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.     

高分辨气相色谱/质谱法分析新闻纸和复印纸中的二恶英

随机选择空白的和经过印刷的新闻纸和复印纸,粉碎后用溶剂提取并经多步色谱柱纯化,采用同位素稀释、高分辨气相色谱/高分辨质谱(HRGC/HRMS)联用技术分析了其中的二含量。结果表明:新闻纸中二的总量高于复印纸,但是毒性当量却低于复印纸;经过印刷的新闻纸和复印纸中二的含量均高于空白的新闻纸和复印纸;新闻纸和复印纸中相同二异构体的含量 是不同的。空白的和经过印刷的新闻纸中二毒性当量分别为0.48 ng/kg和0.61 ng/kg,而空白的和经过印刷的复印纸中二恶英毒性当量分别为0.74 ng/kg和0.79     

高分辨气相色谱/质谱法分析新闻纸和复印纸中的二噁

随机选择空白的和经过印刷的新闻纸和复印纸 ,粉碎后用溶剂提取并经多步色谱柱纯化 ,采用同位素稀释、高分辨气相色谱 /高分辨质谱 (HRGC/HRMS)联用技术分析了其中的二含量。结果表明 :新闻纸中二的总量高于复印纸 ,但是毒性当量却低于复印纸 ;经过印刷的新闻纸和复印纸中二的含量均高于空白的新闻纸和复印纸 ;新闻纸和复印纸中相同二异构体的含量是不同的。空白的和经过印刷的新闻纸中二毒性当量分别为 0 4 8ng/kg和 0 6 1ng/kg,而空白的和经过印刷的复印纸中二毒性当量分别为 0 74ng/kg和 0 79ng/kg。所有样品中添加的13 C标记的 2 ,3,7,8位取代的二回收率均在 4 9 82 %至 131 34%之间。     

Dioxins in Soil and Fish Samples from a Waste Pentachlorophenol Manufacturing Plant

A method to analyze polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/DFs, dioxins) and coplanar polychlorinated biphenyls in environmental samples is developed and used to determine the levels of PCDD/DFs in 13 soil and 4 fish samples collected around a waste pentachlorophenol manufacturing plant. The 2,3,7,8-tetrachlorodioxin toxic equivalents (TEQ) values range from 0.239 ng-TEQ/g to 1357 ng-TEQ/g in soil samples and 0.041 ng-TEQ/g to 0.247 ng-TEQ/g in fish samples. A broader survey of PCDD/DF contamination in the vicinity around the manufacturing plant is strongly recommended.     

Trace analysis of polychlorinated dibenzo-p-dioxins, dibenzofurans and WHO polychlorinated biphenyls in food using comprehensive two-dimensional gas chromatography with electron-capture detection

Trace analysis of 2,3,7,8-polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and the 12 WHO-PCBs (four non-ortho and eight mono-ortho congeners that have been assigned toxic equivalence factors, TEFs, by the World Health Organisation) was conducted by comprehensive two-dimensional gas chromatography with a micro electron-capture detector (GC x GC-microECD). Four food matrices (fish oil from herring, spiked cows' milk, vegetable oil and an eel extract) were analysed by two GC x GC laboratories, and four GC-HRMS laboratories generated reference values. The two GC x GC laboratories used different column combinations for separating the target analytes. For the first dimension, non-polar DB-XLB and VF-1 columns were used, and for the second dimension, an LC-50 liquid crystalline column with unique selectivity for planar compounds. The congener-specific and total toxic equivalence (TEQ) data obtained using DB-XLB x LC-50 were in good agreement with results obtained by the GC-HRMS laboratories. The WHO-PCB data obtained with the VF-1 x LC-50 combination was also good, but the PCDD/F concentrations were sometimes overestimated due to matrix interferences. GC x GC-microECD using DB-XLB x LC-50 seems to fulfil the European Community requirements of a screening method for PCDD/F and WHO-PCB TEQ in food.     

杭州市冬季大气气溶胶PM2.5中二恶英和多氯联苯的污染特征

2014年1月在杭州市选择5个点位采集大气颗粒物PM_2.5样品,采用同位素稀释高分辨气相色谱/高分辨质谱测定PM_2.5中的二恶英（PCDD/Fs）和多氯联苯（PCBs）,对PM_2.5的污染状况以及PM_2.5中PCDD/Fs和PCBs的污染水平及分布特征进行了研究。PM_2.5的质量浓度范围为85~168 μg/m³,PM_2.5污染较重,但与2004年同期相比明显降低。PM_2.5中PCDD/Fs的毒性当量（TEQ）为0.277~0.488 pg I-TEQ/m³,明显高于2004年同期采集样品。颗粒物中PCDD/Fs以八氯代二苯并-对-二恶英（OCDD）为主,毒性当量主要贡献者为2,3,4,7,8-五氯代二苯并呋喃（2,3,4,7,8-PeCDF）。PM_2.5中PCBs的质量浓度范围为2.9~8.1 pg/m³,二恶英类多氯联苯（DL-PCBs）的毒性当量范围为2.6~6.1 fg WHO-TEQ/m³,污染较低。PCBs在颗粒物中分布以PCB-28为主,但对毒性当量贡献最大的为PCB-126。PCDD/Fs和PCBs的气-固分配特征表现为PCDD/Fs主要分布于颗粒物中,而PCBs主要分布于气相中。     

Micropore-free surface-activated carbon for the analysis of polychlorinated dibenzo-p-dioxins-dibenzofurans and non-ortho-substituted polychlorinated biphenyls in environmental samples

2,3,7,8-Substituted polychlorinated dibenzo-p-dioxins/polychlorinated dibenzofurans (PCDD/Fs) and non-ortho-substituted polychlorinated biphenyls (PCBs) account for almost all of the total toxic equivalents (TEQ) in environmental samples. Activated carbon columns are used to fractionate the samples for GC-MS analysis or bioassay. Micropore-free surface-activated carbon is highly selective for PCDD/Fs and non-ortho-PCBs and can improve the conventional activated carbon column clean-up. Along with sulfuric acid-coated diatomaceous earth columns, micropore-free surface-activated carbon provides a rapid, robust, and high-throughput sample preparation method for PCDD/Fs and non-ortho-PCBs analysis.     

Polychlorodibenzo-p-dioxin and polychlorodibenzofuran levels in cow milk samples, egg samples and meat in Finland

Summary To survey the levels of PCDD/F in Finnish foods, cow milk samples from the largest dairies, egg samples from the major producers and meat from the main slaughterhouses have been analyzed. For twenty whole milk samples supplied in cartons in June 1991 and stored for 14 days in a refrigerator, the total level of toxic equivalents, calculated on a wet weight basis using Nordic toxic equivalents, had a mean of 38 pg TEO/l (range 18 – 78 pg TEQ/l). This corresponds to an average intake of 15 pg TEQ/day for a person consuming 400 ml milk/day on an average. The cow milk analyzed from glass bottles showed a mean of 29 pg TEQ/l (range < 10 – 67 pg TEQ/l). Migration from the carton container and/or other treatment of the milk in connection with the packing process made a 30% contribution to the total PCDD/F content of the cow milk. The results for milk samples in cartons and glass bottles, as expressed on a fat basis were 1.17 pg TEQ/g (range 0.57 – 2.48 pg TEQ/g) and 0.83 pg TEQ/g (range <0.5 – 1.80 pg TEQ/g), respectively. For 20 pooled egg samples the total level of toxic equivalents calculated on whole product basis had a mean of 0.12 pg TEQ/g (range 0.023 – 0.427 pg TEQ/g). This corresponds to an average intake of 3.9 pg TEQ/day for a person consuming 33 g egg per day on an average. For twenty pork and twenty bovine meat samples, the levels of PCDD/F were very low and below the detection limit (<0.5 pg/g fat) with the exception of some detectable traces of 2,3,7,8-Cl₄DF, 1,2,3,4,6,7,8-Cl₇DD and OCDD. These results showed that, at least in summertime when cows are outdoors and eat fresh hay, PCDD/F concentrations in Finnish cow milk are low. This conclusion is supported by the results from the meat samples which showed that beef contained notably low levels of PCDD/F. On some farms chicken feed contains fish products in addition to corn. This is probably the reason why the level of PCDD/F in egg varied but was still reasonably low.     

“永固紫”染料和四氯苯醌中多氯代二苯并二/呋喃的分析

 采用浓硫酸对5种不同来源的四氯苯醌和“永固紫”染料样品进行溶解分散,用甲苯提取和 多层色谱柱纯化,利用同位素稀释法及高分辨气相色谱/质谱 (HRGC/HRMS)联用技术,测定 了其中的多氯代二苯并二/呋喃(PCDD/F)的质量比。结果表明“永固紫”染料及其原料中P CDD/F的质量比异常高,八氯代二苯并二/呋喃(OCDD/F)已达到μg/g级甚至100 μg/g级,七氯代二苯并二/呋喃(HpCDD/F)和六氯代二苯并二/呋喃(HxCDD/F)质量比也达到 ng/g级。所有样 品的毒性当量浓度(TEQ)均已超过10 pg/g的危险水平,是纺织品中PCDD/F的一类重要污染源 。并进一步探讨了PCDD/F的来源。     

Interpretation of CALUX results in view of the EU maximal TEQ level in milk

Analyses of dioxins in food have become increasingly important since the European Commission has enforced maximal toxic equivalent concentration (TEQ) levels in various food and feed products. Screening methodologies are usually used to exempt those samples that are below the maximum permitted limit and that can, therefore, be released to the market. In addition, one needs to select those samples that require confirmation of their dioxin TEQ level. When bioassays are used as screening tools, the interpretation of the obtained results should consider the higher variability and uncertainty associated with them. This paper explores the use of CALUX data as quantitative screening results. The validation of the method for the polychlorinated dibenzo-p-dioxins (PCDD)/F TEQ determination in milk samples is described with emphasis on the decision limit (CC) and the precision of the method. The decision limit amounts to 4.53 pg TEQ/g fat. Repeatability and within-lab reproducibility coefficients of variation are below 30%. The newly introduced parameter CC^* of 1.47 pg TEQ/g fat delimits with CC a range of suspicious results. These data are not significantly different from the maximum limit of 3 pg TEQ/g fat and should be confirmed by a confirmatory analytical method such as HRGC–HRMS.     

Polychlorinated naphthalene concentrations and profiles in cheese and butter,and comparisons with polychlorinated dibenzo-p-dioxin,polychlorinated dibenzofuran and polychlorinated biphenyl concentrations

Polychlorinated naphthalenes (PCNs) are candidates for inclusion in the Stockholm Convention on persistent organic pollutants. PCNs are structurally and toxicologically similar to 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) and its analogues. Intake in food is considered to be an important human exposure pathway for PCNs. In this preliminary study, cheese and butter samples were analysed for PCNs, polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and polychlorinated biphenyls (PCBs) using an isotope dilution gas chromatography high-resolution mass spectrometry method. The aim of this study was to evaluate the PCN concentrations in the cheese and butter samples and to compare them with the PCDD, PCDF and PCB concentrations. The PCN concentrations were 5.6–103 pg g^?1 of wet weight in the seven cheese samples tested and 5.0–199 pg g^?1 of wet weight in the seven butter samples tested. The mass concentrations of lower chlorinated congeners were greater than those of the higher chlorinated congeners. Congeners of CN45/36, CN27/30 and CN33/34/37 were much more abundant than other congeners found in tetrachlorinated PCNs. Congeners of CN51, CN66/67 and CN73 were determined to be the predominant congeners in penta-, hexa- and heptachlorinated homologs, respectively. The PCNs contributed around 5% of the total PCN, PCDD, PCDF and PCB toxic equivalence (TEQ) values. CN73 was found to be the dominant PCN congener and contributed more than 40% to the PCN TEQ value. Congeners CN66/67, CN69 and CN63 were also found at relatively high levels. The PCB congener CB118 was the predominant congener (by mass-based concentration) of the 12 dioxin-like PCBs (dl-PCBs). The PCBs contributed 53.8% of the total TEQ, and congener CB126 contributed more than any other compound that was analysed to the total TEQ. The PCDDs and PCDFs contributed 11.6% and 29.7% of the total TEQ values, respectively.     

Advances in analytical techniques for polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and dioxin-like PCBs

Analytical techniques for the determination of polychorinated dibenzo-p-dioxins (PCDD), polychlorinated dibenzofurans (PCDF) and dioxin-like PCBs (DLPCB) are reviewed. The focus of the review is on recent advances in methodology and analytical procedures. The paper also reviews toxicology, the development of toxic equivalent factors (TEF) and the determination of toxic equivalent quantity (TEQ) values. Sources, occurrence and temporal trends of PCDD/PCDF are summarized to provide examples of levels and concentration ranges for the methods and techniques reviewed.     

A new cleanup method of dioxins in sediment using large volume injection gas chromatography online coupled with liquid chromatography

A new cleanup method was developed and validated for the determination of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) in sediment. The sample extract was first treated with sulfuric acid and then cleaned up by a large volume injection gas chromatography online coupled with liquid chromatography (LVI-GC-LC) system. PCDD/Fs in the extract were separated by a GC column (DB-5), selected cut, cool trapped and transferred to a LC column (alumina). The fraction of PCDD/Fs eluted from the alumina column was collected and concentrated for the instrumental analysis. Under the optimized conditions, LVI-GC-LC method achieved the recoveries of 57–102% for 2,3,7,8-substituted PCDD/Fs, which met the requirements of US Environmental Protection Agency (EPA) Method 1613 and were better than those obtained using the conventional multistep column cleanup method. Meanwhile, compared with the conventional method, the limit of detection (LOD) values of 2,3,7,8-substituted PCDD/Fs cleaned up by LVI-GC-LC method were decreased due to the high-efficiency removal of interferents. These results suggested that the LVI-GC-LC cleanup method was a promising alternative to the multistep cleanup procedure for the determination of dioxins in environmental samples.     

Determination of polychlorinated dibenzo-p-dioxin and dibenzofuran congeners in air particulate and marine sediment standard reference materials (SRMs)

Due to the limited number of environmental matrix certified reference materials (CRMs) with assigned values for natural levels of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), an interlaboratory study was undertaken by the National Institute of Standards and Technology (NIST) and Environment Canada to establish reference concentration values for selected PCDD/Fs in two well-characterized NIST Standard Reference Materials (SRMs): SRM 1649a (Urban Dust) and SRM 1944 (New York/New Jersey Waterway Sediment). Results from 14 laboratories were used to provide reference values for the seventeen 2, 3, 7, 8-substituted PCDD/F congeners, the totals for individual tetra- through hepta-substituted PCDD/F homologues, and the total amount of tetra- through hepta-substituted PCDD/Fs. The mass fractions for the individual 2, 3, 7, 8-substituted congeners range from approximately 0.01 microg/kg to 7 microg/kg dry mass.     

Determination of polychlorinated dibenzo-p-dioxin and dibenzofuran congeners in air particulate and marine sediment standard reference materials (SRMs)

Due to the limited number of environmental matrix certified reference materials (CRMs) with assigned values for natural levels of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), an interlaboratory study was undertaken by the National Institute of Standards and Technology (NIST) and Environment Canada to establish reference concentration values for selected PCDD/Fs in two well-characterized NIST Standard Reference Materials (SRMs): SRM 1649a (Urban Dust) and SRM 1944 (New York/New Jersey Waterway Sediment). Results from 14 laboratories were used to provide reference values for the seventeen 2, 3, 7, 8-substituted PCDD/F congeners, the totals for individual tetra- through hepta-substituted PCDD/F homologues, and the total amount of tetra-through hepta-substituted PCDD/Fs. The mass fractions for the individual 2, 3, 7, 8-substituted congeners range from approximately 0.01 μg/kg to 7 μg/kg dry mass.     

Analysis of polychlorinated dibenzo-p-dioxins, dibenzofurans and dioxin-like polychlorinated biphenyls in vegetable oil samples by gas chromatography-ion trap tandem mass spectrometry

Gas chromatography coupled to ion trap tandem mass spectrometry (CG-MS-MS) has been evaluated for the analysis of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dl-PCBs) in vegetable oil samples containing different concentration levels (0.2-6 pg WHO-TEQ g(-1) for both PCDD/Fs and dl-PCBs) of the 29 toxic congeners of PCDD/F and dioxin-like PCBs. The effect of potential interfering compounds such as polychlorinated naphthalenes (PCNs), polychlorinated biphenyls (PCBs) and polychlorinated diphenylethers (PCDEs) on the quantification of mono-ortho PCBs has been investigated. In addition, the influence of the clean-up procedure on the final determination by CG-MS-MS was studied, showing that the quality of the results depends to a great extent on this analytical step. Quality parameters have been established and good precisions (CV: 3-19%) and low limits of detection for PCDD/Fs (0.04-0.20 pg g(-1) oil) and dl-PCBs (0.08-0.64 pg g(-1) oil) were obtained. The method was validated by a comparison of the CG-MS-MS results with those obtained by GC-HRMS.     

Fractionation of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and planar polychlorinated biphenyls by high performance liquid chromatography on a pyrenyl-silica column

Analytical procedures for the determination of polychlorinated dibenzo-p-dioxins (PCDD), polychlorinated dibenzofurans (PCDF) and non-ortho polychlorinated biphenyls (PCB) require a fractionation step to separate PCDD/F from planar PCB and the bulk of PCB. An HPLC method which achieves the separation of the bulk of PCB (0–6 mL of hexane), mono-ortho PCB (6–8 mL of hexane), non-ortho PCB (8–15 mL of hexane) and PCDD/F (15–50 mL of toluene) on a PYE column (2-(1-pyrenyl) ethyldimethylsilylated silica gel) in a single step without the use of backflush as other authors proposed was developed. The method shows a good accuracy and precision and it is linear in the range studied, e.g from 5.8 to 2420 pg injected in HPLC for TCDD/F, from 28.8 to 12100 pg for PeCDD/F, HxCDD/F, HpCDD/F and from 57.6 to 24200 pg for OCDD/F. It has been successfully applied to the analysis of technical mixtures of PCB (Aroclors), a pine wood sample and several water samples of different origins. Received: 29 November 1998 / Revised: 25 February 1999 / Accepted: 3 March 1999     

Pressurized liquid extraction of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and coplanar polychlorinated biphenyls from contaminated soil

Extraction solvents for pressurized liquid extraction (PLE) used to extract polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans (PCDD/PCDFs), and coplanar polychlorinated biphenyls (Co-PCBs) from contaminated soil were investigated. The PCDD/PCDFs and Co-PCBs in Certified Reference Material: CRM 0422 (Forest soil) were extracted using toluene, n-hexane, acetone, acetone/toluene and acetone/n-hexane (1:1, v/v). Soxhlet extraction was the reference method. Results demonstrated that PLE using mixed solvents produced better analyte recoveries than the single solvents. However, these results were lower than those for Soxhlet extraction. Additional extraction cycles using mixed solvents achieved better recovery results. Mixed solvents and several extraction cycles were necessary for satisfactory extraction of more tightly bound PCDD/PCDFs and Co-PCBs from soil.     

Validation of the CALUX bioassay for the screening of PCDD/Fs and dioxin-like PCBs in retail fish

The chemical-activated luciferase expression (CALUX) assay is a reporter gene assay that detects dioxin-like compounds based on their ability to activate the aryl hydrocarbon receptor (AhR) and thus expression of the reporter gene. In this paper, the CALUX assay was examined for its application in the screening of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dioxin-like PCBs) in retail fish. The sample extracts were cleaned up on a sulfuric acid-silica gel column followed by an activated carbon column, and the AhR activity of the separated PCDD/F and dioxin-like PCB fractions was determined using the assay. The quantitative limit for 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) was 0.98 pg ml(-1) (0.19 pg assay(-1) in the standard curve, corresponding to 0.16 pg g(-1) of CALUX-based toxic equivalency (2,3,7,8-TCDD equivalents) in the tested sample. Recovery tests in which dioxins were added to fish samples resulted in acceptable recoveries (77-117%). The CALUX assay performed well in the analysis of dioxins in fish samples and a comparative study revealed a strong correlation between the CALUX assay and high-resolution gas chromatography-high-resolution mass spectrometry analysis for the determination of PCDD/Fs (r = 0.89) and dioxin-like PCBs (r = 0.91) in retail fish (n = 22). These data revealed that the CALUX assay would be a useful screening method for PCDD/Fs and dioxin-like PCBs in retail fish.     

Fractionation of polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and planar polychlorinated biphenyls by high performance liquid chromatography on a pyrenyl-silica column

Analytical procedures for the determination of polychlorinated dibenzo-p-dioxins (PCDD), polychlorinated dibenzofurans (PCDF) and non-ortho polychlorinated biphenyls (PCB) require a fractionation step to separate PCDD/F from planar PCB and the bulk of PCB. An HPLC method which achieves the separation of the bulk of PCB (0–6 mL of hexane), mono-ortho PCB (6–8 mL of hexane), non-ortho PCB (8–15 mL of hexane) and PCDD/F (15–50 mL of toluene) on a PYE column (2-(1-pyrenyl) ethyldimethylsilylated silica gel) in a single step without the use of backflush as other authors proposed was developed. The method shows a good accuracy and precision and it is linear in the range studied, e.g from 5.8 to 2420 pg injected in HPLC for TCDD/F, from 28.8 to 12100 pg for PeCDD/F, HxCDD/F, HpCDD/F and from 57.6 to 24200 pg for OCDD/F. It has been successfully applied to the analysis of technical mixtures of PCB (Aroclors), a pine wood sample and several water samples of different origins. Received: 29 November 1998 / Revised: 25 February 1999 / Accepted: 3 March 1999