Polybrominated diphenyl ethers in foods from the Region of Valencia: Dietary exposure and risk assessment

Chemosphere 250 (2020) 126247

Contents lists available at ScienceDirect

Chemosphere journal homepage: www.elsevier.com/locate/chemosphere

Polybrominated diphenyl ethers in foods from the Region of Valencia: Dietary exposure and risk assessment
ndez a, b, Leyre Quijano c, Silvia Marín a, d, Pilar Villalba a, d, Olga Pardo a, *, Sandra F. Ferna  a, d Francisca Corpas-Burgos a, Vicent Yusa a

Foundation for the Promotion of Health and Biomedical Research of Valencia Region, FISABIO-Public Health, 21, Avenida Catalunya, 46020, Valencia, Spain ~ oz, Dr. Moliner 50, 46100, Burjassot, Spain Analytical Chemistry Department, University of Valencia, Edifici Jeroni Mun Preventive Medicine and Public Health, Food Sciences, Toxicology and Forensic Medicine Department, University of Valencia, Valencia, Spain d Public Health Laboratory of Valencia, 21, Avenida Catalunya, 46020, Valencia, Spain b c

h i g h l i g h t s
PBDE intake in Spain including representative sampling and foods with limited data.
Salty fish, common in Spain and with high values, was included for the first time.
Derived MOEs from PBDEs dietary exposure do not indicate possible health risk.

a r t i c l e i n f o

a b s t r a c t

Article history: Received 15 October 2019 Received in revised form 13 February 2020 Accepted 15 February 2020 Available online 19 February 2020

Dietary exposure to polybrominated diphenyl ethers (PBDEs) of the population in the Region of Valencia, Spain, was assessed. A group of 320 composite samples of different fatty foods was collected and analyzed, including the following: vegetable oils, and foods of animal origin such as (a) fish and seafood, (b) eggs, (c) milk and dairy products, and (d) meat and meat products. Two scenarios were assumed for left-censored results: lower-bound (LB) and upper-bound (UB). Vegetable oils, and fish and seafood presented the highest content of PBDEs [mean values of 503 and 464 pg g1 wet weight (ww) for total PBDEs, respectively, in the UB]. The dominating congeners were BDE47 in the food categories of fish and seafood, meat and meat products, and vegetable oils, and BDE99 in the categories of eggs, and milk and dairy products. The dietary exposure to PBDEs through consumption of the studied foods by the population in the Region of Valencia was estimated for adults (>15 years of age) and young people (6e15 years of age). Average intake levels (UB scenario) were 1.443 and 3.456 ng kg bw1 day1 for adults and young people, respectively. In a risk-assessment context, the margin of exposure (MOE) for congener BDE47, -99, 153, and 209 (ranged: 30-3E6) indicate that the current dietary exposure to these substances does not pose a risk to human health. © 2020 Elsevier Ltd. All rights reserved.

Handling Editor: Myrto Petreas Keywords: Dietary exposure Dietary intake Polybrominated diphenyl ethers (PBDEs) Risk assessment

1. Introduction A priority of the European Union is the risk assessment of food contaminants which are hazardous to human health. In the last decade, polybrominated diphenyl ethers (PBDEs), a class of brominated flame retardants classified as persistent organic pollutants, have been included among the substances of priority interest. There are 209 theoretical brominated diphenyl ethers (BDE)

* Corresponding author. E-mail address: [email protected] (O. Pardo). https://doi.org/10.1016/j.chemosphere.2020.126247 0045-6535/© 2020 Elsevier Ltd. All rights reserved.

congeners that are numbered according to the International Union of Pure and Applied Chemistry (IUPAC) system. Three commercial technical mixtures (PentaBDE, OctaBDE, and DecaBDE) have been widely used to reduce flammability in many types of commercial and household products, such as plastics, electronics, furniture, textiles, and construction materials (Costa et al., 2016). The use of these mixtures began to spread in the 1970s, and their long-term application has resulted in ubiquitous environmental pollution (Shi et al., 2017). Likewise, due to their lipophilicity, bioaccumulation, and biomagnification characteristics, they have been introduced into the food chain (Piersanti et al., 2015). This, along with the potential adverse effects of PBDE exposure on human

2

O. Pardo et al. / Chemosphere 250 (2020) 126247

health, including thyroid disorders, reproductive health effects, and neurobehavioral and developmental disorders (Kim et al., 2014), has caused public health concern. The publication of the European Commission’s Recommendation 2014/118/EU (EC, 2014), which recommends that Member States perform monitoring programs for the presence of PBDEs in food (including a wide variety of individual foodstuffs reflecting consumption habits to give an accurate estimation of exposure), is evidence of this growing interest. Food and dust have been identified as the two main potential sources of human exposure to PBDEs (Korcz et al., 2017; Pietron and Malagocki, 2017). High-fat content foods of animal origin such as fish, milk, eggs, meat, and animal fats are of particular relevance (Boucher et al., 2018). For populations with non-occupational exposure, the contribution of house dust ingestion was less important than diet intake in most countries (Besis and Samara, 2012). Therefore, dietary estimations are appropriate tools to estimate the exposure to such compounds and to evaluate the potential risk in a population. A recent global review (Boucher et al., 2018) identified Spain as a poor contributor regarding PBDE data (approximately 5%), highlighting the need for Spanish data in order to assess population-specific intake and risk. In addition, new studies for PBDEs exposure and risk assessment are justified because of large gaps. In 2014, the results of a monitoring program on PBDEs in fish and seafood carried out by the Department of Public Health of the Valencian government, Spain, were presented (Pardo et al., 2014). The estimated dietary exposure to these pollutants was also reported. However, only fish and seafood were considered in that study. In contrast, this new study carried out in the Region of Valencia, Spain, combines representative consumption data (from a dietary survey) with the occurrence of PBDEs in foods. A group of 320 composite samples including vegetable oils, and foods of animal origin such as (a) fish and seafood, (b) eggs, (c) milk and dairy products, and (d) meat and meat products were used to derive an improved exposure estimation. While foods of animal origin have been identified as having high PBDE levels, little is known regarding vegetable oils (Boucher et al., 2018). Consequently, these foods were included in the present study. In addition, 13 congeners were measured: 10 BDEs included in the European Commission’s Recommendation 2014/118/EU (EC, 2014) such as BDE28, BDE47, BDE49, BDE99, BDE100, BDE138, BDE153, BDE154, BDE183, and BDE209, along with three additional congeners corresponding to BDE66, BDE139, and BDE155. These three additional congeners were included because BDE66 is one of the most frequently measured congeners (Boucher et al., 2018; EFSA, 2011a), and BDE139 and BDE155 have been detected in OctaBDE and PentaBDE technical products, respectively. Results for congener BDE209 were not previously reported in this region. With this approach, which includes an extensive list of food groups and 13 PBDE congeners, a more comprehensive exposure and risk estimation for PBDEs has

been achieved. The goal of this study is to report the occurrence of PBDEs in foods of animal origin and in vegetable oils from the Region of Valencia and to perform a dietary risk assessment in two population subgroups (adult and young people) from this region, which would help policy makers with risk management.

2. Materials and methods 2.1. Sampling A sampling plan was designed to obtain representative data on concentrations of PBDEs in foods consumed by the population in the Region of Valencia, Spain. Several studies identified foods of animal origin with high lipid content, such as fish and seafood (Kiviranta et al., 2004), eggs (Garcia et al., 2018), milk and dairy products, meat and meat products (Shi et al., 2017), and animal fats as having high PBDE levels. Although it is well known that fish oil supplements usually present high PBDE levels (Boucher et al., 2018), these were not included in our study because none of the participants reported their consumption in the dietary survey. Conversely, due to the lack of data for PBDEs in vegetable oils, we included them to increase our understanding of this food type. As shown in Table 1, a total of 32 food items grouped into 5 categories (vegetable oils, meat and meat products, eggs, milk and dairy products, and fish and seafood) were collected in 2010 by techni
gico de la Industria Alimentaria cians from the Instituto Tecnolo (AINIA, Valencia, Spain). Four sampling periods (JanuaryeMarch, AprileJune, JulyeSeptember, and OctobereDecember) were planned to include seasonal foods. In order to account for variability within each food item, 100 samples per food item were collected (sampling error of 9.8% at a 95.5% confidence interval). Therefore, a total of 3200 food samples were collected. The sampling plan (see Fig. 1) covered urban and rural areas (70% and 30%, respectively) and different geographic regions and seasons. Two fundamental criteria were considered in the design of the sampling plan: the type of establishment and its geographic location. A two-stage design was established: (1) Selection of a random sample of clusters corresponding to different locations or core regions of Valencia (the sample size assigned to each cluster was calculated maintaining proportionality of the represented population). The Region of
n, Valencia is made up of three provinces: Valencia, Castello and Alicante. According to the Valencian Institute of Statistics (2008), the province of Valencia represented 52% of the population of the Region of Valencia, the province of
n 12%, and the province of Alicante 38%. Therefore, Castello 1664, 352, and 1184 samples were taken in different

Table 1 Foods included in the total diet study for PBDEs analyses. Food categories

Food items

Nº Food items

Nº total samples

Nº total of composites (or analysis)

Vegetable oils (Vo) Meat and meat products (Meat)

Olive oil and sunflower oil Chicken, pork, beef, lamb, rabbit, hamburger, sausages, cured ham, cooked ham, cured sausage, foie-grass and offal. Chicken egg Cow milk, cheese, yogurt, custards and smoothies, butter Tuna and albacore tuna, squid and cuttlefish, crustacean, mussel, salmon and trout, hake, sardine and anchovy, swordfish, salty fish, smoked fish, seabream and seabass, canned fish.

2 12

200 1200

20 120

1 5 12

100 500 1200

10 50 120

32

3200

320

Eggs (Egg) Milk and dairy products (Milk) Fish and seafood (Fish)

Total Nº samples/food item ¼ 100. Nº samples/composite ¼ 10.

O. Pardo et al. / Chemosphere 250 (2020) 126247

3

Fig. 1. Distribution of the food sampling conducted in 2010 in the Valencian Region (N ¼ 3200 food samples).


n, and Alilocations from the provinces of Valencia, Castello cante, respectively. (2) Selection using stratified random sampling based on the type of establishment. This included three different supermarket chains (each contributing 30%), which supply food to an important part of the population of the Region of Valencia. The remaining 10% was allocated to local markets. To reduce the number of analyses, a composite sample was obtained by combining 10 samples of the same food item (collected during the same season). The total number of composites (samples analyzed) was 320. Only the edible parts of each food were included in the composites, and the analyses were performed on fresh unprocessed samples. A Thermomix TM-21 food processor was used for the homogenization. These composite samples were kept in high-density polyethylene bags and shipped at approximately 4  C to the laboratory, where they were stored at 20  C until analysis.

2.2. Consumption data For the intake estimation, consumption data were obtained from a dietary survey conducted and validated by the Public Health Directorate of Valencia between 2010 and 2011. Aims, design and methods used in this survey were described in a previous publication (Fullana et al., 2010). Information on food intake was collected through 24 h recalls. A total of 1484 subjects (196 young people from 6 to 15 years of age with a mean body weight (bw) of 43.5 kg and 1288 adults from 16 to 95 years of age with a mean bw of 73.2 kg) were asked, in a face-to-face interview, to recall and describe the types and quantities of all foods and beverages ingested during the previous 24-h period. Body weight was self-

reported. To take into account variations in consumption patterns according to the season, the interviews were carried out during three different periods: JuneeJuly 2010, September-November 2010, and November 2010eFebruary 2011. Although the age groups included broad age ranges, based on the study carried out by the same authors as in this study (Pardo et al., 2014) in which calculated margins of exposure (MOEs) for PBDEs indicated that a health concern was unlikely, these groups were considered suitable to make a first approximation of the PBDE dietary risk assessment of the population studied.
n y Salud v2.0 was used for the evalThe software Alimentacio uation of the 24 h recalls, whose core analyses are based on the Food Composition Tables of the University of Granada (Mataix Verdu, 2009). Then, the data were grouped into 5 food categories and recoded into 32 food items, according to the categorization followed by FOODEX (EFSA, 2011b), commonly used in the literature. The details of the food consumption data (g kg1 bw day1) for the two population sub-groups studied are detailed in Table 2 by Table 2 Consumption (g kg1 bw day1) per food group in the Region of Valencia. Food category

Adults >15 years n ¼ 1288 Mean

P95

Mean

P95

Vegetable oils Meat and meat products Eggs Milk and dairy products Fish and seafood

0.30 1.99 0.35 3.69 0.77

0.86 9.49 1.63 18.73 2.38

0.49 4.50 0.63 9.72 0.84

1.50 22.45 3.46 44.72 3.11

Young people (6e15 years) n ¼ 196

bw: body weight. *n ¼ 1288; mean body weight; 73.2 kg mean bw; range from from 37.1 to 153.2 kg. **n ¼ 196; 43.5 kg mean bw; range from 14.9 to 98 kg.

4

O. Pardo et al. / Chemosphere 250 (2020) 126247

food category. 2.3. Chemical analysis and quality assurance/quality control Analyses were performed in the Public Health Laboratory of Valencia, which operates under the quality assurance system established by ISO/IEC 17025 (ISO/IEC17025:2017). 2.3.1. Standards and solvents All solvents were suitable for PBDE residue analysis. The supplies were obtained from the following companies: Isooctane and sulfuric acid from Panreac (Barcelona, Spain), n-hexane and anhydrous sodium sulfate from Scharlau (Barcelona, Spain), diatomaceous earth from Sigma-Aldrich (Steinheim, Germany), and Strata silica SPE cartridges (500 mg 6 mL1) from Phenomenex (Macclesfield, U.K.). Individual standards of BDE28, BDE47, BDE49, BDE66, BDE99, BDE100, BDE138, BDE139, BDE153, BDE154, BDE155, BDE183, and BDE209, and internal standards of [13C]BDE28, [13C]BDE47, [13C] BDE99, [13C]BDE153, and [13C]BDE209 (each at a concentration of 50 mg mL1, in isooctane) were provided by Wellington Laboratories (Guelph, ON, Canada). The purities of all standards were more than 95%. Two working standard mix solutions (containing 500 or 100 mg mL1 of each individual BDE accordingly), and one mix internal standard solution (with 100 mg mL1 of each internal standard) were prepared by diluting the stock standards in isooctane. Standard solutions containing 0.1e100 ng mL1 of each individual standard and 25 ng mL1 of internal standards were prepared in isooctane and stored in the dark at 20  C. 2.3.2. Sample preparation About 1 kg of sample was scrunched (if necessary) and homogenized (Ultra-Turrax TR-50, Germany). A portion of approximately 200 g of meat and meat products, fish and seafood, milk and dairy products, and egg samples was lyophilized, and an analytical aliquot was fortified with 62.5 mL of the internal standard mix solution (1000 mg mL1) and extracted by pressurized liquid extraction (PLE). 2.3.3. Extraction PBDEs from vegetable oils were extracted using liquid-liquid extraction with hexane. For meat and meat products, fish and seafood, milk and dairy products, and eggs, the analytical aliquot was mixed with diatomaceous earth, and the sample was introduced into a 34 mL stainless-steel cell (containing a cellulose filter in the cell outlet). Once closed, the sample cell was placed in the carousel of an accelerated solvent extraction system (ASE 350, Dionex, Sunnyvale, CA, USA). The extraction conditions were: an oven temperature of 100  C, hexane as the extraction solvent, a 5 min heat-up time under a pressure of 1500 psi, and three static cycles with a static time of 5 min. A volume equivalent to 20% of the extraction cell capacity was selected as the flush volume. After the extraction, a purge using pressurized nitrogen for 1 min was used. The resultant extract (approximately 30 mL) was dried over 5 g of anhydrious sodium sulfate, and the supernatant was collected and evaporated under a gentle nitrogen stream at 40  C to approximately 3 mL with a Turbo Vap 500 (Zymark, Idstein, Germany). 2.3.4. Clean-up The procedure consists of a slightly modified solid-phase extraction (SPE) clean-up method previously developed (Medina et al., 2008, 2009). The 3 mL extract was digested with 1 mL of concentrated H2SO4 (added drop by drop) and vortexed for 1 min.

After, it was centrifuged for 5 min at 4000 rpm. The upper organic phase was collected, and the aqueous acid phase was extracted two more times with 3 mL of new hexane. The final extract was made up to volume into a 10 mL volumetric flask. A 2 mL aliquot was passed through a silica SPE cartridge which had been previously conditioned with 5 mL of hexane. The first 1 mL was discarded, and the following volume was collected and combined with an additional fraction that was obtained by passing 1.5 mL of hexane through the cartridge. The final extract was concentrated to dryness under a gentle nitrogen stream at 40  C and redissolved in 0.250 mL of hexane. 2.3.5. Gas chromatography-tandem mass spectrometry (GC-MSMS) MSeMS analysis was performed on a Finnigan PolarisQ ion trap mass spectrometer (Austin, TX, USA). A heated transfer line was used to connect the mass spectrometer to a Thermoquest Trace GC 2000 (Waltham, MA, USA) gas chromatograph equipped with a Combi Pal autosampler from CTC Analytics AG (Zwingen, Switzerland).). Xcalibur 1.2 was used for data acquisition. The analyses were carried out with a 30 m  0.25 mm i. d., 0.25 mm film thickness TR-5MS capillary column (Thermo), directly connected to a programmed temperature vaporizing (PTV-LV) injector, using He as the carrier gas. The liner used was a PTVeLV 2.75  2 TRC from Thermo Finnigan (Milano, Italy). The oven temperature program was: from 80  C (held for 1.50 min) to 140  C (held for 1.50 min) at a rate of 50  C min1, then to 220  C (held for 1 min) at 20  C min1, after to 280  C (held for 10 min) at 2  C min1, and finally to 300  C (held for 5 min) at 30  C min1. The MS-MS and PTV-LV operating  et al., 2006). conditions were previously optimized (Yusa The confirmation criteria were the following: (1) The intensity of two product ions should be at least three times greater than the base noise of the MS detector. (2) The relative abundance between ions in the sample should be the same as in the standard, with the acceptable deviations described in the legislation (European Commission, 2014). (3) The ratio between the chromatographic retention time of the analyte and that of the internal standard should correspond to that of the calibration solutions, with a tolerance of ±0.5%. Quantification was performed by internal standard calibration using a six-point regression line ranging from 0.1 to 100 ng mL1. 2.3.6. Method validation and analytical quality assurance The analytical method was validated at the low, medium, and high level of the standard calibration curve (corresponding to 0.1, 20, and 100 ng mL1). The limit of quantification (LOQ) was fixed at 10 pg g1 wet weight (ww) for all the BDE congeners, as established in the European Commission’s Recommendation 2014/118/EU (EC, 2014) with the exception of BDE209, for which the LOQ was fixed at 90 pg g1 ww since its analysis is hindered by its inherent properties. The recovery ranged from 85% to 107%, and the coefficient of variation from 7% to 15%. The linear regression model was statistically validated considering that residual values were randomly distributed around the regression line, the p-value of the F-test statistic was <0.05, and the coefficient of determination (R2) was >0.99 (Eurachem Guide, 1998). All test batches were analyzed under quality-assurance protocols, including procedural blanks, spiked samples, and reference materials [SRM 1947 Fish tissue, SRM 1974b Mussel tissue, and SRM1588b Cod liver oil from the National Institute of Standards and Technology (NIST), USA]. The results obtained for these reference materials were in agreement with the certified values (recoveries were between 78% and 112%), and the laboratory participated regularly in laboratory proficiency testing provided by Quasimeme

O. Pardo et al. / Chemosphere 250 (2020) 126247

(Wageningen, The Netherlands) with satisfactory z-scores. 2.4. Occurrence of PBDEs Analytical concentrations were reported as wet weight. 2.5. Dietary exposure assessment The long-term dietary exposure to BDE congeners was assessed because it is relevant to consumers who are repeatedly exposed to these compounds. Dietary exposure to PBDEs in the population was calculated individually for each subject. We used the following P formula: Ei,j (ng kg1 bw day1)¼ ( nk¼1 Ci,k (ng g1) x Lk,j (g day1)/ bwi (kg), where Ei,j is the dietary exposure to BDE congener j of individual i; n is the number of food items considered in this study (n ¼ 32); Ci,k is the consumption of food item k by individual i; Lk,j is the level of BDE j in food item k; and bwi is the body weight of individual i. Exposures were estimated for adults (16e95 years with bw from 37.1 kg to 153.2 kg) and young people (6e15 years with bw from 14.9 kg to 98 kg). Data below the LOQ were treated according to the substitution method recommended by the WHO, widely used in dietary risk assessment (WHO , 1999; EFSA, 2010). Briefly, for each congener, two scenarios were assumed given that all BDE congeners presented a censoring rate higher than 60%: the lower-bound (LB) scenario and the upper-bound (UB) scenario, in which unquantified results were set to zero or to the LOQ value, respectively. 2.6. Risk characterization The European Food Safety Agency (EFSA) suggests a margin of exposure (MOE) approach for the risk characterization of PBDEs from the dietary intake. The Panel on Contaminants in the Food Chain (CONTAM) identified effects on thyroid hormone homeostasis, neurobehavioral functions, and sperm quality and reproductive performance in animal experiments as the critical endpoints, and they derived lower 95% confidence limit for a benchmark response of 10%) (BMDL10) (for the most sensitive effects of BDE47, BDE99, BDE153, and BDE209 (EFSA, 2011a,b). EFSA considers that if a calculated MOE is greater than 2.5, it indicates that a health concern is unlikely (with risk decreasing as the MOE increases). As relevant risk reference guidance values were only available for BDE47, BDE99, BDE153, and BDE209, risk characterization was performed only for these four individual BDE congeners. 3. Results and discussion 3.1. PBDE levels The average concentration (mean) of the 13 BDE congeners in the different food items analyzed are given in Table 3, in which the unquantified results (below the LOQ) were set to the LOQ value (based on the UB approach). As can be seen, the censoring rate is high in this study. However, we have included a large number of congeners (13 in total), some of which are seldomly reported in the literature or are known to be present at low levels. The latter limitation could be exacerbated by the use of composite samples due to potential dilution. It has been reported that when an individual sample with a high value is combined with others with low values, it results in a composite sample that shows falsely low results (EPA , 1995). Another explanation for the high censoring rate could be the analytical LOQ of this study, which although in agreement with that established in the European Commission’s Recommendation 2014/ 118/EU (EC, 2014), might not be sufficient to detect the low levels of

5

the PBDEs present in some food items. The determination of PBDEs in food is still methodologically challenging, especially for the BDE209 congener, which has inherent properties that difficult its analysis. The most contaminated food categories were vegetable oils, and fish and seafood (mean values of 503 and 464 pg g1 ww for total PBDEs, respectively). It is important to highlight the high levels found in vegetable oils since it was recently identified as a food with limited data availability globally (Boucher et al., 2018). These findings agree with the results reported by Babalola et al. (2018), which also found a high concentration of the sum of BDE congeners in edible oil in an adult population in Southwest Nigeria. Regarding fish and seafood, high levels of PBDEs have been previously re
nisseau et al., 2018, ported (EFSA, 2011a,b; Tao et al., 2017; Ve Babalola and Adeyi, 2018; Garcia Lopez et al., 2018). The PBDE concentrations reported in this study indicate a decrease in contamination when compared with data from this region from 2014 (mean values of 482e3456 pg g1 ww compared to 970e3870 pg g1 ww from Pardo et al., 2014). This observation is consistent with the regional decline seen in other areas of Spain
n et al., 2017). (Trabalo Important variations were observed among fish types. Regarding fish and seafood, salty fish contained the highest level (mean values of 1533 pg g1 ww for total PBDEs), probably due to how these products are processed. These processing procedures involve dehydration to a great extent, which, in turn, could lead to the concentration of these contaminants in this type of fish. To our knowledge, this kind of product has not been included in previous studies carried out in Spain, where they are frequently consumed. Dark or oily fish (including smoked fish, sea bream and sea bass, salmon and trout, sardine and anchovy, canned fish, and tuna) also presented high levels of PBDEs (mean values of 654, 502, 587, 395, 210, and 460 pg g1 ww, respectively). Dark or oily fish (including smoked fish, sea bream and sea bass, salmon and trout, sardine and anchovy, canned fish, and tuna) also presented high levels of PBDEs (mean values of 654, 502, 587, 395, 210, and 460 pg g1 ww, respectively) in accordance with previous studies (Leblanc et al., 2006; Boucher et al., 2018). In the national context, similar results were reported in Tarragona, Spain, where salmon had the highest
n et al., 2017). Similarly, concentration of the sum of PBDEs (Trabalo in a previous study carried out in the current study region (Pardo et al., 2014), the highest levels of PBDEs were found in salmon (370e470 pg g1 ww, LB-UB respectively). This PBDE level in salmon (470 pg g1 ww using the UB scenario) was lower than the value reported in the present study (587 pg g1 ww using the UB scenario). Our current study, however, included the determination of BDE209 which had a higher LOQ (90 pg g1 ww), implying an overestimation in the UB scenario. White or lean fish (including hake and swordfish) presented mean values for the sum of all PBDE congeners between 249 and 244 pg g1 ww. These values are higher than those reported by Boucher et al. (2018) (115 pg g1 ww), although a 30-fold variation within the group was detected. Furthermore, other fish products with low lipid content, such as squid and cuttlefish, and crustaceans and mussels, contained low PBDE levels (generally below LOQ for all individual congeners), as previously reported in Spain
n et al., 2017). Contrarily, Boucher et al. (Pardo et al., 2014; Trabalo (2018) identified shellfish as a high PBDE contaminated food, although a 60-fold difference was observed between the lowest and the highest items. The lowest PBDE mean concentrations were found in meat and meat products, milk and dairy products, and eggs (mean values of 224, 247, and 229 pg g1 ww for total PBDEs, respectively), largely due to the high number of unquantified results. The results in meat

6

Table 3 Mean BDE levels (pg g ww1) in the different foods included in this study, UB approach (in which not detected congeners were set to the LOQ value). Food group

Food item

BDE28

BDE47

BDE49

BDE66

BDE99

BDE100

BDE138

BDE139

BDE153

BDE154

BDE155

BDE183

BDE209

Total PBDEs

146 (50%) 64 (70%) 105 54 (100%) 10 (0%) 10 (0%) 25 (40%) 10 (0%) 24 122 (40%) 10 (80%) 10 (0%) 10 (0%) 13 (10%) 10 (0%) 10 (0%) 16 (70%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 20

124 (90%) 37 (20%) 81 11 (10%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 13 (10%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10

10 (0%) 131 (10) 71 15 (70%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 11 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10 (0%) 10

10 10 10 48 27 10 10 10 21 22 10 10 11 16 10 11 10 10 12 10 10 12

10 10 10 11 10 10 10 10 10 20 10 10 10 10 10 10 10 10 14 11 10 11

10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10

10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10

10 10 10 11 10 10 10 10 10 11 10 10 11 10 10 10 10 10 10 10 10 10

76 38 57 11 15 10 10 10 11 10 10 10 10 10 10 10 10 10 10 10 10 10

50 23 37 15 12 10 11 10 11 10 10 10 10 10 10 10 10 10 10 10 10 10

10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10

90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90

556 449 503 371 234 210 211 210 247 355 210 210 212 219 210 211 216 210 217 211 210 224

121 (50%) 230 (80%) 61 (100%) 240 (100%) 231 (80%) 10 (0%)

30 (60%) 77 (80%) 10 (0%) 133 (100%) 16 (70%) 10 (0%)

10 17 10 29 10 10

10 36 48 17 18 10

10 (10%) 12 (10%)

37 (90%) 83 (100%)

11 (40%) 26 (80%)

10 (0%) 10 (20%)

10 (0%) 16 (30%)

11 (70%) 10 (0%)

18 (90%) 10 (0%)

10 (0%) 10 (0%)

10 (0%) 10 (10%)

11 (20%) 29 (80%)

11 (30%) 28 (70%)

10 (0%) 10 (0%)

90 (0%) 90 (0%)

249 (27%) 344 (31%)

21 10 10 10 16 10

1139 (100%) 10 (0%) 17 (100%) 10 (0%) 182 16

63 10 10 10 34 10

34 10 10 10 14 10

10 10 10 10 17 20

52 10 10 10 21 19

17 10 10 10 11 10

10 10 10 10 10 10

19 10 10 10 12 10

30 10 10 10 24 10

33 10 10 10 21 10

15 10 10 10 11 10

90 90 90 90 90 90

1533 (52%) 210 (0%) 217 (18%) 210 (0%) 464 229

Mean (n) pg g ww¡1 Vegetable oils

Milk and dairy products

Meat and meat products

Eggs

10 10 10 10 10 10 10 10 10 17 10 10 10 10 10 10 10 10 11 10 10 11 13 16 47 22 10 10

(0%) (0%) (0%) (0%) (0%) (0%) (0%) (10%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (10%) (0%) (0%)

(30%) (70%) (90%) (100%) (0%) (0%)

(100%) (0%) (0%) (0%)

(80%) (0%) (10%) (0%)

(0%) (40%) (30%) (10%) (0%) (0%)

(60%) (0%) (80%) (0%)

(0%) (0%) (20%) (10%) (0%) (0%) (0%) (80%) (0%) (0%) (20%) (30%) (0%) (20%) (0%) (0%) (10%) (0%) (0%)

(0%) (90%) (100%) (10%) (30%) (0%)

(10%) (0%) (0%) (0%)

15 32 50 22 22 10

(0%) (0%) (10%) (0%) (0%) (0%) (0%) (20%) (90%) (0%) (0%) (10%) (0%)) (0%) (80%) (0%) (50%) (30%) (0%)

(30%) (90%) (100%) (100%) (40%) (0%)

(100%) (0%) (60%) (0%)

10 10 12 10 10 10

(0%) (0%) (10%) (0%) (0%) (0%) (0%) (20%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%)

(50%) (0%) (10%) (10%) (0%) (0%)

(10%) (0%) (0%) (0%)

10 11 10 10 10 10

(0%) (0%) (0%) (0%) (0%) (0%) (0%) (20%) (0%) (60%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%)

(0%) (10%) (10%) (10%) (0%) (0%)

(0%) (0%) (0%) (0%)

11 10 12 10 23 10

(0%) (0%) (30%) (0%) (0%) (0%) (0%) (10%) (0%) (0%) (10%) (10%) (0%) (0%) (0%) (0%) (0%) (0%) (0%)

(10%) (10%) (10%) (20%) (40%) (0%)

(10%) (0%) (0%) (0%)

75 28 24 27 29 10

(70%) (30%) (10%) (10%) (0%) (90%) (0%) (0%) (70%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%)

(80%) (90%) (90%) (90%) (40%) (0%)

(100%) (0%) (0%) (0%)

55 20 11 29 23 10

(40%) (20%) (10%) (10%) (0%) (10%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%)

(80%) (80%) (20%) (100%) (80%) (0%)

(100%) (0%) (0%) (0%)

10 10 10 15 10 10

(0%)) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%)

(0%) (0%) (0%) (10%) (0%) (0%)

(10%) (0%) (0%) (0%)

90 90 90 90 90 90

(0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%) (0%)

(0%) (0%) (0%) (0%) (0%) (0%)

(0%) (0%) (0%) (0%)

460 587 395 654 502 210

(20%) (10%) (21%) (2%) (0%) (11%) (0%) (16%) (18%) (5%) (2%) (5%) (0%) (2%) (16%) (0%) (5%) (2%) (0%)

(31%) (49%) (43%) (52%) (29%) (0%)

In parenthesis: percentage of samples above the LOQ. The analyses were performed on fresh unprocessed samples. The samples were collected during JanuaryeDecember 2010. n: Number of composite samples. LOQ: 90 pg g1 ww. for all the BDE209,10 pg g1 ww for the rest. a Cheese: cured, semi-cured and fresh cheese; Offal: lamb liver, pork kidney and chicken gizzards; Smoked fish: smoked salmon, trout and cod; Canned fish: tuna, mackerel, sardine in vegetal oil; Salty fish: Cod, tuna and anchovy; Crustaceans: prawn crayfish and shrimp.

O. Pardo et al. / Chemosphere 250 (2020) 126247

Fish and fish products

Sunflower oil Olive oil Mean (n ¼ 20) Butter Cow milk Custards and smoothies Cheesea Yogurt Mean (n ¼ 60) Foie-grass Chicken meat Beef meat Rabbit meat Lamb meat Pork Meat Cured ham Cooked ham Cured sausage Fresh sausage Beef hamburger Offala Mean (n ¼ 120) Dark or oily fish Tuna and albacore tuna Salmon and trout Sardine and anchovy Smoked fisha Seabream and bass Canned fisha White or lean fish Hake Swordfish Other fish products Salty fisha Squid and cuttlefish Crustaceana Mussel Mean (n ¼ 120) Chicken egg (n ¼ 10)

O. Pardo et al. / Chemosphere 250 (2020) 126247

(in general close to the LOQ, except for foie gras) are in agreement with those reported by Pietron et al. (2019) in farm animal meat, with mean values ranging from 0.22 to 3.82 pg g1 ww for the individual congeners, except for BDE209 with mean values from 11.7 (in turkey) to 37.2 pg g1 ww (in ostrich). All results in the study by Pietron et al. (2019) were below the LOQ of the present study (set at 10 pg g1 ww except for BDE209, whose LOQ was 90 pg g1 ww). Similarly, Boucher et al. (2018) reported mean PBDE values of 90, 236, 117, and 71 pg g1 ww for red meat, poultry meat, read meat liver and processed meat, respectively and they were all near or below the LOQ set in this study. Foie gras was the meat product with the highest PBDE level (355 pg g1 ww), which was expected due to its high fat content. Low PBDE concentrations in milk could be explained by the inclusion of skimmed dairy products in the composite samples, which could affect the usual profile of BDE congeners. Nevertheless, Boucher et al. (2018) also reported low mean levels (below 100 pg g1 ww) in 44 samples of dairy products (including cheese, cream, ice cream, milk, yogurt, and composite dairy products). In our study, eggs presented mean PBDE levels of 229 pg g1 ww, results that are in line with those reported previously (mean of 110 pg g1 ww) by Boucher et al. (2018). In view of the results obtained, it is evident that even greater efforts must be made to decrease the LOQ in order to detect the low PBDE levels present in some matrices such as meat, milk, or eggs. The average PBDE profiles in the different food categories studied are presented in Fig. 2. The dominating congener in fish and seafood, meat and meat products, milk, and vegetable oils was BDE47 (30%), in accordance with findings in fish from previous
n et al., 2017). In the studies in Spain (Pardo et al., 2014; Trabalo Netherlands (Boon et al., 2016), authors also concluded that BDE47 and BDE100 were important contributors to PBDE levels in fish. Regarding meat products, our results agree with those reported by Pietron et al. (2019), in which BDE47 and BDE99 were identified as important contributors to the total amount of PBDEs. Although some authors (Pietron et al., 2019; Garcia Lopez et al., 2018) identify BDE209 as a congener frequently present in foodstuffs, our study does not draw the same conclusion, probably due to the different LOQs used. It should be taken into account that these contributions in our study have been calculated using the LB levels, whereas in the other studies, they were calculated with the UB levels, and it is well known that this congener has a high LOQ. In meat products, BDE100 highly contributed to the total amount of PBDEs as compared with the results by Garcia Lopez et al. (2018) where BDE47, BDE99, and BDE100 were identified as

7

the most frequently occurring congeners in meat. In milk and dairy products, BDE47 was previously identified as the most important contributor (EFSA, 2011a,b). The dominating congener in eggs was BDE99, in accordance with the results reported by EFSA (2011)a,b. In general, it is well known that the PBDE patterns observed in foodstuffs are dominated by congeners related to the penta-mix formulation (such as BDE47, BDE99, BDE100, BDE153, BDE154, and BDE155), which could reflect the fact that debromination of higher congeners can occur in the environment. This fact has also been confirmed by the data from the present study. 3.2. Evaluation of dietary intake The estimated daily intakes (EDI) among adults and young people in the LB and UB scenarios are presented in Table 4. In the UB scenario, the highest EDIs for adults were BDE209 (0.526 ng kg1 bw day1) and BDE99 (0.095 ng kg1 bw day1), results which are in line with those reported by EFSA (EFSA, 2011a,b), which identified BDE209 as the highest contributor to the total PBDE exposure. However, it should be considered that in the UB scenario, dietary intake of BDE209 is often overestimated. This is because if this congener is not quantified in a sample, the concentration is assumed to be the LOQ, which is 9 times higher for BDE209 than for the rest of the congeners (90 vs 10 pg g1 ww, respectively). For young people, the highest EDIs were for BDE209 (1.345 ng kg1 bw day1) and BDE99 (0.270 ng kg1 bw day1). The EDIs of the sum of PBDEs in the UB scenario for adults and young people were 1.443 and 3.456 ng kg1 bw day1, respectively. This variation can be explained by differences in body weight and food consumption patterns across age groups. In recent years, the dietary intake of PBDEs has been evaluated in multiple studies in the literature. Table 5 includes EDIs of different studies from the literature. Interpreting the differences among exposure assessments should be done with caution due to the different methodologies used (e.g. sampling method, food consumption data, and exposure assessment model), foods analyzed, number of congeners considered, occurrence data reported (LB-UB), and many other factors and/or assumptions that can influence the final exposure estimations. In a national context, based on EDIs previously calculated in the same study region (Pardo et al., 2014), one could conclude that the exposure to PBDEs has increased in adults and young people from this area over time. However, this could be explained because the

Fig. 2. Average BDE profiles in the different food categories studied (percentage of the total, pg/g ww basis). Only those detected congeners were included (LB levels).

8

O. Pardo et al. / Chemosphere 250 (2020) 126247

Table 4 Estimated daily intake (ng kg bw1day1) of PBDEs by the population of the Valencia Region (Spain). Minimum

Median

Maximum

P90

LB

LB

UB

LB

UB

LB

UB

LB

UB

LB

UB

LB

UB

LB

UB

0.000 0.047 0.032 0.046 0.119 0.019 0.000 0.007 0.000 0.070 0.027 0.000 0.000 0.367

0.130 0.173 0.155 0.169 0.240 0.135 0.131 0.129 0.129 0.182 0.153 0.130 1.217 3.073

0.005 0.083 0.040 0.049 0.138 0.027 0.004 0.012 0.002 0.086 0.036 0.000 0.000 0.482

0.144 0.226 0.177 0.194 0.270 0.147 0.145 0.143 0.144 0.208 0.169 0.144 1.345 3.456

0.002 0.087 0.048 0.072 0.169 0.042 0.000 0.019 0.001 0.110 0.042 0.000 0.000 0.592

0.184 0.254 0.227 0.250 0.331 0.185 0.184 0.182 0.182 0.271 0.218 0.182 1.711 4.361

0.099 0.767 0.284 0.222 0.622 0.116 0.110 0.072 0.053 0.296 0.223 0.000 0.000 2.864

0.454 2.452 0.547 0.628 1.016 0.456 0.454 0.454 0.456 0.694 0.535 0.454 4.273 12.873

0.014 0.164 0.076 0.097 0.273 0.062 0.005 0.033 0.003 0.152 0.082 0.000 0.000 0.961

0.257 0.406 0.295 0.322 0.456 0.263 0.257 0.255 0.255 0.349 0.294 0.255 2.393 6.057

0.030 0.299 0.115 0.112 0.399 0.077 0.038 0.042 0.006 0.228 0.103 0.000 0.000 1.449

0.318 0.523 0.366 0.373 0.675 0.315 0.337 0.315 0.315 0.448 0.362 0.315 2.960 7.622

0.080 0.641 0.230 0.178 0.597 0.110 0.091 0.062 0.035 0.262 0.169 0.000 0.000 2.455

0.440 0.960 0.464 0.516 0.970 0.440 0.440 0.440 0.443 0.602 0.513 0.440 4.137 10.805

0.000 0.028 0.019 0.028 0.033 0.011 0.000 0.004 0.000 0.032 0.010 0.000 0.000 0.165

0.052 0.077 0.071 0.081 0.082 0.054 0.053 0.051 0.052 0.077 0.061 0.052 0.484 1.247

0.005 0.063 0.029 0.032 0.043 0.016 0.003 0.006 0.002 0.038 0.016 0.000 0.000 0.253

0.058 0.155 0.081 0.089 0.095 0.059 0.057 0.056 0.057 0.086 0.068 0.056 0.526 1.443

0.001 0.054 0.033 0.045 0.065 0.024 0.000 0.010 0.001 0.050 0.019 0.000 0.000 0.302

0.075 0.121 0.101 0.113 0.128 0.077 0.075 0.073 0.073 0.110 0.088 0.073 0.684 1.791

0.195 1.013 0.342 0.152 0.309 0.152 0.109 0.042 0.053 0.261 0.174 0.007 0.000 2.809

0.264 4.789 0.380 0.332 0.506 0.264 0.264 0.264 0.264 0.368 0.302 0.264 2.479 10.740

0.015 0.163 0.065 0.064 0.100 0.039 0.008 0.017 0.003 0.074 0.038 0.000 0.000 0.586

0.103 0.240 0.142 0.153 0.186 0.107 0.101 0.099 0.101 0.150 0.121 0.099 0.932 2.534

0.032 0.290 0.103 0.081 0.127 0.052 0.024 0.021 0.006 0.092 0.057 0.000 0.000 0.885

0.123 0.526 0.174 0.176 0.222 0.127 0.121 0.117 0.121 0.180 0.139 0.117 1.096 3.239

0.075 0.539 0.186 0.115 0.194 0.082 0.050 0.031 0.035 0.135 0.098 0.002 0.000 1.542

0.160 1.738 0.258 0.213 0.309 0.173 0.156 0.156 0.156 0.230 0.189 0.156 1.463 5.357

UB

Young people (6e15 years) BDE28 0.000 0.005 BDE47 0.000 0.012 BDE49 0.000 0.010 BDE66 0.000 0.013 BDE99 0.000 0.006 BDE100 0.000 0.005 BDE138 0.000 0.005 BDE139 0.000 0.005 BDE153 0.000 0.005 BDE154 0.000 0.009 BDE155 0.000 0.007 BDE183 0.000 0.005 BDE209 0.000 0.051 Total PBDEs 0.000 0.138 Adults >15 years BDE28 0.000 0.000 BDE47 0.000 0.000 BDE49 0.000 0.000 BDE66 0.000 0.000 BDE99 0.000 0.000 BDE100 0.000 0.000 BDE138 0.000 0.000 BDE139 0.000 0.000 BDE153 0.000 0.000 BDE154 0.000 0.000 BDE155 0.000 0.000 BDE183 0.000 0.000 BDE209 0.000 0.000 Total PBDEs 0.000 0.000

Mean

P75

P95

P99

LB: Lower bound; UB: upper bound.

Table 5 Adult’s estimated daily intake (ng kg ww1day1) of BDEs from other recent studies. Country

Dietary exposure

Matrix

References

Japan China France China Italy Spain UK Nigeria China

0.98 (MB) 0.76 (MB) 0.55 (UB) 1.4 (MB) 0.29 (MB) 0.45 (MB) 1.77 (MB) 0.13 (LB) 9.77 (LB)

Sunggyu et al. (2013) Zhang et al. (2013) re at al., 2014 Rivie Gong et al. (2015) Martellini et al. (2016)
n et al. (2017) Trabalo Tao et al. (2017) Babalola et al., 2018 Wang et al. (2019)

Spain

0.48 (LB)-3.46(UB)

Fish Fish, meat, egg, milk, cereal, legumes and nuts, tubers and vegetables TDS Carp Fish and mollusc Fish and shellfish Meat, liver, fish, egg, dairy products Meat products, aquatic foods, dairy products, edible oil, eggs, fruit and vegetable, cereals Eggs, fish and seafood, meat, dairy products, vegetables, fruit and berry, edible oils, soybean and nut, cereals Eggs, fish and seafood, meat and meat products, milk and dairy products, vegetable oils

Present study

LB: Lowerbound; MB: Middle bound; UB: Upperbound TDS: Total Diet Study.

present study includes 13 BDE congeners and a greater number of foods, whereas the previous study included 12 BDE congeners and focused exclusively on fish and seafood. Other studies carried out in
n et al., 2017) estimated a daily intake of 0.45 ng kg1 Spain (Trabalo 1 bw day , but again, only fish and shellfish were analyzed, underestimating the dietary intake of PBDEs. In an international context, several studies have reported dietary intake of PBDEs, but these have included only fish or seafood in their calculations, and they used the middle bound (MB) scenario (see Table 5). High exposure levels were observed if foods from different categories, and especially foods of animal origin, were included in the calculations. For example, the EDI was 0.76 ng kg1 bw day1 in China (Zhang et al., 2013) and 1.77 ng kg1 bw day1 in U.K. (Tao et al., 2017), both in the MB scenario. However, low EDIs of PBDEs were reported in France (0.55 ng kg1 bw day1 in the UB re at al., 2014) and Nigeria (0.13 ng kg1 bw day1 in scenario) (Rivie

the LB scenario) (Babalola et al., 2018), despite having included several types of food. Nevertheless, these results should be considered with caution due to the scenario and the LOQ used for the calculations. The contribution of the different food categories to the mean EDI of the sum of PBDEs was calculated taking into account the PBDE levels in the LB scenario and the mean consumption values by food category. The results for the two populations studied are shown in Fig. 3. In the group of young people, milk and dairy products contributed largely to the exposure (20%), whereas in the adult group, fish and seafood were the most important contributors, a difference which can be explained by the consumption patterns within the age groups. These results are in accordance with those from other studies, which suggest that the most important contribution to PBDE exposure is from fish and seafood in adults (Domingo and Bocio, 2007; Babalola et al., 2018).

O. Pardo et al. / Chemosphere 250 (2020) 126247

9

Fig. 3. Dietary exposure contribution of PBDEs, by food category, in the LB scenario (percentage of the total).

3.3. Risk characterization The MOE can be calculated as the ratio between the lower bound of the benchmark dose (BMDL) and the EDI (EFSA, 2011a,b). Estimated MOEs for the mean and the 95th percentile (P95) of the exposure levels for adults and young people are shown in Table 6. In both scenarios (LB and UB), the calculated MOEs are higher than 2.5 for BDE47, BDE99, BDE153, and BDE209. In conclusion, the estimated MOE values indicate no health concern regarding current dietary exposure in the region of Valencia. Nevertheless, it should be taken into account that for BDE209 the concentration in food was set to the LOQ value since it was not detected in any of the samples. Although EFSA identified a potential health concern for BDE99 in young children (1e3 years) in 2011 (EFSA, 2011a,b), the CONTAM Panel related this finding to the use of UB estimates and the longest reported half-life in humans for the calculation of the dietary intake associated with the body burden at the BMDL10. This could have resulted in an overestimation of the risk for this specific age group. Nevertheless, taking into account that exposure during key developmental stages in infancy is the most damaging, and that this is the time when altered hormone regulation will have the greatest impact (Bramwell et al., 2017), efforts will be made in the future to refine the exposure calculation for this specific population group in order to protect them from possible health risks. 3.4. Study strengths and limitations This study reports the occurrence of a large number of PBDE congeners in foods known to present high PBDE levels (those of animal origin) and in other foods with limited data available (vegetable oils) from the Region of Valencia, using a representative sampling. Salty fish, commonly consumed in Spain, was included for the first time. The risk of dietary exposure to PBDEs was assessed in two population subgroups (adult and young people) from this region, which could help policy makers with risk management. This study increases the contribution of Spain to the global PBDE data available, identified as poor by Boucher et al.

(2018). This type of data is particularly needed to assess population-specific intake and risk. At the same time, it contributes to fill in the existing research gaps regarding PBDE exposure and risk estimates. However, this study also presents a series of limitations. For instance, the effect of cooking or food processing was not considered in the present study, although it is well known to affect the exposure. The fat content of the different food items was not determined since the European Commission’s Recommendation 2014/118/EU (EC, 2014) established the LOQ based on wet weight. In addition, the censoring rate in this study is high. However, it should be taken into account that it includes a large number of congeners (13 in total), some of which are not usually included in research studies and are generally present at very low levels. The possible reasons causing this limitation have been described above. It should also be remarked that a low censoring rate introduces a source of uncertainty in the risk assessment. Specifically, the EDI, and consequently the MOE, for BDE209 was calculated from an exposure level set at the LOQ since this congener was not quantified in any sample. Nevertheless, this risk calculation for BDE209 is valuable as this congener has never been reported in the literature, and it could serve as a good point of departure for future research. 4. Conclusions To our knowledge, this is the first study that has calculated the PBDE intake in a Spanish population considering a wide variety of fatty foods, obtaining a more realistic estimation. In the present study, vegetable oils, and fish and seafood presented the highest content of PBDEs (mean values of 503 and 464 pg g1 ww for total PBDEs, respectively), whereas the lowest PBDE concentrations were found in meat and meat products, milk and dairy products, and eggs (mean values of 224, 247, and 229 pg g1 ww for total PBDEs, respectively). Regarding fish, temporal trends in our region suggest that regulations have greatly influenced the PBDE contamination levels. The dominating congeners were BDE47 in fish and seafood, meat and meat products, and vegetable oils; and BDE99 in eggs, and milk and dairy products. The average EDIs of the sum of PBDEs (UB scenario) for adults

1263941 574324 3231939 1551095

UB LB UB

1.345 2.96 0.526 1.096 0 0 0 0

UB

67 30 168 79

LB

4800 1600 4800 1600

LB

CRediT author statement

9.6 UB

0.144 0.315 0.057 0.121

LB

0.002 0.006 0.002 0.006

UB

637 255 1811 775

LB

30 431 4000 1354 4.2 0.27 0.675 0.095 0.222

UB LB

0.138 0.399 0.043 0.127 761 329 1110 327

UB LB

2072 575 2730 593 172 0.226 0.523 0.155 0.526

UB LB

0.083 0.299 0.063 0.29 Mean P95 Mean P95 Young people Adults

MOE BMDL10 (mg/kg¡1 bw) Estimated intake (ng kg¡1 bw)

BDE-209

MOE Dr,h (ng kg¡1 bw) Estimated intake (ng kg¡1 bw)

BDE-153

MOE Dr,h (ng kg¡1 bw) BDE-99

Estimated intake (ng kg¡1 bw) MOE

and young people were 1.443 and 3.456 ng kg1 bw day1, respectively. Considering EDIs based on the average and P95, the obtained MOEs do not show evidence of risk. Although humans are mainly exposed to PBDEs through diet, it should be highlighted that for the total PBDE daily intake, other external exposure routes such as dust or air must be considered. Lastly, despite the known methodological limitations of determining trace levels of PBDEs in food, the LOQs in our study are low enough to discard possible health risks.

Olga Pardo: Writing- Original draft preparation, Sample Analysis, Validation, Visualization, Investigation, Writing- Reviewing and Editing. Sandra F. Fern
andez: Editing, Validation, Sample Analysis. Leyre Quijano: Data curation, Editing, Writing- Reviewing. Silvia Marín: Conceptualization, Methodology, Software, Sampling. Pilar Villalba: Conceptualization, Methodology, Software, Sampling. : ConcepFrancisca Corpas-Burgos: Statistical analysis. Vicent Yusa tualization, Methodology, Supervision. Declaration of competing interest

Dr,h (ng kg¡1 bw) Estimated intake (ng kg¡1 bw)

BDE-47

PBDE congeners

Subgroup of population

Table 6 Margins of exposure (MOEs) for BDE-47, -99, 153 and 209, for different population groups, based on the lower bound (LB) and upper bound (UB) scenarios of the dietary intake.

nc nc nc nc

O. Pardo et al. / Chemosphere 250 (2020) 126247

1.7

10

No potential conflict of interest was reported by the authors. Acknowledgments This work is part of the Total Diet Study in the Region of Valencia, Spain (EDT-CV). References Babalola, B.A., Adeyi, A.A., 2018. Levels, dietary intake and risk of polybrominated diphenyl ethers (PBDEs) in foods commonly consumed in Nigeria. Food Chem. 265, 78e84. Besis, A., Samara, C., 2012. Polybrominated diphenyl ethers (PBDEs) in the indoor and outdoor environments-a review on occurrence and human exposure. Environ. Pollut. 169, 217e229. Boon, P.E., te Biesebeek, J.D., Leeuwen, S.P.J., van Zeilmaker, M.J., Hoogenboom, L.A.P., 2016. Dietary Exposure to Polybrominated Diphenyl Ethers in the Netherlands. P.O. Box 1j3720 BA Bilthoven. The Netherlands. Boucher, B.A., Ennis, J.K., Tsirlin, D., Harris, S.A., 2018. A global database of polybrominated diphenyl ether flame retardant congeners in foods and supplements. J. Food Compos. Anal. 69, 171e188. Bramwell, L., Harrad, S., Abou-Elwafa Abdallah, M., Rauert, C., Rose, M., Fernandes, A., Pless-Mulloli, T., 2017. Predictors of human PBDE body burdens for a UK cohort. Chemosphere 189, 186e197. ~ iguez, C., NavarreteCosta, O., Lopez-Espinosa, M.J., Vizcaino, E., Murcia, M., In ~ oz, E.M., Grimalt, J.O., Tardon, A., Ballester, F., Fernandez-Somoano, A., 2016. Mun Dietary and household sources of prenatal exposure to polybrominated diphenyl ethers (PBDEs) in the INMA birth cohort (Spain). Environ. Sci. Technol. 50 (11), 5935e5944. Domingo, J.L., Bocio, A., 2007. Levels of PCDD/PCDFs and PCBs in edible marine species and human intake: a literatura review. Environ. Int. 33, 397e405. EFSA, 2010. Management of left-censored data in dietary exposure assessment of chemical substances. EFSA J 8, 1557. EFSA, 2011a. Scientific opinion on polybrominated diphenyl ethers (PBDEs) in food. EFSA J 9, 2156. EFSA, 2011b. Evaluation of the foodex, the classification system applied to the development of the EFSA comprehensive European food consumption database. EFSA 9 (3), 1970. EPA (Environmental Protection Agency), 1995. Volume 1: Composite Sampling. EPA230-R-95-005. European Commission, 2014. Commission Recommendation 2014/118/EU of 3 March 2014 on the monitoring of traces of brominated flame retardants in food. Off. J. Eur. Union L39eL65.
n, R., Quiles, J., Rizk, J., Zubeldia, L., 2010. Fullana, A.M., Jimenez, R., Marín, S., Ramo General Directorate of Public Health. Nutrition Survey in the Valencia Region a 2010-2011, 1 ed. Generalitat Valenciana. Conselleria de Sanitat, Valencia. 97884-482-5866-5. Garcia Lopez, M., Driffield, M., Fernandes, A.R., Smith, F., Tarbin, J., Lloyd, A.S., Christy, J., Holland, M., Steel, Z., Tlustos, C., 2018. Occurrence of polybrominated diphenylethers, hexabromocyclododecanes, bromophenols and tetrabromobisphenols A and S in Irish foods. Chemosphere 197, 709e715. Gong, Y., Wen, S., Zheng, C., Peng, X., Li, Y., Hu, D., Peng, L., 2015. Potential risk

O. Pardo et al. / Chemosphere 250 (2020) 126247 assessment of polybrominated diphenyl ethers (PBDEs) by consuming animalderived foods collected from interior areas of China. Environ. Sci. Pollut. Res. 22, 8349e8358. International Organization for Standardization ISO/IEC 17025, 2017. General Requirements for the Competence of Testing and Calibration Laboratories. Kim, Y.R., Harden, F.A., Toms, L.M., Norman, R.E., 2014. Health consequences of exposure to brominated flame retardants: a systematic review. Chemosphere 106, 1e19. Kiviranta, H., Ovaskainen, M.A.L., Vartiainen, T., 2004. Market basket study on dietary intake of PCDD/Fs, PCBs, and PBDEs in Finland. Environ. Int. 30, 923e932. Korcz, W., Strucinski, P., Goralczyk, K., Hernik, A., Łyczewska, M., Matuszak, M., Czaja, K., Minorczyk, M., Ludwicki, J.K., 2017. Levels of polybrominated diphenyl ethers in house dust in Central Poland. Indoor Air 27, 128e135. Leblanc, J.-C., Volatier, J.-L., Sirot, V., Bemrah-Aouachria, N., 2006. CALIPSO, Fish and Seafood Consumption Study and Biomarker of Exposure to Trace Elements, Pollutants and Omega 3. The General Directorate for Foods of France’s Ministry of Agriculture and Fisheries, AFSSA. the French Food Safety Agency and the French Institute for Agronomy Research INRA. Martellini, T., Diletti, G., Scortichini, G., Lolini, M., Lanciotti, E., Katsoyiannis, A., Cincinelli, A., 2016. Occurrence of polybrominated diphenyl ethers (PBDEs) in foodstuffs in Italy and implications for human exposure. Food Chem. Toxicol. 89, 32e38.
n de alimentos. Universidad de Granada. Mataix Verdú, J., 2009. Tabla de composicio
s, T., Lo
pez, F.J., Hern
Medina, C.M., Pitarch, E., Portole andez, F., 2008. Determination of PBDEs in human breast adipose tissues by gas chromatography coupled with triple quadrupole mass spectrometry. Anal. Bioanal. Chem. 390, 1343e1354.
s, T., Lo
pez, F.J., Hern
Medina, C.M., Pitarch, E., Portole andez, F., 2009. GC-MS/MS multi-residue method for the determination of organochlorine pesticides, polychlorinated biphenyls and polybrominated diphenyl ethers in human breast tissues. J. Separ. Sci. 32 (12), 2090e2102. Pardo, O., Beser, M.I., Yusa, V., Beltran, J., 2014. Probabilistic risk assessment of the exposure to polybrominated diphenyl ethers via fish and seafood consumption in the Region of Valencia (Spain). Chemosphere 104, 7e14. Piersanti, A., Tavoloni, T., Bastari, E., Lestingi, C., Romanelli, S., Saluti, G., Moretti, S., Galarini, R., 2015. Polybrominated diphenyl ethers in mussels (Mytilus galloprovinciallis) collected from Central Adriatic sea. Mar. Pollut. Bull. 101 (1), 417e421. Pietron, W., Pajurek, M., Mikolajczyk, S., Maszewski, S., Warenik-Bany, M., Piskorska-Pliszczynska, J., 2019. Exposure to PBDEs associated with farm animal meat

11

consumption. Chemosphere 224, 58e64. Pietron, W.J., Malagocki, P., 2017. Quantification of polybrominated diphenyl ethers (PBDEs) in food. A review. Talanta 167, 411e427. Shi, Z., Zhang, L., Zhao, Y., Sun, Z., Zhou, X., Li, J., Wu, Y., 2017. Dietary exposure assessment of Chinese population to tetrabromobisphenol-A, hexabromocyclododecane and decabrominated diphenyl ether: results of the 5th Chinese Total Diet Study. Environ. Int. 229, 539e547. Sunggyu, L., Sunmi, K., Eunkyo, K., In-Seok, L., Gyuyeon, C., Hai-Joong, K., Jeongim, P., Jeong, Jae L., Sooran, C., Su Young, K., Suungjoo, K., Sungkyoon, K., Kyungho, C., Hyo-Bang, M., 2013. Polybrominated diphenyl ethers (PBDEs) in breast milk of Korea in 2011: current contamination, time course variation, influencing factors and health risks. Environ. Res. 126, 76e83. Tao, F., Abou-Elwafa Abdallah, M., Ashworth, D.C., Douglas, P., Toledano, M.B., Harrad, S., 2017. Emerging and legacy flame retardants in UK human milk and food suggest slow response to restrictions on use of PBDEs and HBCDD. Environ. Int. 105, 95e104.
n, L., Vilavert, L., Domingo, J.L., Pocurull, E., Borrull, F., Nadal, M., 2017. HuTrabalo man exposure to brominated flame retardants through the consumption of fish and shellfish in Tarragona County (Catalonia, Spain). Food Chem. Toxicol. 104, 46e56.
nisseau, A., Bichon, E., Brosseaud, A., Vaccher, V., Lesquin, E., Larvor, F., Durand, S., Ve Dervilly-Pinel, G., Marchand, P., Le Bizec, B., 2018. Occurrence of legacy and novel brominated flame retardants in food and feed in France for the period 2014 to 2016. Chemosphere 207, 497e506. Wang, J., Zhao, X., Wang, Y., Shi, Z., 2019. Tetrabromobisphenol A, hexabromocyclododecane isomers and polybrominated diphenyl ethers in foodstuffs from Beijing, China: contamination levels, dietary exposure and risk assessment. Sci. Total Environ. 666, 812e820. WHO (World Health Organisation), 1999. Environmental Healt Criteria 210, Principles for the Assessment of Risk to Human Health from Exposure to Chemicals. WHO, Geneva, Switzerland. , V., Pardo, O., Pastor, A., de la Guardia, M., 2006. Microwave-assisted extraction Yusa of polybrominated diphenyl ethers and polychlorinated naphthalenes concentrated on semipermeable membrane devices. Anal. Chim. Acta 565, 103e111. Zhang, L., Li, J., Zhao, Y., Li, X., Wen, S., Shen, H., Wu, Y., 2013. Polybrominated diphenyl ethers (PBDEs) and indicator polychlorinated byphenyls (PCBs) in foods from China: levels, dietary intake, and risk assessment. J. Agric. Food Chem. 61, 6544e6551.