Marine Mammal Blubber Research Articles

Distribution Patterns Suggest Biomagnification of Halogenated 1′-Methyl-1,2′-Bipyrroles (MBPs)

The halogenated 1'-methyl-1,2'-bipyrroles (MBPs) are a suite of marine natural products that have been detected in marine mammals worldwide. Although their concentrations are similar to persistent organic pollutants that biomagnify, such as 2,2',4,4',5,5'-hexachlorobiphenyl (CB-153), it is notyet clearthat these natural products also biomagnify. Here we analyze MBPs and CB-153 isolated from the blubber and liver of marine mammals stranded on the eastern coast of Massachusetts. Four odontocete species (Delphinus delphis, Lagenorhynchus acutus, Phocoena phocoena, and Globicephala melas) and two pinniped species (Halichoerus grypus and Phoca groenlandica) were sampled. MBPs were present in all odontocetes, but not detected in pinnipeds; CB-153 was detected in every species. MBP patterns indicative of biomagnification were found, including age-dependent concentration increases and reduced concentrations in adult females. Also explored were the similarities and differences with CB-153, the effects of nutritional state on contaminant distribution, and the maternal transfer of blubber-based organic contaminants.

Identification and quantification of new polybrominated dimethoxybiphenyls (PBDMBs) in marine mammals from Australia

Marine mammals from Queensland, Australia, are bioaccumulating elevated concentrations of a range of polybrominated natural products. In this study, we detected three new polybrominated dimethoxybiphenyls (PBDMBs) in the blubber of selected marine mammal samples which were identified as 2,6′-dimethoxy-3,3′,5-tribromobiphenyl (2,6′-diMeO-BB 36), 2,2′-dimethoxy-3,3′-dibromobiphenyl (2,2′-diMeO-BB 36), and 6,6′-dimethoxy-3,3′-dibromobiphenyl (6,6′-diMeO-BB 11). These three PBDMBs are structurally related to the known natural product 2,2′-dimethoxy-3,3′,5,5′-tetrabromobiphenyl (2,2′-diMeO-BB 80). In the first part of this study, 2,2′-diMeO-BB 80 was photochemically debrominated under UV irradiation. This resulted in seven of eight possible mono- to triBDMBs as debromination products. In the second part of this study, the structure of all PBDMBs debromination products was investigated. This was supported by synthesis of two diBDMB and one triBDMB via bromination and subsequent methylation of 2,2′-biphenyldiol. Structures of the remaining PBDMBs were tentatively assigned by considering the retention times, mass spectra and amounts formed during UV irradiation of 2,2′-diMeO-BB 80 . In the third part of this study, blubber of marine mammals from Australia was analysed for PBDMBs using gas chromatography in combination with electron ionization mass spectrometry (GC/EI-MS) in the selected ion monitoring mode. In these samples, 2,2′-diMeO-BB 80 was found at concentrations of 200–1800 ng g −1 lipid weight (lw). The latter represents the highest concentration reported for this compound in environmental samples. 6,6′-diMeO BB 11, 2,2′-diMeO BB 36, and 2,6′-diMeO BB 36 were present at ∼7 ng g −1 lipids, or 0.43–1.5% of diMeO-BB 80. No further PBDMBs were detected in the samples. The di- and triBDMBs identified in marine mammal blubber have not been reported as natural products. They may represent either new natural products or transformation products of 2,2′-diMeO-BB 80.

Stratification, Composition, and Function of Marine Mammal Blubber: The Ecology of Fatty Acids in Marine Mammals

Abstract This study of vertical fatty acid profiles, based on analysis of 58 fatty acids sampled at 3-mm intervals throughout the blubber column of a model marine mammal, the ringed seal (Pusa hispida), revealed three chemically distinct layers. The average depths of the outer and inner layers were quite consistent (approximately 1.5 and approximately 1 cm, respectively). Consequently, the middle layer varied greatly in thickness, from being virtually absent in the thinnest animals to 2.5 cm thick in the fattest. The relative consistencies of the thickness and composition of the layers as well as the nature of the fatty acids making up each layer support the generally assumed function of the various layers: (1) the outer layer is primarily structural and thermoregulatory, (2) the inner layer is metabolically active with a fatty acid composition that is strongly affected by recent/ongoing lipid mobilization/deposition, and (3) the middle layer is a storage site that contracts and expands with food availability/consumption. The remarkable dynamics of the middle layer along with the discrete pattern of stratification found in the vertical fatty acid profiles have important implications for methodological sampling design for studies of foraging ecology and toxicology based on analyses of blubber of marine mammals.

Congener pattern of hexabromobiphenyls in marine biota from different proveniences

Blubber of seals, harbour porpoises and fish originating from the North Sea, the Baltic Sea, as well as coastal waters of Iceland and North America were analyzed for residues of polybrominated biphenyls. According to GC/ECNI-MS, hexabromobiphenyls (hexaBBs) dominated in the samples, followed by pentabromobiphenyls (pentaBBs) and heptabromobiphenyls (heptaBBs), whereas octabromobiphenyls (octaBBs) were only detected in selected samples and nonabromobiphenyls (nonaBBs) and PBB 209 were not found at all. Eleven peaks of hexaBBs—two peaks might have been each composed of two congeners, respectively—were detected in the different samples. The hexaBB pattern in samples of Iceland was a mixture of the pattern determined in samples from North America and continental Europe. In samples from the European continent, the major congener in technical hexabromobiphenyl, PBB 153, accounted for 11–37% of the hexaBBs. Other prominent isomers were PBB 155 (7.1–64%) and PBB 154 (11–33%), followed by PBB 133 as well as PBB 132 and/or PBB 146. Most of these PBBs were determined for the first time in marine biota and must be metabolites of higher brominated PBBs since they were not present in any technical product. PBB 153 was dominant in blubber of marine mammals from North America (92–96%). Blubber of harbour porpoises and also seal milk showed more peaks, which is indicative of a lower capability of biotransforming PBBs. It is concluded from this and other studies that PBB residues in North America are mainly originating from technical hexabromobiphenyl, while the bulk of the PBB residues in the European marine samples originated from technical octa- and decabromobiphenyl. Finally, it is evident from this study that a global ban of PBBs should not only be restricted to hexabromobiphenyls, but also needs to include higher brominated mixtures.

Cytochrome P4501A1 expression, polychlorinated biphenyls and hydroxylated metabolites, and adipocyte size of bottlenose dolphins from the Southeast United States

Persistent organic pollutants (POPs) bioaccumulate in blubber of marine mammals. Therefore, it is important to understand the structure and dynamics of blubber layers and how they affect the accumulation of POPs and subsequent biochemical responses. We used established histological and immunohistochemical methods to document the structure of bottlenose dolphin ( Tursiops truncatus) blubber and to assess the expression of cytochrome P4501A1 (CYP1A1) in skin-blubber biopsies of dolphins sampled in the waters off Charleston, SC (CHS) ( N = 38), and Indian River Lagoon, FL (IRL) ( N = 36). CYP1A1 expression was strongest and most frequent in capillary endothelial cells and was stratified in blubber; the greatest CYP1A1 staining was in the deepest layer. CYP1A1 expression in deep blubber and 2,3,7,8-TCDD Toxic Equivalents measured in the entire blubber were significantly higher in dolphins from CHS as compared to those from IRL. Adipocyte size was associated with the extent of CYP1A1 expression. Male dolphins with smaller adipocytes from CHS and IRL had higher levels of CYP1A1 expression in deep blubber. In CHS females, CYP1A1 expression in vascular endothelial cells varied with reproductive status. CYP1A1 expression in the deep layer was highest in simultaneously pregnant-lactating dolphins, and these dolphins had the smallest adipocytes in deep blubber. In all dolphins, CYP1A1 expression in the deep blubber layer was positively related to concentrations of hydroxylated PCBs (OH-PCBs) in plasma. In summary, redistribution of AHR agonists from blubber into the circulatory system may enhance PCB metabolism and production of OH-PCBs by induction of CYP1A1 in hepatocytes and, possibly, by induction of CYP1A1 in endothelial cells of the deep blubber. The OH-PCBs thus formed have the potential to interfere with thyroid hormone homeostasis.

Concentrations of Polychlorinated Biphenyls (PCBs), Polychlorinated Dibenzo-p-dioxins and Furans (PCDD/Fs), and Polybrominated Diphenyl Ethers (PBDEs) as Functions of Sample Depth in Killer Whale (Orcinus orca) Blubber

AbstractConcentrations of polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs), and polybrominated diphenyl ethers (PBDEs) were examined as a function of depth in killer whale (Orcinus orca) blubber samples. Lipid-normalized concentrations of PCBs, PCDD/Fs, and PBDEs did not display significant variation with depth in three distinct blubber layers (outer, central, and inner). Significantly more variation in contaminant concentrations were observed with depth on a wet weight basis for the killer whale sample. The current study indicates that non-invasive microdart biopsy sampling methods commonly used for monitoring contaminants in marine mammals yield representative details on contaminant burdens for chlorinated and brominated aromatic compounds in marine mammal blubber, regardless of the quantity and type of blubber sampled, provided that lipid normalization is performed on resulting analytical determinations.

Effects of organohalogen pollutants on haematological and urine clinical–chemical parameters in Greenland sledge dogs ( Canis familiaris)

Seven West Greenland sledge dog bitches ( Canis familiaris) and their three pups were fed 50–200 g of contaminated West Greenland minke whale ( Balaenoptera acutorostrata) blubber, and in a control cohort eight sister bitches and their five pups were fed a similar amount pork fat. Blood plasma and urine clinical–chemical parameters were measured and compared between the bitches and pups form the control and exposed cohorts. Based on existing reference intervals, Arctic mammals may have blood clinical–chemical endpoint levels that differ from comparable species at lower latitudes. The cortisol:creatinine ratio, protein:creatinine ratio, alkaline phosphatase, cholesterol and inorganic phosphate were significantly highest (ANCOVA: all p < 0.05 ) in the pup generation. The cortisol:creatinine ratio, cholesterol, lactate dehydrogenase and creatinine kinase were significantly higher (ANCOVA: all p < 0.05 ) in the control group, while glucose was significantly highest (ANCOVA: p < 0.05 ) in the exposed group. Furthermore, the blood cholesterol levels indicate that exposure via the diet to marine mammal blubber has a preventive effect on the development of cardiovascular diseases. We therefore suggest that the consumption of contaminated Arctic marine blubber impacted liver and kidney function in adult and pup sledge dogs.

Monobromo and higher brominated congeners of the marine halogenated natural product 2,3,3′,4,4′,5,5′-heptachloro-1′-methyl-1,2′-bipyrrole (Q1)

The marine halogenated natural product 2,3,3′,4,4′,5,5′-heptachloro-1′-methyl-1,2′-bipyrrole (Q1) is widely distributed in the environment. In this study, we screened samples which have previously been found to contain remarkably high residues of Q1 (blubber of marine mammals from Australia, samples from Antarctica, human milk from the Faroe Island) for the additional presence of mixed chlorinated and brominated congeners. Using GC/ECNI-MS, all samples tested were positive and many contained four out of five possible bromohexachloro congeners (BrCl 6-MBPs), five out of 14 possible dibromopentachloro congeners (Br 2Cl 5-MBPs), five of 21 possible tribromotetrachloro-congeners (Br 3Cl 4-MBPs), as well as several higher brominated congeners. About 20 heptahalo congeners of Q1 are described for the first time in the scientific literature. Isomers eluted within about one minute, respectively. Hence it is possible, that the peak clusters identified may be composed of more, co-eluting congeners. Similarities in the GC/ECNI-MS mass spectra with polychlorinated biphenyls (PCBs) were addressed. We also suggest an acronym system similar to that in use for polychlorinated biphenyls that may simplify the use of this substance class in scientific papers. In the samples from Australia, BrCl 6-MBPs and Br 2Cl 5-MBPs amounted for 7–27.5% and 0.4–4.2% of Q1, respectively whereas Br 3Cl 4-MBPs and higher brominated MBPs were found in the range of <1% of Q1 or less.

Identification of highly brominated analogues of Q1 in marine mammals

Three novel halogenated organic compounds (HOCs) have been identified in the blubber of marine mammals from coastal New England with the molecular formulae C9H3N2Br6Cl, C9H3N2Br7, and C9H4N2Br5Cl. They were identified using high and low resolution gas chromatography mass spectrometry (GCMS) and appear to be highly brominated analogues of Q1, a heptachlorinated HOC suspected to be naturally produced. These compounds were found in Atlantic white sided dolphin (Lagenorhynchus acutus), bottlenose dolphin (Tursiops truncatus), common dolphin (Delphinus delphis), Risso's dolphin (Grampus griseus), harbor porpoise (Phocoena phocoena), beluga whale (Delphinapterus leucas), fin whale (Balaenoptera physalus), grey seal (Halichoerus grypus), harp seal (Phoca groenlandica) and a potential food source (Loligo pealei) with concentrations as high as 2.7μg/g (lipid weight). The regiospecificity of C9H3N2Br6Cl is suggestive of a biogenic origin. Debromination of C9H3N2Br6Cl may be significant in the formation of C9H4N2Br5Cl.

Concentrations of selected persistent organochlorine contaminants in store-bought foods from northern Alaska

Objectives. We address marine and terrestrial mammal blubber, liver, muscle, kidney, heart, tongue, maktak and maktaaq (epidermis and blubber from bowhead, beluga whales, respectively), and fish muscle and livers, as commonly consumed tissues in subsistence communities across northern Alaska in the context of organochlorine (OC) contamination of store-bought foods. Human exposure to contaminants from biota, as part of a subsistence diet, has been superficially evaluated in numerous studies (focused on liver and blubber), but are limited in the type of tissues analyzed, and rarely consider the contaminants in the alternatives (i.e., store-bought foods).Study Design. Concentrations from published literature on selected persistent organochlorine contaminants (OCs) in eight tissues of the bowhead whale and other biota (1) were compared to store-bought foods evaluated in this study.Results. As expected, store-bought foods had lower concentrations of OCs than some tissues of the marine mammals (especially blubber, maktak, and maktaaq). However, blubber is rarely eaten alone and should not be used to give consumption advice unless considered as a portion of the food item (i.e., maktak). This study indicates that the store-bought food alternatives have detectable OC concentrations (e.g., < 0.01 to 22.5 ng/g w.w. for hexachlorobenzene) and, in many cases, have greater OC concentrations than some subsistence food items. Many wildlife tissues had OC concentrations similar to those quantified in local store-bought food.Conclusions. Switching from the traditional diet to western store-bought foods will not always reduce exposure to OCs. However, raw blubber-based products are clearly more contaminated with OCs due to lipid content. A detailed profile of traditional/country foods and western foods consumed by subsistence communities of northern Alaska is required to address chronic exposure in more detail for the diverse sources of foods (subsistence use and commercially available) and the widely varying concentrations of contaminants reported therein. This should be combined with biomonitoring people dependent upon subsistence foods. Further assessment of essential and non-essential elements, emerging contaminants (e.g. brominated flame retardants), etc. should be conducted in order to improve our understanding of the differences and similarities between wildlife and store-bought foods. (Int J Circumpolar Health 2005; 64(4):303–313)

Human exposure to contaminants in the traditional Greenland diet

The traditional diet is a significant source of contaminants to people in Greenland, although contaminant levels vary widely among species and tissue from very low in many to very high in a few. Our study has included cadmium, mercury, selenium, polychlorinated biphenyls (PCB), dichlorophenyltrichloroethane (DDT), chlordane, hexachlorocyclohexanes (HCH), chlorobenzenes, dieldrin and toxaphene in the major species and tissues consumed by Greenlanders. In general, the levels of these are very low in terrestrial species and in muscle of many marine species. High organochlorines concentrations are typically found in blubber of marine mammals and high metal levels in liver and kidney of seals and whales. In this study, the mean intakes of cadmium, chlordanes and toxaphene significantly exceed ‘acceptable/tolerable intakes’ (ADI/TDI) by a factor between 2.5 and 6. Mean intakes of mercury, PCB and dieldrin also exceed ADI/TDI by up to approximately 50%. However as these figures are mean intakes and as variation in both food intake and contaminant levels is large, the variation of contaminant intake among individuals is also large, and some individuals will be exposed to significantly higher intakes. The mean intakes of DDT, HCH and chlorobenzenes are well below the ADI/TDI values, and it seems unlikely that the TDI for these contaminants normally is exceeded in the Greenland population. The evaluation of contaminant intake in this study points to seal muscle, seal liver, seal kidney, seal blubber and whale blubber as the dominant contributors of contaminants in the traditional diet. Levels in liver from Greenland halibut, snow crab, king eider, kittiwake, beluga and narwhal and kidney of beluga and narwhal are also high but were, with the exception of toxaphene in Greenland halibut liver, not important sources in this study, because they were eaten in low quantities. A way to minimize contaminant intake would be to avoid or limit the consumption of diet items with high contaminant levels. If we assume a traditional diet composition in this study without fish liver, bird liver, seal liver, seal kidney, seal blubber, whale liver, whale kidney and whale blubber, the intake of all contaminants would be below the TDIs for these. This will result in a reduction of the intake of the amount of traditional food of only 24–25%, and it is not likely that this changed diet will result in deficiency of minerals, vitamins or other nutritional compounds.

Human exposure to contaminants in the traditional Greenland diet

The traditional diet is a significant source of contaminants to people in Greenland, although contaminant levels vary widely among species and tissue from very low in many to very high in a few. Our study has included cadmium, mercury, selenium, polychlorinated biphenyls (PCB), dichlorophenyltrichloroethane (DDT), chlordane, hexachlorocyclohexanes (HCH), chlorobenzenes, dieldrin and toxaphene in the major species and tissues consumed by Greenlanders. In general, the levels of these are very low in terrestrial species and in muscle of many marine species. High organochlorines concentrations are typically found in blubber of marine mammals and high metal levels in liver and kidney of seals and whales. In this study, the mean intakes of cadmium, chlordanes and toxaphene significantly exceed ‘acceptable/tolerable intakes’ (ADI/TDI) by a factor between 2.5 and 6. Mean intakes of mercury, PCB and dieldrin also exceed ADI/TDI by up to approximately 50%. However as these figures are mean intakes and as variation in both food intake and contaminant levels is large, the variation of contaminant intake among individuals is also large, and some individuals will be exposed to significantly higher intakes. The mean intakes of DDT, HCH and chlorobenzenes are well below the ADI/TDI values, and it seems unlikely that the TDI for these contaminants normally is exceeded in the Greenland population. The evaluation of contaminant intake in this study points to seal muscle, seal liver, seal kidney, seal blubber and whale blubber as the dominant contributors of contaminants in the traditional diet. Levels in liver from Greenland halibut, snow crab, king eider, kittiwake, beluga and narwhal and kidney of beluga and narwhal are also high but were, with the exception of toxaphene in Greenland halibut liver, not important sources in this study, because they were eaten in low quantities. A way to minimize contaminant intake would be to avoid or limit the consumption of diet items with high contaminant levels. If we assume a traditional diet composition in this study without fish liver, bird liver, seal liver, seal kidney, seal blubber, whale liver, whale kidney and whale blubber, the intake of all contaminants would be below the TDIs for these. This will result in a reduction of the intake of the amount of traditional food of only 24–25%, and it is not likely that this changed diet will result in deficiency of minerals, vitamins or other nutritional compounds.

DETERMINING BLUBBER FATTY ACID COMPOSITION: A COMPARISON OF IN SITU DIRECT AND TRADITIONAL METHODS

AbstractFatty acids (FAs) are used to make inferences about the foraging behavior and diets of free‐ranging marine mammals. However, several methods are currently available for determining the FA composition of blubber and these methods may produce different results. We compared in situ direct transesterification methods, where a small amount of tissue is sampled, with more traditional methods involving prior lipid extraction of the entire sample of interest. Using gray seal (Halichoerus grypus) and beluga whale (Delphinapterus leucas) blubber, we found that when the direct in situ method was used on a 2‐mg sample of blubber, the resulting FA profile differed significantly from that produced when traditional full‐extraction methods were employed. Regardless of where the small spot sample was taken within the blubber depth, it was not representative of the entire blubber FA composition, as blubber is non‐homogeneous throughout its depth. We also modified the in situ direct method to allow analysis of the entire blubber layer. Results of this full‐layer direct method compared quite favorably with traditional extraction methods and may provide a reasonable alternative for analyses. Although application of our full‐layer direct method will require further verification in certain marine mammal blubber samples, we conclude that the large differences obtained when using the direct method are not a consequence of the chemical method itself. Rather, they arise from non‐representative sampling of the blubber FA composition.

Radiocarbon evidence for a naturally produced, bioaccumulating halogenated organic compound.

Halogenated organic compounds (HOCs) such as 1,1'-dimethyl-3,3',4,4'-tetrabromo-5,5'-dichloro-2,2'-bipyrrole (DBP-Br4Cl2) and heptachloro-1'-methyl-1,2'-bipyrrole (Q1) have been detected worldwide, sometimes at high levels in Antarctic air, seabird eggs, the blubber of marine mammals, and, most notably, even human milk. To date, it has been difficult to determine whether these compounds are natural products or derived from industrial synthesis. Molecular-level 14C analysis of these compounds is particularly appealing because most industrial compounds are manufactured from petrochemicals (14C-free) and natural compounds should have "modern" or "contemporary" 14C levels. To investigate the source of DBP-Br4Cl2, we isolated 600 microg of this compound (150 microg of carbon) from marine animal extracts by employing gel permeation chromatography, Florisil column chromatography, and two-dimensional preparative capillary gas chromatography. The purified DBP-Br4Cl2 was split into two samples (75 microg of carbon each) and analyzed by accelerator mass spectrometry for 14C content. The delta14C values were -449 percent per thousand and -467 percent per thousand, corresponding to conventional 14C ages of 4740 and 5000 years before present (BP), respectively. The presence of detectable 14C in the DBP-Br4Cl2 strongly points to at least a natural or biogenic source. However, these delta14C values for DBP-Br4Cl2 are more depleted than expected for a recently synthesized natural product. Several explanations are discussed, but additional samples

Lipid-soluble and water-soluble arsenic compounds in blubber of ringed seal ( Pusa hispida)

High concentrations of arsenic were observed in the blubber of ringed seals ( Pusa hispida) in our previous study. To better understand the arsenic accumulation in blubber of marine mammals, arsenicals in the blubber of ringed seal were characterized using high performance liquid chromatography–inductively coupled plasma mass spectrometry (HPLC–ICPMS). The most predominant water-soluble arsenical in the blubber was dimethylarsinic acid (DMA), in spite of the predominance of arsenobetaine in other tissues. Lipid-soluble fraction was hydrolyzed under mild (tetraethylammonium hydroxide (TEAH) hydrolysis) and strong (NaOH hydrolysis) conditions, and then an aliquot of hydrolysate was injected onto HPLC–ICPMS. Both TEAH-labile and TEAH-stable/NaOH-labile lipid-soluble fractions contained precursors of DMA. These results suggest that the blubber might be the pool of DMA and DMA-containing precursors in ringed seals.

Non-polar halogenated natural products bioaccumulated in marine samples. I. 2,3,3 ′,4,4 ′,5,5 ′-Heptachloro-1 ′-methyl-1,2 ′-bipyrrole (Q1)

This presentation adds new spectroscopic and analytical data on the natural product Q1 that was recently identified by synthesis as 2,3,3 ′,4,4 ′,5,5 ′-heptachloro-1 ′-methyl-1,2 ′-bipyrrole. Solid state magic angle spinning 13C NMR data of Q1 is presented as an option for structural proof. Furthermore, the UV spectrum of neat Q1 (absorption maximum at 223 nm) was recorded and, with NMR spectroscopic data, confirmed a twisted bipyrrole ring system. A quantitative standard of Q1 was prepared which allowed to correct previous concentration estimates relative to the electron capture detector response factor of trans-nonachlor. As a result, the actual Q1 response was only 0.65 ± 15% of the response factor of trans-nonachlor. Therefore, actual Q1 levels are about 50% higher than the previous estimates. With this result the highest (corrected) Q1 concentration determined to date in the blubber of marine mammals from Australia is 14 mg/kg lipid. Analysis of Q1 and trans-nonachlor in specimens from the German North Sea coast suggests that harbor seals are more able to metabolize Q1 than harbor porpoises. Finally, we calculated that 79 congeners of Q1 (i.e. lower chlorinated 1 ′-methyl-1,2 ′-bipyrroles) are theoretically possible and present their structures.

Sponge halogenated natural products found at parts‐per‐million levels in marine mammals

Several unknown, abundant brominated compounds (BCs) were recently detected in the blubber of dolphins and other marine mammals from Queensland (northeast Australia). The BCs were interpreted as potential natural products due to the lack of anthropogenic sources for these compounds. This study investigated whether some of the BCs accumulated by diverse marine mammal species are identical with natural BCs previously isolated from sponges (Dysidea sp.) living in the same habitat. Isolates from sponges and mollusks (Asteronotus cespitosus) were compared with the signals detected in the mammals' tissue. Mass spectra and gas chromatography retention times on four different capillary columns of the isolates from sponges and mammals were identical in all respects. This proves that the chemical name of the compound previously labeled BC-2 is 4,6-dibromo-2-(2',4'-dibromo)phenoxyanisole and that the chemical name of BC-11 is 3,5-dibromo-2-(3',5'-dibromo,2'-methoxy)phenoxyanisole. Using a quantitative reference solution of BC-2, we established that the concentrations of the brominated metabolites found in the marine mammals are frequently >1 mg/kg. The highest concentration (3.8 mg/kg), found in a sample of pygmy sperm whale (Kogia breviceps), indicates that BC-2 is a bioaccumulative, natural organohalogen compound. This is supported by the concentrations of the BCs in our samples being equal to the highest concentrations of anthropogenic BCs in any environmental sample. The quantitative determination of BC-2 in blubber of marine mammals from Africa and the Antarctic suggests that BC-2 is widespread. These results are direct proof that marine biota can produce persistent organic chemicals that accumulate to substantial concentrations in higher trophic organisms.

Global distribution of halogenated dimethyl bipyrroles in marine mammal blubber.

Four halogenated dimethyl bipyrroles (HDBPs), hypothesized to be naturally produced, were quantitated in marine mammal blubber from a number of species obtained from various locations worldwide. HDBPs were found in samples from all locations studied. Concentrations of total HDBPs (SigmaHDBPs) ranged from 0.4 ng/g lipid weight in ringed seals (Phoca hispida) from the White Sea to 2,540 ng/g lipid weight in Dall's porpoise (Phocoenoides dalli) from the northwestern North Pacific Ocean. At their highest levels, SigmaHDBPs made up 11% of the total quantitated organohalogen body burden of adult male Dall's porpoises. In two beluga (Delphinapterus leucas) data subsets, it was found that males contained significantly higher concentrations of SigmaHDBPs than females. No significant effects of age or sex on SigmaHDBPs were observed in harbor seal (Phoca vitulina) and bowhead whale (Balaena mysticetus) data subsets. The geographical distribution of concentrations did not resemble that of the ubiquitous anthropogenic organohalogen, polychlorinated biphenyl congener CB-153. Higher concentrations of HDBPs and different patterns of congeners were observed in samples from Pacific as opposed to non-Pacific Ocean influenced environments. Concentrations of HDBPs in beluga from the Arctic and St. Lawrence River were similar. Their high abundance in north Pacific Ocean biota and widespread occurrence suggest that HDBPs undergo extensive transport from a source located primarily in the Pacific Ocean. Evidence from HDBP congener patterns indicates that both ocean currents and atmospheric transport likely play a role in the movement of HDBPs. These results imply that HDBPs and anthropogenic organohalogens have different sources and support the natural production hypothesis.

A GC/ECNI-MS Method for the Identification of Lipophilic Anthropogenic and Natural Brominated Compounds in Marine Samples

GC/ECNI-MS in both the full scan and selected ion monitoring modes was applied to study the patterns of brominated compounds in the blubber of marine mammals from a variety of locations. The adipose tissue of one polar bear was also studied. Occurrence of three major residues originating from the use of brominated flame-retardants (PBDE 47, 99, and 100) and several potential naturally occurring brominated compounds was studied in the samples. A series of three major natural brominated compounds (BC-1, BC-2, BC-3) recently identified in dolphins from Australia was included in this study. Two further brominated compounds, BC-10 and BC-11, were studied for the first time. ECNI-MS full scan spectra were obtained for BC-3 and BC-10. A natural mixed halogenated compound (MHC-1) and an unknown brominated compound (UBC-1) were investigated as well. Evidence for the natural production of these secondary metabolites and their bioaccumulation in higher organisms as well as analytical protocols for their detection in the environment are presented. Some of these naturally occurring compounds may be misinterpreted as anthropogenic brominated compounds. In ECNI-MS, brominated compounds are usually identified by the detection of the fragment ions m/z 79 ([79Br]-) and m/z 81 ([81Br]-). In this work, it is shown that monitoring of additional ion traces corresponding to [Br2]- (160 type), [HBr2]- (161 type), [BrCl]- (116 type), and [HBrCl]- (117 type) fragment ions allows distinguishing between different classes of brominated compounds. This technique was used to demonstrate that UBC-1 is neither a PBDE nor a PBB congener, whereas a second mixed halogenated compound (MHC-2) was identified as a result of the ECNI-MS response at m/z 114/116. Studies on blubber extracts of marine mammals from four continents resulted in the detection of significant differences in the global distribution of brominated compounds. Our results suggest that naturally occurring organobromines are more abundant than anthropogenic brominated compounds in several regions of the world.

An efficient clean-up method for the GC–MS determination of methylsulfonyl-PCBs/DDEs extracted from various marine mammal tissues

Existing methods for analyzing methylsulfonyl-PCBs in animal tissues require multiple laborious clean-up steps prior to quantitation. In this paper, we report a new, efficient method for the isolation and determination of methylsulfonyl-PCB metabolites in the blubber, liver, kidney, lung, and milk from mammals of marine origin using: solvent extraction, GPC fractionation, and clean-up on an adsorption chromatography column prior to analysis by gas chromatography–mass spectrometry. The method is rapid and can be completed in about half the analysis time required by most other reported methods. The method exhibits excellent analyte recoveries (89–95%), and good reproducibilities with CV's ranging from 3–12% depending on analyte concentration. Detection limits are approximately 1 ng/g lipid. The method is further validated by comparing its quantitative results to that of an independent reference analytical method for several different marine mammal blubber samples. Finally, the method is applied to profile the distribution of methylsulfones among the various tissue types studied for a single bottle-nose dolphin ( Tursiops truncatus) sample. In addition, the extent of their transplacental transfer from mother to unborn fetus for several pilot whale ( Globicephala melaena) mother-fetus pairs is examined.