Perfluorinated alkylated substances and brominated flame retardants in serum of the Czech adult population

Accepted Manuscript Title: Perfluorinated alkylated substances and brominated flame retardants in serum of the Czech adult population ˇ a Author: Lenka Sochorov´a Lenka Hanzl´ıkov´a Milena Cern´ ˇ Anna Drg´acˇ ov´a Alena Fialov´a Andrea Svarcov´a Tom´asˇ Grambliˇcka Jana Pulkrabov´a PII: DOI: Reference:

S1438-4639(16)30114-6 http://dx.doi.org/doi:10.1016/j.ijheh.2016.09.003 IJHEH 12970

To appear in: Received date: Revised date: Accepted date:

30-6-2016 15-8-2016 5-9-2016

ˇ a, Milena, Please cite this article as: Sochorov´a, Lenka, Hanzl´ıkov´a, Lenka, Cern´ ˇ Drg´acˇ ov´a, Anna, Fialov´a, Alena, Svarcov´ a, Andrea, Grambliˇcka, Tom´asˇ, Pulkrabov´a, Jana, Perfluorinated alkylated substances and brominated flame retardants in serum of the Czech adult population.International Journal of Hygiene and Environmental Health http://dx.doi.org/10.1016/j.ijheh.2016.09.003 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Perfluorinated alkylated substances and brominated flame retardants in serum of the Czech adult population Lenka Sochorováa,*, Lenka Hanzlíkováa, Milena Černáa, Anna Drgáčováa, Alena Fialováa, Andrea Švarcováb, Tomáš Grambličkab, Jana Pulkrabováb a

National Institute of Public Health, Šrobárova 48, 100 42 Prague, Czech Republic

b

University of Chemistry and Technology, Prague, Faculty of Food and Biochemical Technology,

Department of Food Analysis and Nutrition, Technická 3, 166 28 Prague, Czech Republic

*Corresponding author. Tel.: +420 267082268 E-mail address: [email protected]

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Abstract Persistent organic pollutants, such as perfluorinated alkylated substances (PFASs) and brominated flame retardants (BFRs) are widespread in the environment and most of them are bioaccumulated in wildlife and humans. The present study is the first investigation to reveal the PFAS and BFR levels of serum samples in the adult population of the Czech Republic. Altogether, 300 serum samples from blood donors in four cities were examined. In all samples 19 PFASs and 33 BFRs, including some of their metabolites, were targeted. The analyses were performed using ultra high performance liquid chromatography coupled with tandem mass spectrometry or gas chromatography with mass spectrometry (according to the type of analyte). PFASs, with the carbon chain length C6 and higher, dominated in all samples. Perfluorooctanesulfonate (PFOS; median: 2.43 ng/mL), perfluorooctanoic acid (PFOA; median: 0.756 ng/mL), perfluorodecanoic acid (PFDA; median: 0.145 ng/mL) and perfluorohexanesulfonate (PFHxS; median: 0.184 ng/mL) were detected in 100 % of samples. Perfluorononanoic acid (PFNA; median: 0.325 ng/mL) and perfluoroundecanoic acid (PFUdA; median: 0.058 ng/mL) in 99.7% and 96.0 % of samples, respectively. We observed statistically significant associations (p<0.05) between selected PFAS concentrations and the locality, gender, age of donors and education level. None of the BFRs was detected above the LOQ in more than 9 % of the samples. The most frequently detected representatives of this group were congeners of polybrominated diphenyl ethers, namely BDE-47 (in 8.7 %; range: 0.496-5.44 ng/g lipid weight (l.w.)), BDE-99 (in 6.0 %; range: 0.706 – 9.46 ng/g l.w.), BDE-153 (in 7.3 %; range: 0.736-6.44 ng/g l.w.) and BDE-209 (in 7.0 %; range: 13.7 -2693 ng/g l.w.). Highlights 

The first study in the Czech Republic to measure PFASs and BFRs in serum samples

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PFOS, PFOA, PFDA and PFHxS were the most abundant compounds

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BFRs were only detected in 9 % of the samples

Keywords Human biomonitoring, persistent organic pollutants, perfluorinated alkylated substances, brominated flame retardants, Czech Republic Introduction Over the last decade, persistent organic pollutants (POPs) such as perfluorinated alkylated substances (PFASs) and brominated flame retardants (BFRs) have received attention all over the world due to their widespread occurrence, potential toxicity, persistence and bioaccumulation in the environment (Antignac et al., 2013; Fromme et al., 2015; Kärrman et al., 2007). A number of studies have revealed that these substances have been found in various environmental matrices, in wildlife and humans (Lyche et al., 2015; Pinney et al., 2014; Pulkrabová et al., 2009; Schröter-Kermani et al., 2013; Toms et al., 2009b; Wang et al., 2011; Wu et al., 2015). Perfluorooctanesulfonate (PFOS) and perfluorooctanoic acid (PFOA) represent two of the most studied compounds from the PFASs group which are typically reported in environmental studies and have been widely used for many industrial and consumer applications such as surface treatments for fabrics, carpets or paper packaging (Axmon et al., 2014). Contrary to the classic POPs, including BFRs, PFASs do not tend to accumulate in lipids, but rather bind to protein (Fromme et al., 2015; Melzer et al., 2010; Pulkrabová et al., 2009). The BFRs of most concern are polybrominated diphenyl ethers (PBDEs), hexabromocyclododecanes (HBCDs) and tetrabromobisphenol A (TBBPA). They are integrated into potentially flammable materials such as electronic devices, upholstery or textiles to prevent fire hazard (Casas et al., 2013; Covaci et al., 2011; Lankova et al., 2013). Both PFASs and BFRs contain compounds that are considered endocrine disruptors (Kadar et al., 2011; Legler, 2008; Lyche et al., 2015; Wan et al., 2013). 3

Human biomonitoring is an important tool for assessing human exposure to emerging environmental pollutants. As the data on Czech population exposure to BFRs and PFASs are very limited, these compounds were first included as new biomarkers into the ongoing Czech Human Biomonitoring system in 2014 (CZ-HBM) (Černá et al., 2016, this issue). Hitherto, only few studies on BFRs in human milk (Kazda et al., 2004) and human adipose tissue (Pulkrabová et al., 2009) and PFASs in human milk (Lankova et al., 2013) are available. But no comprehensive studies dealing with these issues have been conducted in this region. The main objective of the present study was to assess current levels of PFAS and BFR in the Czech adult population and the potential effects of certain factors (gender, age, locality, etc.) on serum concentration. Material and methods Samples collection A total of 300 serum samples were collected in 2015 from the blood donors in four cities Prague, Ostrava, Liberec and Ţďár nad Sázavou (Fig.1). These locations were chosen for regional diversities in terms of population and urbanization. Prague is the most densely populated area in the Czech Republic with the highest level of traffic. Ostrava is third biggest city of the Czech Republic and is the most industrialized area. Liberec and Ţďár nad Sázavou are smaller urban residential areas with number of inhabitants approximately around 100 000. All participants (mean age 40.8) were residents in the represented area for at least 3 years and also completed a detailed questionnaire to provide information on their lifestyle factors. The main characteristics of this population are reported in Table 1. The study protocol was approved by the Ethical Committee on the National Institute of Public Health in Prague. Participants received written information and signed appropriate informed consent. Sample preparation

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The donors donated 9 mL of whole blood at the Transfusion Center. The serum was separated by centrifugation at 3000×g for 20 min and thereafter transferred into methanol-rinsed glass tubes. Frozen sera were shipped to laboratories and stored at -20 °C until analysis. Analyses were performed in accredited laboratories according to standard operation procedures in compliance with external quality assurance and control. Analytical method The accredited procedure used for the analysis of human blood serum samples is described in detail in our earlier study (Švarcová et al., 2016); therefore, there is only a brief summary of procedure steps in the following paragraphs: The sample (3 g of serum) was mixed with 3 mL of acetonitrile and 6 mL of diethyl ether/n-hexane (9:1, v/v) and shaken for 2 minutes. The extraction step was repeated twice with adding 3 mL diethyl ether/n-hexane (9:1, v/v). The combined extract of the upper layer (11 mL) was purified after evaporation using solid phase extraction (SPE) on silica column (0.5 g), the target analytes were eluted by 6 mL of the mixture n-hexane:dichloromethane (3:1, v/v). The eluate was dissolved after solvent evaporation in 200 µl of isooctane. This fraction was analyzed for 16 PBDEs and other 6 BFRs (hexabromobenzene (HBB), pentabromoethylbenzene (PBEB), pentabromotoluene (PBT), bis(2,4,6-tribromophenoxy) ethane (BTBPE), octabromo-1-phenyl-1,3,3-trimethylindan (OBIND), decabromodiphenyl ethane (DBDPE)) using a Agilent 7890A GC chromatograph (Agilent Technologies, USA) coupled to a triple quadrupole mass spectrometer Agilent 7000B MS (Agilent Technologies) operated in the negative chemical ionization mode (NCI). Further details about GC-MS method are described in detail by Kalachova et al., 2013. The lower acetonitrile layer was shaken for 2 min. after addition of further 5 mL of acetonitrile and 5 mL of deionized water, sodium chloride (1 g) and magnesium sulphate (4 g). The organic phase was evaporated and the residues were dissolved in 0.25 mL of 5

methanol. The last step of this extraction was filtration through a nylon filter (0.22 µm) and transfer into a vial for the LC analysis on an Acquity Ultra-Performance LC system (Waters, USA) hyphenated with a tandem quadrupole mass spectrometer XEVO TQ-S (Waters) operated in the multiple reaction monitoring mode (MRM) with electrospray in the negative ionization mode (ESI). Further details about the LC-MS method are described in detail by (Lankova et al., 2015). QA/QC To control the background contamination by target compounds, a procedural blank sample (deionized water instead of serum) was prepared together with each batch of twenty samples. The determined concentration of BFRs and PFASs in each procedural blank was subtracted from the result of the respective samples. The repeatabilities (expressed as relative standard deviations, RSD %) and recoveries (REC %) of the complete method for BFRs and PFASs were obtained from six replicate analyses of spiked blank human serum, the spike concentrations were 0.2 and 1 ng/g lipid weight (l.w.) for BFRs; 0.07 and 0.33 ng/mL for PFASs and OH-BDEs and 0.3 and 1.5 ng/mL for HBCDs, TBBPA and brominated phenols. The average recoveries of targeted BFRs and PFASs were in the range of 80–109 % and 71– 110 %, respectively, with repeatabilities below 20 % and LOQs ranging from 0.1 to 2.5 ng/g l.w. - for BFRs and 0.01 to 0.3 ng/mL for PFASs. The LOQs were estimated as the lowest calibration standard which provided a signal-to-noise ratio (S/N) higher than 10 and the second MS/MS transition/ion (if available) had to provide an S/N ratio higher than 3. For compensation of unexpected influence of matrix or loss of target analytes, validation experiments were performed with the addition of isotopically labeled surrogates. Statistical analyses All statistical analyses were conducted using SPSS 23.0. (SPSS Inc., Chicago, IL). Spearman’s rank correlation was applied to assess the relationships between PFASs levels in 6

serum. To ensure precision of this data we used the natural logarithm of PFASs. Differences among the serum PFAS levels in different groups were evaluated using the Analysis of variance (ANOVA) or t-test, respectively. The four-way ANOVA model containing factors of locality, gender, age group, and educational level was used to investigate the association between biomarkers of internal exposure to PFASs. Statistical significance was set as α=0.05 (two-side). Concentrations lower than the LOQ were assigned a value of half of the LOQ. Lipid content determinations An enzymatic lipid determination of the serum samples was carried out by a clinical laboratory in Prague. Two types of lipids (triglycerides and total cholesterol) were measured in serum sub-samples. The method according to Phillips et al. was used to estimate the total lipid concentrations (Bernert et al., 2007; Phillips et al., 1989). The total lipid content was used for calculation of BFRs concentrations. Results and discussion The main results are summarized in Table 2 (PFASs) and Table 6 (BFRs), the following paragraphs are focused separately on these two groups of contaminants, since their sources are independent and there is typically no correlation between their concentrations. PFASs Out of nineteen PFASs, six were not detected in any of the analyzed samples, thirteen could be measured in concentrations above the LOQ in at least one sample and only six PFASs were quantifiable in more than 50 % of samples (Table 2). Thus, further statistical analyses were focused on these six substances (PFOA, PFNA, PFDA, PFUdA, PFHxS and PFOS) with a detection frequency from 96.0 to 100 %. The highest median concentration was observed for PFOS (2.43 ng/mL), followed by PFOA (0.756 ng/mL), PFNA (0.325 ng/mL), PFHxS (0.184 ng/mL), PFDA (0.145 ng/mL), and PFUdA (0.058 ng/mL) (Fig. 2). For comparison, serum 7

levels of selected PFAS so far reported in different countries are illustrated in Table 3. Concentrations of PFAS in samples examined in our study were relatively lower than those reported in other countries (Calafat et al., 2007, 2006; De Felip et al., 2015; Hsu et al., 2013; Ingelido et al., 2010; Kannan et al., 2004; Kärrman et al., 2009; Li et al., 2013; Lindh et al., 2012; Nøst et al., 2014; Olsen et al., 2012, 2003; Salihovic et al., 2015; SchröterKermani et al., 2013; Toms et al., 2014; Vassiliadou et al., 2010). However, there are some limits affecting comparability (such as year of collection of samples or specific participant group) and differences in the study design or analytical techniques. Correlation between individual PFASs Significantly (p<0.001) positive correlations between various PFAS concentrations in serum were found in the present study. Spearman’s rank correlation coefficients among PFOA, PFNA, PFDA, PFUdA, PFHxS, and PFOS for all samples are listed in Table 4. The most strongly correlated PFASs were PFDA and PFNA (ρ=0.87), followed by PFDA and PFUdA (ρ=0.86) and then PFHxS and PFOS (ρ=0.83). The weakest correlation (ρ=0.37) was among PFOA and PFUdA. The widespread correlations among PFAS concentrations suggest common exposure sources of these substances. However, possible sources of exposure and pathways are not clearly understood and further research is needed to identify the factors that contribute in these chemical concentrations. The influence of the sampling locality Median concentrations exhibited an evident statistically significant associations between localities and all six substances, namely PFOA (p<0.001); PFNA (p<0.001); PFDA (p<0.001); PFUdA (p<0.001); PFHxS (p=0.002) and PFOS (p<0.001). The association with locality also remained significant after adjusting for other factors (gender, age group and educational level); see Table 5. The highest concentrations were mostly observed in Ţďár nad Sázavou (Fig. 3), although at present we have no explanation for these differences. Therefore 8

more studies are necessary in the Czech Republic in order to find the relevant sources and exposure pathways. Differences in serum PFAS levels among regions were also previously reported in studies from the USA or Korea (Calafat et al., 2006; Cho et al., 2015). The influence of gender Concentrations of selected PFASs in serum collected from males were slightly, but significantly lower than in females, namely for PFOA (median: 0.731 vs. 0.803 ng/mL; p=0.001), PFNA (median: 0.316 vs. 0.333 ng/mL; p=0.028) and PFDA (median: 0.140 vs. 0.152 ng/mL; p=0.039). The association with gender also remained significant after adjusting for other factors (Table 5). Our findings are partly inconsistent with many studies worldwide. Current studies on gender differences in levels of PFAS among the non-occupational exposed population often report higher PFAS concentrations in males than in females (Calafat et al., 2007; Ericson et al., 2007; Fromme et al., 2007; Hsu et al., 2013; Ingelido et al., 2010; Toms et al., 2009a; Yeung et al., 2006), or no gender-related differences (Kannan et al., 2004; Kärrman et al., 2006; Olsen et al., 2004). These inconsistencies might be due to differences in the study design and lifestyle of the participants. Another possible explanation is also our specific group of participants, because female blood donors naturally can donate smaller quantities of blood and less frequently than males. The influence of age The correlation between the PFAS serum levels with age in the various age groups (18-29; 30-39; 40-49; 50-65) was not consistent. Statistically significant age-dependent differences were noted only for PFOA (p=0.006) and PFHxS (p=0.044). Serum concentrations of PFOA showed higher concentrations in the group of participants aged 50-65 (median: 0.924 ng/mL) in comparison with participants aged 18-29 9

(median: 0.840 ng/mL; p>0.05), 30-39 (median: 0.755 ng/mL; p>0.05) and 40-49 (median: 0.626 ng/mL; p=0.006). For PFHxS, the concentrations in the 30-39 age-group (median: 0.167 ng/mL) and 4049 (median: 0.186 ng/mL) were significantly lower than those in the 50-65 age-group (median: 0.248 ng/mL), with p-values of 0.019 and 0.009, respectively. Non-significant lower concentrations were also observed for participants aged 18-29 (median: 0.172 ng/mL). The association with age groups also remained significant after adjusting for other factors (Table 5). Moreover, a significant positive association was also obtained for PFOS (p=0.005) with similar tendency across age group as PFHxS. Some authors reported association between selected PFASs (in particular PFOS and PFOA) and age (Fromme et al., 2007; Cho et al., 2015), while others report no significant association of PFAS concentrations with age (Guo et al., 2011; Midasch et al., 2006; Olsen et al., 2003; Vassiliadou et al., 2010; Yeung et al., 2006). However, more monitoring data are needed to explain the differences across studies. The influence of level of education Levels of PFOA, PFNA, PFHxS and PFOS in serum samples increased with level of education; however, these differences were not significant, except for PFOA (p=0.019). After adjusting for other factors, level of education showed certain influence on the borderline of significance also for other PFASs (Table 5). Previous human biomonitoring studies have shown only an association between the higher levels of PFOA and more educated individuals (Calafat et al., 2007; De Felip et al., 2015; Melzer et al., 2010), and similarly for PFOS (De Felip et al., 2015; Melzer et al., 2010). Other PFASs are not frequently included in biomonitoring studies. Other factors 10

No statistically significant relationships were observed between PFAS concentrations and other various socio-demographical parameters, including smoking habits. Consumption of different food groups was mostly not associated with higher levels of PFAS, but there were some exceptions. Higher dairy product and milk intake was associated with slightly increased PFOA, PFNA, PFDA and PFOS levels, but it was statistically significant only for PFOA (p=0.039). Relationships between PFASs and consumption of fish, meat and poultry were generally weak with the exception of slightly but significantly higher PFUdA levels (p=0.013, p=0.010, respectively). BFRs No BFRs were detected at levels above the LOQ in more than 9 % of serum samples, even when using highly-sensitive analytical methods. Results are shown both as wet weight (nanograms per milliliter of serum) and lipid adjusted (nanograms per gram of lipid weight) in Table 6. The most frequently detected representatives of this group were congeners of PBDEs, namely BDE-47 (in 8.7 %), BDE-99 (in 6.0 %), BDE-153 (in 7.3 %) and BDE-209 (in 7.0 %). Although there are many studies on PBDE concentrations in human serum, the contribution of individual congeners is quite variable in reported studies. In recent studies from Germany (Fromme et al., 2009) and Greece (Kalantzi et al., 2011), the most commonly found were BDE-47, BDE-99, BDE-100 and BDE 153, with a detection frequency for these 4 congeners ranging 49-94 % and 26-74 %, respectively. BDE-47, BDE-100, BDE-153 and BDE-154 were predominant congeners in a study from the United Kingdom (Thomas et al., 2006), with a number of samples above LOD between 68-99 %. In the present study, detection frequency of the most PBDEs was much lower than these studies. Other BFRs, such as hexabromocyclododecane (HBCDD), novel brominated flame retardants (NBFRs), hydroxylated metabolites of PBDEs and tetrabromobisphenol A 11

(TBBPA) were mostly present at levels below the LOQ, which is in accordance with many other studies from various countries (Axmon et al., 2014; Darnerud et al., 2015; Fromme et al., 2016; Jakobsson et al., 2002; Kalantzi et al., 2011; Karlsson et al., 2007; Rawn et al., 2014; Zhou et al., 2014; Zhu et al., 2009). Conclusion In this study, we investigated for the first time ever the levels of PFAS and BFR in human serum samples, obtained from 300 blood donors in the Czech Republic. The results clearly show the frequent occurrence of PFASs in the Czech adult population, with PFOA, PFNA, PFDA, PFUdA, PFHxS and PFOS being the most abundant. Overall, PFOS concentrations were the highest. Compared to PFASs, BFRs were quantifiable in only 9 % of analyzed serum samples. The highest concentrations were observed for PBDEs, especially for BDE-209. Exposure to PFASs is evidently highly variable worldwide and even within individual countries. However, levels of these compounds in the Czech Republic are relatively lower than those reported in other studies worldwide. Therefore, continuous monitoring is essential for determining the trend of exposure in the Czech population to emerging environmental chemicals over time. Acknowledgements This research was funded by the Ministry of Health. We gratefully acknowledge the donors for collaborating with the study and voluntarily donating blood samples.

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17

Fig.1: Sampling sites in the Czech Republic

18

Fig.2: Concentrations of PFAS in human serum samples

19

ng/mL of serum

Fig.3: Median values of individual PFAS in four sampling localities 5.00 4.50 4.00 3.50 3.00 2.50 2.00 1.50 1.00 0.50 0.00 Ţďár nad Sázavou

PFOA

PFNA

Prague

PFDA

Liberec

PFUdA

PFHxS

Ostrava

PFOS

20

Table 1: Sample size characteristics of adult participants Parameter Sex men women Age 18-29 30-39 40-49 50-65 Education primary secondary tertiary Locality Praha Liberec Ostrava Ţďár nad Sázavou

N

%

171 129

57.0 43.0

43 99 92 66

14.3 33.0 30.7 22.0

74 125 100

24.8 41.8 33.4

76 73 76 75

25.3 24.3 25.3 25.0

21

Table 2: Content of PFASs in serum samples (ng/mL) Detection frequency (%) ND ND ND 45.0 100 99.7 100 96.0 47.3 36.3 4.33 20.0 100 100 2.33 ND ND ND ND = not detected; NC = not calculated PFBA PFPeA PFHxA PFHpA PFOA PFNA PFDA PFUdA PFDoA PFTrDA PFTeDA PFBS PFHxS PFOS PFDS PFOSA N-MeFOSA N-EtFOSA

LOQ 0.013 0.013 0.013 0.013 0.013 0.013 0.013 0.013 0.013 0.013 0.013 0.006 0.006 0.004 0.006 0.007 0.004 0.004

Range
GM NC 0.716 0.300 0.141 0.055 NC NC NC NC 0.171 2.29 NC -

Median NC 0.756 0.325 0.145 0.058 NC NC NC NC 0.184 2.43 NC -

22

Table 3: Median concentration of selected PFASs in human plasma/serum

Country

Year

Sample type

Age (years )

Na

Referenc e

Concentration (ng/mL) PFO A

PFN A

PFD A

PFUd A

PFHx S

PFO S

Europe 13.4

Salihovic et al., 2015 De Felip et al., 2015 Nøst et al., 2014 SchröterKermani et al., 2013 Lindh et al., 2012 Ingelido et al., 2010 Vassiliad ou et al., 2010 Kannan et al., 2004

20012004

Plasma

Italy

2011/20 12

Serum

470479c

2041

1.55

-

-

-

-

2.43

Norway

2007

Serum

52d

4050

3.1

1.5

0.8

1.3

1.9

33

Germany

2010

Plasma

18

2029

3.2

0.86

3.7

Poland

2002/20 03

Serum

190d

2537e

4.84

1.19

0.38


1.18

18.5

Italy

2008

Serum

230

2065

3.59

-

-

-

-

6.31

Greece

2009

Serum

182

1589

2-4f

-

-

-

-

7.515f

Belgium

19982004

Serum/plas ma

20

1963

4.1

-

-

-

1.3

17.2

1.43

0.37

0.19

0.26

0.05

1.47

Fu et al., 2014

4.3

0.8

-

-

3.3

9.4

Toms et al., 2014

3.22

-

-

-

-

8.52

1.0

-

-

-

-

6.47


-

-

-


1.2

Sweden

Asia/Australia China 2011

1006

133

2010/20 11

Serum

24

g

Taiwan

2011

Serum

59

China

-

Serum

64

Afghanist an

2007

Serum

55

JapanOsaka 2004 JapanMiyagi North America

3.33

0.71

0.33

-

2.08

(LPFOS)

b

Serum

Australia

70

Serum

10c

USA

2010

Plasma

600

USA

2003/20

Serum

209

0-80 0(60> ) 1965 1953 2.565

3475

2069 12-

(LOQ=0 .5)

(LOQ=0. 5)

30.7

1.9

6.9

2.5

3.2

31.7

4.56

1.93

2.7

0.92

5.1

14.2

2.44

0.83h

0.27 h

0.14 h

1.34 h

8.30

h

3.9 h

1.0 h

-

-

1.9 h

20.7

h

Hsu et al., 2013 Li et al., 2013 Hemat et al., 2010 Kärrman et al., 2009

Olsen et al., 2012 Calafat

23

04

4

(60> )

h

et al., 2007
24

Table 4: Values of Spearman´s rank correlation coefficient (ρ) for PFASs in serum samples from Czech adult population PFOA

PFNA

PFDA

PFUdA

PFHxS

PFOS

PFOA

-

0.79

0.60

0.37

0.74

0.65

PFNA

0.79

-

0.87

0.68

0.73

0.75

PFDA

0.60

0.87

-

0.86

0.61

0.71

PFUdA

0.37

0.68

0.86

-

0.44

0.55

PFHxS

0.74

0.73

0.61

0.44

-

0.83

PFOS

0.65

0.75

0.71

0.55

0.83

-

25

Table 5: P-values of factors locality, gender, age group, and educational level of four-way ANOVA model Locality

Gender

Age group

Education level

PFOA

<0.001

<0.001

0.020

0.068

PFNA

<0.001

0.001

0.101

0.035

PFDA

<0.001

0.003

0.363

0.115

PFUdA

<0.001

0.048

0.545

0.088

PFHxS

<0.001

0.428

0.013

0.041

PFOS

<0.001

0.337

0.005

0.079

26

Table 6: Content of BFRs in serum samples

6-OH-BDE 47 4´-OH-BDE 49 2´-OH-BDE 68 6´-OH-BDE 99 TBBPA 2,4-DBP 2,4,6-TBP PBP α-HBCD β-HBCD γ-HBCD BDE 28 BDE 47 BDE 49 BDE 66 BDE 85 BDE 99 BDE 100 BDE 153 BDE 154 BDE 183 BDE 196 BDE 197 BDE 203 BDE 206 BDE 207 BDE 209 PBT PBEB HBB BTBPE OBIND DBDPE ND = not detected

LOQ (ng/mL) 0.004 0.004 0.004 0.004 0.333 0.364 0.333 0.067 0.036 0.036 0.036 0.005 0.005 0.005 0.005 0.005 0.005 0.005 0.005 0.005 0.005 0.005 0.005 0.005 0.100 0.100 0.100 0.010 0.010 0.010 0.010 0.200 0.200

Detection frequency (%) ND ND ND ND 0.67 ND 0.33 0.67 0.33 0.33 ND ND 8.67 ND ND ND 6.00 0.67 7.33 ND 2.67 1.33 3.33 0.67 0.33 0.33 7.00 ND ND ND ND ND ND

Range (ng/mL)
Range (ng/g l.w.)
27