Halogenated Organic Contaminants Research Articles

Thin-layer chromatography coupled with HPLC-DAD/UHPLC-HRMS for target and non-target determination of emerging halogenated organic contaminants in animal-derived foods

In this study, a simple, solvent-saving, and sensitive method was established using high-performance liquid chromatography coupled with high-resolution mass spectrometry to quantitively determine 16 emerging halogenated organic contaminants (HOCs), including polyhalogenated carbazoles, halogenated phenols, and tetrabromobisphenol-A analogs, and to qualitatively identify non-target HOCs in animal-derived food samples. The sample extracts were physically frozen to remove most lipids and further purified by thin-layer chromatography according to the targets polarity. The method detection limit and method quantification limit of 16 HOCs were in the range of 0.003–9.168 and 0.010–30.601 ng·g−1 dry weight, respectively. The recoveries were within 69.1–111.0 %, the intra/inter-day precisions were 0.1–6.1 % and 0.1–6.7 %, and the matrix effects were between −12.1 and 10.8 %, all within the acceptable range. Finally, 16 HOCs were detected in nine actual samples in range of not detected–307.22 ng·g−1 dry weight. Moreover, five bromides and two chlorides were identified by using non-target analysis in animal-derived foods.

Halogenated organic contaminants of concern in urban-influenced waters of Lake Ontario, Canada: Passive sampling with targeted and non-targeted screening

Passive samplers are useful tools for monitoring hydrophobic, persistent, and potentially bioaccumulative contaminants in the environment. In this study, low density polyethylene passive samplers were deployed in urban-influenced and background nearshore freshwaters of northwestern Lake Ontario and analyzed for a broad range of both legacy halogenated organic contaminants (HOCs) and halogenated flame retardants (HFRs). Non-targeted analysis was conducted for screening additional halogenated substances. For most compounds, concentrations were greatest in the industrialized Hamilton Harbour and more generally at sites that have stronger influences of wastewater effluent discharges and stormwater run-off through rivers and creeks. Polychlorinated biphenyls (PCBs) remain the dominant class of HOCs in water, with dissolved-phase concentrations ranging from 10 to 4100 pg/L (ΣPCBs), followed by polybrominated diphenylethers (ΣPBDEs; 14–960 pg/L) and the organochlorine pesticides (OCPs; 22–290 pg/L). Several non-PBDE brominated flame retardants (nBFRs) and chlorinated Dechlorane-related compounds were detected, with hexabromocyclododecanes (ΣHBCDD; sum of 3 diastereoisomers) the most abundant (1.0–21 pg/L). Non-targeted screening of samples by high resolution mass spectrometry using Kendrick mass defect plots for data analysis indicated that several other halogenated compounds were present in waters at relatively high abundances compared to the flame retardants, based on semi-quantitative estimates. These included methyl-triclosan, four halogenated anisoles (2,4,6-tribromoanisole, dimethyl-trichloroanisole, pentachloroanisole, and pentachlorothioanisole), and pentachloro-aniline. Dissolved-phase methyl-triclosan was estimated to contribute up to approximately 40% of the summed target HOC concentrations. Polyethylene passive samplers provided an excellent medium for both non-targeted screening of HOCs not currently included in monitoring programs and tracking brominated and chlorinated chemicals slated for reductions in uses and emissions through international (Stockholm Convention) and binational (Great Lakes) agreements.

Contaminant Exposure Linked to Cellular and Endocrine Biomarkers in Southern California Bottlenose Dolphins.

Cetaceans in the Southern California Bight (SCB) are exposed to high levels of halogenated organic contaminants (HOCs), which have previously been linked to impaired reproductive health and immune responses. We used a combination of molecular tools to examine the potential physiological impacts of HOC exposure in two bottlenose dolphin ( Tursiops truncatus) ecotypes in the SCB. We quantified 25 HOCs in the blubber of 22 biopsies collected from males between 2012 and 2016. We then analyzed genome-wide gene expression in skin using RNA-sequencing and measured blubber testosterone to compare HOC exposure with cellular and endocrine biomarkers. We found high levels of HOCs in both ecotypes with significantly higher total polychlorinated biphenyls(PCBs), polybrominated diphenyl ethers (PBDEs), tris(4-chlorophenyl)methanol(TCPMOH), and chlordane-related compounds in the coastal ecotype versus the offshore ecotype. We found evidence of PBDE bioaccumulation in both ecotypes, however, the pattern of bioaccumulation or endocrine disruption for other HOCs was different between the ecotypes, suggesting potential endocrine disruption in the coastal ecotype. We also observed correlations between HOCs and gene coexpression networks enriched for xenobiotic metabolism, hormone metabolism, and immune response that could indicate cellular effects from HOC exposure. By integrating measurements of HOC load with both transcriptome profiling and endocrine biomarkers, our approach provides insight into HOC exposure and potential impacts on wild cetacean health in southern California.

The effect of long-range transport, trophic position and diet specialization on legacy contaminant occurrence in great skuas, Stercorarius skua, breeding across the Northeast Atlantic

High levels of halogenated organic contaminants (HOCs) have been found in the marine predatory seabird great skua (Stercorarius skua) from breeding colonies in the Northeastern Atlantic, with large unexplained inter-colony variation. The present study aimed at analyzing if the HOCs occurrence in breeding great skuas in remote colonies was explained by local baseline food web exposure determined by long-range transport, or by ecological factors such as diet specialization and relative trophic position in the breeding area. The occurrence of organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), and polybrominated diphenyl ethers (PBDEs) was analyzed in plasma of 204 adult great skuas collected over two years (2008 and 2009) and 5 colonies across the North-Atlantic from Shetland to Svalbard. The ΣHOCs levels in plasma ranged across two orders of magnitude, from 40 to 7600 ng/g (wet weight) and differed significantly across the great skua colonies. The variation in contaminant occurrence among colonies did not reflect long-range transport through a latitudinal or remoteness gradient, as the second northernmost colony (Bjørnøya), had the highest contaminant concentrations. No latitudinal or remoteness gradient was evident in the contaminant pattern among the colonies. The contaminant levels increased significantly with increasing δ15N values, and regurgitated pellets of undigested prey suggested that great skuas with higher δ15N values had a higher proportion of bird prey in their diet, mostly seabirds. In contrast, great skuas from colonies with lower δ15N and lower contaminant level fed mostly on fish. The enrichment of δ13C increased with decreasing δ15N and lower contaminant levels. Therefore, individual behavior of great skuas, such as migration strategies and diet specialization, rather than long-range transport and thus baseline food web exposure, explain among and within colony variance in contaminant occurrence.

Effects of contaminant exposure and food restriction on hepatic autophagic lysosomal parameters in Herring Gull (Larus argentatus) chicks

Lysosomal autophagic responses, such as lysosomal membrane stability, neutral lipids (NL), lipofuscin (LF), and malondialdehyde (MDA) levels, are valuable measures of cellular early-onset effects induced by environmental stress factors, such as contaminant exposure and fasting. In this study, these parameters were analysed and related to levels of halogenated organic contaminants (HOCs) in 40 Herring Gull (Larus argentatus) chicks. Chicks were experimentally exposed to HOCs through diet and went through a period of nutrient deprivation at the end of the experiment. HOC exposure and fasting were conducted separately and in combination. NL storages were depleted, and lysosomal membranes were destabilised after HOC exposure and nutrient deprivation. These responses were not related specifically to one type of stress or the extent of the treatment. No synergistic or additive effects from the combination of HOC exposure and fasting were observed. LF accumulated, and MDA levels increased as a result of fasting, but were unaffected by HOC exposure. LF accumulation was strongly associated with the percent weight change in the chicks. Large weight loss was associated with high LF levels, and slight weight gain was associated with low LF levels. Hence, food deprivation affected all the measured parameters, and HOC exposure decreased NL levels and lysosomal membrane stability in HG chick liver. Furthermore, autophagic lysosomal parameters have frequently been applied as biomarkers of cellular health status in previous studies of marine and terrestrial invertebrates, and this study suggests that these parameters may be good candidates for biomarkers of cellular health status in seabirds as well.

Inhibition of Thyroid Hormone Sulfotransferase Activity by Brominated Flame Retardants and Halogenated Phenolics

Many halogenated organic contaminants (HOCs) are considered endocrine disruptors and affect the hypothalamic-pituitary-thyroid axis, often by interfering with circulating levels of thyroid hormones (THs). We investigated one potential mechanism for TH disruption, inhibition of sulfotransferase activity. One of the primary roles of TH sulfation is to support the regulation of biologically active T3 through the formation of inactive THs. We investigated TH sulfotransferase inhibition by 14 hydroxylated polybrominated diphenyl ethers (OH BDEs), BDE 47, triclosan, and fluorinated, chlorinated, brominated, and iodinated analogues of 2,4,6-trihalogenated phenol and bisphenol A (BPA). A new mass spectrometry-based method was also developed to measure the formation rates of 3,3'-T2 sulfate (3,3'-T2S). Using pooled human liver cytosol, we investigated the influence of these HOCs on the sulfation of 3,3'-T2, a major substrate for TH sulfation. For the formation of 3,3'-T2S, the Michaelis constant (Km) was 1070 ± 120 nM and the Vmax was 153 ± 6.6 pmol min(-1) (mg of protein)(-1). All chemicals investigated inhibited sulfotransferase activity with the exception of BDE 47. The 2,4,6-trihalogenated phenols were the most potent inhibitors followed by the OH BDEs and then halogenated BPAs. The IC50 values for the OH BDEs were primarily in the low nanomolar range, which may be environmentally relevant. In silico molecular modeling techniques were also used to simulate the binding of OH BDE to SULT1A1. This study suggests that some HOCs, including antimicrobial chemicals and metabolites of flame retardants, may interfere with TH regulation through inhibition of sulfotransferase activity.

Effect of reduced food intake on toxicokinetics of halogenated organic contaminants in herring gull (Larus argentatus) chicks

The aim of the present study was to investigate how contaminant exposure and reduced food intake affect tissue distribution and biotransformation of halogenated organic contaminants (HOCs) in Arctic seabirds using herring gull (Larus argentatus) as a model species. Herring gull chicks were exposed for 44 d to cod liver oil containing a typical mixture of contaminants. Following exposure, food intake was reduced for a one-week period in a subgroup of the chicks. Polyclorinated biphenyls, organochlorine pesticides, and brominated flame retardants, as well as a wide range of hydroxy, methyl sulfone, and methoxy compounds were measured in liver, brain, and plasma samples. Additionally, phase I biotransformation enzyme activities and phase I and II messenger ribonucleic acid (mRNA) expression were investigated in the liver, brain, or both. Both contaminant exposure and reduced food intake had an increasing effect on the concentrations of HOCs and their metabolites. The HOC exposure and reduced food intake also led to increased 7-ethoxyresorufin-O-deethylation (EROD) activity, whereas mRNA expression of the biotransformation enzymes increased only following the reduced food intake. Tissue distribution of HOCs and their metabolites was not affected by either contaminant exposure or reduced food intake. In conclusion, the results indicate that biotransformation capacity and formation of HOC metabolites increase during reduced food intake. This finding supports the hypothesis that reduced food intake increases the susceptibility of Arctic animals to the effects of lipophilic HOCs.

Halogenated organic contaminants (HOCs) in sediment from a highly eutrophicated lake, China: Occurrence, distribution and mass inventories

Halogenated organic contaminants (HOCs) including 16 polybrominated diphenyl ethers (PBDEs) and 37 polychlorinated biphenyls (PCBs) were determined in 49 surfacial sediments from Chaohu Lake, a highly eutrophicated lake, China. PBDEs were detected in almost samples with the range of the total concentration (defined as Σ16PBDEs) from 0.84 to 86.6ngg−1. Compared with the occurrence of PBDEs in Pearl River Delta and Yangtze River Delta in China, lower percentage of BDE-209 over the concentration of Σ16PBDEs was inferred by the high-volume application of penta-BDE mixture product for local domestic furniture purpose. The total concentration of 37 PCBs (Σ37PCBs) ranged from 0.05 to 3.36ngg−1 with the most detection of PCB-1, -4, -52 and -71. Both the concentrations of Σ16PBDE and Σ37PCB poorly correlated with total organic carbon (TOC), suggesting the significant contribution of phytoplankton organic carbons to sediment TOC. The contamination by PBDEs and PCBs in western region of the lake was significantly more serious than in eastern lake. Our findings about the higher residues of PBDEs and PCBs in sediments at the estuary of Nanfei River compared to the other estuaries also supported the conclusion that urban area (Hefei city) was the main source of PBDEs and PCBs. The comparison with the concentration of HOC in the present study with those in other lacustrine sediments around the world suggested the contamination by PBDEs in Chaohu Lake is at middle of the global concentration range, whereas PCBs is at low end of the global range which could be elucidated by local economic development and historical usage of PBDEs and PCBs. The mass inventories of HOCs in the lake were estimated at 561 and 38kg, which corresponds to only 0.000006% and 0.0001% of these global historical produce volumes, respectively.

Halogenated organic contaminants and their correlations with circulating thyroid hormones in developing Arctic seabirds

Thyroid hormones are essential for normal growth and development and disruption of thyroid homeostasis can be critical to young developing individuals. The aim of the present study was to assess plasma concentrations of halogenated organic contaminants (HOCs) in chicks of two seabird species and to investigate possible correlations of HOCs with circulating thyroid hormone (TH) concentrations. Plasma from black-legged kittiwake (Rissa tridactyla) and northern fulmar (Fulmarus glacialis) chicks were sampled in Kongsfjorden, Svalbard in 2006. The samples were analyzed for thyroid hormones and a wide range of HOCs (polychlorinated biphenyls (PCBs), hydroxylated (OH−) and methylsulphoned (MeSO−) PCB metabolites, organochlorine pesticides (OCPs), brominated flame retardants (BFRs), and perfluorinated compounds (PFCs)).Concentrations of HOCs were generally low in kittiwake and fulmar chicks compared to previous reports. HOC concentrations were five times higher in fulmar chicks compared to in kittiwake chicks. PFCs dominated the summed HOCs concentrations in both species (77% in kittiwakes and 69% in fulmars).Positive associations between total thyroxin (TT4) and PFCs (PFHpS, PFOS, PFNA) were found in both species. Although correlations do not implicate causal relationships per se, the correlations are of concern as disruption of TH homeostasis may cause developmental effects in young birds.

Relationship between persistent halogenated organic contaminants and TCDD-toxic equivalents on EROD activity and retinoid and thyroid hormone status in northern fulmars

We investigated whether the hepatic cytochrome P450 1A activity (measured as 7-ethoxyresorufin- O-deethylase (EROD)) and plasma thyroid hormone and liver retinoid concentrations were explained by liver and blood levels of halogenated organic contaminants (HOCs) in free-ranging breeding northern fulmars ( Fulmarus glacialis) from Bjørnøya in the Norwegian Arctic. Hepatic EROD activity and liver levels of 2,3,7,8-tetrachlorodibenzo- p-dioxin toxic equivalents (TEQs) were positively correlated, suggesting that hepatic EROD activity is a good indicator for dioxin and dioxin-like HOC exposure in breeding northern fulmars. There were not found other strong relationships between HOC concentrations and hepatic EROD activity, plasma thyroid or liver retinoid concentrations in the breeding northern fulmars. It is suggested that the HOC levels found in the breeding northern fulmars sampled on Bjørnøya were too low to affect plasma concentrations of thyroid hormones and liver levels of retinol and retinyl palmitate, and that hepatic EROD activity is a poor indicator of polychlorinated biphenyl (PCB) and pesticide exposure.

Biotransformation of PCBs in Arctic seabirds: Characterization of phase I and II pathways at transcriptional, translational and activity levels

Arctic seabirds are exposed to a wide range of halogenated organic contaminants (HOCs). Exposure occurs mainly through food intake, and many pollutants accumulate in lipid-rich tissues. Little is known about how HOCs are biotransformed in arctic seabirds. In this study, we characterized biotransformation enzymes in chicks of northern fulmars ( Fulmarus glacialis) and black-legged kittiwakes ( Rissa tridactyla) from Kongsfjorden (Svalbard, Norway). Phase I and II enzymes were analyzed at the transcriptional, translational and activity levels. For gene expression patterns, quantitative polymerase chain reactions (qPCR), using gene-sequence primers, were performed. Protein levels were analyzed using immunochemical assays of western blot with commercially available antibodies. Liver samples were analyzed for phase I and II enzyme activities using a variety of substrates including ethoxyresorufin (cytochrome (CYP)1A1/1A2), pentoxyresorufin (CYP2B), methoxyresorufin (CYP1A), benzyloxyresorufin (CYP3A), testosterone (CYP3A/CYP2B), 1-chloro-2,4-nitrobenzene (CDNB) (glutathione S-transferase (GST)) and 4-nitrophenol (uridine diphosphate glucuronyltransferase (UDPGT)). In addition, the hydroxylated (OH-) polychlorinated biphenyls (PCBs) were analyzed in the blood, liver and brain tissue, whereas the methylsulfone (MeSO 2-) PCBs were analyzed in liver tissue. Results indicated the presence of phase I (CYP1A4/CYP1A5, CYP2B, and CYP3A) and phase II (GST and UDPGT) enzymes at the activity, protein and/or mRNA level in both species. Northern fulmar chicks had higher enzyme activity than black-legged kittiwake chicks. This in combination with the higher ΣOH-PCB to parent PCB ratios suggests that northern fulmar chicks have a different biotransformation capacity than black-legged kittiwake chicks.

Organochlorine and PBDE Concentrations in Relation to Cytochrome P450 Activity in Livers of Forster’s Terns (Sterna forsteri) and Caspian Terns (Hydroprogne caspia), in San Francisco Bay, California

We measured halogenated organic contaminants (HOCs) [polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs), and dichloro-diphenyl-trichloroethane (DDT)] and P450 [e.g., ethoxyresorufin-O-dealkylase (EROD)] stress in livers from Caspian tern (Hydroprogne caspia) adults and Forster's tern (Sterna forsteri) adults and chicks in San Francisco Bay (SFB). Penta BDEs and tetra PBDEs composed 46-66% of SigmaPBDE in terns. PCB homologues di, tri, penta, hexa, and hepta composed 93-95% of SigmaPCBs and p'p-DDE composed 82-98% of all SigmaDDTs. We found similar concentrations of SigmaPBDEs [mean micrograms per gram wet weight (ww) +/- standard error = 0.4 +/- 0.1], SigmaPCBs (5.9 +/- 1.6), and SigmaDDTs (0.6 +/- 0.1) among species, sexes, and regions. However, concentrations were higher in Forster's tern adults than chicks (SigmaPBDEs = 0.4 +/- 0.1 and 0.1 +/- 0.1; SigmaPCBs = 7.08 +/- 2.4 and 2.4 +/- 1.4; SigmaDDTs = 0.5 +/- 0.1 and 0.1 +/- 0.1; respectively), and there was a nonsignificant trend of elevated SigmaPBDEs and SigmaPCBs for adult Forster's terns in the Central South Bay and Lower South Bay portions of SFB. Combined Forster's tern and Caspian tern SigmaDDTs bioaccumulated similarly to selenium, but not mercury, and there was a nonsignificant but positive trend for SigmaPBDEs and SigmaPCBs bioaccumulation with mercury. P450 protein activity was higher in adult Forster's terns than Caspian terns, higher in Central South Bay than in Lower South Bay, and higher in adult Forster's terns than in chicks.

Halogenated organic contaminants and mercury in northern fulmars ( Fulmarus glacialis): levels, relationships to dietary descriptors and blood to liver comparison

The northern fulmar ( Fulmarus glacialis) is an interesting candidate for the study of patterns and levels of halogenated organic contaminants (HOCs) since they accumulate high levels of certain HOCs. In the present study we characterized a suite of established and novel HOCs in northern fulmars breeding on Bjørnøya in the Norwegian Arctic. A comparison between blood and liver HOC levels was made, and the levels were related to the ratios of heavier to lighter stable isotopes of nitrogen 15N/ 14N (δ 15N) and carbon 13C/ 12C (δ 13C) in muscle. A significant difference in congener patterns between blood and liver was found. The δ 13C was not related to HOCs, neither in liver nor in blood. Weak correlations were found between δ 15N and liver HOC levels. The 2,3,7,8-tetrachlorodibenzo- p-dioxin toxic equivalents (TEQs) in northern fulmars were well above thresholds for reproductive effects in seabirds.

Modern and historical fluxes of halogenated organic contaminants to a lake in the Canadian arctic, as determined from annually laminated sediment cores

Two annually laminated cores collected from Lake DV09 on Devon Island in May 1999 were dated using 210Pb and 137Cs, and analyzed for a variety of halogenated organic contaminants (HOCs), including polychlorinated biphenyls (PCBs), organochlorine pesticides, short-chain polychlorinated n-alkanes (sPCAs), polychlorinated dibenzo- p-dioxins and dibenzofurans (PCDD/Fs), and polybrominated diphenyl ethers (PBDEs). Dry weight HOC concentrations in Lake DV09 sediments were generally similar to other remote Arctic lakes. Maximum HOC fluxes often agreed well with production maxima, although many compound groups exhibited maxima at or near the sediment surface, much later than peak production. The lower than expected HOC concentrations in older sediment slices may be due to anaerobic degradation and possibly to dilution resulting from a temporary increase in sedimentation rate observed between the mid-1960s and 1970s. Indeed, temporal trends were more readily apparent for those compound classes when anaerobic metabolites were also analyzed, such as for DDT and toxaphene. However, it is postulated here for the first time that the maximum or increasing HOC surface fluxes observed for many of the major compound classes in DV09 sediments may be influenced by climate variation and the resulting increase in algal primary productivity which could drive an increasing rate of HOC scavenging from the water column. Both the fraction ( F TC) and enantiomer fraction (EF) of trans-chlordane (TC) decreased significantly between 1957 and 1997, suggesting that recent inputs to the lake are from weathered chlordane sources. PCDD/Fs showed a change in sources from pentachlorophenol (PeCP) in the 1950s and 1960s to combustion sources into the 1990s. Improvements in combustion technology may be responsible for the reducing the proportion of TCDF relative to OCDD in the most recent slice.