Polychlorinated biphenyls and -naphthalenes in pine needles and soil from Poland – Concentrations and patterns in view of long-term environmental monitoring

Chemosphere 67 (2007) 1877–1886 www.elsevier.com/locate/chemosphere

Polychlorinated biphenyls and -naphthalenes in pine needles and soil from Poland – Concentrations and patterns in view of long-term environmental monitoring Barbara Wyrzykowska a,b, Nobuyasu Hanari b, Anna Orlikowska a, Ilona Bochentin a, Pawel Rostkowski a, Jerzy Falandysz a,*, Sachi Taniyasu b, Yuichi Horii b, Qinting Jiang b,c, Nobuyoshi Yamashita b a

Department of Environmental Chemistry and Ecotoxicology, University of Gdan´sk, 18 Sobieskiego Street, PL 80-952 Gdan´sk, Poland b National Institute of Advanced Industrial Science and Technology (AIST), EMTECH, Tsukuba, Japan c Department of Biology and Chemistry, City University of Hong Kong, Hong Kong Accepted 26 May 2006 Available online 4 January 2007

Abstract Pine needles were selected as cost effective and easy collectable matrices suitable for long-term monitoring of the lower troposphere pollution with polychlorinated biphenyls and polychlorinated naphthalenes. The fingerprints of PCNs and PCBs in the top layers of agricultural soils were used for determination of point sources of pollution for terrestrial ecosystems. The new idea based on the use of nonaand decachlorinated isomers fingerprint as an additional tool suitable for the identification of potential point sources of pollution with PCBs, seemed to be a capable tool to identify contamination of soil and ambient air related to former manufacturing and the use of highly chlorinated technical PCB preparations.  2006 Elsevier Ltd. All rights reserved. Keywords: PCBs; PCNs; Fingerprinting; Bioindicator; Pine needles; Soils; Poland

1. Introduction The fate of chemicals in environment and their toxicity are mostly ruled by its physicochemical properties. Polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs), polychlorinated biphenyls (PCBs), polychlorinated naphthalenes (PCNs) and many other man-made organohalogenated compounds have been reported to be persistent in the environment (Puzyn and Falandysz, 2003; Noma et al., 2004a,b,c, 2005a,b). In general, these compounds have a very low water solubility, high n-octa-

* Corresponding author. Address: Department of Environmental Chemistry and Ecotoxicology, University of Gdan´sk, 18 Sobieskiego Street, PL 80-952 Gdan´sk, Poland. Tel.: +48 58 3450372; fax: +48 58 3450472. E-mail address: [email protected] (J. Falandysz).

0045-6535/$ - see front matter  2006 Elsevier Ltd. All rights reserved. doi:10.1016/j.chemosphere.2006.05.078

nol-water partition coefficients, and a low vapor pressure and tend to bioaccumulate and moreover are biomagnified within food-chains. For compounds with a well known potency to elicit Ah receptor-mediated mechanism of toxicity as PCDDs, PCDFs, non-ortho and mono-ortho PCBs toxic equivalency factors (TEFs) were assigned many years ago. In the near future TEFs for the other dioxin-like compounds such as polychlorinated naphthalenes (PCNs), polychlorinated dibenzothiophenes (PCDTs), polychlorinated trans-azobenzenes (PCt-ABs), polychlorinated trans-azoxybenzenes (PCt-AOBs) and many others might be evaluated. Since the relative potencies (REPs) of some of individual PCNs are related to 2,3,7,8-TCDD, so that the pattern of toxicity of PCNs resembles dioxin-like mechanism, and importantly the REPs of PCNs are similar to TEFs of mono-ortho PCBs (Blankenship et al., 2000; Villeneuve et al., 2000; Falandysz and Puzyn, 2004a).

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The levels of PCBs, PCDDs and PCDFs are monitored in many of the European countries within several national and international inventories including those forced by the UNEP programs and that of the European Environmental Agency, while data on dioxins are rare of Poland (Falandysz, 1999; Lassen et al., 2003). Some other dioxin-like compounds i.e. PCNs are not so extensively studied (Falandysz, 1998, 2003). Since two technical PCB and one technical PCN formulation in limited quantity were both manufactured and used in a few applications in Poland, and some other could be imported, in this study dioxin-like PCBs and PCNs were considered as group of chemicals notably contributing to TEQs of Polish agricultural soil and by means of analysis of pine needles as passive sampler – to air. Manufacture of PCBs started in 1929, as the beginning of their commercial use, and continued until 1990s with above 1.5 million metric tons produced (HELCOM, 2001). Although home production of PCBs did not contributed significantly to the world market, the amount of total Polish PCB formulations – Chlorofen and Tarnol – was estimated as 1700 metric tons. Additionally not identified quantities of PCB formulations from Soviet Union (Sovol) and Czechoslovakia (Delor) were imported to Poland before 1971. The presence of commercial PCB mixtures made in Germany (Clophen), France (Pyralene), and Italy (Phenoclor) in Polish installations and imported technical goods cannot be excluded (Falandysz, 1999; Falandysz and Szymczyk, 2001). Noteworthy highest concentrations of PCBs are reported for organic matter rich soils of northern latitudes – where higher deposition and persistence is possible due to climate conditions. In addition almost all historical emissions of PCBs occurred between 30 and 60 of northern hemisphere and as a result pattern of most recent emissions could be shifted northwards relative to the historical cumulative usage of these compounds (Meijer et al., 2003). Moreover PCBs are continuously formed and released to the environment during many anthropogenic processes (Falandysz, 1999; Lundgren, 2003). Polychlorinated naphthalenes are also widespread environmental pollutants that originate mainly from technical formulations formerly used for many industrial applications. Manufacture of PCNs dates from the beginning of 20th century and its highest rate of used was during 1930– 1960 and total global production of those compounds was estimated as 150 000 tonnes (Falandysz, 1999; Lundgren, 2003). Even though highly hydrophobic, semi-volatile, thermally stabile, with low flammability and with 15 congeners reportedly eliciting Ah receptor-mediated mechanism of toxicity, PCNs are still relatively less studied than other dioxin-like compounds (Falandysz, 2003). Recently published data on the historical profiles of PCNs in dated sediments cores showed that concentrations of those compounds decreased since middle 1980s (Horii et al., 2004; Yamashita et al., 2000a), hence very few data on their long-term distribution in the lower troposphere exist. PCNs were reported at elevated concentrations in a single sample of air collected from an urban and industrialized areas of

Poland (220 pg m 3), but low in a few of surface sediment (Falandysz et al., 1996; Jaward et al., 2004). A somewhat elevated levels of PCNs could be of course associate with ongoing using of PCNs containing products, but rather its formation in thermal processes as well as form volatilization from past uses in Europe should be considered as a potential source. Remarkably it has been reported that PCN are impurities of technical preparations of PCBs but also as unlawfully used in recent days (Yamashita et al., 2000b, 2003; Falandysz et al., 2004b; Taniyasu et al., 2005). In the view of mentioned facts, despite the analysis of routinely monitored PCDDs and PCDFs, authors decided to pay special attention to the historical implications of manufacturing and use of technical polychlorinated biphenyl formulations, as a source of dioxin-like PCBs to soil and air, as well as parallel investigate concentrations of PCNs as potential contributors to TEQs of agricultural soil and air. In the case of PCBs in this study congener-specific pattern of nonaand decaCB, with regards to usually analyzed di- to octaCBs, was considered as one of the tools to identify certain highly chlorinated technical formulations formerly used, as a potential source of pollution with PCBs. The very little studied before isomer-specific pattern of nona- and decaCBs varied in technical mixtures investigated (Wyrzykowska et al., 2006), and consequently this fingerprint could be considered as one of the tools to identify highly chlorinated Chlorofen technical formulation formerly produced and used in Poland, as a potential source of pollution with PCBs. Since preliminary investigations have revealed that pine needles can absorb various organohalogenated compounds from the ambient air (Hanari et al., 2004a,b), in this study they were selected as a cost effective and easy collectable environmental matrix suitable for long-term monitoring of the lower troposphere pollution with dioxin-like compounds. Simultaneously to pine needles, soil samples were investigated to determine interrelationships between concentration and pattern of PCNs and PCBs in pine needles and soil. Soils are natural sinks for persistent and lipophilic compounds such as dioxins received via different pathways of which the most important are atmospheric deposition, application of sewage sludge or composts, spills, erosion from nearby contaminated areas. Soil is a typical accumulating matrix with a long memory – dioxin-like compounds inputs received in the past will remain and, due to the POPs’ very long half-lives in soils, there is hardly any clearance and additionally due to adsorption to the organic carbon of the soil they will remain relatively immobile (AEA, 1999), therefore soil was selected as a environmental matrix to investigate the status of Polish terrestrial environments with regards to contamination with dioxin-like compounds. 2. Materials and method 2.1. Sampling The one-year-old needles of Scots pine (Pinus sylvestris L.) and soil samples were collected at several locations in

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Fig. 1. Location of the sampling sites and former production sites of technical PCBs formulations.

Poland in October 2002, both with regards to rural areas as well as those with heavy industrialisation ratio and heavily populated, including areas of former production of Polish technical PCBs formulations (Fig. 1). All samples were collected from the trees at a height of 1.5–2 m above the ground at a distance at least 200 m from the nearest road. The samples immediately after collection were packed in a solvent washed aluminum foil and packed in polyethylene bags. In the laboratory pine needles were initially cut into pieces about 3 cm in length, lyophilized and stored at 20 C until analysis. Prior to extraction the samples were homogenized in 300 ml of toluene. Agricultural soil samples were collected from corresponding locations, packed in polyethylene bags, deep-frozen at 20 C. Prior to analysis, the soil samples were freeze-dried, homogenized and sieved (1 mm mesh). 2.2. Chemicals The internal standard mixtures used were 13C-labeledPCDDs/Fs (NK-LCS-P, Wellington Laboratories, Ontario, Canada) and 13C-labeled PCBs (EC-4937, CIL, USA). All solvents used were of PCBs grade (Wako Chemicals, Japan). 2% (w/w) KOH–silica gel, 22% (w/w) H2SO4–silica gel, 44% (w/w) H2SO4–silica gel, 10% (w/w) AgNO3–silica gel and anhydrous sodium sulfate were of analytical grade (Wako Chemicals). Silica gel 60 (230–400 mesh) and basic alumina (70–230 mesh) were of analytical grade (MERCK, Germany). All materials and chemicals were treated as follows: the glassware was solvent-wash, dried and baked overnight at 250 C. The glass wool was Soxhlet-washed in n-hexane and dried using vacuum aspirator. The timber filters (ADVANTEC, Japan) were baked at 300 C for 12 h, while

anhydrous sodium sulfate, silica gel and alumina were baked at 500 C for 24 h. 2.3. Extraction The extraction of pine needles was performed in Soxhletextractor using a two-step procedure (initially needles were extracted using 300 ml of toluene and then followed by 50% methyl chloride in methyl alcohol – each step of extraction proceed for 7 h). After extraction, chlorophyll was removed by filtration of extract through a layer of silica gel. Soil samples were extracted in a two-step procedure with mixture of acetone and hexane (1:1, v/v) and subsequently with toluene in accelerated solvent extraction (ASE), system (ASE-200, DIONEX Co.). Before concentration extracts were filtered through anhydrous sodium sulfate to remove traces of water. The solvents were carefully evaporated to 5 ml using a rotary evaporator under vacuum pressure and finally to 3 ml with stream of nitrogen purged gently at 40 C (Hanari et al., 2004a,b; Horii et al., 2004). 2.4. Clean-up and fractionation Obtained extracts was first subjected to multi-layer silica gel clean-up. Pre-clean up was performed by passing extract through several layers of silica gel packed in a glass column (300 mm length · 20 mm i.d.) in a descending order, as follows: silica gel (0.8 g), 2% (w/w) KOH–silica gel (3 g), silica gel (0.8 g), 44% (w/w) H2SO4–silica gel (4 g), 22% (w/w) H2SO4–silica gel (4 g), silica gel (0.8 g), 10% (w/w) AgNO3–silica gel (8 g) and anhydrous sodium sulfate (5 g) packed on the top. The column was prewashed with 200 ml of n-hexane. Analytes were eluted with

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the next portion of n-hexane (200 ml). The effluent was carefully concentrated to 0.5 ml and adjusted with n-hexane to 1 ml. The analytes were further cleaned-up and fractionated using activated basic alumina column chromatography. Alumina (10 g) was activated at 130 C for 12 h and packed into glass column (300 mm length · 12mm i.d.) with anhydrous sodium sulfate layer (2 g) on the top. Analytes were eluted with n-hexane (20 ml, fraction 1), 0.5% methyl chloride in n-hexane (50 ml, fraction 2) and 50% methyl chloride in n-hexane (120 ml, fraction 3). All fractions were concentrated to 200 ll under a gentle stream of nitrogen. The two-dimensional HPLC – high performance liquid chromatography with porous graphite carbon column (HypercarbHPLC) and high performance liquid chromatography with pyrenyl silica column (PYE-HPLC) – were then performed to separate dioxins and dioxin-like compounds, including planar PCBs and dioxin like PCNs in pine needles and soil samples. Firstly solutions were fractionated by HypercarbHPLC – high performance liquid chromatography with porous graphitic carbon column (Table 1 panel a). Chromatography using activated carbon can separate chlorinated aromatic hydrocarbons on the basis of molecular planarity and to some extent on the degree of chlorination. Planar aromatic structures can reach closer to the carbon surface and therefore interact more strongly to what causes a longer retention time for PCBs with more planar structures (Lundgren, 2003). The 150 ll aliquots of fractions 2 and 3 obtained from alumina layer step was injected to Hypercarb-HPLC column which was forward eluted using 50% methyl chloride in n-hexane (fraction 2-1/3-1; 20 ml, 0-8 min) and back flushed using toluene (fraction 2-2/3-2; 50 ml, 8-28 min) (Table 1 panel a). First fraction contained most of PCBs, including mono- and di-ortho substituted PCBs, second fraction contained non-ortho PCBs and PCNs. The effluent

from Hypercarb-HPLC was spiked with 30 ll of isooctane and carefully microconcentrated to 30 ll and adjusted with n-hexane to 100 ll. The 50 ll of toluene fractions was further subjected for additional sub-fractionation step using a PYE-HPLC – HPLC with pyrenyl silica column (Table 1 panel b). The HPLC amino-columns separate the analytes according to the condensed number of aromatic rings, and it has been shown that the mechanism of separation is mainly due to a charge transfer between the lone pair of electrons on the stationary phase nitrogen and the p-electron cloud of the solute polyaromatics (Lundgren, 2003). The analytes from the PYE-HPLC column were collected into four sub-fractions: 2-2-1/3-2-1 (9.5 ml, 0–9.5 min), 22-2/3-2-2 (2.5 ml, 9.5–12 min), 2-2-3/3-2-3 (12 ml, 1224 min) and 2-2-4/3-2-4 (51 ml, 24–75 min) using 10% methyl chloride in n-hexane (0–27 min) and methyl chloride (27–75 min) as a mobile phase, respectively (Table 1 panel b). Each sub-fraction was spiked with 10 ll of 13C1,2,3,4-TeCDD/F (syringe spike) and microconcentrated to 100 ll under a gentle stream of nitrogen. The first fractions – 2-2-1 and 3-2-1 – contained non-ortho PCBs. PCNs were separated into all PYE-HPLC fractions, both with regards to the chlorination number and position of chlorine in the aromatic ring, which importantly enabled us to solve co-elution problems of some dioxin-like PCNs. The detailed information on PYE-HPLC fractionation are described elsewhere (Hanari et al., 2004a,b; Horii et al., 2004). 2.5. Analysis The standard of equivalent mixture of Kanechlors 300, 400, 500 and 600 (1:1:1:1, v/v) (GL Science, Japan) was used for identification and quantification of individual dito octachlorinated congeners, while the standard of

Table 1 Fractionation by two-dimensional high performance liquid chromatography – parameters of Hypercarb-HPLC (panel a) and PYE-HPLC (panel b) HPLC system Panel a Column Injector Flow Detector Effluent

SHIMADZU LC-10AD Hypercarb, Hypersil Co., stationary phase – porous graphitic carbon (100 mm length · 4.6 mm i.d., 7 lm grain size; keystone scientific, Bellefonte, PA, USA) RHEODYNE 7125, loop: 200 ll, injection volume: 150 ll 2.5 ml min 1 UV 254 nm 50% Methyl chloride/n-hexane (20 ml) (0–8 min) Toluene (50 ml) (back flushed) (8–28 min) Toluene (50 ml) (back flushed) (washing) (28–48 min) 50% Methyl chloride/n-hexane (25 ml) (washing) (48–58 min) Hewlett–Packard 1100

Panel b Column Injector Flow Detector Effluent

PYE, stationary phase – pyrenyl silica (250 mm length · 4.6 mm i.d., 5 lm grain size; Nacalai Tesque Inc., Japan) RHEODYNE 7725i, loop: 200 ll, injection volume: 50 ll 1.0 ml min 1 UV 254 nm 10% Methyl chloride/n-hexane (0–27 min) Methyl chloride (27–90 min) 10% Methyl chloride/n-hexane (90–105 min) (washing)

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equivalent mixtures of CB #206, 207, 208 and 209 (1:1:1:1, v/v) (Accu Standard, New Haven, CT, USA) was used for nona- and decachlorinated congeners, respectively. For the quantification of coplanar PCBs standard solution containing non-, mono- and di-ortho PCBs was used (EPA 1168PAR, Wellington Laboratories Inc., Ontario, Canada). For quantification of PCN congeners standard mixture of mono- to octachlorinated naphthalenes was used (PCN-MXB, Wellington Laboratories Inc., Ontario, Canada). The concentrations of PCBs and PCNs were determined by the HRGC/HRMS analysis using an Hewlett–Packard HP6890 GC interfaced with JEOL JMS-700D HRMS (Japan) operated in an electron impact (38 eV and 500 lA current) selected ion monitoring (SIM) mode at resolution R > 10000 MU (10% valley). For total PCBs analysis (but excluding coplanar congeners) GC was equipped with DB-1 capillary column (0.25 lm film thickness; 30 m length · 0.25 mm, J&W Scientific, Folsom, CA, USA) and for coplanar PCBs and PCNs analysis GC was equipped with DB-17 capillary column. The column oven temperature was programmed from 70 C (1 min) to 200 C at rate of 15 C min 1, and then to 270 C min 1 at rate of 4 C min 1, with a final hold time of 15 min for coplanar PCBs and for total PCBs from 70 C (1 min) to 180 C at rate of 15 C min 1, and then to 210 C min 1 at rate of 1.5 C min 1, and to 280 C min 1 at rate of 15 C min 1 with a final hold time of 25 min. For total PCNs parameters were respectively – from 70 C (1 min) to 180 C at rate of 15 C min 1, and then to 270 C min 1 at rate of 10 C min 1, with a final hold time of 10 min. Temperature of ion source and transfer line temperature was 270 C. Carrier gas (helium) flow rate was 1.5 ml min 1. In the case of coplanar PCBs and PCNs 2 ll of the extract was injected under splitless mode, while for total PCBs injection volume was 1 ll. Data acquisition was controlled by Microsoft Windows based MStation data system, while for identification and quantification JEOL DioK data system v. 2.02 (JEOL co.) software was used.

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2.6. Quality assurance/quality control Quality assurance/quality control (QA/QC) was performed by the analysis of matrix spikes, matrix spikes duplicates and procedural blanks (with each set of samples procedural blank was routinely analyzed). The samples of technical preparations were spiked with 13C-labeled internal standard solution of coplanar PCBs (including non- and mono-ortho PCBs; EC-4937, CIL, USA) prior to fractionation. The samples of pine needles prior to extraction were spiked with internal standards mixture of 13 C-labeled PCBs (EC-4937) and 13C-labeled PCDDs/Fs (NK-LCS-P). 13C-labeled PCDDs/Fs recovery standards were necessary to control recoveries of the further target compounds i.e. 2,3,7,8 substituted PCDDs/Fs (first results were reported – Bochentin et al., 2004; Wyrzykowska et al., 2005) and also into some extent useful for controlling of recovery in PCN containing fractions (since 13C-labeled PCNs recovery standards were not used in the present study). All samples after fractionation were syringe spiked with13C-labeled 1,2,3,4-TeCDD/F (NK-IS-E, Wellington Laboratories Inc., Ontario, Canada). The peaks were identified by retention times compared to standards if signal to noise (S/N) ratio was grater than 3 (S/N > 3) and were quantified if target/qualifier ion ratios were within ±15% of the theoretical values. The recovery rates were 107 ± 13.3 (13C-labeled PCDDs/Fs) and 101 ± 7.9 (13Clabeled PCBs) for pine needles and 93.5 ± 4.6 and 97.2 ± 8.3 for soils, respectively. The reported values are not recovery corrected. 3. Results and discussion 3.1. PCBs Total PCB concentrations of pine needles were in the range from 2.7 to 50 ng/g wet weight, with coplanar PCB concentrations as of 0.21–1.54 pg TEQ/ g wet weight (Table 2).

Table 2 Total PCBs (ng/g wet weight) and dioxin-like PCBs (pg TEQ/g wet weight) concentration of pine needles from Poland 1c

1d

2a

2b

4b

5b

7a

7c

8a

11a

0.047 0.43 1.11 0.85 1.15 0.59 0.055 0.0015 0.00020

0.13 0.94 1.24 1.58 1.15 0.34 0.063 0.0062 0.0012

0.011 0.41 2.14 1.38 2.02 1.21 0.17 0.0029 0.0018

0.095 3.74 12.7 4.25 7.64 3.63 0.36 0.0073 0.0039

0.12 3.73 12.6 4.30 7.68 3.59 0.39 0.0055 0.0037

0.084 0.65 0.72 0.71 0.43 0.14 0.013 0.0018 0.0010

0.12 1.14 1.85 1.26 0.95 0.33 0.040 0.0011 0.0011

0.12 1.06 1.95 1.23 0.95 0.37 0.046 0.0020 0.0012

0.15 2.55 9.08 4.92 10.73 5.71 0.680 0.013 0.0016

0.26 3.84 13.1 6.78 15.7 8.56 1.06 0.019 0.0032

4.2

5.5

7.3

32

32

2.7

5.7

5.7

34

50

0.30

0.28

0.21

12a

13a

ng/g w.w. 2CB 3CB 4CB 5CB 6CB 7CB 8CB 9CB 10CB P PCBs

0.067 0.78 3.32 2.46 4.68 1.81 0.27 0.013 0.0055 13

0.048 0.78 3.25 2.17 4.63 2.41 0.34 0.013 0.0061 14

pg TEQ/g w.w. P Dioxin-like PCBsa a

0.38

0.14

1.20

0.70

1.07

1.53

For the calculation of TEQs WHO TEFs (Van den Berg et al., 1998) for non-ortho and mono-ortho PCBs were used.

1.11

1.13

1.54

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The compositional pattern of PCBs homologue groups observed in pine needles shows that samples collected at sites somehow related to the production and use of Polish technical PCB formulations (Fig. 1) showed the highest abundance of highly chlorinated groups of 6CB, 7CB and 8CB (i.e. sampling site 11a, 12a, 13a – Fig. 2a). These homologue groups were reported to be predominant in the compositional profile of highly chlorinated technical PCB mixture Chlorofen (reportedly used as lubricant in hydraulic and other systems in mining) (Falandysz et al., 1992, 2004b; Wyrzykowska et al., 2006; Ishikawa et al., 2006). Elevated concentrations of total PCBs were in addition found in the samples from other industrialized and heavily populated areas of central Poland (e.g. 4b, 8a), while PCB levels in pine needles collected from rural sites of Poland (e.g. 7a) were almost 10 times lower than from other regions. Nona- and decachlorinated biphenyls were detected in all samples of the Polish pine needles showing a similar tendency in a concentrations as measured for total PCBs, with highest abundances of nona- and decaCB found in pine needles sampled in the south-eastern region

of Poland. Similarly as observed for the total PCBs levels, in sample from vicinity of Olsztyn (7a) neighboring poorly developed areas of Poland, the concentrations of nona- and decaCB was the smallest (Table 2a, Fig. 2a). Noteworthy samples from relatively pristine areas were demonstrating the different ratio between 9CBs:10CBs (approximately 1:1), which was opposite to the tendency of much more abundant 9CBs homologue group – if compared with 10CB – observed in samples form industrialized areas (Fig. 3a). In the soil samples the total concentration of PCBs ranged from 0.67 to 8.3 ng/g dry weight (with the concentration of coplanar PCBs as 0.077–0.42 pg TEQ/g dry weight) (Table 3) and though were noticeably lower than the concentrations observed for pine needles represented somehow a similar tendency both in concentration, homologue group pattern and nona- and decachlorinated biphenyls fingerprint, with a slightly higher proportion of highly chlorinated homologue groups in soil, and remarkably higher absolute concentrations of 9CB and 10CB (almost up to 10 times higher) (Fig. 3). As a comment to this fact it is

13a

13a

12a

12a

11a

11a

8a

8a

7c

7c

7a

7a

5b

5b

4b

4b

2b

2b

2a

2a

1d

1b

1c

1a 0%

20% 2CB

3CB

40% 4CB

5CB

60% 6CB

7CB

80% 8CB

9CB

100%

0%

10CB

20% 2CB

3CB

40% 4CB

5CB

60% 6CB

7CB

80% 8CB

9CB

100% 10CB

Fig. 2. Polychlorinated biphenyl homologue groups pattern in pine needles (a) and agricultural soils (b) from Poland.

a

b 160

20 18

140

16 120 12

pg/g d.w.

pg/g w.w.

14

10 8

100 80 60

6 40 4 20

2 0

0 1c

1d

2a

2b

4b

5b 9CB

7a 10CB

7c

8a

11a 12a 13a

1a

1b

2a

2b

4b

5b 9CB

7a

7c

8a

10CB

Fig. 3. 9CB and 10CB fingerprint in pine needles (a) and agricultural soils (b) from Poland.

11a 12a 13a

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Table 3 Total PCBs (ng/g dry weight) and dioxin-like PCBs (pg TEQ/g dry weight) concentrations in soils from Poland 1a

1b

2a

2b

4b

5b

7a

7c

8a

11a

12a

13a

<0.0008 0.065 0.14 0.18 0.51 0.31 0.068 0.0062 0.0055

0.027 0.22 0.57 0.16 0.33 0.41 0.035 0.0034 0.0029

0.0075 0.18 0.42 0.33 0.63 0.36 0.083 0.011 0.010

0.0019 0.11 0.33 0.25 0.71 0.46 0.16 0.034 0.043

0.026 1.09 0.57 0.43 0.91 0.46 0.11 0.020 0.020

0.0034 0.083 0.14 0.091 0.16 0.15 0.032 0.0046 0.0033

0.018 0.25 0.39 0.66 0.42 0.25 0.090 0.015 0.0073

0.0095 0.11 0.18 0.13 0.24 0.16 0.062 0.0071 0.0049

0.011 0.44 1.58 0.82 2.12 1.91 1.22 0.13 0.070

0.037 0.70 0.98 0.35 0.60 0.51 0.27 0.054 0.021

0.0049 0.15 0.52 0.33 1.04 1.30 1.13 0.15 0.048

1.8

2.0

2.1

3.6

0.67

2.1

0.90

8.3

3.5

4.7

0.054

0.26

0.18

0.14

0.081

0.077

0.10

0.42

0.20

0.17

ng/g d.w. 2CB 3CB 4CB 5CB 6CB 7CB 8CB 9CB 10CB P PCBs

<0.0008 0.081 0.45 0.38 0.94 0.55 0.11 0.011 0.013 2.5

1.3

pg TEQ/g d.w. P Dioxin-like PCBsa a

0.22

0.25

For the calculation of TEQs WHO-TEFs (Van den Berg et al., 1998) for non-ortho and mono-ortho PCBs were used.

worth mentioning that apparently environmental levels of lighter PCB congeners are less well-correlated with usage estimates than of heavier congeners, since lighter congeners tend to travel further from their initial release point, whereas highly chlorinated congeners tend to remain more closer to their sources regions and resemble more accurately the historical usage pattern (Meijer et al., 2003). Additionally nonaCB and decaCB have the lowest evaporation rate in 25 C among all biphenyl homologues – 3.5 · 10 6 and 8.5 · 10 7 g m 2 h 1, respectively (Erickson, 2001), consequently – they should remain in the contaminated soils for the longest time and reflect pollution from evaporative sources for the longest time. The results of this study also seem to predestinate nona- and decachlorinated biphenyls as a sensitive tool for the identification of certain point sources of pollution with PCBs, especially in the case of soil. 3.2. PCNs The concentration of PCNs ranged from 170 to 920 pg/g wet weight (Table 4) with dioxin-like PCNs at concentra-

tions from 0.03 to 0.11 pg TEQ/g wet weight. Correspondingly to PCBs levels the highest concentrations were found in the samples form industrial and heavily populated areas of Poland, with the highest concentration observed for samples nearest to the areas of production and intense use of technical PCBs preparations (mining industry in Silesia region). However, it should be noted that PCNs concentration were more uniform if compared with PCBs both in pine needles and soil (Table 4). Furthermore, in the homologue pattern no clear tendency was observed (Fig. 4). In the soil PCNs concentrations ranged from 350 to 1100 pg/g dry weight (0.023–0.15 pgTEQ/g wet weight) (Table 5). Interestingly a higher proportion of lower chlorinated PCN homologue groups was found in soil if compared with pine needles, what can suggest separate source of this compounds to soil. Based on published data on PCNs recognized as the impurities of PCBs technical preparation (Yamashita et al., 2000b; Falandysz et al., 2004b; Taniyasu et al., 2005) it can be concluded that Chlorofen could have played role as a source of polychlorinated

Table 4 Polychlorinated naphthalenes in pine needles from Poland (pg/g wet weight) 1c

1d

2a

2b

4b

5b

7a

7c

8a

11a

12a

13a

104.7 123.3 100.4 41.02 38.25 154.7

44.4 45.2 23.9 5.96 11.2 33.8

71.9 84.3 42.1 17.3 26.40 128.4

125.9 93.9 38.0 12.9 19.3 95.7

117.3 129.9 41.6 13.0 17.1 59.0

168.6 110.0 41.8 7.68 6.98 27.7

68.9 78.7 34.7 6.38 8.45 24.3

106.1 80.0 47.1 6.78 8.83 23.2

167.8 163.9 62.9 17.5 29.5 135.1

234.0 241.0 37.7 9.7 17.7 72.8

460.8 309.2 34.6 11.6 19.1 88.5

168.2 164.8 42.3 17.2 22.1 101.3

560

170

370

390

380

360

220

270

580

610

920

520

0.11

0.07

0.09

0.03

0.04

0.04

0.11

0.06

0.07

0.09

pg/g w.w. 3CN 4CN 5CN 6CN 7CN 8CN P PCNs

pg TEQ/g w.w. P Dioxin-like PCNsa a

0.15

0.04

For dioxin like PCNs REPs (Blankenship et al., 2000; Villeneuve et al., 2000) were rounded to value of either 1 or 5 according to approach used to deriving TEFs by WHO (Van den Berg et al., 1998).

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B. Wyrzykowska et al. / Chemosphere 67 (2007) 1877–1886

13a

13a

12a

12a

11a

11a

8a

8a

7c

7c

7a

7a

5b

5b

4b

4b

2b

2b

2a

2a

1d

1b 1a

1c 0%

20% 3CN

40% 4CN

5CN

60% 6CN

7CN

80%

100%

0%

20% 3CN

8CN

40% 4CN

5CN

60% 6CN

7CN

80%

100%

8CN

Fig. 4. Polychlorinated naphthalene homologue groups pattern in pine needles (a) and agricultural soils (b) from Poland.

Table 5 Polychlorinated naphthalenes in soils from Poland (pg/g dry weight) 1a

1b

2a

2b

4b

5b

7a

7c

8a

11a

12a

13a

227.7 202.21 93.64 31.47 12.19 18.45

649.2 366.9 57.54 27.06 8.40 19.64

179.2 113.4 23.95 9.25 7.48 25.54

195.2 179.7 32.16 14.27 8.20 29.96

247.2 253.5 22.58 7.04 3.61 4.25

162.2 108.15 36.88 11.25 5.97 25.89

243.2 145.4 21.08 6.28 4.21 13.63

250.4 102.3 18.86 5.30 4.60 31.76

183.4 96.52 16.12 14.50 12.75 37.30

203.5 199.8 28.59 10.86 6.08 6.35

199.5 185.5 62.01 20.95 13.79 90.39

408.7 466.6 172.4 37.13 13.88 34.52

590

1100

360

460

540

350

430

410

360

460

570

1100

0.042

0.056

0.040

0.048

0.031

0.023

0.064

0.057

0.084

0.15

pg/g d.w. 3CN 4CN 5CN 6CN 7CN 8CN P PCNs

pg TEQ/g d.w. P Dioxin-like PCNsa

0.14

0.098

a

For dioxin like PCNs REPs (Blankenship et al., 2000; Villeneuve et al., 2000) were rounded to the value of either 1 or 5 according to the approach used to derive TEFs by WHO (Van den Berg et al. 1998).

naphthalenes to the local environment, but this role should not be overestimated. Other sources – as an incineration processes or leakages from landfills – should not be forgotten and the impact of former use of polychlorinated naphthalene technical formulation should be considered as well, e.g. based on available data on congener-specific composition of PCN technical formulations (Falandysz et al., 2000; Noma et al., 2004d), some Halowaxes (HW1000, 1001, 1031, 1099) can be considered as a source of lowly chlorinated PCNs for soils (probably leakages from unprotected landfills/dump sites where PCN containing wastes are stored), while evaporative emissions from PCB former production and use (i.e. Chlorofen) as well as the use of Halowax 1051 could be a source of highly chlorinated naphthalenes for pine needles. Isomer-specific analysis (data not shown) shown also that 4CN – #44 and 5CN – #54 considered to be among the combustion markers (Hanari et al., 2004b; Helm and Bidleman, 2003) were detected in pine samples 4b, 5b, 8a, 13a, while in samples 1c, 1d, 2a, 2b, 4b, 5b, 7c, 8a, 12a and 13a congener #54 was identified, while in soil those congeners were detected only in few samples: #54 in 1a, 8a, 12a, 13a, and #44 only

in 2b. Nevertheless authors have started investigations on solid waste (furnace bottom ash) from household combustion processes, which improperly used or disposed could be a source of dioxin-like compounds for the soil. In conclusion it should be emphasized that study results confirm sustainability of pine needles as a biomatrix for long-term monitoring of an ambient air pollution – considered as a potential passive sampler for dioxin-like compounds in air, they reflected not only current pollution sources but also historical production, storage and use. Both in case of the PCBs and PCNs concentrations found in pine needles were higher if compared to the soil levels (only in the case of nona- and decachlorinated biphenyls the trend was opposite). This fact could be explained mostly by the presence of waxes on the surface of pines which reported effectively sequestering airborne lipid soluble compounds (Kylin et al., 1994; Hellstro¨m et al., 2004). Generally – both in the case of PCBs and PCNs – elevated concentrations are mostly linked to urbanized source areas. With respect to concentrations – also pattern of homologue groups varied between sampling sites. It should be noted that in the case of PCBs patterns found in soil and

B. Wyrzykowska et al. / Chemosphere 67 (2007) 1877–1886

in pine needles are rather analogous and in industrialized areas are obviously influenced by former production and use of technical PCB formulations. In the case of PCNs their technical formulations also seem to be a possible source of those compounds to the environment, but the former PCBs technical formulation production and use, as well as industrial and combustion-related sources should not be forgotten. For PCNs patterns found in soil were more likely connected to use of PCN technical formulations while in pines also seem to be influenced by many additional sources of those compound (PCBs technical formulations, industrial and combustion processes). Additionally from the findings of this study the fingerprint of nona- and decachlorinated biphenyls seem to be a promising tool to identify production and use of highly chlorinated technical PCB formulation as a source of those compounds to the environment – nona- and decachlorinated biphenyl concentrations and patterns in pine needles and soil from Poland still resemble the historical production and usage of Chlorofen. Further studies should be undertaken to improve the understanding of complex array of factors (air-surface exchange, seasonal and spatial differences, atmospheric transport and fate, life span of the foliage) controlling PCBs and other persistent organic pollutants concentrations in air and pine needles and differences of their behavior in the soil/air media. Acknowledgements This study was supported partly by the Japanese Society of the Promotion of Science (JSPS), Tokyo, Japan. The authors are very grateful to Prof. J. P. Giesy (Michigan State University) and Dr. K. Kannan (Wadsworth Center, New York State Department of Health) and Mr. T. Okazawa (AIST) for their support and useful advices. The authors are also grateful also to Dyrekcja Generalna Laso´w Panstwowych, Warszawa, Poland, and to Dyrekcje Regionalne Laso´w Panstwowych for kind cooperation. References AEA, 1999. Compilation of EU Dioxin Exposure and Health Data. Summary Report. EC DG Environment. DETR, October 1999. Blankenship, A., Kannan, K., Villalobos, S., Villeneuve, D., Falandysz, J., Imagawa, T., Jakobsson, E., Giesy, J., 2000. Relative potencies of Halowax mixtures and individual polychlorinated naphthalenes (PCNs) to induce Ah receptor-mediated responses in the rat hepatoma H4IIE-Luc cell bioassay. Environ. Sci. Technol. 34, 3153–3158. Bochentin, I., Falandysz, J., Orlikowska, A., Hanari, N., Wyrzykowska, B., Yamashita, N., 2004. Dioxin-like compounds (PCDDs, PCDFs and coplanar PCBs) in pine needles from Poland. Organohalogen Compd. 66, 2214–2221. Erickson, M.D., 2001. PCB properties, uses, occurrence and regulatory history. In: Robertson L.W. and Hansen L.G. (Eds.), PCBs. Recent Advances in Environmental Toxicology and Health effects. The University Press of Kentucky, 2001, pp. XI–XXX. Falandysz, J., 1998. Polychlorinated naphthalenes: an environmental update. Environ. Pollut. 101, 77–90. Falandysz, J., 1999. Polychlorinated biphenyls (PCBs) in the environment: chemistry, analysis, toxicity, concentrations and risk assessment (in

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