Heinz Rüdel, Jens Nowak, Josef Müller Fraunhofer Institute for Molecular Biology and Applied Ecology (IME) - Schmallenberg
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1 HBCD diastereomer levels in fish and suspended particulate matter from European freshwater and estuary sites - environmental quality standard compliance and trend monitoring Heinz Rüdel, Jens Nowak, Josef Müller Fraunhofer Institute for Molecular Biology and Applied Ecology (IME) - Schmallenberg Mathias Ricking Department of Earth Sciences, Hydrogeology, Geochemistry - Free University of Berlin 1 Markus Quack, Roland Klein Institute for Biogeography - University of Trier Contact: heinz.ruedel@ime.fraunhofer.de
2 2 Hexabromocyclododecane (HBCD) three environmental relevant diastereomers (pairs of enantiomers): α-hbcd, β-hbcd, γ-hbcd γ-hbcd is the main component in technical products HBCD is an additive flame retardant applied mainly in extruded and expanded polystyrene foams for building insulation, and to minor extents in back-coats, e.g., for upholstery textiles, and in high impact polystyrene (HIPS) used in electrical / electronic equipment estimated annual demand in Europe: tons (ECHA 2008) in 2013 it was decided to add HBCD to the list of persistent organic pollutants (POPs) under the Stockholm Convention (Annex A - Elimination) with specific exemptions for production and for use in expanded and extruded polystyrene in buildings (up to 5 years) (-)α (+)α (-)β (+)β (-)γ (+)γ source of structures: wikipedia.org
3 3 HBCD monitoring project On base of the EU risk assessment it was expected that noticeable emissions of HBCD originate from the coating process of textiles and to a minor extent from the production of polystyrene foam An emission control program (VECAP) has been implemented by HBCD producers and users during recent years - thus decreasing environmental HBCD concentrations were expected To assess the impact of emission reduction measures, an environmental monitoring project for HBCD was initiated by the HBCD Industry working group Objective: to exemplarily follow temporal and spatial trends for fish and suspended particulate matter (SPM) at six different representative locations across Europe
4 Sampling sites representative European aquatic sampling sites were selected river sites are characterized by emissions from diffuse and point sources (e.g., sewage treatment plants) the Tees site (UK) is downstream of a former HBCD production plant Lake Belau was selected as pristine site without urban emissions annual fish sampling at one site per river/lake ( ) sampling of SPM (3 river sites) and sediment (lake) every two years # # 4 Map based on Google Earth # no sampling
5 HBCD EU Water Framework Directive (WFD) EQS Directive 2013/39/EU of 12 August 2013 HBCD was identified as priority substance; a biota EQS for the sum of HBCD diastereomers was derived: 167 µg/kg wet weight; protection goal: secondary poisoning; biota EQS relate to fish For some priority substances, including HBCD, a long-term trend analysis of concentrations in biota (and/or sediment) has to be implemented 5 Guidance on chemical monitoring of sediment and biota under the WFD, Technical Report Fish species in inland water bodies bream, perch, chub. can be used Requirements for temporal trend monitoring Collection of biota annually at the same time within each year Sampling time should be principally outside the spawning period The same size range of the target species is sampled each year Sampling guidelines are necessary to provide some control over biological variation (e.g., fish length, condition)
6 6 Sampling of fish bioaccumulation indicator: bream (Abramis brama) also used in several monitoring programs: e.g. German Environmental Specimen Bank (ESB) bream is non-migrating and common in European rivers as a bottom feeder, bream is in close contact with sediment (potential HBCD sink) at each site 15 adult breams are caught once a year after spawning and fillets are frozen at -150 C preparation of a composite sample by cryo milling Standard Operating Procedures applied (German ESB) because of the brackish water conditions in the Western Scheldt (NL) additional sampling of sole (Solea solea)
7 7 Sampling of Suspended Particulate Matter (SPM) SPM is sampled as an equivalent to newly formed sediment sampling only every two years - temporal changes are expected to be slower as compared to fish SPM is collected with stainless steel traps: operated from buoys in the open water or as flow-through systems in measuring stations (German ESB SOP) composite sample from four 3-months samplings Lake Belau: sediment cores sampled instead of SPM cores are directly frozen in liquid nitrogen; the upper 1-2 cm layer is cut and freeze-dried previous studies showed that this layer corresponds to the newly formed sediment in one year
8 8 Analytical method - fish samples 13 C-labeled α-, β-, and γ-hbcd as internal standards about 1 g tissue sample is homogenized with acetone and extracted with pentane, followed by a GPC clean-up quantification by HPLC coupled to a triple quadrupole mass spectrometer (Waters); negative electron spray ionization in the multiple reaction monitoring mode limits of quantification (LOQ) for α-, β-, γ-hbcd: 0.1 µg/kg ww (wet weight) or 2-13 µg/kg lw (lipid weight) depending on fat content recovery determination according to SANCO guidelines at three fortification levels; recovery at the lowest level (0.1 µg/kg ww) was 76-91% method accreditation according to ISO/IEC
9 9 Results - diastereomer pattern in bream α-diastereomer predominant - as reported in literature but: in Lake Belau bream γ-hbcd mostly dominated
10 10 Results - comparison of concentrations in bream high differences between sum of HBCD concentrations (as µg/kg lipid weight) at different sites in Europe Box plots with fish data from all years; box is defined by 1. and 3. quartiles; red bar = median
11 11 Results - time series for HBCD in bream Replicate analyses n = 4 n = 1 n = 1 n = 2 n = 2 n = 2 n = 2 # log-linear regression analyses based on the PIA tool developed by Bignert (2007) Rhone: significant decreasing trend # at p = 0.01 for sum of HBCD in bream: -28 % per year (Mann-Kendall-test: significant decrease, p < 0.1)
12 Results - comparison bream / sole (West. Scheldt) α-hbcd predominant in bream, but γ-hbcd in sole HBCD levels in bream are lower than in sole in most years significant decreasing trend # for sum of HBCD in bream: -13 % per year (p = 0.04) sole: -45 % per year (p = 0.05) 12 Replicates n = 2 n = 2 n = : 15 individuals n = 2 n = 2 # log-linear regression analyses based on the PIA tool developed by Bignert (2007)
13 Results time series for HBCD in bream Götaälv: low HBCD levels, surprisingly high level in 2012 Mersey: decreasing HBCD levels In 2012 different diastereomer distribution: 55% γ-hbcd content (other years % α-hbcd) Replicates n = 2 n = 3 n = 2 n = 1 n = 2 n = 3 n = 2 n = 1 13 at these sites no sampling
14 Results time series for HBCD in bream Lake Belau: large variations 2007/8; clearly lower HBCD levels in the period ; γ-hbcd predominant Replicates n = 4 n = 15 # n = 2 n = 2 n = 2 n = 2 n = 2 14 # 2008: 15 individual fish
15 15 Results time series for HBCD in bream Highest HBCD levels in bream in comparison to the other European sites were found at the Tees site Location downstream of a former HBCD production plant After massive flood events decrease of levels in 2013 Replicate analyses n = 4 n = 1 n = 1 n = 1 n = 2 n = 2 n = 2
16 16 Comparison with fish Environmental Quality Standard # downstream of a former HBCD plant; EQS was exceeded in previous years EQS (Directive 2013/39/EU): 167 µg/kg wet weight (fish) EU Member States shall implement the EQS by December 2018 Aim: achieving compliance with EQS by December 2027 Environmental monitoring data for bream in 2013 [µg/kg wet weight] Comparison to HBCD fish EQS Rhone (FR) 3.8 Scheldt (NL) 1.7 Götaälv (SE) 2.0 Mersey (UK) 38.8 factor below EQS Tees (UK) # 75.0 Lake Belau (DE) 4.1 µg/kg
17 17 Results - comparison of SPM/sediment and fish SPM/sediment data data normalized to total organic carbon (TOC) content increasing HBCD levels in SPM from Tees and Lake Belau sediment (preliminary conclusion, only 3 SPM/sediment campaigns up to now) different trends for HBCD levels in Lake Belau sediment and fish Ratio of concentrations of sum HBCD SPM/bream about 0.4 Ratio of concentrations of sum HBCD SPM/bream about 0.1-9
18 18 Results - comparison of SPM and fish data comparison for the sites Rhone and Western Scheldt different diastereomers patterns for SPM from both sites SPM/fish trends are similar (preliminary conclusion, 3 SPM campaigns) in the Western Scheldt SPM has a relatively higher HBCD level Ratio of concentrations of sum HBCD SPM/bream about Ratio of concentrations sum HBCD of SPM/bream about 1
19 19 Conclusions the EU Environmental Quality Standard (EQS) for fish was not exceeded in the year 2013 at all chosen sampling sites all over Europe; however, between 2007 and 2012 the levels in the fish from the Tees river were above the EQS for HBCD (site downstream a former production plant) monitoring data reveal decreases of HBCD concentrations in fish at four sites; at two sites - Rhone, Western Scheldt - decreases are significant and preliminary SPM trend data also show a decrease of HBCD levels between 2008 and 2012 at these sites at two sites increasing levels of HBCD in SPM / sediment were detected (preliminary conclusions, only 3 samplings up to now) observed decreases of HBCD levels in fish seem to be consistent with the implementation of emission control measures by HBCD producers and users
20 20 Acknowledgement The authors thank the European Flame Retardant Association - EFRA, PlasticsEurope Association, and European Extruded Polystyrene Insulation Boards Association - EXIBA for funding of the project More information: H. Rüdel, J. Müller, M. Quack, R. Klein (2012): Monitoring of hexabromocyclododecane diastereomers in fish from European freshwaters and estuaries. Environ. Sci. Pollut. Res. 19,
21 Results - individual fish from two sites HBCD levels in individual fish are varying largely: max/min ratio is 506 for bream and 66 for sole analyses of composite samples were confirmed: at two sites γ-hbcd was predominant in fish µg/kg lipid 5,000 4,000 3,000 2,000 1,000 α-hbcd β-hbcd γ-hbcd individual bream, Lake Belau A B C D E F G H Iindividual J K sole, L Western M N Scheldt O ,000 2,500 α-hbcd β-hbcd 2,000 γ-hbcd 1,500 µg/kg lipid 1, A B C D E F G H I J K L M N O
22 22 Comparison with literature data Sawal et al. 2011: Brominated flame retardants in bream from six rivers and a lake in Germany. Organohalogen Comp. 73, the authors also investigated bream muscle investigation of archived samples from the German Environmental Specimen Bank, covering major German rivers and Lake Belau data for the period , no information on trends from Sawal et al Comparison with our study (2009 data) Tees: ng/g Rhone: 531 ng/g Western Scheldt: 51 ng/g Lake Belau: 11 ng/g
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