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Bromochloro alkanes (BCAs) are a class of flame retardants similar in structure to polychlorinated alkanes (PCAs), which are the major component of short-chain chlorinated paraffins (SCCPs) listed as Persistent Organic Pollutants under the Stockholm Convention. BCAs have recently been detected for the first time in environmental samples. Due to the complete lack of commercially available analytical standards, no method for quantifying BCAs has been reported to date. In this study, 16 custom-synthesised standards with mixed bromine and chlorine halogenation and carbon chain lengths ranging from C 10 to C 17 were characterized by liquid chromatography and Orbitrap high-resolution mass spectrometry and used to assess the applicability of pattern deconvolution quantification strategies for BCAs in indoor dust. Br 1-9 and Cl 1-8 BCAs were detected as [M + Cl] - adduct ions among the C 10 to C 17 standards, as well as numerous PCA homologues. After applying correction factors to account for the presence of PCAs in the standards, triplicate fortification experiments using varied halogenation composition and concentration determined an average measurement accuracy of 81% over the carbon chain lengths studied and coefficient of variance ≤20% between replicates. Overall, approximately 89% of the ΣBCA concentrations quantified in the fortification trials met the European Union Reference Laboratory's accuracy acceptability criteria recommended for PCAs, between 50 and 150%. Application of the BCA pattern deconvolution quantification procedure to seven representative indoor dust samples from the United States of America revealed a low correlation between the homologue distribution in the samples and the prototype standards (R 2  ≤ 0.40), which precluded reliable quantification. This study indicates that pattern deconvolution is an appropriate strategy for quantifying BCAs in environmental samples, but that a large set of appropriate mixture standards will be required before more reliable estimates of BCA concentrations can be achieved in indoor dust., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Chlorinated paraffins (CPs) are synthetic organic compounds of growing environmental and social concern. Short-chain chlorinated paraffins (SCCPs) were listed under the Stockholm Convention on Persistent Organic Pollutants (POPs) in 2017. Further, in 2021, medium-chain chlorinated paraffins (MCCPs) were proposed to be listed as POPs. We investigated SCCP and MCCP amounts and homolog profiles in four wild fish species from Bahía Blanca Estuary, a South Atlantic Ocean coastal habitat in Argentina. SCCPs and MCCPs were detected in 41% and 36% of the samples, respectively. SCCP amounts ranged from <12 to 29 ng g -1 wet weight and <750-5887 ng g -1 lipid weight, whereas MCCP amounts ranged from <7 to 19 ng g -1 wet weight and <440-2848 ng g -1 lipid weight. Amounts were equivalent to those found in fish from the Arctic and Antarctic Oceans and from some North American and Tibetan Plateau lakes. We performed a human health risk assessment and found no direct risks to human health for SCCP or MCCP ingestion, according to present knowledge. Regarding their environmental behavior, no significant differences were observed among SCCP amounts, sampling locations, species, sizes, lipid content, and age of the specimens. However, there were significant differences in MCCP amounts across species, which could be attributed to fish size and feeding habits. Homolog profiles in all fish were dominated by the medium-chlorinated (Cl 6 and Cl 7 ) CPs and shorter chain length CPs were the most abundant, with C 10 Cl 6 (12.8%) and C 11 Cl 6 (10.1%) being the predominant SCCPs and C 14 Cl 6 (19.2%) and C 14 Cl 7 (12.4%) the predominant MCCPs. To the best of our knowledge, this is the first study on the presence of CPs in the environment in Argentina and the South Atlantic Ocean. CP occurrence in the environment, particularly in the food chain, promotes the need for further research on their occurrence and behavior, and the impact of CPs in marine ecosystems in Argentina., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Technical chlorinated paraffins (CPs) are produced via radical chlorination of n-alkane feedstocks with different carbon chain-lengths (∼C 10 -C 30 ). Short-chain CPs (SCCPs, C 10 -C 13 ) are classified as persistent organic pollutants (POPs) under the Stockholm Convention. This regulation has induced a shift to use longer-chain CPs as substitutes. Consequently, medium-chain (MCCPs, C 14 -C 17 ) and long-chain (LCCPs, C > 17 ) CPs have become dominant homologues in recent environmental samples. However, no suitable LCCP-standard materials are available. Herein, we report on the chemical synthesis of single-chain C 18 -CP-materials, starting with a pure n-alkane and sulfuryl chloride (SO 2 Cl 2 ). Fractionation of the crude product by normal-phase liquid-chromatography and pooling of suitable fractions yielded in four C 18 -CP-materials with different chlorination degrees (m Cl,EA  = 39-52%). In addition, polar side-products, tentatively identified as sulfite-, sulfate- and bis-sulfate-diesters, were separated from CPs. The new single-chain materials were characterized by LC-MS, 1 H-NMR and EA. LC-MS provided Relative retention times for different C 18 -CP homologues and side-products. Mathematical deconvolution of full-scan mass spectra revealed the presence of chloroparaffins (57-93%) and chloroolefins (COs, 7-26%) in the four single-chain C 18 -CP-materials. Homologue distributions and chlorination degrees were deduced for CPs and COs. 1 H-NMR revealed chemical shift ranges of mono-chlorinated (δ = 3.2-5.3 ppm) and non-chlorinated (δ = 1.0-3.2 ppm) hydrocarbon moieties. The synthesized C 18 -single-chain standard materials and respective spectroscopic data are useful to identify and quantify LCCPs in various materials and environmental samples. CP- and CO-distributions resemble the ones of existing SCCP and MCCP reference materials and technical mixtures. Furthermore, these materials now allow specific studies on the environmental fate and the transformation of long-chain chloroparaffins and chloroolefins., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Chlorinated paraffins (CPs) are complex mixtures, which consist of thousands of individual compounds with no dominant representative. Consequently, knowledge on structure and environmental relevance of individual CP congeners is poor. Similarly to the synthesis of individual CPs, the generation of less complex CP mixtures that can be thoroughly analyzed may be used to overcome some drawbacks of the highly complex technical CP mixtures. Here, we present a new synthesis approach to generate such simple CP mixtures by decarboxylation of polyunsaturated fatty acids followed by saturation of the double bonds by chlorination. Specifically, α-linolenic acid (18:3Δ9,12,15) was decarboxylated to heptadecatriene. The resulting raw product was chlorinated with SO 2 Cl 2 . Purification by column chromatography led to a main fraction consisting of four major peaks originating from hexachloroheptadecane (C 17 H 30 Cl 6 ) isomers (∼80% of the total peak area) along with ∼20 low abundant by-products, according to gas chromatography with electron capture negative ion mass spectrometry. In the same way, decarboxylation and subsequent chlorination of other polyunsaturated fatty acids may lead to further simple CP mixtures with other chain lengths. Although these simple CP mixtures cannot fully reflect the various structural features present in technical mixtures they could be beneficial for transformation studies because changes in the CP pattern can easily be noted which is in contrast to technical CP mixtures. Such simple CP mixtures could also be used in toxicity tests which are difficult to perform with technical CP mixtures because of their high complexity., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Chlorinated paraffins (CPs) are high production volume chemicals currently produced and used in higher quantities than any other medium-size polyhalogenated compound (class). In addition, the composition of industrial CP mixtures is highly complex and poorly understood. In this article, we searched in the literature for the beginning of the chlorination of alkanes and how this substance class developed from niche applications to unmatched quantities in various industrial applications. Also, an estimation was made on the theoretical variety of chloroparaffins and the possible complexity of industrial CP mixtures. These data may explain why little is known about CPs although the production volume throughout the industrial generation was virtually always higher than the one of PCBs and has continued to increase after the ban of the latter., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The determination of chlorinated paraffins (CPs) has posed an intractable challenge in analytical chemistry for over three decades. The combination of an as yet unspecifiable number (tens - hundreds of thousands) of individual congeners in mass produced commercial CP mixtures and the steric interactions between them, contrive to defy efforts to characterise their residual occurrences in environmental compartments, food and human tissues. However, recent advances in instrumentation (mass spectrometric detectors and nuclear magnetic resonance), combined with interlaboratory studies, have allowed a better insight into the nature of the conundrums. These include the variability of results, even between experienced laboratories when there is insufficient matching between analytical standards and occurrence profiles, the poor (or no) response of some instrumentation to some CP congener configurations (multiple terminal chlorines or < four chlorines) and the occurrence of chlorinated olefins in commercial mixtures. The findings illustrate some limitations in the existing set of commercially available standards. These include cross-contamination of some standards (complex CP mixtures), an insufficient number of single chain standards (existing ones do not fully reflect food/biota occurrences), lack of homologue group standards and unsuitability of some configurationally defined CP congeners/labelled standards (poor instrument response and a smaller likelihood of occurrence in commercial mixtures). They also indicate an underestimation in reported occurrences arising from those CPs that are unresponsive during measurement. A more extensive set of standards is suggested and while this might not be a panacea for accurate CP determination, it would reduce the layers of complexity inherent in the analysis., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Environmentally relevant halogenated natural products (HNPs) are frequently similarly high concentrated in marine biota as major anthropogenic persistent organic pollutants (POPs). The lack of widely available reference standards, however, hampers the in-depth research of several HNPs. For instance, (1R,2S,4R,5R,1'E)-2-bromo-1-bromomethyl-1,4-dichloro-5-(2'-chloroethenyl)-5-methylcyclohexane (MHC-1), which is produced by species referred to the red seaweed Plocamium cartilagineum has not yet been synthesized due to its complex structure and stereochemistry. For this reason, we aimed to establish a method for fast isolation of mg-amounts of MHC-1 from its natural producer based on countercurrent chromatography (CCC). Five biphasic solvent systems were tested and finally, the solvent system acetonitrile/n-hexane/toluene (9:9:2, v/v/v) was selected for the separations due to its suitable partition coefficient of MHC-1 (K U/L  = 0.52). n-Hexane extracts of dried P. cartilagineum were directly injected into the CCC system. Four subsequent CCC runs from three samples of Plocamium cartilagineum (two from Heligoland, Germany and one from Brittany, France) could be performed with high reproducibility. Together, the main fraction provided ~16 mg MHC-1 in a purity of >97% according to GC/FID, GC/ECNI-MS and NMR analysis. This amount could be used to prepare ~1600 quantitative standard solutions of MHC-1. The high MHC-1 content in the seaweed sample collected at Brittany indicated that this area was another hotspot of MHC-1., (Copyright © 2021. Published by Elsevier Ltd.)

The photolytic chlorination of n-alkanes in presence of sulfuryl chloride (SO 2 Cl 2 ) was explored to produce new standard materials. Five mixtures of chlorinated tetradecanes were synthesized with chlorination degrees (m Cl,EA ) varying from 43.7% to 59.4% (m/m) based on elemental analysis. Chlorine-enhanced negative chemical ionization mass spectrometry (CE-NCI-MS) forcing the formation of chloride-adduct ions [M+Cl] - was applied to characterize these materials which all contained tetra-to deca-chlorinated paraffins. Deconvolution of respective mass spectra revealed the presence of chlorinated olefins (COs). CO levels were highest in materials, which were exposed longest. All synthesized materials also contained two classes of polar impurities, tentatively assigned as sulfite- and sulfate-diesters with molecular formulas of C 14 H 28-x O 3 SCl x (x = 1-4) and C 14 H 28-x O 4 SCl x (x = 3-6), respectively. MS data were in accordance with the proposed structures but further work is needed to deduce their constitutions. These compounds are thermolabile and were not detected with GC-MS methods. We could remove these sulfur-containing impurities from the CPs with normal-phase liquid chromatography. In conclusion, single-chain CP materials were synthesized via chlorination of n-alkanes with sulfuryl chloride, but these materials contained reactive side products which should be removed to gain non-reactive and stable CP materials suitable as standards and for fate and toxicity studies., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Chlorinated paraffins (CPs) are complex mixtures of polychlorinated n-alkanes of different chain length. Despite their environmental relevance, quantification is still a challenge. Moreover, the individual structures of the molecules in technical CP mixtures are largely unknown. Here, we synthesized 21 and studied 29 single chain length CP mixtures ranging from C 10 - to C 17 -CPs with different chlorine contents between 41.8% and 62.6% Cl and analyzed them by means of nuclear magnetic resonance spectroscopy (NMR). Discrete ranges of chemical shifts were observed in one-dimensional 1 H and 13 C NMR spectra. Two dimensional heteronuclear single quantum coherence spectroscopy (HSQC) enabled to assign nine substructures. These measurements also verified the presence of [-CCl 2 -] groups and chlorine atoms on terminal carbons in highly chlorinated (>59% Cl) mixtures. 1 H NMR spectra of different chain length and the same degree of chlorination looked virtually the same. However, with increasing degree of chlorination the observed patterns in the spectra were slightly shifted downfield. Based on these findings, a calculation mode was developed to estimate the chlorine content (%) of the single chain length CP mixtures. The results agreed well (<0.9% mean deviation) with parallel measurements by elemental analysis., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Brominated 2-phenoxyanisoles (2-methoxydiphenyl ethers, 2-MeO-BDEs) are a class of halogenated natural products, produced by algae and sponges. Especially two tetrabrominated isomers, i.e. 2'-MeO-BDE 68 (BC-2) and 6-MeO-BDE 47 (BC-3), have also been frequently determined in environmental and food samples. In addition, 2-MeO-BDEs are under discussion as metabolites of polybrominated diphenyl ethers (PBDEs). In this study, we synthesized the backbone 2-methoxydiphenyl ether and brominated it to an average degree of four bromine substituents. The reaction mixture only contained one major product (∼90%) along with three further MeO-BDEs and ∼5% hydroxylated BDEs. In all likelihood, the HO-BDEs were formed in a side reaction by cleavage of the methoxy group. The major MeO-BDE was identified as 6'-methoxy-2,3',4,4'-tetrabromodiphenyl ether (6'-MeO-BDE-66). The HO-BDEs were separated by KOH/n-hexane partitioning, and the resulting 2-MeO-BDEs were fractionated by means of high-speed counter-current chromatography (HSCCC). Due to the excellent enrichment facilities of HSCCC, some 15 MeO-BDEs, mainly present at traces only, could be detected in 26 fractions, and eight of them could be characterized by nuclear magnetic resonance spectroscopy (NMR). Only two of the compounds--2'-MeO-BDE 68 and 6-MeO-BDE 123--had been characterized as natural products while the prominent halogenated natural product 6-MeO-BDE 47 was not detected at all in the reaction product. The "non-natural" 2-MeO-BDEs may be useful internal standards in trace analysis., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The 2,3,3',4,4',5,5'-heptachloro-1'-methyl-1,2'-bipyrrole (Q1, MBP-79) and further halogenated 1'-methyl-1,2'-bipyrroles (MBPs) are a class of marine natural products repeatedly detected in seafood and marine mammals from all over the world. Only Q1 is currently commercially available as reference standard and the full synthesis of mixed brominated-chlorinated compound is rather complicated. For this reason, synthetic Q1 (240 mg) was transferred into bromine-containing MBPs by UV-irradiation in the presence of bromine. Bromine, which rapidly vanished from the solutions, was renewed during the reaction in order to generate higher amounts of Br-containing MBPs. A total of ∼150 mg Q1 was transferred after ∼10 min irradiation with high amounts of Br(2) to give 30.5mg BrCl(6)-MBPs along with lower proportions of Br(2)Cl(5)-, Br(3)Cl(4)-, Br(4)Cl(3)- and traces of Br(5)Cl(2)-MBPs. Longer UV-irradiation in the presence of Br(2) even allowed for the detection of Br(6)Cl-MBPs and traces of Br(7)-MBP. However, this reaction also provided some unknown by-products. A sample stored in the dark and later in in-door light (no UV irradiation) also eliminated Q1 after 76 d in favour of heptahalogenated MBPs with up to three bromine substituents. The irradiation products were separated on silica, and fractions containing only Q1 and BrCl(6)-MBPs were then further fractionated by non-aqueous RP-HPLC. A pure isolate of the major BrCl(6)-MBP (∼1.5mg) was characterized by GC/MS and (13)C NMR to be 2-bromo-3,3',4,4',5,5'-hexachloro-1-methyl-1,2'-bipyrrole (Br-MBP-75). Partial GC enantioseparation of the axially chiral Br-MBP-75 was achieved on a β-PMCD column. A full enantioseparation was managed by enantioselective HPLC using a NUCLEOCEL DELTA S column. Low amounts of pure BrCl(6)-MBP enantiomers could be trapped., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

For decades, toxaphene had been used as a major chloropesticide. Degradation of the multicomponent mixture in the environment was mainly reported to be due to anaerobic dechlorination and hydrodechlorination. Little was known about oxidative transformation processes and the potential hydroxylated metabolites were not available as standard compounds. For this reason we synthesized hydroxylated polychlorobornanes by the UV-induced photochlorination of 2-endo-bornyl acetate with sulfuryl chloride followed by hydrolysis of the acetate moiety. The released polychlorinated 2-endo-hydroxybornanes were slightly higher chlorinated the longer the reaction was maintained. After 8h, the main products were pentachlorinated hydroxybornanes followed by hexa- and heptachlorinated homologues. Traces of octachlorinated hydroxybornanes were also observed. The GC/ECNI-MS spectra of the products were characterized by the molecular ions and the [M-Cl]⁻ fragment ions. The molecular ions of the polychlorinated hydroxybornanes are isobaric with those of polychlorinated biphenyls. E.g. hexachlorohydroxybornanes (C₁₀H₁₂Cl₆O) and hexachlorobiphenyls (C₁₂H₄Cl₆) show the molecular ion at m/z 358. Based on fractionation experiments on silica with the synthesis products it might be possible that OH-CTTs if present in samples will elute into a more polar fraction usually discarded or not collected. Both problems might explain why these compounds have not been more frequently described in the scientific literature., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Enantioselective analyses of the chiral brominated flame retardant 2,3-dibromopropyl-2,4,6-tribromophenyl ether (DPTE) are the focus of this work. High performance liquid chromatography (HPLC) equipped with a column that had a chiral stationary phase consisting of a modified cellulose derivative allowed for the fractionation of the enantiomers of DPTE. The enantiomeric excess was 98.2% for enantiomer 1 and 99% for enantiomer 2. Polarimetric measurements verified that the first eluting enantiomer originated from (-)-DPTE and the second peak originated from (+)-DPTE. Two gas chromatographic columns allowed for the direct enantioresolution of DPTE. The elution order of DPTE enantiomers was the same as observed in the chiral HPLC system ((-)-DPTE before (+)-DPTE). The best enantioseparation was achieved on a Chirasil-DEX CB column, which was used to analyze the enantiomer fractions of DPTE in blubber and brain samples of hooded seals (Cystophora cristata) and harp seals (Phoca groenlandica) from the Barents and Greenland Seas. Analyses were carried out by means of gas chromatography/electron capture negative ion mass spectrometry operated in the selected ion monitoring (GC/ECNI-MS-SIM) mode. In both matrices, only minute deviations from the racemate were observed (maximum +/-3% excess of (-)-DPTE). However, the samples from the Barents Sea were either racemic or showed a slight excess of (+)-DPTE (up to 2.5%), whereas all samples from the Greenland Sea contained a slight excess (up to 4%) of (-)-DPTE.

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 200-1800ngg(-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 approximately 7ngg(-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.

The halogenated natural product previously named mixed-halogenated compound 1 (MHC-1) was isolated from the red seaweed Plocamium cartilagineum harvested in Helgoland, Germany. A total of 1.9 mg of pure MHC-1 was obtained from 1g air-dried seaweed. The 1H and 13C NMR data matched those reported for a natural monoterpene isolated from this species. Thus, the structure of MHC-1 was established to be (1R,2S,4R,5R,1'E)-2-bromo-1-bromomethyl-1,4-dichloro-5-(2'-chloroethenyl)-5-methylcyclohexane. Moreover, the isolated monoterpene proved to be identical with the compound previously detected in marine mammals and fish from different locations. In addition we examined two samples of P. cartilagineum from Ireland and from the Antarctic; however MHC-1 was only present at low levels. Not only the concentrations were lower but also the pattern of polybrominated compounds differed from MHC-1. A calibrated solution of MHC-1 was used to determine correct concentrations from samples where previously only estimates existed relative to the gas chromatography-electron capture detector (GC/ECD) response of trans-chlordane, which underrated the MHC-1 concentrations by more than factor 2. The highest MHC-1 concentration determined to date in marine mammals is 0.14 mg kg(-1) blubber. Significantly higher MHC-1 concentrations were determined in farmed fish with up to 2.2 mg kg(-1) lipids. The samples with high concentrations of MHC-1 have in common that they were collected in proximity of the natural habitats of P. cartilagineum.

Polybrominated biphenyls (PBBs) have been introduced as flame retardants in 1970. Despite decreasing application rates since the mid-1970s, PBB residues are still reported in the environment. Furthermore, environmental PBB residues often do not match the PBB pattern in technical products. To get insights into the structures of environmentally-relevant PBBs, the congener patterns of technical hexabromobiphenyl (THBB), octabromobiphenyl (TOBB), synthesized PBB 209 were compared to PBB residues in a cod liver sample from the Baltic Sea. The most relevant PBB congeners in Baltic cod liver were not present in the technical products and therefore most likely metabolites. For this reason, TOBB and HPLC-fractions obtained from this technical product were incubated with super-reduced cyanocobalamine. Reductive debromination was found to be the predominant transformation process. Bromine substituents in ortho-positions proved to be more recalcitrant, and several of the unknown PBBs were tri- and tetra-ortho substituted congeners. Furthermore, the key-PBBs determined in Baltic cod liver were formed during this process. The most important hexabromobiphenyl in Baltic cod liver was identified as PBB 155 by parallel synthesis. PBB 155 which was not detected in the technical PBB product analyzed was suggested as an indicator PBB congener suited to decide whether PBB residues originate from the previous use of THBB (low relative abundance of PBB 155) or TOBB/TDBB (high relative abundance of PBB 155). The latter scenario was found to be valid for the Baltic cod liver sample. Thus, PBBs in the Baltic Sea appeared to originate from partially-weathered residues of PBB 209.

The marine halogenated natural product 2,3,3',4,4',5,5'-heptachloro-1'-methyl-1,2'-bipyrrole (Q1) has been detected in high-trophic level biota throughout the world. In this study we UV-irradiated Q1 in order to produce hexahalogenated 1'-methylbipyrroles (Cl(6)-MBPs). Q1 was transformed with half-lives of <5 min. Already after 5 min, all of the five existing Cl(6)-MBPs (H1-H5) were detected in the irradiated sample. Only one Cl(6)-MBP (2,3,3',4',5,5'-hexachloro-1'-methyl-1,2'-bipyrrole (MBP-77, H2) has been previously described in the literature. H5 was identified as 2,3,3',4,4',5'-hexachloro-1'-methyl-1,2'-bipyrrole (MBP-75) by a specific fragment ion detected by GC/ECNI-MS. Fractionations of the irradiation mixture by reversed-phase HPLC followed by (1)H NMR analysis led to the structure of H4, i.e. 2,3,3',4,4',5-hexachloro-1'-methyl-1,2'-bipyrrole (MBP-74). H1 and H3 showed virtually identical (1)H NMR data. Therefore, it could not determined which of either isomers is 2,3,3',4,5,5'-hexachloro-1'-methyl-1,2'-bipyrrole (MBP-76) and which is 2,3,4,4',5,5'-hexachloro-1'-methyl-1,2'-bipyrrole (MBP-78). In addition, two pentachloro-MBPs (P1 and P3) could be traced back to MBP-62 and MBP-69. Cl(6)-MBPs were analyzed in whale blubber from Australia and skua adipose tissue from Antarctica. The marine mammals contained all Cl(6)-MBPs except for the most abundant in the irradiation experiment. The concentrations of the Cl(6)-MBPs amounted to 0.04-1.76% of the concentration of Q1. The highest concentrations of Cl(6)-MBP isomers in the biota samples were found for MBP-76, MBP-77, and MBP-78. These congeners appeared to be the most lipophilic ones owing to the highest retention time in RP-HPLC. Nevertheless, it remained unclear whether the Cl(6)-MBPs were actual halogenated natural products or environmental metabolites of 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 (BrCl6-MBPs), five out of 14 possible dibromopentachloro congeners (Br2Cl5-MBPs), five of 21 possible tribromotetrachloro-congeners (Br3Cl4-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, BrCl6-MBPs and Br2Cl5-MBPs amounted for 7-27.5% and 0.4-4.2% of Q1, respectively whereas Br3Cl4-MBPs and higher brominated MBPs were found in the range of <1% of Q1 or less.

In this study we compared the contribution of individual congeners and the ratios of stable carbon isotopes of two technical toxaphene products. The former US-American product Toxaphene was from 1978 and the East-German product Melipax from 1979. Both technical products showed the known complexity in GC/ECD measurements. Contributions of 24 peaks to each of the technical products were determined by gas chromatography in combination high resolution electron capture negative ion mass spectrometry (GC/ECNI-HRMS). The percentages of the compounds studied in the technical mixtures ranged from approximately 0.05% to approximately 2.5% but showed some individual differences. 2,2,5,5,8,9,9,10,10-nonachlorobornane (B9-1025 or P-62) was identified as a major congener in both mixtures. 2-Endo,3-exo,5-endo,6-exo,8,8,10,10-octachlorobornane (B8-1413 or P26) and 2-endo,3-exo,5-endo,6-exo,8,8,9,10,10-nonachlorobornane (B9-1679 or P-50) were found at similar concentration in both technical products. Identical amounts of Melipax or Toxaphene were combusted to CO2 in an element analyzer and their delta13C values were determined relative to the international standard Vienna PeeDee belemnite (VPDB). The mean delta13C values of both products varied by 2.8% (determined at two different locations) which is roughly one order of magnitude more than the precision obtained in repetitive analyses of the individual products. Thus, both investigated products could be unequivocally distinguished by stable isotope ratio mass spectrometry (IRMS). IRMS analyses may thus be a suitable tool for tracing back toxaphene residues in environmental and food samples to the one or both of the products.

Several identified and potential natural brominated bioaccumulative compounds were studied in this work. 4,6-dibromo-2-(2('),4(')-dibromo)phenoxyanisole (BC-2) previously detected in Australian marine mammals and isolated from sponges, was synthesized. Two byproducts (a tetrabromo isomer and a tribromo congener) were investigated as well. The byproducts of the synthesis were not identified in the environmental samples investigated. Previously described natural brominated compounds (BC-1, BC-2, BC-3, BC-10, BC-11, MHC-1) and anthropogenic brominated diphenyl ethers (BDE-47, BDE-99, BDE-100, BDE-154) were detected in a sample of human milk. The sample was from a woman from the Faeroe Islands who frequently consumed fish as well as whale blubber and meat. The most abundant compound originated from the natural tetrabromo phenoxyanisole BC-3 which may have a 3:1 distribution of bromine on the two phenyl units. This sample also accumulated a dibromochloroanisole, as well as a previously unknown mixed halogenated compound (MHC-X) and an unknown, most likely aromatic brominated compound. Co-elutions on a DB-5 column were found for BDE-99 and BC-11 as well as BDE-154 and the unknown brominated compound. This suggests that quantification of these two compounds has to be carried out carefully.Two samples of lower trophic level, namely Baltic cod liver and Mexican mussel tissue, were investigated as well. The cod liver samples contained BDE congeners but also abundant signals for the natural 2,3,3('),4,4('),5,5(')-heptachloro-1(')-methyl-1,2(')-bipyrrole Q1 and tribromoanisole (TBA). The mussel sample contained Q1, TBA, another halogenated anisole, BC-1, BC-2, and BC-3, as well as additional, potential natural brominated compounds in the elution range of tribromophenoxyanisoles.

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.

Levels and distribution of eight compounds of technical toxaphene (CTTs) were determined in different marine species (seals, cetaceans, birds, and fish). The eight CTTs included six commercially available and two chlorobornanes prepared in our lab. These congeners were present in all investigated samples. In agreement with earlier studies, the octachlorobornane B8-1413 (P-26) and the nonachlorobornane B9-1679 (P-50) were the most abundant congeners in most of the samples. In seal blubber, B8-1413 (P-26) and B9-1679 (P-50) contributed with up to approximately 80% (Weddell seal) to the sum of the eight CTTs. In seals from the northern hemisphere the nonachlorobornane was more abundant while in those from the southern hemisphere (Antarctic and Namibia), the octachlorobornane B8-1413 (P-26) usually appeared at higher concentrations. Depending on the species the contribution of the other congeners varied significantly. B9-1025 (P-62) ranged from 2-20%, B8-1412 was found at 4-25% with highest contribution in birds, and B8-2229 (P-44) was found at 5-15%. The remaining three congeners B7-1453, B8-1414 (P-40), and B8-1945 (P-41) were lower abundant except B8-1414 (P-40) which was found at high contribution in liver and kidney of birds. The sum of the eight CTTs ranged from 4 microg/kg to 1.4 mg/kg, depending on the species and region. In most of the seal blubber samples, PCBs and DDT were more abundant (factor 2-20) but Antarctic Weddell seals showed higher CTT levels than PCBs and DDT.

Enantiomer separation of chiral toxaphene components in biological samples was studied by application of different chiral stationary phases based on modified cyclodextrins. Several pairs of enantiomers were resolved on permethylated beta-cyclodextrin (beta-PMCD), among them 2-endo,3-exo,5-endo,6-exo,8,8,9,10-octachlorobornane (B8-1412), which was not enantiomerically resolved on tert-butyldimethylsilylated beta-cyclodextrin (beta-BSCD). The latter column was applied to determine the enantiomer ratios (ERs) of 2-endo,3-exo,5-endo,6-exo,8,8,10,10-octachlorobornane (B8-1413 or P-26) in brain tissue of three seal species. The ER of B8-1413 (P-26) in brain was virtually racemic as well as those of the two persistent and chiral components of technical chlordane, 1-exo,2,2,4,5,6,7,8,8-octachloro-3a,4,7,7a-tetrahydro-4,7-metha noindane (trans-nonachlor III or MC 6) and 1-exo,2-endo,3-exo,4,5,6,8,8-octachloro-3a,7,7a-tetrahydro-4,7- methanoindane (U82). In contrast, B8-1412 and 2-exo,5,5,8,9,9,10,10-octachlorobornane (B8-2229 or P-44) were significantly enantiomerically enriched in several samples of high trophic level biota. 2,2,5,5,8,9,9,10,10-Nonachlorobornane (B9-1025 or P-62), a chlorobornane metabolisable by seals and the presumable precursor of B8-2229 (P-44), was also enantiomerically enriched in seal blubber. These results confirm the assumption that some less persistent toxaphene components may be significantly degraded in biological samples. Enantioselective gas chromatography provides the information that such a degradation is happening by the characteristic change of the ratio of the two enantiomers in the respective tissues.

Polychlorinated bornanes, the main components of Toxaphene, are bioconcentrated in aquatic organisms to a high extent. However, up to this time no bioconcentration tests with individual chlorinated bornanes in aquatic organisms have been performed. Therefore, the bioconcentration factors (BCFs) of seven selected persistent chlorinated bornane congeners which are regularly found in aquatic organisms, were predicted from their n-octanol/water partition coefficients (log Kow). Furthermore, these BCF values were compared with the measured bioaccumulation factors (BAFs) in zooplankton and different fish species from the aquatic environment.

A fast and effective sample clean-up procedure for the quantitation of chlorinated hydrocarbons in seal blubber and cod livers is presented. Lipophilic sample ingredients are extracted by application of microwave energy. Microwave-assisted extraction (MAE) was performed with ethyl acetate and cyclohexane as solvent. Without exchange of the solvent, the organochlorine compounds are separated from matrix coextractives by gel permeation chromatography. Traces of matrix remainders were separated on deactivated silica prior to GC/ECD analysis. In this study we focused on quantification of PCB 153, PCB 138, PCB 180, HCB, and p,p'-DDE. In seal blubber, the recovery rates for these organochlorines were > 90% for the complete sample clean-up procedure. The standard deviation of the overall-method was within 5%. MAE of cod livers required softer conditions. After optimization of the MAE the organochlorine levels in cod liver were almost identical with those determined with another independent clean-up method.

The chiral stationary phase beta-TBDM (35% heptakis(6-O-tert.-butyldimethylsilyl-2,3-di-O-methyl)-beta-cyc lod extrin in OV-1701) was applied to study the enantioenrichment of alpha-HCH, oxychlordane, and PCB 149 in blubber of two seal species from Iceland by gas chromatography with electron capture detection. The examined harbour seals (Phoca vitulina) and grey seals (Halichoerus grypus) showed alpha-HCH enantiomeric ratios (ER) > 1. The ER of PCB 149 was comparable in the two species from Iceland but for oxychlordane ER < 1 was observed in harbour seals while the oxychlordane ER in grey seals was > 1. In blubber of Weddell seals (Leptonychotes Weddelli) from the Antarctic we determined an alpha-HCH ER < 1 which is in contrast to the Icelandic seal species and a sample from Lake Baikal.

A precise quantification of toxaphene residues of environmental samples by gas chromatography/ electron capture detection (GC/ECD) requires the separation of the bulk of the polychlorinated biphenyls (PCBs) from the compounds of technical toxaphene (CTT) fraction. For this reason, a PCB/CTT group separation on silica was developed. B8-1413 (Parlar #26) and B7-515 (Parlar #32) eluted as first and last out of ten important CTT standards, and can be used to determine the elution volume of the CTT fraction on silica. GC/ECD quantification of CTTs was possible after separation of PCBs on 8.0 g activated silica eluted with 48 mL n-hexane followed by quantitative elution of CTTs with n-hexane/toluene (65:35; v/v). This method is a compromise between separation efficiency and consumption of material. Finally, eight CTTs were quantified in cod liver samples from Iceland and the Baltic Sea.

The enantiomeric ratios of alpha-HCH were determined by chiral gas chromatography in blubber of marine mammals from regions of the northern hemisphere (North Sea, Baltic Sea, Arctic and Iceland). Cetaceans (harbour porpoises and white-beaked dolphins) showed a preferential accumulation of (+)-alpha-HCH. In blubber of harbour seals, grey seals and harp seals (+)-alpha-HCH was also more abundant than (-)-alpha-HCH. Hooded seals formed an exception with a (+/-) enantiomeric ratio of alpha-HCH < 1.