Biomonitoring of polycyclic aromatic hydrocarbons in urban and industrial environments of the Western Black Sea Region, Turkey

Biomonitoring of polycyclic aromatic hydrocarbonsin urban and industrial environments of the WesternBlack Sea Region, Turkey

Hasan Çabuk & Muhammet Samet Kılıç &

Muhammet Ören

Received: 25 April 2013 /Accepted: 1 October 2013 /Published online: 9 October 2013# Springer Science+Business Media Dordrecht 2013

Abstract This research was carried out in the cities ofZonguldak and Eregli, which have been characterized asurban and industrial environments of the Western BlackSea Region, Turkey, in order to assess the contaminationof polycyclic aromatic hydrocarbons (PAHs) usingmosses as biomonitors. The methodology involved thecollection of moss samples (Hypnum cupressiforme),ultrasonic extraction with dichloromethane, cleanup usingsilica gel and analysis by liquid chromatography withultraviolet detection. The total PAH concentrations rangedfrom 78.1 to 1693.5 ng g−1 in Zonguldak and from 15.2 to275.1 ng g−1 in Eregli. The total PAH concentration inEregli was about six times lower than that in Zonguldak,revealing the importance of switching from coal to naturalgas in residential heating. The diagnostic ratios and thecorrelation analysis have indicated that coal combustionand traffic emissions were the major PAH sources at bothsites. The contour maps were constructed for the determi-nation of spatial distributions of total PAHs, and it wasshown for Zonguldak as well as for Eregli that the PAHpollutionwasmuchmore predominant in highly populated

regions. Moving away from the city centres, a gradualdecrease in PAH pollution rates was observed.

Keywords Polycyclic aromatic hydrocarbons .

Biomonitoring .Hypnum cupressiforme .

Coal combustion . Zonguldak . Eregli

Introduction

Polycyclic aromatic hydrocarbons (PAHs) are the wide-spread environmental contaminants formed during incom-plete combustion of organic energy sources such as coal,oil, gas and wood, and are of special interest due to theirtoxicity, carcinogenicity and mutagenicity (Ravindra et al.2008). Once released into the atmosphere, PAHs arepartitioned between the gas and particle phases,transported over long distances, removed from the atmo-sphere by dry or wet deposition, and therefore accumulat-ed in soil, water and vegetation (Wagrowski and Hites1997; Harmens et al. 2013). A wide variety of plantmaterials and species such as leaves, grass, pine needles,mosses and lichens have been used as indicators to eval-uate PAH contamination levels in cities and heavily pol-luted industrial regions (Prajapati and Tripathi 2008;Cabrerizo et al. 2012; Holoubek et al. 2000; Ratola et al.2012). A number of studies have shown that concentra-tions of PAHs in plant tissues well reflected their ambientair concentrations (Lodovici et al. 1998; Alfani et al. 2001;Müller et al. 2001). In addition, the European legislationhas recommended the use of bioindicators for assessingthe impact of PAHs on ecosystems (EU 2004).

Environ Monit Assess (2014) 186:1515–1524DOI 10.1007/s10661-013-3470-2

H. Çabuk (*) :M. S. KılıçDepartment of Chemistry, Faculty of Arts and Sciences,Bülent Ecevit University,67100 Zonguldak, Turkeye-mail: [email protected]

M. ÖrenDepartment of Biology, Faculty of Arts and Sciences,Bülent Ecevit University,67100 Zonguldak, Turkey

Mosses are particularly useful for monitoring pur-poses due to their large surface area, high capacity toabsorb large organic molecules and their abilities toobtain water and nutrients from the air. These proper-ties make them ideal for evaluating pollutant levels inthe air over a monitored period of time (Lim et al.2006). Concentrations and distribution patterns ofPAHs in various moss species have been reported,including Hylocomium splendens (Migaszewski et al.2009; Zechmeister et al. 2006), Pleurozium schreberi(Dolegowska and Migaszewski 2011; Viskari et al.1997), Hypnum cupressiforme (Holoubek et al. 2000;Ötvös et al. 2004; Foan et al. 2010), Hypnum plumaefor-mae (Liu et al. 2005), Tortula muralis (Gerdol et al. 2002)and Sanionia uncinata (Cabrerizo et al. 2012). There aretwo basic ways to use mosses in biomonitoring studies,active and passive monitoring. In active biomonitoring, themoss species collected from its relatively unpolluted hab-itat are placed inmoss bags, transferred to amonitoring siteand exposed to ambient air over a period of time. Passivemonitoring involves the collection of moss species atvarious distances from the source and the analysis ofaccumulated pollutants in the samples. Due to simplicityand economy, the method of passive biomonitoring withvarious moss species has most often been used to screenatmospheric PAH pollution in urban and industrial areas.

This study presents the results of a biomonitoringstudy of atmospheric PAH deposition by analysingmosssamples from the cities of Zonguldak and Eregli, locatedin the Western Black Sea Region, Turkey. The principalaims of this investigation were to determine the concen-trations, spatial distributions and potential sources ofPAHs in the studied sites along with the comparison ofthe results with respect to site characteristics. The aim ofthe present study was also to test the hypothesis thatdomestic coal usage releases more PAHs than industrialcoal combustion.

Experimental

Study area and sampling

The study was performed in the cities of Zonguldakand Eregli, located in the Western Black Sea Region inTurkey. Zonguldak has a population of about 109,000inhabitants living in an area of 631 km2. The city isknown as a centre for coal mining and characterized asresidential urban area where the coal is mainly used as

fuel for domestic and central heating. The city of Eregliis situated 50 kmwest of the Zonguldak and has an areaof 782 km2 with a population of more than 102,000inhabitants. The city has the largest iron and steel plantin Turkey. For residential heating in the city, naturalgas is commonly utilized, while coal burning occursrarely. The climate of the Western Black Sea Region ispredominantly temperate and rainy, and the averageannual temperature is 13–15 °C. The annual rainfallis about 1,000–1,500 mm.

The moss samples were collected in Zonguldak andEregli from 17 and 21 sampling points, respectively.The sampling points were recorded with a global po-sitioning system (GPS) device. The map of the sam-pling points is shown in Fig. 1. The moss species, H.cupressiforme, was selected due to its widespread dis-tribution in the studied area. Sampling was completedduring a relatively dry period from June to August2012. The sampling points were at least 300 m frommain roads or villages and 100 m from minor roads andbuildings. When necessary, in more densely populatedareas, these distances were reduced to 100 m and 50 m,respectively. The mosses were air-dried at room tem-perature (16–22 °C), cleaned from all dead materialand attached litter by hand, and stored frozen (−18 °C)in polyethylene bags prior to PAH analysis.

Sample extraction and cleanup

A 5 g of homogenized moss sample was extracted twicewith 200 mL of dichloromethane in an ultrasonic bath(Elma T 660, Germany) for 30min at room temperature.The extracts were combined and concentrated to 1 mLusing a rotary evaporator (Büchi R-200, Switzerland).Interfering compounds in the concentrated extracts wereremoved by liquid–solid chromatography containing 2 gactivated silica gel and 1 g anhydrous Na2SO4. Thefraction containing PAHs was collected by eluting thecolumn with 20 mL of n-pentane/dichloromethane (1:1,v/v). The eluate was concentrated to about 1 mL using arotary evaporator and was evaporated to almost drynesswith a gentle stream of nitrogen. The residue was finallydissolved in 0.25 mL of acetonitrile. The extract, afterpassing through a 0.22-μm filter (Membrane Solutions,USA), was analysed.

All of the solvents used in the experiments were ofanalytical grade. Dichloromethane (>99.9 %) and n-pen-tane (>99.5 %) were purchased from Merck (Darmstadt,Germany). Anhydrous Na2SO4, silica gel (28–200 mesh)

1516 Environ Monit Assess (2014) 186:1515–1524

and high-performance liquid chromatography (HPLC)-grade acetonitrile (>99.9 %) were obtained from Sigma–Aldrich (Steinheim, Germany).

Instrumental analysis

The chromatographic analysis was performed using aThermo Finnigan HPLC system (San Jose, USA)consisting of a P1000 pump, a AS3000 automatic injectorsystem, a SCM 1000 degasser and a UV1000 detector.The system was controlled by a Specta System ControllerSN 4000 and a software package Chromquest 4.0. Theseparation was performed by means of a PhenomenexMax-RP column (250×4.6-mm i.d., 4.0 μm) protectedby a C18 guard column (4×3-mm i.d., Phenomenex). Agradient elution program was optimized by using themobile phases of acetonitrile and water (Direct-Q3 waterpurification system, Millipore, USA). The separation wasperformed at room temperature with a constant flow rateof 1 mL min−1 by employing the elution program asfollows: 0–5 min acetonitrile/water 70:30 (v/v) and thena linear gradient elution from 70% acetonitrile at 5 min to100% acetonitrile at 25 min, followed by isocratic elutionwith acetonitrile for 5 min. Finally, 10 min was necessaryin re-establishing the initial conditions. The detectionwavelength was set to 254 nm.

Quality assurance

The HPLC system was calibrated using an external stan-dardmethodwith QTMPAHmix (47930 Supelco), whichcontained naphthalene (NAP), 2-bromonaphthalene(2-BNAP), acenaphthylene (ACY), acenaphthene (ACE),fluorene (FLU), phenanthrene (PHE), anthracene (ANT),pyrene (PYR), fluoranthene (FLT), benzo(a)anthracene(BaA), chrysene (CHY), benzo(b)fluoranthene (BbF),benzo(a)pyrene (BaP), dibenzo(a,h)anthracene (DahA),indeno(1,2,3-cd)pyrene (IcdP) and benzo(g,h,i)perylene(BghiP). Five concentrations of each PAH ranging from20 to 500 ng mL−1 were prepared by diluting the QTMPAH standard solution. The calibration standards of PAHswere analysed by HPLC–UV, and a calibration curve foreach PAH standard was obtained by plotting the concen-tration against the area. Linearity was observed with thesquare of correlation coefficients (R2) ranging from 0.9947to 0.9996. The recovery experiments were carried out byspiking the pre-cleaned moss samples with two differentconcentrations of PAH standard solutions (100 and250 ng mL−1). The spiked samples as well as the unspikedcontrols were analysed in duplicate. The recoveries werecalculated from the differences in the total amounts of eachPAH between the spiked and unspiked samples. The re-coveries obtained were as follows: PHE, 78 %; ANT,54 %; PYR, 64 %; FLT, 69 %; BaA, 77 %; CHY, 69 %;

Fig. 1 Map of the sampling sites in Zonguldak and Eregli, Turkey

Environ Monit Assess (2014) 186:1515–1524 1517

BbF, 73 %; BaP, 75 %; DahA, 74 %; IcdP, 73 %; andBghiP, 72%. The recoveries of NAP, ACY, ACE and FLUwere below 45%, owing to their high volatility and partialloss during the evaporation step; therefore, these com-pounds have not been considered in the results presentedin this study. The results of low recoveries for the lowestmolecular weight PAHs in different matrices were reportedin the literature (Mantis et al. 2005; Miege et al. 2003;Barranco et al. 2003). All the sample concentrations werecorrected based on these recovery values. The repeatabilitywas studied for the three replicate analyses of the spikedsamples at concentration levels of 100 and 250 ngmL−1 ofeach PAH. The relative standard deviations (RSDs) weresatisfactory, ranging from 2 to 8 % for all the compounds.The method detection limit (MDL) of each PAH obtainedfrom the calculations using theHPLC–UVresults based onS/N=3 ranged from 0.06 ng g−1 (ANT) to 1.92 ng g−1

(DahA). The procedural blanks were also performed peri-odically to check for interference and contamination aris-ing from the solvents, glassware and filters. The concen-trations of PAHs in the blank samples were no more thanthe detection limits.

Results and discussion

PAH concentrations in mosses

The arithmetic mean plus standard deviation (SD), geo-metric mean and median values for the concentrations ofPAHs in the moss samples collected from the cities ofZonguldak and Eregli are given in Table 1. All the resultswere expressed on a dry weight basis. The total PAHconcentrations ranged from 78.1 to 1,693.5 ng g−1 with amean value of 467.0 ng g−1 in Zonguldak and from 15.2to 275.1 ng g−1 with a mean value of 83.3 ng g−1 inEregli. Themost abundant PAHs in Zonguldakwere BbF(22%), FLT (14%), CHY (12%) and PHE (10%), whilein Eregli, the most abundant PAHs were BbF (20 %),FLT (13%), PHE (12%) and BghiP (11%). In general, itis expected that the sites located in an industrial environ-ment have the highest contamination levels when com-pared to the urban and residential ones. In contrast, it wasfound that Zonguldak had much higher concentrationsthan Eregli: about 6 orders of magnitude for total PAHsand 4 to 8 orders of magnitude for individual PAHs. Thedifference of PAH concentrations between Zonguldakand Eregli was found to be statistically significant(Student's t test, P<0.01). The rapid and irregular

urbanization and, in particular, un-processed coal usagefor residential heating during the winter months canexplain why the PAH concentrations is so high inZonguldak. In earlier studies, the particle and gas phaseconcentrations, seasonal variations and sources of PAHsin the atmosphere of Zonguldak were examined in detailfor the period of May 2007 and April 2008. The concen-trations of PAHs decreased from May at the end of theresidential heating season and increased again fromSeptember at the start of the residential heating season.The results obtained from factor analysis in combinationwith diagnostic ratios revealed that coal combustion wasthe major pollutant source for atmospheric PAHs inZonguldak (Akyüz and Çabuk 2008; Akyüz and Çabuk2010). Currently, there is an attempt in Zonguldak toreplace coal with natural gas for heating purposes, whichis expected to be an effective way of reducing PAHemission. In recent years, the fact that Eregli switchedfrom coal to natural gas caused improvement of its airquality. Another possible reason for obtaining lower PAHconcentrations in Eregli might be that the iron–steelfactories operating in the region throughout the year havea fairly new technology and made large investments toreduce emissions. Some previous studies also pointed outthat domestic coal usage releases more PAHs than indus-trial coal combustion (Xu et al. 2006; Ravindra et al.2008). The burning of coal at small scale causes incom-plete combustion at low temperature, while the industrialutilization occurs with very high thermal efficiency.

The percentage of three- to four- and five- to six-ringPAHs in themoss samples for both cities was 53 and 47%,respectively. In general, the atmospheric PAHs with threearomatic rings are associated primarily with the gas phase,four-ring PAHs are distributed almost equally between theparticle and gas phases, and five- and six-ring PAHs arealmost completely associated with the particle phase.(Cincinelli et al. 2007; Tasdemir and Esen 2007; Akyüzand Çabuk 2010). The findings implied that the mosssamples comprised together of light PAHs (three to fourrings) and heavy PAHs (five to six rings) supplied by bothgas and particle phase depositions. Ötvös et al. (2004)reported that in the moss samples (H. cupressiforme) col-lected from different locations in Hungary, heavy PAHs(having more than three rings) accounted for less than 1 %of the total PAHs. Dolegowska and Migaszewski (2011)found the percentage of the five+six-ring hydrocarbons tobe about 30% in the mossesH. splendens and P. schreberifrom the Kielce area, Poland. Liu et al. (2005) as well asFoan et al. (2010) also noted lower concentrations of high-


molecular PAHs in the mosses collected from southernChina and northern Spain, respectively. In comparisonwith the mentioned PAH ring profiles, the present studyexhibited higher levels of heavy PAHs, revealing the im-portance of combustion sources in the studied area. HeavyPAHs (especially five- and six-ring PAHs) often comefrom coal and fuel combustion and coke manufacturing(Augusto et al. 2009).

The total PAH concentrations obtained for Zonguldak(78–1,694 ng g−1) and Eregli (15–275 ng g−1) werecompared with the published data for different countries,although it should be noted that the moss species,biomonitoring techniques and site characteristics weredifferent. Total PAH concentrations at both sites weregenerally found to be lower than those in some other

countries as reported by Ötvös et al. (2004) for Hungary(1,567–10,450 ng g−1) and by Migaszewski et al.(2009) for Poland (473–2,970 ng g−1) and Alaska(80–3,390 ng g−1), whereas the total PAH concentrationswere similar to those reported by Liu et al. (2005) forSouth China (310–1,340 ng g−1), by Foan et al. (2010)for Spain (86–372 ng g−1), Viskari (2000) for Finland(115–404 ng g−1) and Krommer et al. (2007) for Austria(120–730 ng g−1).

Possible sources of PAHs

The concentration ratios of some specific PAHs haverecently become an efficient tool for identifying andassessing emission sources in different environmental

Table 1 Concentrations of individual PAHs in moss samples collected from the cities of Zonguldak and Eregli in Turkey

PAHs Zonguldak (ng g−1 dry weight) Eregli (ng g−1 dry weight)

N Arithmetic mean±SD Geometric mean Median N Arithmetic mean±SD Geometric mean Median

PHE 17 48.9±50.6 34.4 29.3 21 9.8±4.3 8.9 7.9

ANT 17 8.54±9.76 5.62 4.31 21 1.5±1.6 1.1 0.9

PYR 16 47.8±54.6 29.9 33.7 19 8.4±4.1 7.5 7.1

FLT 17 64.5±67.5 46.1 43.7 19 10.6±5.9 9.0 8.5

BaA 17 27.1±29.2 17.6 20.5 21 6.1±11.9 3.6 3.0

CHY 17 55.2±60.7 37.9 32.5 21 6.8±6.3 4.7 4.8

BbF 17 104.0±110.4 75.5 71.5 21 16.3±12.6 13.1 12.0

BaP 17 31.8±34.8 22.6 20.5 21 7.2±9.1 4.0 3.3

DahA 9 4.11±2.15 3.69 3.61 – ND ND ND

IcdP 17 39.2±31.9 30.8 33.8 21 8.1±8.2 5.9 6.1

BghiP 17 40.7±35.7 31.9 33.2 21 9.4±11.9 6.4 5.5

∑PAH 17 467.0±464.1 341.8 300.7 21 83.3±61.8 66.7 54.6

ND not detected, SD standard deviation

Fig. 2 Plot of FLT/(FLT+PYR) versus IcdP/(IcdP+BghiP)


media such as air, water, sediment, soil and biomonitoringmaterials. IcdP/(IcdP+BghiP), FLT/(FLT+PYR),ANT/(ANT+PHE), BaP/BghiP and BaA/(BaA+CHY)are the most frequently evaluated ratios in the literature(Mandalakis et al. 2005; Maliszewska-Kordybach et al.2008; Wang et al. 2010; Dolegowska and Migaszewski2011). These ratios distinguish PAH pollution originatingfrom petroleum products, petroleum combustion or bio-mass or coal burning. However, the PAH diagnostic ratios

should be used with caution, as their values may changeduring the environmental fate of these compounds(Tobiszewski and Namiesnik 2012). ANT and BaP areexpected to be degraded more easily than their isomersduring transportation due to their higher reactivity. Liuet al. (2005) found that the average ratio of PHE/ANTwas 20 in moss and 12 in the atmosphere. This findingattested to the continued photodegradation of ANT in themoss. DeNicola et al. (2011) as well as Ratola et al. (2012)

Table 2 Correlation matrix of the individual PAH compounds in Zonguldak

PAH Number PHE ANT PYR FLT BaA CHY BbF BaP DahA IcdP BghiP TPAH

PHE 17 1.000

ANT 17 0.943a 1.000

PYR 16 0.986a 0.931a 1.000

FLT 17 0.977a 0.985a 0.974a 1.000

BaA 17 0.839a 0.965a 0.834a 0.930a 1.000

CHY 17 0.918a 0.991a 0.909a 0.973a 0.977a 1.000

BbF 17 0.762a 0.907a 0.746a 0.854a 0.962a 0.933a 1.000

BaP 17 0.876a 0.964a 0.885a 0.952a 0.981a 0.981a 0.948a 1.000

DahA 9 0.553 0.628 0.611 0.607 0.646 0.648 0.673b 0.682b 1.000

IcdP 17 0.794a 0.912a 0.802a 0.888a 0.970a 0.939a 0.962a 0.967a 0.643 1.000

BghiP 17 0.747a 0.889a 0.748a 0.845a 0.956a 0.922a 0.987a 0.950a 0.730b 0.984a 1.000

TPAH 17 0.919a 0.986a 0.916a 0.974a 0.979a 0.993a 0.948a 0.989a 0.647 0.959a 0.943a 1.000

TPAH total PAH concentrationa Correlation is significant at the 0.01 level (two-tailed)b Correlation is significant at the 0.05 level (two-tailed)

Table 3 Correlation matrix of the individual PAH compounds in Eregli

PAH Number PHE ANT PYR FLT BaA CHY BbF BaP IcdP BghiP TPAH

PHE 21 1.000

ANT 21 0.569a 1.000

PYR 19 0.755a 0.348 1.000

FLT 19 0.839a 0.163 0.775a 1.000

BaA 21 −0.006 0.018 −0.021 0.032 1.000

CHY 21 0.522b 0.354 0.395 0.567a 0.735a 1.000

BbF 21 0.493b 0.244 0.447b 0.520b 0.832a 0.926a 1.000

BaP 21 0.494b 0.289 0.439 0.657a 0.419 0.744a 0.660a 1.000

IcdP 21 0.263 0.100 0.249 0.348 0.865a 0.807a 0.886a 0.730a 1.000

BghiP 21 0.137 0.029 0.181 0.205 0.948a 0.792a 0.891a 0.553a 0.954a 1.000

TPAH 21 0.513b 0.266 0.465b 0.567a 0.803a 0.938a 0.975a 0.782a 0.934a 0.895a 1.000

TPAH total PAH concentrationa Correlation is significant at the 0.01 level (two-tailed)b Correlation is significant at the 0.05 level (two-tailed)


found very high PHE/ANT values in Quercus ilex leavesand pine needles that could have resulted from the fasterdecay of ANT. Hence, the FLT/(FLT/PYR) andIcdP/(IcdP/BghiP) ratios are more representative to esti-mate PAH emission sources than ANT/(ANT/PHE) andBaA/(BaA/CHR), which are particularly sensitive tophotodegradation (Tobiszewski and Namiesnik 2012).

Figure 2 represents the IcdP/(IcdP+BghiP) ratio ver-sus the FLT/(FLT+PYR) ratio, one of the most usedplots in literature (Maliszewska-Kordybach et al. 2008;Demircioglu et al. 2011; Dolegowska and Migaszewski2011). As can be seen in Fig. 2, the results indicated thatthe diagnostic ratios exhibited similar distribution pat-terns for both sites. The IcdP/(IcdP+BghiP) ratio above0.5 is related to pyrolytic processes, particularly tocoal/wood/grass combustion, while the ratio below 0.5is an indicator of liquid fuel combustion. The ratios ofIcdP/(IcdP+BghiP) obtained in this work were in therange of 0.44 to 0.58 with a mean value of 0.51 inZonguldak and in the range of 0.38 to 0.58 with a meanvalue of 0.50 in Eregli, implying the importance of coalcombustion and vehicle emission in the study area. Ithas been reported that FLT/(FLT+PYR) ratios below0.40 imply the prominence of unburned petroleum(petrogenic sources), ratios from 0.40 to 0.50 suggest

the combustion of liquid fossil fuels (vehicle and crudeoil), whereas ratios larger than 0.50 are characteristic forgrass, wood, or coal combustion (Yunker et al. 2002). Inthis study, the ratios of FLT/(FLT+PYR) were in therange of 0.55 to 0.76 with a mean value of 0.61 inZonguldak and in the range of 0.43 to 0.70 with a meanvalue of 0.55 in Eregli, indicating the predominantinfluence of coal combustion in both sites.

In addition to diagnostic ratios, the relationships be-tween individual PAHs give an idea whether they alloriginate from the same source or not. In the literature,correlation analysis has beenwidely used to investigate therelationships between individual PAHs in moss samples(Galuszka 2007; Dolegowska and Migaszewski 2011) aswell as in other environmental matrices (Wu et al. 2005;Chung et al. 2007; Rajput and Lakhani 2009). In this study,the relationships between individual PAHs were investi-gated by Pearson's correlation analysis (SPSS 13.0). Thecorrelation coefficients (r) obtained for Zonguldak andEregli are shown in Tables 2 and 3. The correlation coef-ficients were marked in the table to indicate the signifi-cance levels (P<0.01 or <0.05).

In Zonguldak, not surprisingly, all individual PAHs,except DahA, were well correlated (P<0.01) with eachother, suggesting that PAHs originated from similar

Fig. 3 Spatial distribution of the total PAHs in Zonguldak


emission sources, and they were being influenced by thesame local conditions until they have deposited onmoss. As DahA was detected only in a small numberof moss samples, the correlations between DahA andother PAHs remained weak. The findings are in agree-ment with those of a previous study (Akyüz and Çabuk2010), showing that the local combustion sources inZonguldak determine the atmospheric concentrationsof PAHs, and other potential sources could not be dif-ferentiated due to high dominance of local emissions. InEregli, there was a strong relationship (P<0.01) be-tween high molecular PAHs (BaA, CHY, BbF, BaP,IcdP and BghiP), which indicated significant contribu-tions from combustion sources. High molecular weightparent PAHs is a typical characteristic of a combustionorigin (Augusto et al. 2009). Low molecular PAHsexhibited weak as well as insignificant correlations with

each other, indicating the impact of the mixed sources ofPAHs in the study area. It can be concluded that the mainsource of the PAHs in Eregli is pyrogenic, but othersources also seem to have made minor contributions.

Spatial distribution of PAHs

Spatial distribution of total PAHs was mapped usingMapInfo Professional 7.0 with Vertical Mapper 2.1.The distribution maps for Zonguldak and Eregli areshown in Figs. 3 and 4, respectively. In both Zonguldakand Eregli, the PAH pollution was much more predom-inant in highly populated regions. When moving awayfrom the city centres, a gradual decrease in PAH pollutionwas observed. The highest PAH concentrations inmosseswere observed in Zonguldak at sites Z10 and Z11, with avalue of 1,622 and 1,693 ng g−1, respectively. The

Fig. 4 Spatial distributionof the total PAHs in Eregli


majority of the buildings at these sites are heated by smallcoal stoves, especially in low-income level families, in-stead of a centralized heating system. The populationliving at these sites prefers to use un-processed coal dueto its low cost. Since its quality is very low, a largequantity of coal has to be burned to obtain sufficientheating, which increases PAH emissions significantly.High concentrations of 301, 435, 480, 506 and407 ng g−1 were also found at sites Z3, Z4, Z5, Z6 andZ7, respectively. These sites are close to the main roads,thus the high PAH concentrations found at these sitesmight be associated with emissions from gasoline anddiesel combustion. In Eregli, the sites E1, E2, E3 andE15, located in the proximity of the iron–steel plant, werethe most contaminated sites with the total PAH concen-trations of 157, 172, 275 and 147 ng g−1, respectively. Inthe steel and iron industries, PAHs are released from cokemanufacturing, sintering, iron making, casting, moldporing and cooling, and steel making (Yang et al.2002). In addition to industrial emissions, domesticheating and vehicular traffic also contributed to PAHpollution in Eregli, because the residential settlementwas largely established around the iron–steel plant.

Conclusions

In summary, the use of mosses as a biomonitoringmethod allowed a real comparison in terms of PAHcontamination between urban and industrial sites. Thetotal PAH concentrations at the urban sites were aboutsix times higher than those at the industrial sites,revealing the influence of coal usage in domesticand central heating. The results also indicated thereducing effect of switching from coal to natural gasin residential heating. Evaluation of the diagnosticratios and the correlation analysis suggested that coalcombustion and traffic emissions were the major PAHsources at both sites. The spatial distribution of PAHsindicated relatively high concentrations in denselypopulated sites. When moving away from the citycentres, a gradual decrease in PAH pollution rateswas observed. Therefore, the contribution of localsources seems dominant, rather than long-range at-mospheric transport.

Acknowledgments The author wishes to thank the BülentEcevit University (2012-10-03-07) for the opportunity and sup-port to carry out this research.

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Biomonitoring of polycyclic aromatic hydrocarbons in urban and industrial environments of the Western Black Sea Region, Turkey