter samples from new European Union member states and a candidate country: analytical quality control, results and certain PCB-specific aspects.

Milk and milk products have shown to be good indicator samples for the contamination of persistent organic pollutants (POPs) along the food chain. To gather information on whether exposure to dioxins and PCBs might cause a problem in countries about to join the European Union in 2004 or later, a study for evaluation of dioxin and levels of PCBs in 16 butter samples from eight countries (Cyprus, Czech Republic, Estonia, Lithuania, Poland, Romania, Slovenia and Slovakia) was performed. Comprehensive quality assurance/quality control (QA/QC) was requested. For this, eight quality control samples were included (in comparison to long-term mean, mean recovery for WHO-PCDD/F-TEQ of 97.9% with a CV of 3.0%, mean recovery for WHO-PCB-TEQ of 100.4% with a CV of 3.3%). Additionally, all butter samples were run as duplicates resulting in a confidence region of 95% statistical certainty of 4.6% for WHO-PCDD/F-TEQ and of 3.5% for WHO-PCB-TEQ. All samples except one from Romania were in the range of 0.21-0.59 pg WHO-PCDD/F-TEQ/g fat (upper bound), all samples except one from Romania and one from Estonia in the range of 0.32-0.82 pg WHO-PCB-TEQ/g fat (upper bound) and all samples except one from Romania and one from Estonia in the range of 0.57-1.23 pg sum WHO-TEQ/g fat (upper bound). The maximum values were found in samples from Romania (0.98 pg WHO-PCDD/F-TEQ/g fat; 1.75 pg WHO-PCB-TEQ/g fat) and Estonia (0.26 pg WHO-PCDD/F-TEQ/g fat; 1.62 pg WHO-PCB-TEQ/g fat). As a conclusion, all samples except one from Romania and one from Estonia were in the range of the actual low background contamination for PCDD/F and dioxin-like PCBs. The samples with elevated concentrations were below the EU action and maximum levels which have been valid since 2002, or will be applied from November 2006. In all samples except in one from Estonia, the contribution of dioxin-like PCBs to sum WHO-TEQ was 47-68% which reflects the usual range in Europe. In one sample from Estonia this contribution was 86% which points to a particular yet unknown PCB source. Thirty eight PCB congeners were determined allowing a detailed discussion of the relative contribution of individual congeners to the total PCB concentration. Correlation between PCB 153 and WHO-PCB-TEQ varied considerably between samples from different countries. Major tetra- or pentachlorinated mono-ortho PCBs without assigned TEFs were PCBs 60, 66, 74, and 110.

PCDD/Fs, dioxin-like PCBs and marker PCBs in eggs of peregrine falcons from Germany.

Thirty one egg samples of peregrine falcons were collected in different regions of Germany (South-West, North and East) and analysed for PCDD/Fs, PCBs and marker PCBs. Altogether, 37 PCB congeners were determined, allowing a more detailed discussion of different aspects of the relative importance of these individual congeners. For comparison with dioxin data from other studies, differences between toxic equivalency factors (TEF) have to be taken into consideration. The results of this study are discussed on the basis of WHO-TEFs for birds published in 1998. All samples showed high levels of PCDD/Fs (median of all samples: 368 pg WHO-PCDD/F-TEQ/g fat; range 137-1453) and even higher levels of PCBs (median 967 pg WHO-PCB-TEQ/g fat; range 243-7482). The upper range of these levels reach concentrations found at the beginning of the Belgian dioxin crisis when a significant reduction in hens' egg hatchability was observed. The relative contribution of PCBs to the total TEQ is on average 71% (range 55-92%) with a tendency to higher values in South-West Germany compared to different sites in East Germany or one site in northern Germany. Within a region, a wide range of contamination can be found. Even in the same cities (Stuttgart and Ludwigshafen), results derived from eggs collected in the same year (2003) varied by factors 5-10. These findings show the high degree of variation occurring even within the same kind of biological matrix. Therefore, to allow for comparison of regions or time trends, a considerable number of samples has to be analysed.

Breast milk dioxins in Hong Kong and Pearl River Delta.

There are no previous reports from South China on chemically determined polychlorinated dibenzo-para-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxin-like polychlorinated biphenyls (PCBs) in human breast milk expressed as World Health Organization (WHO) toxic equivalents (TEQs). In a 2002-2003 WHO exposure study, 13 pools of breast milk comprising samples from 316 primiparous women in Hong Kong in 2002 were analyzed by gas chromatography/mass spectrometry for 29 PCDD/F and dioxin-like PCB congeners. Total WHO-TEQs ranged from 8.97 to 16.7 pg/g fat (weighted mean, 12.9 pg; weighted median, 13.4 pg). Variations in TEQs included positive associations with age (R2 = 0.73, p < 0.0005), higher consumption of dairy products and seafood, and lower TEQs in overseas mothers and ever-smokers. Congener profiles indicated geographic specificity of exposure in Hong Kong, mainland China, and overseas Asian countries, including higher proportions of PCB-TEQs (overseas) and PCDF-TEQs (mainland China). The median TEQs of PCDD/Fs (8.69 pg/g fat) and PCBs (4.73 pg/g fat) in Hong Kong were highest among the five Asian Pacific countries but lower than the levels for at least half of the European countries that participated in the WHO study. However, future international studies should incorporate mother's age in the design of the pooling strategy to allow standardization by other exposure factors and valid comparisons among different countries. The findings allow support for the WHO breast-feeding advisory. Trends in human dioxin levels in the region cannot yet be determined, and rigorous controls are needed to reduce emissions of dioxins and human exposure in mainland China.

The use of potassium dichromate and ethyl alcohol as blood preservatives for analysis of organochlorine contaminants.

The "gold standard" for preserving and shipping of human tissue samples for analysis of organochlorine contaminants is freezing. This method can be difficult, costly if using heavy dry ice for shipping, and often unfeasible, especially in less developed countries where electricity and dry ice are frequently rare or absent. Therefore, it is essential that more convenient and practical methods for preservation of blood samples are found. As an alternative to freezing, there have been studies employing potassium dichromate as a preservative for human or cow's milk or ethyl alcohol preservation of blood for dioxin analysis. In this study, four methods were compared to investigate the effectiveness of ethyl alcohol and potassium dichromate as blood preservatives for analysis of dioxins, dibenzofuran, and polychlorinated biphenyl (PCB) congeners. Samples of whole blood from a Dallas, Texas hospital were collected and pooled. Freezing, ethyl alcohol in two concentrations (20% and 40% per volume of sample), and potassium dichromate were used for blood preservation. The blood samples containing potassium dichromate or ethyl alcohol were stored and sent to ERGO laboratory for dioxin analysis and comparison with results from the frozen sample, which was kept frozen at all times until analyzed. This study suggests that potassium dichromate is a suitable alternative to freezing for preservation of whole blood for dioxin, dibenzofuran, and PCB measurements when either lipid or wet weight based results are reported. Potassium dichromate tablets were very easy and convenient to use--two 100 mg tablets (with a dichromate content of about 33 mg each) were added to each bottle containing 65 ml of blood. However, ethyl alcohol at 20% and 40% concentration under the conditions of this pilot study and the analytical method employed did not appear to provide satisfactory preservation when lipid based results are given or when the fat content has to be determined (wet or whole weight). Further research with a larger number of samples, inclusion also of other groups of persistent organic pollutants such as organochlorine pesticides or brominated flame retardants, a longer duration of storage time, and at temperatures greater than US or German room temperature is indicated in order to recommend the routine use of potassium dichromate as preservative for whole blood intended for dioxin, dibenzofuran, and PCB analysis.

Food as a source of dioxin exposure in the residents of Bien Hoa City, Vietnam.

Recently, elevated dioxin levels, over 5 parts per trillion (ppt) 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), from Agent Orange was reported in 95% of 43 selected residents of Bien Hoa City, a city in southern Vietnam near a former air base used for Agent Orange-spraying missions. Agent Orange herbicide, contaminated with TCDD, was sprayed in Vietnam between 1962 and 1971 primarily for use as a defoliant. Typical blood TCCD levels are 2 ppt in Vietnamese, but levels are as high as 413 ppt in Bien Hoa City. Elevated TCDD was found in children born many years after Agent Orange spraying ended and in immigrants from non-Agent Orange-sprayed parts of Vietnam, which documented new exposures. Extremely elevated soil TCDD samples, over 1 million ppt, and elevated TCDD in sediment were found in some nearby areas such as Bien Hung Lake. The primary route of intake of almost all dioxins in humans is food. However, in our prior studies in Bien Hoa, food was unavailable for dioxin analysis so the route of intake was not confirmed. In the 1970s, while Agent Orange was still being sprayed, elevated human milk TCDD levels as high as 1850 were detected in milk from Vietnamese people living in Agent Orange-sprayed areas where consumption of fish was high. Furthermore, also in the 1970s, elevated TCDD levels (up to 810 ppt) were found in fish and shrimp from the same area as the milk donors. In the 1980s, we found elevated TCDD and also other organohalogen levels in human tissue, pork, fish, a turtle, and a snake in Southern Vietnam. For these reasons, we recently collected food from Bien Hoa and analyzed it for dioxins, polychlorinated biphenyls (PCBs), DDT and its metabolites, and other organochlorines. We found marked elevation of TCDD, the dioxin characteristic of Agent Orange, in some of the food products, including ducks with 276 ppt and 331 ppt wet weight, chickens from 0.031-15 ppt wet weight, fish from 0.063-65 ppt wet weight, and a toad with 56 ppt wet weight. Usual TCDD levels in food are less than 0.1 ppt. Total TEQ for ducks was from 286-343 ppt wet weight or 536 ppt and 550 ppt lipid; for chickens from 0.35-48 ppt wet weight or 0.95-74 ppt lipid, for fish from 0.19-66 ppt wet weight or 3.2 ppt and 15,349 ppt lipid, and the toad was 80 ppt wet weight and 11,765 ppt lipid. Interestingly, this study did not find elevated levels of TCDD in the pork and beef samples. Clearly, food, including duck, chicken, some fish, and a toad, appears responsible for elevated TCDD in residents of Bien Hoa City, even though the original Agent Orange contamination occurred 30-40 years before sampling. Elevated levels of PCBs and DDT and its metabolites were found in some food samples. Furthermore, measurable levels of hexachlorocyclohexanes (HCH) and hexachlorobenzene (HCB) were found in a wide range of measurable levels. All of the 11 dioxin-like PCBs measured and presented plus 6 dioxins in addition to TCDD and 10 dibenzofurans contributed to the total dioxin toxicity (TEQ). However, when elevated, TCDD frequently contributed most of the TEQ. Thirty-six congeners from 7 classes of chemicals were measured in each of the 16 specimens providing a total of 576 congener levels.

Are Vietnamese food exports contaminated with dioxin from Agent Orange?

In this study the levels of dioxins and dioxinlike compounds, dibenzofurans, and polychlorinated biphenyls (PCBs) were measured in food exported from Vietnam to the United States. In the past, some Vietnamese fish was found to be contaminated with dioxin, 2,3,7,8-TCDD, or TCDD, from Agent Orange. U.S. commercial catfish farmers recently claimed that exported Vietnamese food, especially catfish, was contaminated with dioxin from Agent Orange and was thus a matter of concern to the U.S. Congress. In 2001, twenty-two exported Vietnamese food samples (mostly fish) were purchased in the United States and Laos to evaluate possible contamination with dioxin from Agent Orange. Dioxin, dibenzofuran, and coplanar PCB congeners were measured by high-resolution gas chromatography-high-resolution mass spectrometry by laboratories in Germany and Canada certified for dioxin analysis by the World Health Organization (WHO). In the current study, only low levels of dioxins and dioxinlike compounds were present in all exported Vietnamese food analyzed, despite recent findings of elevated levels of one dioxin, 2,3,7,8-TCDD, from Agent Orange exposure in some Vietnamese individuals, presumably due to food consumption. It appears unlikely that exported Vietnamese food is, in general, contaminated with large amounts of dioxin (TCDD) from Agent Orange or other sources. Further food sampling from areas of heavy Agent Orange spraying in Vietnam is warranted to map out where the source of TCDD contamination of food may be originating.

Dioxin, dibenzofuran, and polychlorinated biphenyl (PCB) levels in food from Agent Orange-sprayed and nonsprayed areas of Laos.

During the Vietnam War, a phenoxy-herbicide mixture called Agent Orange, which was contaminated with 2,3,7,8-tetrachlorodibenzodioxin, or TCDD, was used primarily for destruction of forest and other foliage in order to prevent enemy troop movement and protect American and allied troops and military bases in the south of Vietnam. Smaller areas of Laos and Cambodia were also sprayed with Agent Orange between 1962 and 1971 from fixed-wing aircraft. In 2001, 28 food samples consisting of meat, fish, and dairy products were collected in sprayed and nonsprayed areas of Laos and analyzed for dioxins, dioxinlike dibenzofurans, and selected polychlorinated biphenyl (PCB) congeners by high-resolution gas chromatography/high-resolution mass spectrometry at laboratories in Germany and Canada. Low levels of dioxin and other dioxinlike substances were found in Laotian food, similar to levels present in recent exported Vietnamese food to United States. Vietnam is geographically adjacent to Laos. Total dioxin toxic equivalent (TEQ) levels were similar in samples from sprayed and non-sprayed areas, ranging from 0.009 to 0.851 pg/g or parts per trillion (ppt) in sprayed Sepone, and from 0.022 to 0.537 pg/g or ppt wet weight in non sprayed Vientiane. However, the Laotian fish samples from the Agent Orange-sprayed area had, on average, a higher proportion of total TEQ from TCDD (31.7% vs. 9.4%) compared to the nonsprayed area. Some other food items, duck eggs and pork liver from Sepone, also had severalfold higher TCDD levels than similar food samples from Vientiane, 0.029 vs. 0.011 pg/g and 0.070 vs. 0.004 pg/g wet weight, respectively. There were no substantial differences in levels of dibenzofuran and PCB congeners. In general, elevated TCDD levels were not found in Laotian food samples. It is possible that dioxin-contaminated areas, or "hot spots," exist in Laos as they do in Vietnam, although they have not yet been identified.