Accepted Manuscript
Risk assessment of benzene in food samples of Iran's market
Ali Heshmati, Sabah Ghadimi, Amin Mousavi Khaneghah, Francisco J. Barba, Jose
M. Lorenzo, Fatemeh Nazemi, Yadolah Fakhri
PII:
S0278-6915(18)30114-5
DOI:
10.1016/j.fct.2018.02.043
Reference:
FCT 9614
To appear in:
Food and Chemical Toxicology
Received Date: 8 December 2017
Revised Date:
1 February 2018
Accepted Date: 18 February 2018
Please cite this article as: Heshmati, A., Ghadimi, S., Mousavi Khaneghah, A., Barba, F.J., Lorenzo,
J.M., Nazemi, F., Fakhri, Y., Risk assessment of benzene in food samples of Iran's market, Food and
Chemical Toxicology (2018), doi: 10.1016/j.fct.2018.02.043.
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Risk assessment of benzene in food samples of Iran’s market
Ali Heshmati a, Sabah Ghadimi a,**, Amin Mousavi Khaneghah b,*, Francisco J. Barba
a
, Jose M. Lorenzo d, Fatemeh Nazemia, Yadolah Fakhri e
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c,***
Nutrition Health Research Center, Hamadan University of Medical Sciences, Hamadan,
Iran
b
Department of Food Science, Faculty of Food Engineering, State University of Campinas
Paulo, Brazil
Nutrition and Food Science Area, Preventive Medicine and Public Health, Food Sciences,
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c
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(UNICAMP), Rua Monteiro Lobato, 80. Caixa Postal: 6121, CEP: 13083-862 Campinas, Sao
Toxicology and Forensic Medicine Department, Faculty of Pharmacy, Universitat de València,
Avda. Vicent Andrés Estellés, s/n, 46100 Burjassot, València, Spain
d
Centro Tecnológico de la Carne de Galicia, rúa Galicia n° 4, Parque Tecnológico de
Galicia, San Cibrao das Viñas, 32900 Ourense, Spain
e
Student Research Committee, Department of Environmental Health Engineering, School
*,**, ***
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of Public Health, Shahid Beheshti University of Medical Sciences, Tehran, Iran;
Corresponding authors
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mousavi@fea.unicamp.br (Amin Mousavi Khaneghah)
s.ghadimi@edu.umsha.ac.ir (Sabah Ghadimi)
Francisco.Barba@uv.es (Francisco J. Barba)
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ABSTRACT
The concentration of benzene in ninety-eight collected food and drink samples (carbonated
beverage, fruit juice, pickle, lime juice, mayonnaise and salad dressing, 16 samples from each)
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from Iran local markets were investigated using gas chromatography equipped with flame
ionization detector (GC-FID). Moreover, the correlation of benzene concentration with sodium
benzoate and ascorbic acid concentrations was assessed. Benzene concentration in carbonated
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beverages, fruit juices, pickle, lime juices, mayonnaise and salad dressing were 3.57±1.70,
5.17±3.63, 4.37±2.24, 4.99±0.54, 1.38±0.87 and 1.47±0.83 µg/L, respectively, being in all cases
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below the acceptable limit (10 µg/L) proposed by the World Health Organization (WHO) as a
reference for drinking water. Benzene concentration exceeded the maximum tolerable limit in
12.5% of carbonated beverages and fruit juices (two samples from each). A significant
correlation coefficient between the concentrations of benzene, sodium benzoate, and ascorbic
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acid in fruit juices, and levels of benzene, sodium benzoate in carbonated beverage samples was
observed. To characterize the risk of exposure of urban and rural consumers of Iran (female in
age group of 15-24 and male in age group of 25-64) to benzene in food and drink samples the
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Margin of Exposure (MoE) approach was applied revealing there is no severe concern regarding
benzene intake through assessed food and beverages samples in Iran.
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Keywords: Benzene; Ascorbic Acid; Sodium Benzoate; Carbonated Beverage; Risk Assessment
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1.
Introduction
Genotoxic and carcinogenic effects of benzene as a dangerous chemical contaminant have been
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documented for several years (Khalade et al., 2010) and International Agency for Research on
Cancer (IARC) classified benzene as a Group 1 human carcinogen (IARC 1987). Acute
occupational exposure to benzene may cause dizziness and narcosis (characterized by headache,
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confusion, drowsiness, tremors, and loss of consciousness). Moreover, chronic exposure to
benzene can result in a decrement in the production of both red and white blood cells from bone
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marrow in humans and consequently in aplastic anemia (Mohammadyan et al., 2016). Moreover,
inhalation exposure to benzene might cause childhood leukemia, a significant form of childhood
cancer (Vinceti et al., 2012).
Although the most common way of exposure to benzene is inhalation, other routes such as
consumption of benzene-contaminated foods and beverages should be listed. The occurrence of
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benzene in food can be correlated with contaminated raw materials, contamination during
formulation and processing (e.g. smoking), migration of benzene from packaging materials into
the product and contact of final product with contaminated areas (Arisseto et al., 2013;
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Lachenmeier et al., 2010; Salviano dos Santos et al., 2015). In addition, benzene might be
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produced during cooking and irradiation due to thermal degradation of food components as well
as food additives (Salviano dos Santos et al., 2015; Vinci et al., 2012).
Salts of benzoic acid such as sodium benzoate and potassium benzoate are widely used as
preservatives agents in the food formulations (Montesinos-Herrero et al., 2016). Likewise,
ascorbic acid is utilized to delay oxidative reactions due to its antioxidant activity (Karatoprak et
al., 2016; Tundis et al., 2017). It has been reported that the decarboxylation of sodium benzoate
in the presence of ascorbic acid resulted in the production of benzene (Casado et al., 2011;
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Gardner, and Lawrence 1993). This reaction can be accelerated by transition metals including
Cu+2 and Fe+3, light as well as heating (Erickson & Doyle 2017; Loch et al., 2016).
Despite the fact that, no maximum limits for benzene in foods or beverages have been
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established, the acceptable level of benzene in potable water was considered for foods or
beverages. According to World Health Organization (WHO), the United States Environmental
Protection Agency (US EPA), and the European Council the maximum acceptable level of
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benzene in drinking water was set at 10, 5 and 1 µg/L, respectively (EC 1998; USEPA 1985;
WHO 2004).
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The occurrence of benzene in a variety of beverages, especially soft drinks has been
investigated (Arisseto et al., 2013; FSAI 2008; Ibolya et al., 2012; Kharat et al., 2016; Morsi, et
al., 2012; Sanchez et al., 2013; Yang et al., 2010). Based on the previous studies, benzene
concentration in food was reported to be low. Therefore, it appears the administration pathways
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play a minor role in benzene exposure (Smith et al., 2010; Vinci et al., 2012).
For the safety assessment of contaminants such as benzene that are both genotoxic and
carcinogenic, MOE (margin of exposure) value should be determined. MOE is calculated as a
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ratio of BMDL10 (benchmark dose lower confidence limit) and contaminant exposure (EFSA
2009; EFSA 2012; WHO 2005). In this regard, BMDL10 of 17.6 mg/kg bw/d derived from
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female rat Zymbal gland carcinoma can be used to calculate MOE (Smith et al., 2010; Vinci et
al., 2012). Based on the previously calculated BMDL10, MOE values ranging from 400, 000 to 2,
000, 000 for dietary exposure of benzene was estimated (Smith et al., 2010). MOE values higher
than 10, 000 can be considered as “low concern” for human health (EFSA 2005).
Based on our knowledge, there is no study regarding the determination of benzene level in
food and beverage products from Iran´s market. Therefore, two main objectives are considered:
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1) to determine benzene concentration in carbonated beverage, fruit juice, pickle, mayonnaise
and salad dressing, and lime juice samples collected from Iran market; and 2) to assess the risk
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associated for Iran's population exposure to benzene via food and beverages consumption.
Materials and methods
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2.1. Samples
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The current cross-sectional study was conducted from May to July 2016 by collecting ninety-six
samples (6 categories of food, 16 samples from each group) including carbonated beverage, fruit
juice, pickle, lime juice, mayonnaise and salad dressing were collected from retails and local
markets in Hamadan province (Iran). Samples were chosen from different production batches
and produced by various manufactures in all over Iran and distributed in Hamadan and were
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stored at 4 ºC before analysis (maximum 2 weeks). Precautionary measures were carried out to
avoid sample contamination prior the analysis.
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2.2. Chemicals and reagents
Ascorbic acid (99.5%) was purchased from Sigma-Aldrich (St Louis, MO, USA). The used high-
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quality water was prepared by a Milli-Q system (Millipore, Billerica, Massachusetts, USA).
Benzoic acid (99.5% pure), benzene (99.9% pure), sodium salicylate (99.5% pure) and HPLCgrade acetonitrile (99.9% pure), ammonium acetate (98% pure), methanol, acetic acid glacial
(100% pure) and other chemicals were obtained from Merck (Darmstadt, Germany).
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2.3. Measurement of benzene
Benzene measurement was carried out according to the recommended procedure by Casado et al.
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(2011) with minor modifications. A Varian CP-3800 gas chromatography equipment (Kyoto,
USA), with a flame ionization detector and split/splitless injector, was used. Chromatographic
separations were performed on a DB5 capillary column (40 m × 0.18 mm, 0.18 µm film
thickness) (Agilent Technologies, Palo Alto, CA, USA). Nitrogen (99.99% pure) was used as the
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carrier gas at initial flow-rate of 1.4 and 40 mL/min–1, respectively. The original column oven
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temperature was set at 40 °C (1 minute); afterward increasing 10 °C/min–1 to 150 °C (held for 3
min) and then programmed to raise 10 °C min–1 to a final temperature of 200 °C, which took
place for 5 min. The injector and detector temperature were kept at 300 ºC and 280 °C,
respectively. The split ratio was performed at 1:100.
The samples containing solid and/or fat were gently homogenized. Subsequently, the borate
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buffer (0.8 M, pH 10) as antifoam agent, was incorporated. In the case of liquid samples, fat
homogenization was carried out by shaking for 30 s and degassing of carbonaceous drinks was
performed by sonicating 15 mL of the soft drink for 1.5 min or until all carbon dioxide bubbles
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disappeared. Afterward, 5 mL of filtrated liquid sample (for pickle, 5 mL of brine) or 5 g of solid
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sample (dressing salad and mayonnaise) was poured into a vial of 10 mL. The pH was adjusted
to 10 by addition of NaOH solution (30%). After incorporation of NaCl (one gram), the vial was
sealed and heated at 80 °C for 5 min to extract benzene and its transfer into the headspace. Then,
2 µL of sample headspace was injected into the GC-FID. The identification of positive benzene
samples by GC-FID was confirmed by using Headspace-Solid Gas Chromatography/Mass
Spectrometry (HS/GC-MS) (Agilent Technologies, Palo Alto, USA) according to previous
procedure (Casado et al., 2011).
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2.4. Measurement of ascorbic acid content
The ascorbic acid concentration was determined by a redox titration with potassium iodate in the
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presence of starch indicator (Suntornsuk, Gritsanapun, Nilkamhank, & Paochom, 2002). Briefly,
in a conical flask, 25 mL of sample was added to 25 mL of sulfuric acid solution (2 mol/L), 50
mL of distilled water and 3 mL of starch indicator solution (0.5%). This mixture was titrated
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with iodine solution (0.1 mol/L) until a blue-black color remained, which indicate the titration
endpoint and the formation of the starch-iodine complex. A blank titration was performed on
2.5. Sodium benzoate determination
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each sample.
After sample preparation (as was mentioned before), 200 µL sodium salicylate (internal
standard) (1000 mg/kg) were added to 10 g of the solid food samples and 10 mL of liquid
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samples. Afterward, samples were diluted with deionized water to 50 mL. The mixture was
shaken for 5 min and centrifuged at 700 × g for 15 min. After that, 2 mL of the upper layer was
diluted to 10 mL with the mobile phase and then filtered through a 0.45 µm cellulose acetate
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syringe filter. For the measurement, 20 µL of filtered sample was injected into the HPLC
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(Zamani Mazdeh et al., 2014).
A Waters HPLC apparatus (Milford, MA, USA) containing UV detector and equipped
with C18 analytical column (250 mm × 4.6 mm, i.d., 5 µm) was used for the benzoic acid
measurement. The mobile phase consisted of 80% aqueous ammonium acetate buffered at
pH=4.2 with 20% of acetonitrile, at a flow rate of 1 mL/min. The benzoic acid was determined at
a wavelength of 225 nm.
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2.6. Performance of analytical methods
Before analysis benzene in the collected samples, a calibration curve was constructed. Firstly,
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benzene (100 µL) was dissolved in 5 mL dimethylformamide (DMF) to obtain a stock solution.
Then, five calibrating solutions (1, 5, 10, 15 and 20 µL/L) of the stock solution were prepared.
The correlation coefficient of 0.9993 was obtained for the mentioned curve. The signal-to-noise
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ratios for LOD and limit of quantification (LOQ) were 3 and 10, respectively. LOD and LOQ for
benzene were determined as 0.11 and 0.36 µg/L, respectively. The precision of analysis
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procedure was 1.7%. Moreover, the LOD and the LOQ values for benzoic acid were identified as
0.51 and 1.6 mg/kg, respectively. Mean recoveries of benzoic acid in the analyzed samples were
95.5% (alcoholic soft drink), 98.7% (fruit juice), 92.51% (pickle), 94.82% (lime juice) and
88.95% (mayonnaise and dressing salad). For quantification, a calibration curve with standard
solutions of benzoic acid was plotted. The linear regression equation was obtained, and the
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determination coefficient was calculated (R2 = 0.9994).
2.7. Exposure assessment of benzene
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Estimation of hazard risk of benzene through food substances and beverage was carried out by a
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deterministic approach (Richardson 1996). Benzene intake was estimated by considering upper
bound concentrations of this contaminant by Equation 1 (USEPA, 2015; WHO, 2009):
: DDE = IR × C / BW
Equation (1)
Where DDE is daily dietary exposure to benzene (mg/kg bw); IR, ingestion rate of food and
beverage (kg/day); C, the concentration of benzene (µg/L) and Bw, body weight in the males and
females age groups (kg).
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The consumption per capita of soft drink, fruit juice, pickle, lime juice and mayonnaise
and dressing salad in Iran was estimated as 115, 24, 6.2, 9.5, 8.2 g/day, respectively. For
determining the consumption of relevant foodstuffs and beverages, 450 peoples complete a food
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frequency questionnaire form. Two different population of each sex were considered: youth (1524 years) and adult (25-64 years) group. Food frequency questionnaire consisted 40 item of
specific food consumed in Iran. Risk assessment was done for 15-24 and 25-64 age groups for
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males and females in the urban and rural of Iran. The BW in the 15-24 and 25-64 age groups for
urban males is 69.24, and 75.64 kg and females are 58.90 and 69.90 kg, respectively. Also, BW
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in the 15-24 and 25-64 age groups for rural males is 63.51, and 70.07 kg and females are 55.96
and 64.80 kg, respectively (Asgari et al., 2007). The MOE values of benzene intake were
calculated by Equation 2:
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Equation 2: MOE = BMDL10 / DDE
Where, MOE is Margin of Exposure (Units); BMDL, Benchmark Dose Lower Confidence Limit
(mg/kg bw/day) (USEPA 2003) and DDE daily dietary exposure to benzene (ng/kg bw). In this
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study, the BMDL is based on the human research, therefore can provide an even higher level of
safety evidence. For samples with benzene concentration lower than the limit of detection
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(LOD), in order to calculate the dietary benzene exposure lower, medium and upper bound,
should be included (Vinci et al., 2012).
2.8. Statistical analysis
SPSS (version 16:0) software (SPSS Inc.; Chicago, IL, USA) was used to analyze data. The
findings were presented as a mean ± standard deviation. To compare the mean concentrations of
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benzene in the samples with the maximum acceptable level of this contaminant in drinking water
(10, 5 and 1 µg/L according to WHO, USEPA, and the European Council, respectively), a one-
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sample t-test was applied.
Results and discussion
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As can be seen in Table 1, the mean benzene concentrations in carbonated cola, orange and
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lemonade flavored beverage were 2.11±0.48, 4.20±1.78 and 4.25±1.70 µg/L, respectively.
Benzene levels of two out of 16 carbonated beverage samples were higher than the maximum
permissible content in water by the WHO (10 µg/L). Cola flavored beverage had the lower
benzene level followed by orange flavored and lemonade flavored beverage. Although ascorbic
acid was not detected in any of the samples (Table 2), all samples contained sodium benzoate in
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the range of 44.00-86.02 mg/kg, which was lower than the maximum level established by Iranian
National Standardization Organization (ISIRI) for these products: 150 mg/L (ISIRI, 2016).
The mean concentrations of benzene in orange, peach, pineapple and mango juices were
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6.97±3.61, 3.86±0.48, 8.97±1.79 and 0.89±0.84 µg/L, respectively (Table 1). In general, benzene
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level of fruit juice ranged between <LOD and 11.20 µg/L, with a mean of 5.17±3.63 µg/L. Two
samples (12.5% of all juices) of fruit juice (one sample of orange juice and one sample of
pineapple juice) contained benzene at a higher level than the acceptable limit by WHO (10
µg/L). In Iran, the addition of sodium benzoate into fruit juice has been forbidden. In our study
sodium benzoate was detected in the range of 14.70-64.10 mg/kg in fruit juice samples (Table 2).
The correlation coefficient between benzene and sodium benzoate (0.682) and benzene and
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ascorbic acid (0.503) in fruit juice samples was recognized as positive, strong and significant
(Table 3).
Based on previous investigations, the nonalcoholic beverages contained the highest benzene
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concentrations (<0.01-23 µg/L) among the food products such as cheese, eggs, alcoholic
beverages, fish, and fruits (Salviano dos Santos et al., 2015). The investigated occurrence of
benzene in food and beverage samples in several countries is summarized in Table 4.
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Techakriengkrai and Lertborwornwong (2013) reported that 13 (27.08%) out of 48 Thai
commercial non-alcoholic beverages contained benzene, the range from 5.47 to 16.91 µg/L, and
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4 samples (8.33%) were above the WHO drinking acceptable water limit of 10 µg/L (10.2216.91 µg/L). The significant positive correlation between benzene concentrations and the level of
iron and copper in the soft drink was reported by Lachenmeier et al. (2010).
All pickle samples contained benzene lower than the maximum limit accepted by WHO (10
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µg/L). However, 2 (40%) out of 5 pickled vegetable mixture and 4 (80%) out of liteh (mixture of
vegetables including eggplant, celery, basil, tarragon, black caraway, pepper, garlic, salt and
vinegar that are grinded and pickled) samples contained more benzene than the allowable limit
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established by USEPA (5 µg/L). The mean concentrations of sodium benzoate and ascorbic acid
in a pickle, both occurred naturally in pickles, were 39.63±31.26 and 7.95±2.74 mg/kg,
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respectively. In several countries, food additives (including sodium benzoate and other benzoate
salts) are incorporated into pickles to extend their shelf-life. However, these additives have been
being forbidden in Iran for providing the goals as mentioned earlier in pickle production (ISIRI,
2016).
During storage of fermented vegetables, sodium benzoate concentration was constant or
might be partly reduced as reported by Casado et al. (2011). These authors also argued that
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ascorbic acid content may be decreased or decomposed due to packaging conditions, vegetable
matrix and rate of lactic acid bacteria growth. Also, antioxidants such as polyphenols might
reduce the rate of benzene formation in fermented vegetables due to their hydroxyl and peroxyl
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radical scavenging activities (Casado et al., 2011). According to Kharat et al. (2016), 11 out of
24 Indian pickle samples contained benzene, and the highest value was related to lime pickle
(4.36 µg/L). This sample also had the highest level of benzoic acid and the lowest hydroxyl
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scavenging activity. Although the authors observed that pickles contained higher levels of
benzene level was reported.
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ascorbic and benzoic acid and pro-oxidant components such as copper and iron, low or no
All mayonnaise and salad dressing samples had benzene content below 10 mg/kg, with a
mean of 1.38 and 1.47 µg/L, respectively. In this study, the highest concentration of sodium
benzoate concentration was found in mayonnaise (479.38±127.35 mg/kg), followed by salad
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dressing (446.53±97.33 mg/kg). The use of preservatives is a conventional approach to inhibit
microbial growth in sauces (Depree & Savage 2001). According to Institute of Standards &
Industrial Research of Iran, the addition of sodium benzoate (maximum permissible
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concentration: 750 mg/kg) into mayonnaise and salad dressing is allowed (ISIRI 2014). Our
results showed a low level of ascorbic acid in mayonnaise (1.67±0.71 mg/kg) and salad dressing
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(1.32±0.61 mg/kg).
Despite the high content of sodium benzoate in mayonnaise and salad dressing, no significant
correlation between this preservative and benzene concentration was observed (Table 3). The
information about the occurrence of benzene in mayonnaise and salad dressing samples remained
deficient. However, our findings were in close agreement with a previous study conducted by
Vinci et al. (2012) who reported benzene contamination of sauce samples collected in Belgian
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market in a range of 1.71-6.35 µg/L (average of 1.18 µg/L). It seems that the formation of
benzene in food and drink products assessed in the current study had various mechanisms and
pathways beside the reaction through sodium benzoate and ascorbic acid.
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All lime juice samples contained benzene levels lower than 10 µg/L. Although lime juice
naturally contained sodium benzoate and ascorbic acid, no significant correlation among
concentrations of both (sodium benzoate and ascorbic acid, as well as benzene levels, was
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observed (Table 3). In this regard, lime fruit is rich in ascorbic acid, although the concentration
of this vitamin depends on various pre-harvest factors including genotype, climatic conditions,
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cultural practices, fruit maturity and postharvest management (Magwaza et al., 2017).
To make a detailed judgment about the health risk of consuming benzene through food and
beverages, we have estimated DDE and MOE in the age groups of males and females (Table 5).
In general, an MOE of 10,000 or higher, when according to BMDL from an animal study, can be
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considered a low carcinogenic risk (EFSA 2005).
The range of DDE values of benzene in food and beverage for 15-24 age groups urban males
were from 1.74E-07 to 5.93E-06 mg/kg bw-d; 25-64 age groups urban males 1.59E-07 to 5.43E-
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06 mg/kg bw-d; 15-24 age groups urban females 2.05E-07 to 6.97E-06 mg/kg bw-d; and 25-64
age groups urban females 1.72E-07 to 5.87E-06 mg/kg bw-d (Table 5). The lowest and highest
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DDE in all age groups consumers was related to dressing salad and soft drink, respectively
(Table 5).
The range of DDE values of benzene in food and beverage for 15-24 age groups rural males
were from 1.90E-07 to 6.46E-06 mg/kg bw-d; 25-64 age groups rural males 1.72E-07 to 5.86E06 mg/kg bw-d; 15-24 age groups rural females 2.15E-07 to 7.34E-06 mg/kg bw-d; and 25-64
age groups rural females 1.86E-07 to 6.33E-06 mg/kg bw-d (Table 5). Also in the rural
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consumers, lowest and highest DDE in all age groups consumers was related to dressing salad
and soft drink, respectively (Table 5).
In addition, benzene intake through assessed food and beverage was lower than the minimum
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risk value for chronic oral exposure to this contaminant (0.0005 mg/kg bw/day) established by
US Public Health Service Agency for Toxic Substances and Disease Registry (ATSDR 2007)
benzene (0.0005 mg/kg bw/day) (ATSDR 2007).
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DDE in all age groups urban and rural consumers were lower than safe limit of DDE of the
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The range of MOE values of benzene in food and beverage for 15-24 age groups urban males
were from 202, 000 to 68 900,,000; 25-64 age groups urban males 221, 000 to 7,530,000; 15-24
age groups urban females 172,000 to 5,860,000; and 25-64 age groups urban females 204 000 to
6,960,000 (Table 5). The lowest and highest MOE in all age groups consumers was related to
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dressing salad and soft drink, respectively.
The range of MOE values of benzene in food and beverage for 15-24 age groups rural males
were from 186,000to 6,320, 000; 25-64 age groups rural males 205,000 to 6,980,000; 15-24 age
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groups rural females 164,000to 5,570,000; and 25-64 age groups rural females 189,000 to
6,450,000 (Table 5). Also in the rural consumers, lowest and highest MOE in all age groups
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consumers was related to soft drink and dressing salad, respectively.
The rank order for the risk of food and beverage based on MOE in the urban and rural
consumers was dressing salad > mayonnaise sauce > lime juice > pickle > fruit juice > soft drink
(Figure 1).
MOE in the 25-64 age groups males in urban and rural were higher than 15-24 age groups.
Therefore, there are in the lower risk of benzene than 15-24 age groups (Figure 1). MOE for all
age groups in the urban and rural were higher than 10,000 (Figure 1). Therefore, consumers are
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in the safe range induced by benzene intake from beverage and food ingestion. Since males have
higher BW than females, their MOE is higher than females. Therefore, males are at a lower risk
of benzene carcinogenicity. On the other hand, rural consumers have lower body weight.
and beverage were considered equal in urban and rural areas.
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Therefore, MOE in the rural was lower than urban consumers. In this study, consumption of food
Therefore, to carry out a more accurate risk assessment between rural and urban consumers, it
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is necessary to determine the per capita consumption of food and beverage separately.
The current study findings were in agreement with previous results reported by Vinci et al.
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(2012) who observed MOE values for benzene ranged from 140,000 to 880,000 in the market
foods. Similarly, Lachenmeier et al. (2010) noted that MOE mean value for benzene in infant
carrot juice was in the range of 320,000 to 130,000,000. On the other hand, the utilization of
alternative food preservative instead of sodium benzoate could reduce benzene levels as
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suggested by many authors (Lachenmeier et al., 2010; Morsi et al., 2012; Salviano dos Santos et
Conclusions
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al., 2015).
This study is the first report evaluating the benzene contamination in food and drink products
marketed in Hamadan (Iran). Benzene concentration in 12.5% of carbonated soft drink and fruit
juice samples (two samples from each), exceeded the maximum tolerable limit (10 µg/L)
established by WHO for drinking water. Moreover, in fruit juice and non-alcohol beverage
samples, a significant correlation coefficient among benzene, sodium benzoate, and ascorbic acid
was observed. MOE values indicated there is not a health threat for benzene intake. In all age
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groups of females, higher health risk due to consumption the foods contain benzene were
reported. The constant monitoring of available food products in the market as well as calculating
people's cumulative benzene exposure, besides the establishment of related legal limitations in
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food and beverages, is strongly recommended. Also, products containing high benzene levels
Conflict of interest
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Authors declare that they have no conflict of interest.
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should be reformulated, mainly to replace sodium benzoate by other food preservatives.
Acknowledgments
This study was supported by Hamadan University of Medical Science, Hamadan, (Iran). Amin
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Mousavi Khaneghah gratefully acknowledges the support of CNPq-TWAS Postgraduate
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Table 1. Average values and ranges of benzene in food and beverages consumed in Iran.
Range of concentration N(%)b
Food subcategory
Na
Carbonated beverage
Cola - flavored
1.76-2.64
2.11±0.48
1 (20%)
2.64-4.20
4.20±1.78
4 (66.7%)
0
1.76-6.16
4.25±1.70
6 (37.5%)
2 (12.5%)
1.76-6.6
3.57±1.70
>10 µg/L
5
0
4 (80%)b
0
Orange- flavored
5
0
2 (40%)
2 (40%)
Lemonade - flavored
6
0
2 (33.3%)
Total Carbonated beverage
16
0
8 (50%)
Orange
4
0
Peach
4
0
Pineapple
4
0
Total Fruit juice
16
Pickled vegetable mixture
5
Pickled cucumber
Total Pickle
Lime juice
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4
3 (75%)
0
3 (75%)
1(25%)
2.64-11.20
6.97±3.61
4 (100%)
0
0
3.17-4.30
3.86±0.48
0
3 (75%)
1 (25%)
6.30-10.10
1 (25%)
8.97±1.79
c
0.89±0.84
0
0
<LOD-2.00
3 (18.8%) 5 (31.2%)
6 (37.5%)
2 (12.5%)
<LOD-11.20
5.17±3.63
0
3 (60%)
2 (40%)
0
3.52-8.81
4.93±2.29
5
0
1 (20%)
4 (80%)
0
3.52-8.81
5.98±2.01
6
0
6 (100%)
0
0
1.76-3.76
2.55±0.89
16
0
10 (62.5%) 6 (37.5%)
0
1.76-8.81
4.37±2.24
16
0
9 (56.2%)
0
4.04-5.84
4.99±0.54
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Mango
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5-10 µg/L
Liteh
Lime juice
1 (20%)
1-5 µg/L
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Pickle
Meand ±SD (µg/L)
>1 µg/L
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Fruit juice
Range (µg/L)
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Food category
7 (43.8%)
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Mayonnaise
16
5 (31.2%) 11 (68.8%) 0
0
0-2.92
1.38±0.87
Salad dressing
Salad dressing
16
3 (18.8%) 13 (81.2%) 0
0
<LOD-2.88
1.47±0.83
a
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Mayonnaise
N: number of samples; bN(%): number of samples in the range of concentration (% of samples in the range of concentration); cLOD: limit of detection,
mean o all samples.
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Ascorbic acid (mg/kg)
Mean ±SD
Range
Mean±SD
Range
56.01±14.54
44.00-77.01
0
0
5
61.24±8.68
44.03-72.01
0
0
Lemonade - flavored
6
77.50±8.16
65.90-86.02
0
0
Total Carbonated beverage
16
66.88±13.19
44.00 -86.02
0
0
Orange
4
56.52±9.90
42.00-64.10
135.66±23.77
100.80-153.84
Peach
4
27.17±20.61
14.70-58.02
40.76±30.92
22.05-87.01
Pineapple
4
49.36±12.78
30.40-58.20
98.72±25.57
60.80-116.40
Carbonated beverage
Cola - flavored
5
Orange- flavored
4
16.95±1.25
16.01-18.70
67.80±5.01
64.01-74.80
16
37.50±20.31
14.70-64.10
85.73±42.12
22.05-153.84
5
14.12±7.86
6.70-23.10
4.72±0.92
3.90-6.00
5
20.44±12.84
7.20-36.10
7.74±1.24
6.30-9.60
Pickled cucumber
6
76.89±8.17
65.40-85.20
10.81±0.56
10.10-11.40
Total Pickle
6
39.63±31.26
6.70-85.2
7.94±2.74
3.90-11.40
16
70.48±13.43
47.60-96.30
15.52±3.32
10.57-21.13
Total Fruit juice
Pickle
Mix pickled vegetable
Lime juice
Lime juice
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Liteh
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Mango
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Na
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Food subcategory
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Sodium benzoate (mg/kg)
Food category
Fruit juice
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Table 2. Average values and ranges of sodium benzoate and ascorbic acid in food and beverages consumed in Iran.
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Mayonnaise
16
479.38±127.35
330.01-660.02
1.67±0.71
0.88-3.96
Salad dressing
Salad dressing
16
446.53 ±97.33
330.00-660.00
1.32±0.61
0.38-2.01
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N: number of samples;
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Mayonnaise
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Table 3. Correlation coefficient between benzene value and sodium benzoate, and between benzene value and ascorbic acid concentration.
Correlation of benzene with
Ascorbic acid
Correlation coefficient
0
0.503
-0.377
0.402
-0.262
-0.131
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P-value
0.003
0.004
0.053
0.149
0.729
0.916
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Carbonated beverage
Fruit juice
Pickles
Lime juice
Mayonnaise
Salad dressing
Sodium benzoate
Correlation coefficient
0.697
0.682
-0.491
0.378
0.094
-0.029
AC
C
Food category
P-value
0
0.047
0.150
0.123
0.326
0.628
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Table 4. Contamination incidence of benzene in food products in others countries
Food
N
Benzene concentration ( µg/ L)
3
Carbonated soda beverages
199
Non-alcoholic and alcoholic
beverages
Carbonated soda beverages
124
118
124
Thailand
China
Techakriengkrai and Lertborwornwong
2013
Yang et al. 2010
77% samples <1
9% samples >5
47 samples <1
20 samples >1.1–17.6
1.1–3.67
USA
Nyman et al. 2007
Canada
Cao et al. 2007
Italy
Fabietti et al. 2001
Canada
Cao et al. 2007
Brazil
Arisseto et al. 2013
Romania
Ibolya et al. 2012
Egypt
Morsi et al. 2012
Brazil
Sanchez et al. 2012
4.36±0.82
India
Kharat et al. 2016
green apple(0.70 – 40.14 ng/mL)
orange color carbonated soft drink (0.9 – 25 ng/mL)
Lemon (0.77 – 33.45 ng/mL)
Egypt
Mor et al. 2012
77
4 samples >5
2 samples >10
1 sample = 23
<0.08 - 10.84
15
Lemon (0.67)
75
Orange (3.25)
green apple (0.70 – 40.14)
orange color carbonated soft drink (0.9 – 25)
Lemon (0.77 – 33.45)
9
Cola (0.8±0.2 µg/L )
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Beverages
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Cola (1.55)
Soft drink beverage
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Alcoholic beverages
27.08% samples ranged 5.47–16.91 (where 8.33%
samples >10)
2.4 in one sample
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Reference
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Non- alcoholic beverages
Country
AC
C
Lemon juice (0.8±0.3)
Citric fruit juice(<LOD)
Pickles
24
Soft drinks
75
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Rural
15-24
DDE
MOE
(mg/kg-d)
25-64
DDE
MOE
(mg/kg-d)
Females
15-24
DDE
MOE
(mg/kg-d)
25-64
DDE
MOE
(mg/kg-d)
221 000
6.97E-06
172 000
5.87E-06
204 000
1.74E-06
691 000
2.23E-06
538 000
1.88E-06
639 000
1 270 000
8.67E-07
1 380 000
1.11E-06
1 080 000
9.38E-07
1 280 000
2 690 000
4.09E-07
2 930 000
5.25E-07
2 280 000
4.43E-07
2 710 000
6 340 000
1.73E-07
6 920 000
2.23E-07
5 390 000
1.88E-07
6 400 000
1.74E-07
6 890 000
1.59E-07
7 530 000
2.05E-07
5 860 000
1.72E-07
6 960 000
6.46E-06
186 000
5.86E-06
205 000
7.34E-06
164 000
6.33E-06
189 000
2.07E-06
581 000
1.87E-06
640 000
2.35E-06
512 000
2.03E-06
592 000
1.03E-06
1 160 000
9.35E-07
1 280 000
1.17E-06
1 020 000
1.01E-06
1 190 000
Concentration
Benzene
(µg/L)
3.57
115
5.93E-06
202 000
5.43E-06
Fruit juice
5.47
24
1.90E-06
633 000
Pickle
4.37
15
9.47E-07
Lime juice
4.99
6.2
4.47E-07
Mayonnaise
sauce
Dressing
salad
Soft drink
1.38
9.5
1.89E-07
1.47
8.2
3.57
115
Fruit juice
5.47
24
Pickle
4.37
15
EP
TE
D
Food or
beverage
category
Soft drink
AC
C
Urban
Daily Intake
(g/d)
M
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Males
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Table 5. Risk characterization of exposure of urban and rural consumers of Iran to benzene through food and beverage
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4.99
6.2
4.87E-07
2 460 000
4.42E-07
2 720 000
5.53E-07
2 170 000
4.77E-07
2 510 000
Mayonnaise
sauce
Dressing
salad
1.38
9.5
2.06E-07
5 810 000
1.87E-07
6 410 000
2.34E-07
5 120 000
2.02E-07
5 930 000
1.47
8.2
1.90E-07
6 320 000
1.72E-07
6 980 000
5 570 000
1.86E-07
6 450 000
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C
EP
TE
D
M
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SC
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Lime juice
2.15E-07
AC
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Highlights
Benzene mean value in food samples was below the acceptable limit
•
Benzene-sodium benzoate-ascorbic acid (AA) relationship in food was investigated
•
Significant correlation between benzene, sodium benzoate, and AA in fruit juices
•
Dietary exposure to benzene could not threat consumer health in Iran
•
It is suggested setting legal limitation for benzene in food and beverage.
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The authors do not declare any conflicts of interest.