Increased levels of dioxin-like substances in adipose tissue in patients with deep infiltrating endometriosis.

STUDY QUESTION: Are the levels of biologically active and the most toxic dioxin-like substances in adipose tissue of patients with deep infiltrating endometriosis (DIE) higher than in a control group without endometriosis? SUMMARY ANSWER: DIE patients have higher levels of dioxins and polychlorinated biphenyls (PCBs) in adipose tissue compared with controls without endometriosis. WHAT IS KNOWN ALREADY: Some studies have investigated the levels of dioxin-like substances, in serum samples, in patients with endometriosis, with inconsistent results. STUDY DESIGN, SIZE, DURATION: Case-control study including two groups of patients. The study group (DIE group) consisted of 30 patients undergoing laparoscopic surgery because of DIE. In all patients, an extensive preoperative work-up was performed including clinical exploration, magnetic resonance imaging (MRI) and transvaginal sonography. All patients with DIE underwent a confirmatory histological study for DIE after surgery. The non-endometriosis control group (control group), included the next consecutive patient undergoing laparoscopic surgery in our center due to adnexal benign gynecological disease (ovarian or tubal procedures other than endometriosis) after each DIE patient, and who did not present any type of endometriosis. PARTICIPANTS/MATERIALS, SETTING, METHODS: During the surgical procedure 1-2 g of adipose tissue from the omentum were obtained. Dioxin-like substances were analyzed in adipose tissue in DIE patients and controls without endometriosis. MAIN RESULTS AND THE ROLE OF CHANCE: The total toxic equivalence and concentrations of both dioxins and PCBs were significantly higher in patients with DIE in comparison with the control group (P < 0.05), mainly due to the significantly higher values of the two most toxic dioxins (2,3,7,8-tetrachlorodibenzo-p-dioxin [2,3,7,8-TCDD] and 1,2,3,7,8-pentachlorodibenzo-p-dioxin [1,2,3,7,8-PeCDD]) (P < 0.01 for each compound). The levels of furan 2,3,4,7,8-PeCDF were statistically higher in the DIE group compared with controls. Only four congeners of PCBs had toxic equivalence values and concentrations that were statistically higher in patients with DIE, but these included the most toxic and carcinogenic PCB-126 (PCB-114 P < 0.05; PCB-156 P < 0.05; PCB-189 P = 0.04; PCB-126 P < 0.01). LIMITATIONS, REASONS FOR CAUTION: Since few patients were recruited, the study is only exploratory. Our results need to be confirmed in larger and more heterogeneous population studies since environmental and even genetic factors involved in determining dioxins and PCBs widely vary in different countries. Furthermore, the strict eligibility criteria used may preclude generalization of the results to other populations and the surgery-based sampling frame may induce a selection bias. Finally, adipose tissue was obtained only from the omentum, and not from other adipose tissue of the body. WIDER IMPLICATIONS OF THE FINDINGS: Our results suggest a potential role of dioxin-like substances in the pathogenesis of DIE. Further studies are warranted to confirm our findings. STUDY FUNDING/COMPETING INTERESTS: None. TRIAL REGISTRATION NUMBER: Not applicable.

PCDD/Fs and DL-PCBs in feeding fats obtained as co-products or by-products derived from the food chain.

Among the tasks included in the "Quality and safety of feeding fats obtained from co-products or by-products of the food chain" Project, supported by the European Union and included in the 6th Framework Program, a number of fats and oils collected as co- or by-products from the food chain were selected for the determination of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and 'dioxin-like' polychlorinated biphenyls (DL-PCBs). In the majority of the cases these samples are currently employed as feed ingredients. Nevertheless, additional fats, which are forbidden for feedstuff purposes were also considered in this study. In general terms, fats and oils were classified taking into account their nature and the processes applied to obtain these co- or by-products. PCDD/F and DL-PCB levels were evaluated in a first group of samples composed of fish oils, animal fats and lecithins. As expected, fats and oils with an animal origin presented higher concentrations, expressed in pg WHO-TEQ/g, compared to the levels found in vegetable samples like lecithins. The category of fish oils had the highest values for both PCDD/Fs and the sum of PCDD/Fs and DL-PCBs, with some samples showing levels above the maximum established at the present legislation related to the presence of PCDD/Fs and DL-PCBs in animal feed [Commission Directive 2006/13/EC of 3 February 2006 amending Annexes I and II to Directive 2002/32/EC of the European Parliament and of the Council on undesirable substances in animal feed as regards dioxins and dioxin-like PCBs. Official Journal of the European Communities L32, 44-53]. In a second group, fats and oils with a more complex composition obtained from different transformation processes or even mixtures of fats were considered; thus, acid oils from chemical refining, acid oils from physical refining, recycled cooking oils, oils extracted from exhausted bleaching earths, hydrogenated by-products, fatty acids calcium soaps and miscellaneous fats were analyzed. The data revealed a significant variability in PCDD/F and DL-PCB levels, expressed in pg WHO-TEQ/g, in these products. It has to be pointed out that the analyses of fats and oils belonging to these categories of products were sometimes difficult due to the complexity of the samples. In terms of legislation most of these samples cannot be easily included into one of the specific categories of substances intended for feedstuff purposes that are regulated in Commission Directive 2006/13/EC.

Occurrence of polychlorinated naphthalenes, polychlorinated biphenyls and short-chain chlorinated paraffins in marine sediments from Barcelona (Spain).

Polychlorinated naphthalenes (PCNs), short-chain chlorinated paraffins (SCCPs) and polychlorinated biphenyls (PCBs) were analysed in marine sediment samples collected from the coastal area of Barcelona (Spain) and near of a submarine emissary coming from a waste water treatment plant located at the mouth of the Besòs River (Barcelona). An integrated sample treatment based on Soxhlet extraction followed by a simple clean-up with Florisil and graphitized carbon cartridge was employed. Gas chromatography coupled to ion-trap tandem mass spectrometry (GC-MS/MS) and gas chromatography-mass spectrometry in electron capture negative ionization mode, were used for PCN and SCCP determinations, respectively, while for PCB analysis gas chromatography with electron capture detection (GC-ECD) was used. The method developed provided low limits of detection (0.001-0.003 ng g(-1) dry weight (dw) for PCNs, 1.8 ng g(-1) for SCCPs and 0.006-0.014 ng g(-1)dw for PCBs) and good run-to-run precisions (lower than RSD 8%) for the analysis of sediment samples. Concentration levels ranging from 0.17 to 3.27 ng g(-1)dw for PCNs, between 0.21 and 1.17 microg g(-1)dw for SCCPs, and from 2.33 to 44.00 ng g(-1) (dw) for PCBs, were found in the coastal sediments, while for samples collected near to the submarine emissary higher levels (from 2.02 to 6.56 ng g(-1)dw for PCNs, between 1.25 and 2.09 microg g(-1)dw for SCCPs and from 22.34 to 37.74 ng g(-1)dw for PCBs) were obtained. The results obtained provide new data about the occurrence of PCN and SCCP in the coastal area of Barcelona.

Occurrence and bioaccumulation study of PCDD and PCDF from mineral feed additives.

Extraction of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) was evaluated in sepiolite matrices. Soxhlet extraction using different extracting strategies and acid treatment were tested. Acid treatment or Soxhlet extraction using a mixture of toluene:ethanol as solvent allowed to reach the minimum requirements for recovery rates. However, Soxhlet extraction using a mixture cyclohexane:toluene as extracting solvent did not allow to comply with these minimum requirements. Significant differences were obtained in TEQ units when acid treatment was applied in comparison to Soxhlet extraction. This fact can be explained because the use of drastic acid conditions allows removing strongly adsorbed analytes which can be uniquely extracted after a total destruction of the crystalline. On the contrary, Soxhlet extraction was not able to destroy the structure of sepiolite and as a consequence the PCDDs/Fs remain adsorbed in the internal structure of the mineral. A bioaccumulation study was also conducted to evaluate the transference of PCDDs/Fs from the sepiolite into the animal when fed with feed containing sepiolite. Four groups of chickens were exposed through diet to a control feed, feed with 3% w/w sepiolite, spiked feed contaminated with PCDDs/Fs and feed containing contaminated kaolinitic clay. Livers of animals were analyzed throughout the exposure period. Results of this trial showed that the performance of broilers was not affected by the presence of dioxins at levels tested, and chickens did not show any abnormal behaviour. Dioxins intentionally added to the diet were significantly absorbed and accumulated in liver, whereas the PCDDs/Fs from sepiolite were not available for chickens since livers from broilers fed 3% sepiolite presented similar WHO-TEQ values than those from broilers fed control diet.

Long-term monitoring programme of polychlorinated dioxins and polychlorinated furans in ambient air of Catalonia, Spain (1994-2015).

Reductions in concentrations of persistent organic pollutants were observed for polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) in a comparison of two consecutive decades ending in December 31, 2015 (n = 413) and compiled from a 48-station long-term air-monitoring network in Catalonia, Spain. Reductions were observed in geometric mean concentration of total PCDD/Fs, decreasing at a rate of 11% yr-1. Results reflect the relative success of improved pollution control measures at industry and traffic locations. At traffic and industrial locations, PCDD/Fs were reduced by around 68% (from 70 to 22 fg I-TEQ/m3) and 66% (from 57 to 19 fg I-TEQ/m3), respectively. The highest concentration of 1196 fg I-TEQ/m3 was observed during the first decade at an industrial location. The highest concentration over the second decade was at a background location during winter (674 fg I-TEQ/m3). Winter observations are generally higher than those recorded in summer, as evident in an 8-year period of moderate PCDD/F levels (<250 fg I-TEQ/m3, n = 143). PCDD/F congener profiles were identified has having less chlorinated PCDD/F, indicating the presence of nearby combustion sources at selected locations. More long-term sampling campaigns, combined with network optimization and integration into a meteorological model, can offer a tool for future PCDD/Fs control measures in not only Catalonia - but also applicable to other areas.

Polychlorinated dibenzo-p-dioxins, dibenzofurans and dioxin-like PCBs in commercialized food products from Colombia.

Polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and dioxin-like polychlorinated biphenyls (dl-PCBs) are commonly known as dioxins and are the most toxic members of persistent organic pollutants (POPs) because present a variety of health effects especially as promoting agent of growing and transformation of cancer cells. They are bio-accumulate in humans primarily via the diet, specifically by ingestion of foods that have high lipid content which are generally associated with foods of animal origin such as oils and fats and with fishery and dairy products. In Colombia the Ministry of Health and Social Protection which is the entity responsible for surveillance food conditions, has established maximum levels for dioxins and dl-PCBs in oils from animal and vegetable origins. Oils of vegetable and animal origin represent an appreciable intake in the country thus the presence of dioxins and dl-PCBs in these materials is a matter of concern because they can bioaccumulate in fat. In this contribution the levels of PCDD/Fs and dl-PCBs in olive, soybean, fish oil, butter and shrimp consumed in Colombia were determined using HRGC-HRMS and were compared with the maximum levels permitted in oil samples according to both the Colombian and European regulations. WHO-TEQ concentrations for PCDD/Fs and dioxin like PCBs ranged from 0.24 to 1.710pgWHO-TEQ PCDD/Fg(-1) of fat and from 0.050 to 3.000pgWHO-TEQ PCBg(-1) of fat, respectively. As expected, fish oils and shrimp present the highest WHO-TEQ PCDD/Fs and dl-PCBs values followed by butter and soybean oil sample, while the olive oil shows the lowest levels. In general, the vegetable oils show levels below the limits established by both the Colombian and European regulations. The levels from soybean oil found in this study were slightly higher than the threshold established both by the Commission Regulation European Union (EU) and the Colombian legislation, while fish oils showed concentrations above the European regulations. Furthermore, shrimp exhibited values below the maximum concentration levels established by the EU and Colombian regulation.

Quadrupole mass spectrometer operating in the electron-capture negative ion mode as detector for comprehensive two-dimensional gas chromatography.

A recently introduced rapid-scanning quadrupole mass spectrometer (qMS) with an electron-capture negative ion (ECNI) option, the Perkin-Elmer Clarus 500, was tested as a detector for comprehensive two-dimensional gas chromatography (GC x GC). The parameters influencing the data acquisition rate in the scan mode, such as scan time and inter-scan delay, and in the selected ion monitoring mode, such as dwell time and inter-channel delay, were evaluated. In the scan mode, good-quality mass spectra covering a range of 300 Da can be obtained at an acquisition rate of 23 Hz; in selected ion monitoring, an acquisition rate of 90Hz can be achieved when monitoring a single ion. Compared with electron ionisation, the use of electron-capture negative ionisation causes no extra peak broadening. As applications, mixtures of polychlorinated n-alkanes (PCAs), polybrominated diphenyl ethers (PBDEs) and polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) were analysed. The separation of PCAs based on their number of chlorine substituents was confirmed for the first time by using GC x GC-ECNI qMS in the scan mode and a significantly improved limit of detection was achieved for BDEs (10-150 fg injected) and CDD/Fs (10-700 fg injected) in the selected ion monitoring mode.

Characterization of polychlorinated n-alkanes using comprehensive two-dimensional gas chromatography--electron-capture negative ionisation time-of-flight mass spectrometry.

Comprehensive two-dimensional gas chromatography with electron-capture negative ionization time-of-flight mass spectrometry (GC x GC-ECNI-TOF-MS) is used to study the composition and characteristics of short-, medium- and long-chain polychlorinated n-alkane (PCA) mixtures. Distinct ordered structures, which enable the highlighting and interpretation of group and sub-group separations are observed when using a DB-1 x 007-65HT column combination. The analysis of a number of, mutually rather different, technical mixtures and 35 individual standard compounds provides information on the role of chlorine substitution (number of substituents as well as their position), the contribution of carbon versus chlorine atoms to analyte volatility, i.e. GC x GC behaviour, and the influence of the chain length of the carbon skeleton. Two dust samples are analyzed to illustrate the practical usefulness of the proposed procedure.

Microwave-assisted extraction versus Soxhlet extraction for the analysis of short-chain chlorinated alkanes in sediments.

Microwave-assisted extraction (MAE) was evaluated as a possible alternative to Soxhlet extraction for analysing short-chain chlorinated alkanes (commonly called short-chain chlorinated paraffins, SCCPs) in river sediment samples, using gas chromatography coupled to negative chemical ionisation mass spectrometry. For MAE optimisation, several extraction parameters such as solvent extraction mixture, extraction time and extraction temperature were studied. Maximum extraction efficiencies for SCCPs (90%) and for 12 polychlorinated biphenyl (PCB) congeners (91-95%) were achieved using 5 g of sediment sample, 30 ml of n-hexane-acetone (1:1, v/v) as solvent extraction, and 15 min and 115 degrees C of extraction time and temperature, respectively. Activated Florisil was used to clean-up the extracts, allowing highly selective separation of SCCPs from other organic contaminants such as PCBs. MAE was compared with a conventional extraction technique such as Soxhlet and good agreement in the results was obtained. Quality parameters of the optimised MAE method such as run-to-run (R.S.D. 7%) and day-to-day precision (R.S.D. 9%) were determined using spiked river sediment samples, with LODs of 1.5 ng g(-1). This method was successfully applied to the analysis of SCCPs in river sediment samples at concentrations below the ng g(-1) level. O 2004 Elsevier B.V. All rights reserved.

Polychlorinated dibenzo-p-dioxins, dibenzofurans, biphenyls, paraffins and polybrominated diphenyl ethers in marine fish species from Ebro River Delta (Spain).

The results of a surveillance programme on the determination of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs) and short chain chlorinated paraffins (SCCPs) in marine fish and shellfish species which are fished, commercialised and consumed in the Ebro River Delta area (NE, Spain) are presented. The study included the analysis of five marine fish species (sardine, gilthead sea bream, conger, eel and flounder) and three shellfish species (murex, carpet shell and mussel) collected in 2012 in five fishing harbours near to this area. WHO-TEQ concentrations for PCDD/Fs and dioxin like PCBs (dl-PCBs) ranged from 0.03 to 0.31 pg WHO-TEQ2005PCDD/Fg(-1)wetweight (ww) and from 0.02 to 3.15 pg WHO-TEQ2005PCBg(-1)ww, respectively. All levels were below the maximum concentrations established by the EU Regulation. The PCBs and PCDD/Fs accumulation pattern found in the samples analysed showed a distribution typically reported for marine samples. For marker PCBs and PBDEs, concentration levels ranging from 929 to 57494 pg g(-1)ww and from 36.2 to 827 pg g(-1)ww were obtained, respectively, meanwhile for SCCPs levels were between 3.1 and 141 ng g(-1)ww. Finally, the trends in the levels of PCDD/F and dl-PCBs found from 2006 to 2012 in fish and shellfish species were studied. A slight decrease of PCDD/F and dl-PCB concentrations was found since 2006.

Evolution of PCDD/Fs and dioxin-like PCBs in the general adult population living close to a MSW incinerator.

It is well known that municipal solid waste incinerators (MSWI) produce dioxins and furans. The aim of this study was to assess the serum levels of compounds with dioxin-like activity such as polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and dioxin-like PCBs after the commissioning of a MSWI plant. In 2006 and 2008 serum samples were obtained from 322 and 326 volunteers, respectively, from the general population, some living in the vicinity of the plant and others, as a control group, living further away from the MSWI plant and out of the path of prevailing winds. Samples for sets of 20 individuals, grouped by age and sex, were pooled, obtaining 16 pooled samples per year. No significant differences were observed in PCDD/Fs as a function of area of residence or proximity to the plant over the time of the study, with levels of WHO-TEQ/g lipid nearby and far from the MSWI being 26.9 and 20.0pg, respectively, in 2006 and 20.8 and 26.4pg, respectively, in 2008. There was a significant decrease in the concentration of non-ortho PCBs in the nearby areas, whilst there was a significant increase in the concentration of mono-ortho PCBs, both in the nearby areas and in those further away from the MSWI. Age was significantly associated with PCDD/Fs and dioxin-like PCBs, but sex showed no significant association. Overall, this study found that there was no increase in the levels of compounds with dioxin-like activity in a population living close to an MSWI, in agreement with the literature reviewed, for plants of recent construction using state-of-the-art technology.

Suitability of selective pressurized liquid extraction combined with gas chromatography-ion-trap tandem mass spectrometry for the analysis of polybrominated diphenyl ethers.

A one-step extraction and clean-up method using pressurized liquid extraction (PLE) (selective PLE) combined with gas chromatography-ion-trap tandem mass spectrometry (GC-ITMS-MS) was evaluated for the analysis of polybrominated diphenyl ethers (from tri- to hepta-PBDEs) at low concentrations in fish and shellfish samples. To this end, the performance of an on-line PLE extraction/clean-up method and of a classical Soxhlet extraction and clean-up method using a multi-layer modified silica column were compared. The two sample treatment methods provided similar results, although an important reduction in the sample treatment time (40 min per sample) was achieved using the selective PLE method. In addition, the suitability of the PLE combined with GC-ITMS-MS method was evaluated by comparing the results obtained in the analysis of fish samples with those obtained by gas chromatography-high resolution mass spectrometry (GC-HRMS). Good agreement between both techniques was obtained with differences between the mean values of less than 16%. The selective PLE method coupled to GC-ITMS-MS produced accurate results for PBDE determination with low limits of detection (1.0-16.8 pg g(-1) wet weight) and quantification (3.1-51 pg g(-1) wet weight) as well as good precision (RSD<16%). This method has been applied to the analysis of PBDEs in fish and shellfish samples collected at fish markets in Catalonia (NE Spain).

Serum levels of polychlorinated dibenzodioxins and dibenzofurans and PCBs in the general population living near an urban waste treatment plant in Biscay, Basque Country.

Levels of PCDD/Fs and dioxin-like PCBs were measured in 16 pooled samples of serum from a total of 322 adults in the general population, to coincide with the start-up of a new municipal solid urban waste treatment plant in Biscay, Basque Country (Spain). Two hundred and eighty-three individual serum samples were also obtained, in which the most common PCBs (28, 52, 101, 118, 138, 153 and 180) were quantified. The samples were taken from four geographical zones: two from the metropolitan area of Bilbao, located less than 2 km from the plant and with high traffic density (Zones E1 and E2), a third located 5 km from the plant in an urban area of Bilbao, also with high traffic density (Zone C1) and the fourth located 20 km from the plant, in a municipality with minimal industrial activity and low traffic density (Zone C2), the latter two being out of the path of the prevailing winds. The median levels of dioxins+furans were similar by zone: E1=24.3, E2=27.3, C1=21.3, C2=18.8 pg g(-1) lipid (p=0.362); by sex: 20.2 vs. 22.6 pg g(-1) lipid in men and women (p=0.328); and by age: 20.8 vs. 21.3 pg g(-1) lipid in subjects aged 20-44 and 45-69 (p=0.505). No detectable levels of PCBs 52 and 101 were found. Significant differences by zone were found only for PCB 180 (p=0.041), with higher values in Zone C2, the zone with the lowest presumed contamination levels. Dioxin-like PCBs (p<0.001) and the most common PCBs (138, 153, 180) (p<0.001) were both statistically associated with age, higher values being found in the 45-69 age group.

Analysis of polychlorinated n-alkanes in environmental samples.

Polychlorinated n-alkanes (PCAs), also known as chlorinated paraffins (CPs), are highly complex technical mixtures that contain a huge number of structural isomers, theoretically more than 10,000 diastereomers and enantiomers. As a consequence of their persistence, tendency to bioaccumulation, and widespread and unrestricted use, PCAs have been found in aquatic and terrestrial food webs, even in rural and remote areas. Recently, these compounds have been included in regulatory programs of several international organizations, including the US Environmental Protection Agency and the European Union. Consequently, there is a growing demand for reliable methods with which to analyze PCAs in environmental samples. Here, we review current trends and recent developments in the analysis of PCAs in environmental samples such as air, water, sediment, and biota. Practical aspects of sample preparation, chromatographic separation, and detection are covered, with special emphasis placed on analysis of PCAs using gas chromatography-mass spectrometry. The advantages and limitations of these techniques as well as recent improvements in quantification procedures are discussed.