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Journal of Toxicology and Environmental Health, Part A: Current Issues Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/uteh20

Determination of Polycyclic Aromatic Hydrocarbon (PAH) Content and Risk Assessment From Edible Oils in Korea a

b

Bomi Kang , Byung-Mu Lee & Han-Seung Shin

a

a

Department of Food Science and Biotechnology and Institute of Lotus Functional Foods Ingredients, Dongguk University–Seoul, Seoul, Korea b

Division of Toxicology, College of Pharmacy, Sungkyunkwan University, Suwon, Gyeonggi-do, Korea Published online: 24 Oct 2014.

To cite this article: Bomi Kang, Byung-Mu Lee & Han-Seung Shin (2014) Determination of Polycyclic Aromatic Hydrocarbon (PAH) Content and Risk Assessment From Edible Oils in Korea, Journal of Toxicology and Environmental Health, Part A: Current Issues, 77:22-24, 1359-1371, DOI: 10.1080/15287394.2014.951593 To link to this article: http://dx.doi.org/10.1080/15287394.2014.951593

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Journal of Toxicology and Environmental Health, Part A, 77:1359–1371, 2014 Copyright © Taylor & Francis Group, LLC ISSN: 1528-7394 print / 1087-2620 online DOI: 10.1080/15287394.2014.951593

DETERMINATION OF POLYCYCLIC AROMATIC HYDROCARBON (PAH) CONTENT AND RISK ASSESSMENT FROM EDIBLE OILS IN KOREA Bomi Kang1, Byung-Mu Lee2, Han-Seung Shin1 1

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Department of Food Science and Biotechnology and Institute of Lotus Functional Foods Ingredients, Dongguk University–Seoul, Seoul, Korea 2 Division of Toxicology, College of Pharmacy, Sungkyunkwan University, Suwon, Gyeonggi-do, Korea Polycyclic aromatic hydrocarbons (PAH) content and a risk assessment from consumption of Korean edible oils were investigated. Liquid–liquid extraction and gas chromatography–mass spectroscopy were used to measure eight PAH in edible oils commonly consumed in Korea. The total average PAH concentration was 0.548 µg/kg from edible oils and the content of the 8 PAH was lower than 2 µg/kg, which is the maximum tolerable limit reported by the commission regulation. The contents of the eight PAH were converted to exposure assessment and risk characterization values. Dietary exposure to PAH from edible oils was 0.025 ng-TEQBaP /kg/d, and margin of exposure (MOE) was 4 × 106 , which represents negligible concern. Although PAH were detected from edible oils in Korea, their contribution to human exposure to PAH is considered not significant.

are reliable toxicity and occurrence indicators of PAH in oils have been confirmed. The International Agency of Research on Cancer (IARC) also classified PAH based on toxicity. IARC classifies known carcinogens in humans (i.e., Group 1) and a number of PAH are considered to be probable (Group 2) or possible (Group 2B) carcinogens to humans. Benzo[a]pyrene (BaP) was reclassified from Group 2A (probably carcinogenic to humans) to Group 1 (carcinogenic to humans), and chrysene (CRY) was reclassified from Group 3 (not classifiable in humans) to Group 2B (possibly carcinogenic to humans). Benzo[a]anthracene (BaA) was reclassified from Group 2A to Group 2B. Figure 1 lists the eight PAH used in this study, along with their classes defined by IARC and their chemical properties (IARC, 2010). PAH formation results from incomplete combustion or pyrolysis of organic matter

Polycyclic aromatic hydrocarbons (PAH) include more than 100 different compounds that have two or more fused aromatic rings (Guillen et al., 1994). PAH are environmental pollutants present in air, water, soil, and sediments. Due to their lipophilic nature, oil and fats are a principle contamination source of PAH. PAHs are mostly formed by incomplete combustion of organic matter as a consequence of a series of natural and anthropogenic processes (Kitts et al., 2012; Artur et al., 2013). Sixteen priority PAH were defined by the European Food Safety Authority (EFSA). More than 16 PAH, which are potentially mutagenic and carcinogenic to humans, are formed at parts per billion (ppb) levels during cooking under conditions of high temperature (EFSA, 2008). PAH are classified as large organic compounds that are present as pollutants in edible oils. Eight priority PAH and four PAH that

Address correspondence to Han-Seung Shin, Department of Food Science and Biotechnology, Dongguk University–Seoul, 26, 3-Ga, Pil-dong, Chung-gu, Seoul, 100-715, Korea. E-mail: [email protected] 1359

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FIGURE 1. Chemical structures of 8 PAH (including BaA, benzo[a]anthracene; CRY, chrysene; BbF, benzo[b]fluoranthene; BkF, benzo[k]fluoranthene; BaP, benzo[a]pyrene; DahA, dibenzo[a,h]anthracene; BghiP, benzo[g,h,i]perylene; IcdP, indeno[1,2,3-cd]pyrene) found in edible oils.

(Chang et al., 2006), and PAH act as carcinogens, mutagens, and genotoxins by binding to DNA (CCFAC, 2005; EFSA, 2008; Rojas et al., 2004). Edible oils may contain PAH and thus be highly contaminated (Dennis et al., 1991). In terms of variable conditions, a number of parameters are assessed during the technological processing of edible oils, such as heating time and temperature, fuel used, distance from the heat source, and impact of smoke on the product. In accordance with PAH in edible oils, some countries (Spain, Italy, Portugal, and Greece) have established limits for the concentration of the following eight heavy PAH: benzo[a]anthracene, chrysene, benzo[b] fluoranthene, benzo[k]fluoranthene, benzo[a] pyrene, dibenz[a,h]anthracene, benzo[ghi] perylene, and indeno[1,2,3-cd]pyrene. A maximum limit value of 2 ppb for each single PAH and 10 ppb for the sum of 8 heavy PAH was established in edible oils. Several manufacturing processes generate PAH that can contaminate edible oils such as olive, soybean, cottonseed, sesame, fruit, or seeds of plants. Further, PAH generated by environmental polluting of raw vegetables during processing also contaminates edible oils via air, soil, and water (Lawrence and Weber, 1984). Fats typically undergo washing, drying, and roasting, distribution, milking, bottling, and packaging processes, and PAH may be generated from roasting and heat-treatment processes, such as evaporation and drying. Therefore, carcinogenic and genotoxic PAH need to be monitored

in edible oils in the Korean market. (D’Mello et al., 2003). A limited number of studies examined the usefulness of early biological markers of effect of genotoxic compounds such as PAH (Lee et al., 2002). The aim of this study was to measure the content of genotoxic and carcinogenic PAH including benzo[a]anthracene (BaA), chrysene (CRY), benzo[b]fluoranthene (BbF), benzo[k]fluoranthene (BkF), benzo[a] pyrene (BaP), dibenzo[a,h]anthracene (DahA), benzo[g,h,i]perylene (BghiP), and indeno[1,2, 3-cd]pyrene (IcdP) in edible oils in the Korean market and to perform a public health risk assessment.

MATERIALS AND METHODS Reagents and Materials N,N-Dimethylformamide (N,N-DMF), acetonitrile, methanol, ethanol, n-hexane, and dichloromethane were obtained for gas chromatography–mass spectrophotometry (GC-MS) analyses and purchased from Burdick & Jackson (Muskegon, MI). Water was acquired from a Milli-Q water purification system (Billerica, MA). Eight PAHs, namely, BaA (benzo[a]anthracene), CRY (chrysene), BbF (benzo[b]fluoranthene), BkF (benzo[k]fluoranthene), BaP (benzo[a]pyrene), DahA (dibenzo[a,h]anthracene), BghiP (benzo[g,h,i]perylene), and IcdP (indeno[1,2,3c,d]pyrene), were used as standards and benzo[a]pyrene-d12 was used as an internal

DETERMINATION OF POLYCYCLIC AROMATIC HYDROCARBONS

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standard. All of these PAH were purchased from Supelco (Bellefonte, PA). Sodium sulfate (Na2 SO4 ) for dehydration was purchased from Junsei (Chuo-ku, Tokyo, Japan). Sep-Pak silica cartridges, supplied by Waters (Milford, MA), were used as solid-phase extraction (SPE) columns for purification. Polytetrafluoroethylene (PTFE) membrane filters (0.45 µm) were purchased from Macherey-Nagel (Düren, NRW, Germany).

Sample Preparation for Analysis In total, 201 edible oils including 60 olive oil, 5 sesame oil, 19 soybean oil, 8 corn oil, 21 canola oil, 57 grapeseed oil, 19 sunflower oil, and 12 rich bran oil samples were analyzed in this study. All samples were purchased at Korean markets and stored at 5◦ C. Sample preparation procedures were modified according to Rey-Salgueiro et al. (2009) and Kishikawa et al. (2003). A 10-g sample was weighed, deposited in a separatory funnel, and spiked with 1 ml 100 µg/kg benzo[a]pyrened12 . Then 50 ml of N,N-DMF:H2 O (9:1, v/v) and n-hexane were added, and the mixture was shaken. The –-hexane layer solution was extracted twice with 25 ml N,N-DMF:H2 O (9:1, v/v) by shaking and purifying. A 100-ml aliquot of sodium sulfate solution and 50 ml nhexane were added to the N,N-DMF:H2 O (9:1, v/v) layer and shaken, and the n-hexane layer was transferred to another separatory funnel. Two 35-ml aliquots of n-hexane were added and then two 40-ml aliquots of deionized H2 O were added. The resulting solution was dehydrated with anhydrous sodium sulfate (15 g), and concentrated under reduced pressure on a rotary evaporator at 35◦ C to a final volume of 2 ml. The cleaned-up samples were preconditioned with 10 ml dichloromethane and 20 ml n-hexane with an a SPE cartridge activated in advance, and eluted with 5 ml n-hexane and 15 ml n-hexane–dichloromethane (3:1) mixture. The solution was concentrated to near dryness under a gentle stream of nitrogen gas at 37◦ C. The dried residue was redissolved in 1 ml

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dichloromethane and passed through a 0.45µm PTFE membrane filter. An aliquot of 20 µl of this solution was injected into the GC-MS system. Standard Preparation A mixture of 8 PAH priority pollutants from Supelco (Bellefonte, PA) was used for the identification and quantification of the PAH present in the samples and included benzo[a] anthracene (BaA) 50.2 µg/ml, chrysene (CRY) 50.2 µg/ml, benzo[b]fluoranthene (BbF) 20 µg/ml, benzo[k]fluoranthene (BkF) 20 µg/ml, benzo[a]pyrene (BaP) 50.2 µg/ml, dibenzo[a,h] anthracene (DahA) 200.4 µg/ml, benzo[g,h,i] perylene (BghiP) 80.2 µg/ml, and indeno[1, 2,3-cd]pyrene (IcdP) 50.2 µg/ml. This standard mixture was stored at –10◦ C in darkness to avoid photodegradation and volatilization. Stock solutions containing 20 µg/L were prepared by dilution of this standard mix in methylene chloride and stored at –10◦ C in darkness. GC-MS Analysis of PAHs PAH analysis was carried out using an Agilent Technologies 6890N/5975 MSD GCMS apparatus (Santa Clara, CA). Separation of PAH was achieved using an HP-5MS column (30 m × 0.25 mm, ID particle size 0.25 µm) with a gradient elution program. The GC-MS conditions for each PAH are listed in Table 1. PAH analysis was conducted in the selective ion monitoring (SIM) mode to check for the absence of residual contamination. In the SIM mode, the MS gathers data for masses of interest, rather than looking for all masses over a wide range. Because the instrument is set to look for only masses of interest it can be specific for a particular analyte of interest. Typically, two to four ions are monitored per compound, and the ratios of those ions will be unique to the analyte of interest. The BaA and CRY ions were 228, 226, 229 ions, and the quantitative analysis target ion was the 228 ion. BbF, BkF, and BaP were 252, 253, 250 ions, and the quantitative analysis target ion was the 252 ion.

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TABLE 1. Analytical Gas Chromatography–Mass Spectroscopy Analysis Conditions for the Polycyclic Aromatic Hydrocarbons

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Instrument Column Injection type and temperature column temperature program Carrier gas Injection volume MS source temperature/MS quadrupole temperature MS mode

DahA was the 278, 279, 276 ions, and the quantitative analysis target ion was the 278 ion. BghiP and IcdP were the 276, 277, 274 ions, and the quantitative analysis target ion was the 276 ion. Identification and Quantification of PAHs The identification of PAH was shown by comparison of the standards’ retention time and their retention time in the same conditions. Spiking samples of extracts with the standards also confirmed peak identity. A mixed standard stock solution was prepared in methylene chloride from this solution, and 5 working ones, ranging from 1 to 20 µg/L, were used to construct linear regression lines (peak area ratios vs. PAH concentration). Validation Study Calibration curves were obtained using a series of standard solutions containing eight PAH and internal standard at five concentrations. Calibration and validation of 8 PAH were carried out using the internal standard method and a standard mixture of 8 PAH at concentrations of 1, 2, 5, 10, and 20 µg/kg with 100 µg/kg benzo[a]pyrene-d12 . All standard mixtures were injected at 1 µl in triplicate to construct calibration curves. Recovery was assessed by spiking the samples with 100 µg/kg of benzo[a]pyrene-d12 as an internal standard. Data are presented as ranges and mean ± standard deviation. Precision (%) and accuracy (%) were evaluated by repeating the runs three times per day (intraday) and at intervals of 3 d

Agilent Technologies 689ON/5975 MSD HP-5MS 30 m 0.25 mm ID 0.25 µm Splitless, 270 (2 min) → 300 (10/min, hold 10 min) → 310 (10 min, post run) Helium (1.0 ml/min) 1.0 µl 230/150 SIM

(intraday). Precision (%) was expressed as the coefficient of variation. Exposure and Risk Assessment Human exposure to PAH was calculated based on benzo[a]pyrene-d12 concentrations measured in edible oils. This study was made analyzing the exposure of population living in each studied data. Populations were subgrouped by age, sorted by year: 1–2; 3–5; 6–11; 12–18; 19–29; 30–49; 50–64; and above 65. Human exposure was considered for eating foods containing edible oil. Pathways through ingestion of foods and also breastfeeding were not considered due to lack of data for the study area and thus not contributing significantly to the differences in the exposure assessment. The risk assessment is based on following the U.S. Environmental Protection Agency (EPA) guidelines (U.S.EPA, 1993, 2012). This study used toxic equivalent factors (TEF) proposed by Nisbet and Lagoy (1992) to calculated the TEQBaP : TEQBaP =

  Ci × TEFi

(1)

where Ci is PAH congener (i) concentration in samples and TEFi is the BaP relative potency value published for each individual PAH. TEF values were reported by the U.S. Environmental Protection Agency (U.S.EPA, 1984, 1993; Chu and Chen, 1984; Clement Associates, 1988; Nisbet and Lagoy, 1992). To estimate the exposure dose/chronic daily intake (CDI) rates using the BaP equivalent concentration, separate calculations for Korea children and adult exposure to the dominant food products consumed were

DETERMINATION OF POLYCYCLIC AROMATIC HYDROCARBONS

performed following the applicable guidelines using Eq. (2). The following section provides an overview for the PAH risk assessment.

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CDI (ng-TEQBaP/kg/d) =

 Ci × IRi × ED BW × AT

(2)

In Eq. (2), CDI is chronic daily intake (ngTEQBaP/kg/d), Ci is the concentration of PAH in the oils (µg-TEQBaP /kg), IRi is the average daily intake of oil (ingestion rate, [g/d]), BW is the average body weight (kg) at each age (Ministry of Health and Welfare [MHW], 2009), ED is exposure duration (yr), and AT is average exposure time (average time = 80.5 yr; National Statistical Office [NSO], 2009). Margin of exposure (MOE) is calculated on the rate of benchmark dose lower confidence limit and chronic daily intake: MOE =

BMDL CDI

(3)

BMDL is the benchmark dose lower confidence limit (mg/kg bw/d) and CDI is the chronic daily intake (mg/kg bw/d). High MOE values such as those >100 for a no-observed-adverseeffect level (NOAEL)-based MOE or 1000 for a lowest-observed-adverse-effect level (LOAEL)based MOE suggest a low level of concern. As the MOE decreases, the level of concern increases. As with the hazard quotient, it is important to remember that the MOE is not a probabilistic statement of risk. The MOE is the ratio of a NOAEL or LOAEL (U.S.EPA, 2012). The toxicity of individual PAH was assessed by the toxic equivalency factor (TEF) based on the carcinogenicity of BaP. TEF values used in this study were proposed by Nisbet and Lagoy (1992), and the concentration of each of the PAH in the edible oils and TEQBaP values were calculated. BaA and CRY were 0.1, BbF and BkF were 0.1, BaP was 1, DghA was 5, BghiP was 0.01, and IcdP was 0.1. Chronic daily intake (CDI) of BaP from all ages assessed cancer risk due to dietary exposure to BaP and slope factor of BaP (for oral exposure 7.3E +

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0 (mg/kg/d)−1 ) (U.S.EPA, 1993; Lee and Shim, 2007). Statistical Analysis All experiments were performed in triplicate and data are presented as mean and standard deviation; p < .05 was considered significant.

RESULTS Analysis of PAH by GC-MS The linear calibration curves were prepared using five different concentrations (1, 2, 5, 10, and 20 µg/kg) of standard mixtures of PAH. The calibration curve correlated the chromatogram peak area to the concentration expressed as the squared correlation coefficient of determination (R2 ). The linearity of the 8 PAH was exemplified by correlation coefficients of 0.9955–0.9999. The limit of detection (LOD) was from 0.1 to 0.12 µg/kg and the limit of quantification (LOQ) was from 0.16 to 0.38 µg/kg. The details of linearity, LOD, and LOQ are shown in Table 2. The recoveries determined by the peak area of benzo[a]pyrene-d12 were from 73.77 to 100% (Table 3). Intraday and interday accuracy was from 97.26 to 103.9%. Interday precision was from 0.16 to 11.93% and intraday precision was from 0.68 to 9.08% (Table 4). In the European Commission (EC) Regulation number 333/2007, the performance criteria for analyzing BaP showed LOD < 0.3 µg/kg and LOQ < 0.9 µg/kg, with recovery rates of 50–120% (EC, 2007). Purcaro et al. (2007) reported the following values for 6 PAH: LOD, 0.1–0.4 µg/kg; LOQ, 0.04–1.4 µg/kg; recovery, >50%; and .988 < R2 < .998, respectively. In another study, Gemma et al. (2003) performed a method validation for analysis of PAH from oil and margarine, and the calculations were based on the original weight. The mean recovery rate for 15 PAH was 80.2–616% (range, 54.5–113%). The detection limit was 0.2 mg/kg except for acenaphthylene, which was 2 mg/kg. Therefore, our method for

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TABLE 2. LOD and LOQ for PAH Based on the GC/MS Analysis PAH

LOD (µg/kg)

LOQ (µg/kg)

Benzo[a]anthracene Chrysene Benzo[b]fluoranthene Benzo[k]fluoranthene Benzo[a]pyrene Dibenzo[g,h]anthracene Benzo[g,h,i]perylene Indeno[1,2,3-c,d]pyrene

0.10 0.11 0.11 0.09 0.05 0.06 0.12 0.11

0.32 0.33 0.33 0.28 0.16 0.20 0.38 0.35

Note. LOD (limits of detection) defined as 3 × standard deviation of the blank, and at the LOQ (limits of quantification) defined as 10 × standard deviation of the blank. GC/MS is gas chromatography–mass spectroscopy.

TABLE 3. Recovery of Each PAH for Analysis PAH

Recovery (%)

Benzo[a]anthracene Chrysene Benzo[b]fluorathene Benzo[k]fluoranthene Benzo[a]pyrene Dibenzo[g,h]anthracene Benzo[g,h,i]perylene Indeno[1,2,3-c,d]pyrene

92.82 ± 2.92 80.63 ± 1.93 91.61 ± 2.54 85.82 ± 2.31 83.03 ± 2.02 80.14 ± 2.73 80.72 ± 2.83 80.71 ± 3.54

Note. Recovery was evaluated with 5 µg/kg spiked concentrations. Values are mean ± standard deviation (SD) (n = 3).

detecting eight PAH from edible oils appeared to be satisfactory compared with results of previous reports. PAH Contents From Edible Oils The GC-MS chromatograms for the standards, spiked samples, and eight PAH from edible oils are shown in Figure 2. The contents of the 8 PAH from edible oils in Korean market and concentrations of 8 PAH from 201 edible oils are presented in Table 5. Content of 8 PAH from edible oils in Korean market averaged 0.55 µg/kg, which was below the EFSA regulation of BaP content < 2 µg/kg. The PAH contamination profiles of corn oils presented a higher contamination levels of eight PAH than those of other edible oils. Seven PAH, excluding BaP, showed the lowest level (below the LOD) among PAH in the edible oils. The contamination levels from edible oils in Korea are shown

in Table 5. Eight PAH content from 201 edible oils in Korea ranged from 8.78 to 9.72 µg/kg, which were close to those noted previously (Hopia et al., 1986; Kolarovic and Traitler, 1982; Speer et al., 1990). Mean concentrations of BaP from olive oils, sesame oils, soybean oils, corn oils, canola oils, grapeseed oils, rice oils, and sunflower oils were 0.67, 0.55, 0.79, 0.74, 0.48, 0.65, 0.41, and 0.65 µg/kg, respectively. Toxic equivalency factors (TEF) based on the carcinogenicity of BaP from edible oils in Korea are presented in Table 6. In olive oils, mean concentration of BaP was 0.67 µgTEQBaP /kg, 4 PAH was 0.67 µg-TEQBaP /kg, and 8 PAH expressed as BaP was 0.67 µgTEQBaP /kg. In sesame oils, mean concentration of BaP was 0.55 µg-TEQBaP /kg, 4 PAH was 0.55 µg-TEQBaP /kg, and 8 PAH expressed as BaP was 0.55 µg-TEQBaP /kg. In soybean oils, mean concentration of BaP was 0.59 µgTEQBaP /kg, 4 PAH was 0.59 µg-TEQBaP /kg, and the 8 PAH expressed as BaP was 0.59 µgTEQBaP /kg. In corn oils, mean concentration of BaP was 0.74 µg-TEQBaP /kg, 4 PAH was 0.74 µg-TEQBaP /kg, and 8 PAH expressed as BaP was 0.74 µg-TEQBaP /kg. In canola oils, mean concentration of BaP was 0.48 µg-TEQBaP /kg, 4 PAH was 0.48 µg-TEQBaP /kg, and 8 PAH expressed as BaP was 0.48 µg-TEQBaP /kg. In grapeseed oils, mean concentration of BaP was 0.65 µg-TEQBaP /kg, 4 PAH was 0.65 µgTEQBaP /kg, and 8 PAH expressed as BaP was 0.65 µg-TEQBaP /kg. In rice oils, mean concentration of BaP was 0.19 µg-TEQBaP /kg, 4 PAH was 0.19 µg-TEQBaP /kg, and 8 PAH expressed as BaP was 0.19 µg-TEQBaP /kg. In sunflower oils, mean concentration of BaP was 0.65 µgTEQBaP /kg, 4 PAH was 0.65 µg-TEQBaP /kg, and 8 PAH expressed as BaP was 0.65 µgTEQBaP /kg. Exposure and Risk Assessment Exposure assessments of eight PAH content from edible oil consumption in Korea were evaluated. In this study, consumption of a variety of edible oils in Korea for ages 1–2 yr, 3–5 yr, 6–11 yr, 12–18 yr, 19–29 yr,

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TABLE 4. Accuracy and Precision for the Determination of PAH Based on the GC/MS Analysis Intraday (n = 3)

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Concentration (µg/kg) BaA 1 2 5 10 20 CRY 1 2 5 10 20 BbF 1 2 5 10 20 BkF 1 2 5 10 20 BaP 1 2 5 10 20 DahA 1 2 5 10 20 BghiP 1 2 5 10 20 IcdP 1 2 5 10 20

Interday (n = 3)

Accuracy (%)

CV (%)

Accuracy (%)

CV (%)

101.49 101.61 99.68 99.82 99.23

1.76 1.59 2.53 1.27 1.52

100.05 98.73 99.59 98.74 100.53

6.33 2.17 4.11 2.89 2.07

103.32 100.53 98.76 100.80 98.06

4.82 8.37 6.01 2.55 2.57

101.14 99.10 99.93 100.79 99.45

9.14 5.34 0.16 2.46 2.52

99.05 102.45 98.76 100.80 98.06

1.09 8.52 6.01 2.55 2.57

101.83 102.32 99.93 100.79 99.45

2.64 7.77 0.16 2.46 2.52

100.93 100.65 99.11 99.93 100.59

3.42 4.15 8.38 1.15 2.75

103.45 98.54 101.47 97.23 100.15

5.94 3.47 6.85 6.68 0.93

99.75 102.57 101.58 100.49 98.62

2.03 6.69 9.76 9.01 1.52

103.87 101.13 98.84 104.32 99.35

5.92 1.30 2.09 3.92 3.24

97.61 98.56 102.37 99.56 97.82

4.92 1.40 4.03 0.68 3.31

102.49 101.56 104.78 99.98 99.37

3.94 4.84 8.36 2.67 0.97

99.73 99.41 97.70 100.11 98.68

1.71 6.35 2.04 1.51 6.40

100.57 98.03 101.58 100.46 99.89

0.86 9.17 4.22 1.20 2.32

102.87 103.90 98.13 97.26 100.34

2.78 8.42 6.82 2.55 2.79

103.26 99.30 100.22 97.46 99.33

5.20 11.93 2.75 3.51 0.58

Note. Accuracy(%) = [1 – (mean concentration measured-concentration spiked)/concentration] × 100. CV (coefficient of variation, %) = (standard deviation /mean) × 100.

30–49 yr, 50–64 yr, and above 65 yr is subdivided by each age group and established by exposure duration, and body weight for each age group is presented. The results

of exposure assessment and risk characterization are shown in Table 7. Lifetime average chronic daily intake of PAH for total of 1–2 age was 0.003 µg-TEQBaP /kg/d and 95th

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FIGURE 2. Total ion chromatogram of 8 PAH (including BaA, benzo[a]anthracene; CRY, chrysene; BbF, benzo[b]fluoranthene; BkF, benzo[k]fluoranthene; BaP, benzo[a]pyrene; DahA, dibenzo[a,h]anthracene; BghiP, benzo[g,h,i]perylene; IcdP, indeno[1,2,3-cd]pyrene) from standard, spiked samples and food samples. TABLE 5. PAH Contents From the Edible Oils in Korea PAHs (µg/kg) Edible oils

BaA

CRY

BbF

BkF

BaP

DahA

BghiP

IcdP

Olive oil Sesame oil Soybean oil Corn oil Canola oil Grapeseed oil Rice oil Sunflower oil

ND1) ND ND ND ND ND ND ND

ND ND ND ND ND ND ND ND

ND ND ND ND ND ND ND ND

ND ND ND ND ND ND ND ND

0.67 ± 0.39 0.55 ± 0.47 0.79 ± 0.42 0.74 ± 0.40 0.48 ± 0.49 0.65 ± 0.41 0.41 ± 0.38 0.65 ± 0.35

ND ND ND ND ND ND ND ND

ND ND ND ND ND ND ND ND

ND ND ND ND ND ND ND ND

Note. ND, not detected. Mean ± standard deviation of benzo[a]pyrene.

percentile of total 1–2 age was 0.02 µgTEQBaP /kg/d. Lifetime average chronic daily intake of PAH for total of 3–5 age was 0.006 µg-TEQBaP /kg/d and lifetime average chronic daily intake for 95th percentile of total 3–5 age was 0.023 µg-TEQBaP /kg/d. Lifetime average chronic daily intake of PAH for total 6–11 age was 0.01 µg-TEQBaP /kg/d and lifetime average chronic daily intake for 95th percentile of total 6–11 age was 0.036 µg-TEQBaP /kg/d. Lifetime average chronic daily intake of PAH for total

12–18 age was 0.009 µg-TEQBaP /kg/d and lifetime average chronic daily intake of 95th percentile for total 13–18 age was 0.032 µgTEQBaP /kg/d. Lifetime average chronic daily intake of PAH for total 19–29 age was 0.013 µg-TEQBaP /kg/d and lifetime average chronic daily intake of 95th percentile for total 19–29 age was 0.049 µg-TEQBaP /kg/d. Lifetime average chronic daily intake of PAH for total of 30–49 age was 0.020 µgTEQBaP /kg/d and lifetime average chronic daily

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TABLE 6. TEQ Values for PAH From Edible Oils in Korea TEQBaP value (µg-TEQBaP /kg)

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Edible oils TEF Olive oil Sesame oil Soybean oil Corn oil Canola oil Grapeseed oil Rice oil Sunflower oil Average

B[a]A

CRY

B[b]F

B[k]F

B[a]P

D[ah]A

B[ghi]P

I[cd]P

0.10

0.01

0.10

0.10

1.00

5.00

0.01

0.10

Total PAH

Sum of 4 PAH

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 0 0 0 0 0 0 0

0.67 0.55 0.59 0.74 0.48 0.65 0.19 0.65 0.55

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

0.67 0.55 0.59 0.74 0.48 0.65 0.19 0.65 0.55

0.67 0.55 0.59 0.74 0.48 0.65 0.19 0.65 0.55

Note. TEQ, toxicity equivalency. Sum of 4 PAH is sum of B[a]A, CRY, B[b]F, B[a]P. TABLE 7. Results of PAH Exposure From Edible Oils in Total Population as Recorded in All Ages Lifetime average daily intake (µg-TEQBaP /kg/d) Age (yr)

Total

95th percentile of total

1–2 3–5 6–11 12–18 19–29 30–49 50–64 >65

0.003 0.006 0.010 0.009 0.013 0.020 0.009 0.006

0.020 0.023 0.036 0.032 0.049 0.085 0.047 0.037

intake for 95th percentile of total 30–49 age was 0.085 µg-TEQBaP /kg/d. Lifetime average chronic daily intake of PAH for total of 50–64 age was 0.009 µg-TEQBaP /kg/d and lifetime average chronic daily intake of 95th percentile for total 50–64 age was 0.047 µg-TEQBaP /kg/d. Lifetime average chronic daily intake of PAH for total of above 65 age was 0.006 µgTEQBaP /kg/d and lifetime average chronic daily intake of 95th percentile for total above 65 age was 0.037 µg-TEQBaP /kg/d. The risk assessment of PAH from consumption of edible oils in Korea was conducted using the margin of exposure (MOE) (WHO, 2007; KFDA, 2011; Committee on Carcinogenicity of Chemicals in Food, Consumer Products, and the Environment [COC], 2006). MOE was calculated from human exposure analysis from eight PAH content of edible oils in Korea. MOE values of BaP from edible oil consumption in

Korea are presented in Table 8. Average exposure of the whole population to BaP and high exposure sum of 95th percentile for edible oils average exposure of the whole population were 0.069 × 10−6 and 0.03 × 10−6 , respectively. MOE of 4 PAH for average exposure of the whole population was 1.37 × 105 and high exposure sum of 95th percentile for edible oils average exposure of the whole population was 1.37 × 104 . MOE of 8 PAH for average exposure of the whole population was 1.29 × 105 and high exposure sum of 95th percentile for edible oils average exposure of the whole population was 1.3 × 104 . Based on the Committee on Carcinogenicity of Chemicals in Food, Consumer Products, and the Environment (COC, 2006) criteria, MOE levels of PAH from edible oil comsumption in Korea were considered a “negligible concern.” Chronic daily intake of PAH from all ages assessed cancer risk due to dietary exposure to BaP and slope factor of BaP [for oral exposure 7.3E + 0 (mg/kg/d)−1 ] (U.S. EPA, 1993).

DISCUSSION The contents of 8 PAH in 201 edible oils were evaluated from Korean markets. The collective examination from edible oils in Korea for BaP from edible oils indicated that it was 50%, and 0.988 < R2 < 0.998, respectively. Gemma et al. (2003) performed a method validation of PAH from edible oil and margarine. The mean recovery rate for 15 PAH ranged from 54.5 to 113%. The detection limit was 0.2 mg/kg except for acenaphthylene, which was 2 mg/kg. The values of the carcinogenic PAH obtained for the markers appeared to fall within the range reported for markers in other edible oils (Guillen et al., 1994; Sagredos et al., 1988; Zougag et al., 2009; Kim et al., 2013). According to the European Commission Regulation (EC) number 835/2011, the maximum level for BaP of foods was found to be 5 µg/kg and the maximum level for the sum of 4 PAH (BaA, CRY, BbF, BaP) of foods 30 µg/kg. PAH concentration detected in this study satisfied BaP and 4 PAH criteria offered by the European Commission Regulation (EC). Martorell et al. (2010) demonstrated that the concentration of BaP was 0.07 µg/kg, 4 PAH was 0.57 µg/kg, 8 PAH was 0.88 µg/kg in foods, and estimated intake concentration of BaP was 0.14 µg/kg, 4 PAH was 1.45 µg/kg, and 8 PAH was 1.71 µg/kg in foods. In foods samples, mean concentration of BaP was 1.34 µg/kg and 8 PAH was 13.11 µg/kg, and mean concentration of BaP was 0.8 µg/kg and 8 PAH was 7.83 µg/kg reported by Alomirah et al. (2011). Martí-Cid et al. (2008) reported that

concentration of BaP was 0.14 µg/kg, 4 PAH was 1.16 µg/kg, 8 PAH was 1.85 µg/kg in foods, and concentration of BaP was 0.13 µg/kg, 4 PAH was 1.47 µg/kg, 8 PAH was 1.88 µg/kg in foods. As a result, changed carcinogenic levels of individual PAH, besides BaP, were presented similarly with the original data due to TEF values. All samples presented similarly with respect to the 8 PAH concentrations except BaP. Bojanowska and Czerwinski (2010) reported that during thermal treatment of rapeseeds the concentration of PAH increased, but levels of BaP did not exceed the threshold permissible levels. This may result in greater exposure of rapeseed to contamination with PAH, but the levels do not appear to be harmful. Kishikawa et al. (2003) reported that the sum of 12 PAH was 0.99–2.01 µg/kg from edible oils. Lifetime average chronic daily intake average BaP concentration was detected as 0.1 ng-TEQBaP /kg/d and total PAH concentration was 0.02 ng-TEQBaP /kg/d in a British diet study. Polycyclic aromatic hydrocarbons from foods on the Irish market noted that 15 species of PAH including BaA, benzo[a]anthracene, DahA, BbF, benzo[j]fluoeanthene, benzo[k] fluoranthene, benzo[g,h,i]perylene, chrysene, cyclopenta[c,d]pyrene, dibenzo[a,e]pyrene, dibenzo[a,h]pyrene, dibenzo[a,i]pyrene, dibenzo[a,l]pyrene, 5-methyl chrysene, and IcdP were detected (Food Safety Authority of Ireland [FSAI], 2006). Food Survey Information Sheets (2006) also carried out a determination of PAH in edible oils in the United Kingdom. BaP concentrations exceeded the maximum permitted level of 2 µg/kg but 64% of the edible oil samples were below the LOD of BaP. PAH contamination levels were 0.75–1.5 µg/kg in edible oils from the Irish market (FSAI,

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2006). According to WHO/JECFA guidelines (Joint FAO/WHO Expert Committee on Food Additives, 2005), fish and shellfish and smoked products are of negligible concern (MOE > 1,000,000) and meat is negligible concern with action minimizing future exposure (MOE > 100,000). The results of MOE for PAH exposure from edible oils in Korea mean negligible concern in this study. The risk characterization was conducted using the MOE (WHO, 2007; KFDA, 2011; COC, 2006). Risk estimation of PAH exposures is a complex issue. PAH in the environment comprise several hundred compounds, most of which occur together with a large number of other carcinogenic pollutants. In addition, individuals are also exposed to sources of PAH other than environmental, such as from tobacco smoking and food and water ingestion, which all increase the uncertainty in cancer risk assessment (Sofia et al., 2014; Dahshan et al., 2013; Lin et al., 2013; Taxell et al., 2013). The exposure assessment results and MOE of all PAH was 0.025 ngTEQBaP /kg/d and 4,000,000 from consuming edible oils in Korea. The BaP exposure assessment was 0.025 ng-TEQBaP /kg/d and the MOE was 4,000,000 from edible oil consumption. According to the COC (2006), all MOE results were of “negligible concern” with action minimizing future exposure (MOE > 100,000). As a result, according to the exposure assessment and risk characterization, individual PAH are considered to be within safe ranges in edible oil consumption from Korea. PAH carcinogenic potency factor of 7.3E + 0 (mg/kg/d)−1 (U.S. EPA, 1993) was used for this study. Cancer risk due to dietary BaP intake in Koreans is estimated as 0.069 × 10−6 . This result is of “negligible concern” with action minimizing future exposure.

FUNDING This research was supported by a grant (14162MFDS072) from Ministry of Food and Drug Safety in 2014.

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