Abstract
The objective of the present research was to examine 16 compounds of polycyclic aromatic hydrocarbons (PAHs) in grilled meat samples (including grilled beef, kebab, and grilled chicken) sold in Sulaymaniyah, Iraq, using the MSPE-GC/MS method (magnetic solid-phase extraction with gas chromatography-mass spectrometry). The Σ16PAH levels in grilled beef, kebab, and grilled chicken were 7.18, 11.09, and 7.68 µg/kg, respectively. The highest amount of benzo[a]pyrene (BaP) was found in kebab (1.25 ± 0.26 µg/kg), followed by grilled beef (0.75 ± 0.13 µg/kg) and grilled chicken (0.62 ± 0.11 µg/kg). The BaP levels in all the samples were within the European Union (EU) standard limits. The health risk assessment of the PAHs in the samples was performed via the Monte Carlo simulation. The results revealed that the values of hazard quotient (HQ), incremental lifetime cancer risk (ILCR), and margin of exposure (MoE) for children and adults were within the acceptable ranges. Our findings indicate that the consumption of grilled meat does not pose a health risk for Iraqi children and adults.
Keywords: Polycyclic aromatic hydrocarbon, Grilled meat, Kebab, Magnetic solid-phase extraction, Health risk assessment, GC–MS
Subject terms: Environmental sciences, Chemistry
Introduction
Polycyclic aromatic hydrocarbons (PAHs) are organic compounds characterized by two or more fused aromatic rings of carbon and hydrogen atoms1. Over 150 compounds of PAHs have been identified thus far2. For nonsmokers and adults not exposed occupationally, food accounts for more than 90% of PAHs exposure3. PAHs in food may be derived from various sources, including the environment, packaging materials, as well as food processing and cooking (e.g., grilling, smoking, roasting, and frying)4,5. The United States Environmental Protection Agency (USEPA) listed sixteen PAHs as high-priority contaminants according to their incidence and carcinogenicity, including naphthalene (Nap), acenaphthylene (Acy), acenaphthene (Ace), fluorene (Flu), phenanthrene (Phe), anthracene (Ant), fluoranthene (Flt), pyrene (Pyr), benzo[a]anthracene (BaA), chrysene (Chr), benzo[b]fluoranthene (BbF), benzo[k]fluoranthene (BkF), benzo[a]pyrene (BaP), Indeno[1,2,3-cd]pyrene (Ind), dibenzo[a,h]anthracene (DBA), and benzo[g,h,i]perylene (BghiP)6.
In 2005, the European Union (EU) recommended monitoring fifteen priority PAHs, and BaP was selected as an indicator of their occurrence and carcinogenicity7. The European Food Safety Authority (EFSA) stated that Σ4PAH and Σ8PAH are more appropriate measures for assessing PAHs contamination in food compared with BaP. The sum of BaA, Chr, BbF, and BaP is considered Σ4PAH. In contrast, the sum of BaA, Chr, BbF, BaP, BkF, BghiP, DBA, and IcdP is considered Σ8PAH6,8. The estimated average intake of Σ8PAH among European countries is 1.73 mg/day9. According to the International Agency for Research on Cancer (IARC), BaP is carcinogenic to humans (Group 1), whereas BaA, BbF, and Chr are categorized as possibly carcinogenic (Group 2A)10,11. On the other hand, PAHs with 2 to 4 benzene rings are known as light PAHs (LMW), whereas PAH compounds with more than 4 benzene rings are described as heavy PAHs (HMW), which have greater stability and toxicity7. PAHs bind to DNA and proteins in the human body, causing DNA mutations, altering gene expression, and eventually causing carcinogenicity12,13.
Meat is a valuable source of essential nutrients in human diets and is prepared in different culinary forms, such as stews, grilled meats, and processed meats. Meat consumption is significantly affected by the economic situation, consumer lifestyle, and people’s beliefs13–15. Grilled meat, also called kebab, is extremely popular in the Middle Eastern population because of its wide variety, delightful flavour and aroma, as well as its simple preparation16. However, research studies have indicated that cooking kebabs over charcoal or gas flames may pose significant health risks due to the formation of toxic contaminants, especially PAHs17,18. The generation of PAHs in grilled meat can result from the incomplete burning (pyrolysis) of organic matter, particularly fat, at high temperatures (above 200 °C). Moreover, dripping fat from meat on fuel during grilling produces volatile PAHs that rise from smoke and deposit on the meat surface. On the other hand, incomplete fuel combustion under inadequate oxygen can generate more PAHs in grilled meat13,19–21.
In recent years, several research studies in many countries have been conducted to investigate PAHs concentrations in kebabs3,11,14,22,23. In most cases, the techniques used to quantify PAHs are gas chromatography-mass spectrometry (GC–MS) and high-performance liquid chromatography with fluorescence detection (HPLC-FLD), of which GC–MS offers better selectivity and precision than HPLC-FLD. Recently, several sample pretreatment methods have been employed to separate PAHs. Carbon nanotubes (CNTs) with magnetized particles (MP) is an efficient and cost-effective innovative approach11,24. Grilled meat is a popular dish in the Kurdistan Region of Iraq, where charcoal-grilled kebabs are preferred for their more palatable taste. Grilled beef, kebab, and grilled chicken are the most favored among the Iraqi people. Kebab (in Iraq) and koobideh kebab (in Iran) are similar grilled meat dishes, and both are made from minced meat and onions [8]. However, no study has been conducted in Iraq to investigate PAHs in grilled meat. Therefore, the purpose of the present research was to determine 16 PAHs (Nap, Acy, Ace, Flu, Phe, Ant, Flt, Pyr, BaA, Chr, BbF, BkF, BaP, Ind, DBA, and BghiP) in grilled meat samples (grilled beef, kebab, and grilled chicken). Moreover, the health risks of PAHs through grilled meat consumption were evaluated for Iraqi children and adults.
Materials and methods
Regent and chemical
All chemicals were of analytical grade. The PAH standards were obtained from Supelco (Bellefonte, PA, USA). The internal standard was made by dissolving biphenyl in methanol (0.05 µg/mL). Multi-walled carbon nanotubes (MWCNTs) were supplied from Nanoshell (Panchkula, India, L = 5–30 µm and d = 30–60 nm). Biphenyl (as internal standard), potassium hydroxide, methanol, acetonitrile, hydrochloric acid, sodium chloride, and dichloromethane were purchased from Merck Co. (Darmstadt, Germany).
Sample collection
In total, 90 grilled meat samples, including “grilled beef (n = 30)”, “kebab (n = 30)”, and “grilled chicken (n = 30)” were purchased from 15 of the most popular restaurants in Sulaymaniyah, Iraq. Figure 1 illustrates the sample collection regions on the map. The sample collection was conducted in January 2025, and all the samples were grilled over charcoal. After being delivered to the laboratory, the samples were homogenized and maintained in the dark at -25 °C for less than two weeks until further analysis.
Fig. 1.
Map of sample collection regions.
Sample preparation
The functionalizing MWCNTs with hydroxyl and carboxyl groups and magnetization MWCNTs with Fe3O4 were performed according to the method described by Moazzen et al. (2019)25. The sample preparation was carried out using the procedure described in previous studies with minor modifications in the following three steps11,24:
Sample clean-up: First, 5 g of the frozen sample and 1 mL of internal standard were homogenized for 10 min. Then, 7.5 mL of 1 M potassium hydroxide and 7.5 mL of methanol–acetonitrile (70:30, v/v) were added (as an extraction solution), and the solution was sonicated in an ultrasonic bath at 40 °C for 10 min. Next, the solution was centrifuged at 5500 × g for 20 min. The fat of the samples was removed using the freeze lipid filtration method, in which the samples were first frozen at 24 °C for 24 h and then filtered with filter paper (Whatman No. 1) to remove frozen lipids. Ultimately, the pH of the solution was precisely adjusted to 6.5 using 1 M hydrochloric acid.
Analyte adsorption: In the next step, the aqueous layer was moved to another vial. The prepared adsorbent (10 mg, MWCNT-Fe₃O₄) and sodium chloride (0.5 g) were added to the solution and stirred for 5 min. Lastly, samples were kept for 24 h at 40–45 °C.
Desorption of analytes from adsorbent: The supernatant was removed, and then 5 mL of dichloromethane was applied to elute the analytes from the adsorbent, followed by stirring for 5 min. Following that, a magnet was used to collect the magnetic adsorbent on the side of the vial, and then the liquid phase was discarded. Finally, the absorbent was rinsed with deionized water (three times), and the solvent was evaporated using a gentle N₂ gas stream at 30 °C. The residue was redissolved in 50 mL of dichloromethane, and the solution was stirred for 1 min. The solution (1 μL) was injected into the GC–MS.
Sample analysis
The analysis of samples was conducted using Gas Chromatography (Agilent 6890, Palo Alto, CA, USA) equipped with a quadrupole mass spectrometer detector (Agilent 5973) and a capillary column (DB-5ms; 30 m × 0.25 mm × 0.25 µm). The temperature instructions were as follows: the injector temperature was 290 °C, the initial temperature was 70 °C with a 2 min hold, then ramped to 300 °C at 10 °C/min, and subsequently kept at 300 °C for 7 min. The carrier gas was helium (99.999%) with a flow rate of 1 mL per min. The mode of injection was splitless. The ion source and quadrupole temperatures were 230 and 150 °C, respectively. Ultimately, the SIM (selected ion monitoring) mode was utilized, and the electronic beam energy was 70 eV. To determine the amounts of PAHs in the grilled meat, an external calibration curve was constructed for each PAH. The form of straight lines was obtained by injecting extracts of the PAHs at different concentrations in dichloromethane into GC–MS. The extracts of cleaned samples (without PAHs) were prepared that were spiked with different concentrations of the PAHs ranging from 5 to 100 ng/g and 500 ng/g of biphenyl as an internal standard. A triplicate analysis was conducted for recoveries (repeatability and reproducibility). Following that, precision and accuracy were evaluated. The limit of detection (LOD) and limit of quantitation (LOQ) of PAHs were calculated based on signal-to-noise ratios of 3 (S/N = 3) and 10 (S/N = 10), respectively. The limit of detection (LOD), limit of quantification (LOQ), coefficient of estimation (R2), recovery rate, repeatability relative standard deviation (RSD), and equation of calibration curve of 16 PAH compounds were presented in Table 1. Some chromatograms of real samples and standards were shown in Figures S1 and S2.
Table 1.
Limit of detection (LOD), limit of quantification (LOQ), coefficient of estimation (R2), recovery rate, repeatability relative standard deviation (RSD), and equation of calibration curve of PAH compounds.
| PAHs | Method validation data | |||||
|---|---|---|---|---|---|---|
| LOD (µg/kg) | LOQ (µg/kg) | R2 | Recovery (%) | RSD (%) | Equation of calibration curve | |
| Nap | 0.08 | 0.20 | 0.9949 | 102.4 | 6.1 | y = 138.7x + 595.3 |
| Acy | 0.08 | 0.18 | 0.9928 | 96.1 | 4.6 | y = 127.7x—1380.1 |
| Ace | 0.08 | 0.15 | 0.9936 | 97.2 | 11.2 | y = 127.4x—208.3 |
| Flu | 0.12 | 0.15 | 0.9893 | 101.9 | 9.9 | y = 259.9x + 3177.9 |
| Phe | 0.04 | 0.20 | 0.9879 | 98.1 | 7.5 | y = 50.7x + 386.7 |
| Ant | 0.06 | 0.08 | 0.9915 | 96.3 | 10.3 | y = 182.5x + 1868.1 |
| Flt | 0.05 | 0.10 | 0.9974 | 101.3 | 9.6 | y = 31.3x + 396.1 |
| Pyr | 0.08 | 0.12 | 0.9895 | 99.4 | 11 | y = 61.4x—969.0 |
| BaA | 0.05 | 0.15 | 0.9962 | 96.7 | 8.4 | y = 227.2x—1721.6 |
| Chr | 0.05 | 0.12 | 0.9873 | 98.3 | 7.8 | y = 215.6x—1567.0 |
| BbF | 0.05 | 0.12 | 0.9957 | 94.9 | 10.7 | y = 127.3x—208.3 |
| BkF | 0.06 | 0.15 | 0.9885 | 95.5 | 7.9 | y = 27.9x—262.7 |
| BaP | 0.06 | 0.18 | 0.9961 | 103.4 | 6.5 | y = 21.9x—62.5 |
| Ind | 0.08 | 0.18 | 0.9914 | 99.1 | 8.1 | y = 31.3x + 396.1 |
| DBA | 0.08 | 0.20 | 0.9878 | 101.2 | 10.6 | y = 20.9x—61.5 |
| BghiP | 0.08 | 0.20 | 0.9984 | 94.2 | 8.9 | y = 9.3x—38.6 |
Fat measurement
Six samples of each grilled meat type were randomly chosen to determine their fat content, which was measured using the Soxhlet procedure based on the AOAC 991.36 method22,26.
Human health risk assessment
The estimated daily intake (EDI) of oral exposure to PAHs through grilled meat samples was calculated using Eq. 1. The total toxic equivalent quotient (TEQ) of each PAH was computed using Eq. 2. The risk associated with PAHs in grilled meat was assessed using the following approach: hazard quotient (HQ) (Eq. 3), incremental lifetime cancer risk (ILCR) (Eq. 4), and margin of exposure (MoE) (Eq. 5)14,24,27.
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1 |
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2 |
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3 |
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4 |
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5 |
If the HQ value surpasses 1, a non-carcinogenic health risk is expected for individuals, and vice versa. ILCR examined carcinogenic health risks. Cancer risks are unacceptable if the ILCR values exceed 10–4 and negligible if the ILCR values are below 10-6. If the ILCR values ranged from 10–6 to 10–4, individuals can tolerate the cancer risk 24,28. MoE represents the ratio between a toxicological reference point and a dose that results in a low but measurable response. If the MoE values are below 10.000, it is considered a potential concern for public health14,29. To enhance the accuracy of risk assessment by accounting for uncertainties, the Monte Carlo simulation technique was employed in the Oracle Crystal Ball version 11.1.2.4.850 (https://www.oracle.com/crystalball). The Monte Carlo simulation was used to provide a more realistic assessment by minimizing the likelihood of underestimating or overestimating risks30,31. The 95th percentile was considered significant risk, and the trial number was considered 10,000. Table 2 outlines the details of the parameters considered in calculating the health risk assessment associated with PAH from grilled meats consumption.
Table 2.
Description of parameters used to calculate the risk assessment of PAH in kebab sample.
| Parameters | Description | Units | Values | References |
|---|---|---|---|---|
| C | PAHs concentration | mg/kg | Concentrations of PAHs found in the present study | |
| IR | Ingestion rate of kebab | kg/day | Grilled beef = 15.2, kebab = 16.4, grilled chicken = 19.7 | 37 |
| EF | Exposure frequency | days | 350 | 38 |
| ED | Exposure duration | year | Adults = 70, children = 6 | 38 |
| BW | Body wight | kg | Adults = 70, children = 15 | 39 |
| AT | Average time | days | Adults = 25,550, children = 2190 | 38 |
| TEF | Toxic equivalent factors | - | DBA, and BaP = 1; Ind, BkF, BbF, and BaA = 0.1; BghiP, Chr, and Ant = 0.01; Nap, Acy, AcP, Flu, Phe, Flt, and Pyr = 0.001 | 11,40 |
| RfD | Reference dose | mg/kg/day | Nap = 0.02, Ace = 0.06, Flu and Flt = 0.04, Ant = 0.3, Pyr = 0.03, BghiP = 0.0003 | 40,41 |
| SF | Slope factor | mg/kg/day | BaP = 7.3 | 24 |
| BMDL10 | Benchmark dose lower confidence limit at 10% | mg/kg/day | BaP = 0.07, Σ4PAH = 0.34, Σ8PAH = 0.49 | 42 |
Statistical analysis
Data analysis was conducted using SPSS v27. The data were analyzed utilizing the Kruskal–Wallis test. The significance level was p < 0.05.
Results and discussion
Concentrations of PAHs in kebab samples
The levels of PAHs found in the grilled beef, kebab, and grilled chicken samples sold in Sulaymaniyah, Kurdistan region of Iraq, are presented in Table 3. The mean concentrations of Acy, Ace, Flu, Flt, Pyr, BaA, Chr, BbF, and BaP in the grilled beef samples were 0.72 ± 0.10, 0.53 ± 0.12, 1.33 ± 0.11, 1.56 ± 0.23, 1.31 ± 0.12, 0.44 ± 0.15, 0.28 ± 0.11, 0.26 ± 0.07, and 0.75 ± 0.13 µg/kg, respectively. The highest PAHs were found in grilled beef related to Flt (2.00 µg/kg). The average amounts of Nap, Acy, Ace, Flu, Flt, Pyr, BaA, Chr, BbF, BaP, DBA, and BghiP in the kebab samples were 0.22 ± 0.07, 0.76 ± 0.18, 0.57 ± 0.15, 1.40 ± 0.21, 1.29 ± 0.14, 2.41 ± 0.47, 0.69 ± 0.12, 1.46 ± 0.09, 0.45 ± 0.12, 1.25 ± 0.26 0.33 ± 0.10, and 0.26 ± 0.21 µg/kg, respectively. The maximum PAHs were detected in kebab related to Pyr (3.11 µg/kg). The mean levels of Acy, Ace, Flu, Flt, Pyr, Chr, BbF, BkF, BaP, and DBA in the grilled chicken samples were 0.68 ± 0.14, 0.46 ± 0.05, 1.29 ± 0.03, 2.09 ± 0.33, 1.12 ± 0.20, 0.38 ± 0.07, 0.52 ± 0.09, 0.30 ± 0.16, 0.62 ± 0.11, and 0.22 ± 0.09 µg/kg, respectively. The maximum levels of PAHs were found in grilled chicken related to Flt (2.60 µg/kg). The amounts of Phe, Ant, and Ind in all samples were below the detection limits. Furthermore, Nap and BghiP levels in grilled beef and grilled chicken, BaA in grilled chicken, BkF in grilled beef and kebab, and DBA in grilled beef were below the detection limits. The European Commission (EC) established acceptable limits for BaP and Σ4PAH at 5 and 30 µg/kg in meat products, respectively32. The levels of BaP and Σ4PAH determined in our study were within the EC standard.
Table 3.
Concentrations of different PAH compounds in grilled meat samples.
| PAHs | PAHs concentrations in grilled meat samples (µg/kg) | |||||||
|---|---|---|---|---|---|---|---|---|
| Grilled beef | Kebab | Grilled chicken | ||||||
| Mean ± SD | Max | Mean ± SD | Max | Mean ± SD | Max | |||
| Nap | < LOD | < LOD | 0.22 ± 0.07 | 0.30 | < LOD | < LOD | ||
| Acy | 0.72 ± 0.10 | 0.85 | 0.76 ± 0.18 | 1.20 | 0.68 ± 0.14 | 0.89 | ||
| Ace | 0.53 ± 0.12 | 0.72 | 0.57 ± 0.15 | 0.82 | 0.46 ± 0.05 | 0.56 | ||
| Flu | 1.33 ± 0.11 | 1.50 | 1.40 ± 0.21 | 1.83 | 1.29 ± 0.03 | 1.38 | ||
| Phe | < LOD | < LOD | < LOD | < LOD | < LOD | < LOD | ||
| Ant | < LOD | < LOD | < LOD | < LOD | < LOD | < LOD | ||
| Flt | 1.56 ± 0.23b | 2.00 | 1.29 ± 0.14b | 1.55 | 2.09 ± 0.33a | 2.60 | ||
| Pyr | 1.31 ± 0.12b | 1.52 | 2.41 ± 0.47a | 3.11 | 1.12 ± 0.20b | 1.44 | ||
| BaA | 0.44 ± 0.15b | 0.71 | 0.69 ± 0.12a | 0.94 | < LOD | < LOD | ||
| Chr | 0.28 ± 0.11b | 0.53 | 1.46 ± 0.09a | 1.62 | 0.38 ± 0.07b | 0.59 | ||
| BbF | 0.26 ± 0.07a | 0.48 | 0.45 ± 0.12b | 0.64 | 0.52 ± 0.09b | 0.66 | ||
| BkF | < LOD | < LOD | < LOD | < LOD | 0.30 ± 0.16 | 0.58 | ||
| BaP | 0.75 ± 0.13b | 1.00 | 1.25 ± 0.26a | 1.64 | 0.62 ± 0.11b | 0.78 | ||
| Ind | < LOD | < LOD | < LOD | < LOD | < LOD | < LOD | ||
| DBA | < LOD | < LOD | 0.33 ± 0.10 | 0.52 | 0.22 ± 0.09 | 0.40 | ||
| BghiP | < LOD | < LOD | 0.26 ± 0.21 | 0.71 | < LOD | < LOD | ||
| LMW | 2.58 | 2.95 | 2.43 | |||||
| HMW | 4.60 | 8.14 | 5.25 | |||||
| Σ4PAH | 1.73 | 3.85 | 1.52 | |||||
| Σ8PAH | 1.73 | 4.44 | 2.04 | |||||
| Σ16PAH | 7.18 | 11.09 | 7.68 | |||||
Different letters in the row indicate statistically significant differences (P < 0.05).
Jiang et al. (2018) reported that the mean levels of Nap, Ace, Fle, Phe, Ant, Flu, Pyr, BaA, Chr, BbF, BkF, BaP, DahA, IcdP, BghiP in the grilled meat products were 23.9, 9.98, 11.4, 11.1, 2.69, 8.35, 9.23, 0.91, 0.98, 0.43, 0.23, 0.33, 0.14, 0.18, and 0.14 μg/kg, respectively6. Farhadian et al. (2010) examined PAH in grilled meat sold in Malaysia using HPLC-FLD and showed that Flt, BbF, and BaP in the beef kebab (n = 18) was 10.65, 0.32, and 0.37 ng/g, respectively, and in the chicken kebab (n = 18) was 13.55, 0.42, and 1.57 ng/g, respectively 20. Authors in Lebanon detected PAH in charcoal-grilled chicken and demonstrated that the mean concentration of BaA, Chr, BbF, and BaP in samples was 3.47, 5.16, 3.58, and 1.07 μg/kg, respectively33. Another study determined the PAH levels in grilled meats from restaurants in Egypt using the GC–MS method and reported that the concentration of BaP in the grilled beef steak and beef kofta was 0.02 and 3.63 µg/kg23.
In our study, the decreasing order of Σ4PAH in samples was as follows: kebab (3.85 µg/kg) > grilled beef (1.73 µg/kg) > grilled chicken (1.52 µg/kg). The level of Σ8PAH in the grilled beef, kebab, and grilled chicken was 1.73, 4.44, and 2.04 µg/kg, respectively. The decreasing order of total PAH (Σ16PAH) in the sample was as follows: kebab (11.09 µg/kg) > grilled chicken (7.68 µg/kg) > grilled beef (7.18 µg/kg). In the present study, the highest Σ16PAH values were found in kebab, which is in agreement with previous studies 16. Our results were significantly lower than the findings by Alomirah et al. (2011), which examined PAHs in grilled and smoked foods in Kuwait using GC–MS and found that total PAHs in meat kabab and grilled chicken were 241 µg/kg (ranging 40.8–534 µg/kg) and 222 µg/kg (ranging 48.2–342 µg/kg), respectively3. Khalili et al. (2023) conducted a study to measure PAHs by the MSPE-GC/MS technique and indicated that Σ16PAH in the juje kebab and kebab koobideh was 112.9 ± 7.2 and 124.4 ± 9.1 μg/kg11. Gholizadah et al. (2021) reported that the median of total PAHs in kebab koobideh, juje kebab, and kebab barg grilled on charcoal were 167, 89.7, and 145 μg/kg, respectively. Additionally, the median of total PAHs in kebab koobideh, juje kebab, and kebab barg grilled on gas were 96.6, 102, and 94.6 μg/kg, respectively. They concluded that the maximum PAHs formed in kebab cooked on charcoal were dibenzo[a,h]pyrene16. In a study in Turkey, the average concentrations of BaP, Σ4PAH, Σ8PAH, and Σ16PAH in the grilled chicken were < 0.05, 2.13 ± 0.06, 3.68 ± 0.08, and 4.91 ± 0.09 µg/kg, respectively14.
In our study, the increasing order of LMW in kebabs was as follows: grilled chicken (2.43 µg/kg) < grilled beef (2.58 µg/kg) < kebab (2.95 µg/kg). The increasing order of HMW in kebabs was as follows: grilled beef (4.60 µg/kg) < grilled chicken (5.25 µg/kg) < kebab (8.14 µg/kg). In another study, the HMW level in the koobideh sahel, koobideh hatam, koobideh nakhlestan, koobideh khalij fars, juje sahel, juje hatam, juje nakhlestan, and juje khalij fars was 8.3, 7.14, 8.31, 21.95, 0.29, 1.97, 0.97, 0.64 ng/g, respectively; and LMW level in the koobideh sahel, koobideh hatam, koobideh nakhlestan, koobideh khalij fars, juje sahel, juje hatam, juje nakhlestan, and juje khalij fars was 22.73, 77.46, 54.96, 51.64, 119.02, 2.49, 1.78, and 158.5 ng/g, respectively7. Authors in Iran demonstrated that the total PAH, BaP, Σ4PAH, Σ8PAH, LMW, and HMW levels in juje kabab grilled over charcoal were 9.89, 0.86, 1.87, 2.39, 3.37, and 6.25 µg/kg, respectively, and in the kabab koobideh were 13.27, 1.45, 4.72, 5.84, 3.55, and 9.72 µg/kg, respectively. They also detected BaP contents in 4 samples above the EU standard22. Besides the analytical discrepancy, the difference in PAHs levels found in our study and previous research can be explained by several factors, such as the type of meat, fat content of meat, cooking method, time and temperature of cooking, type of fuel, oxygen accessibility, oil dripping on the flame, and distance from the flame3,6,11,18,24.
The fat content in different grilled meat samples is presented in Table 4. The fat content in grilled beef, kebab, and grilled chicken was 1.93 ± 0.3%, 8.12 ± 1.2%, and 2.85 ± 0.9%, respectively. These results are in accordance with findings by Es’haghi Gorji et al. (2016), which show that the fat content in kebab koobideh, kebab barg, and juje kebab was 7.8%, 7.3%, and 3.2%, respectively22. The highest concentration was detected in kebab, which can be attributed to its high fat content. The following processes are suggested for PAHs generation in foods: (a) fat pyrolysis at high temperatures; (b) pouring fat droplets from food being cooked over fire; (c) production of PAHs due to incomplete coal burning and contamination of food by them2.
Table 4.
Fat content in grilled meat samples.
| Sample | Meat type | Fat content (%) |
|---|---|---|
| Grilled beef | Beef | 1.93 ± 0.3 |
| Kebab | Minced beef or lamb (with fat) | 8.12 ± 1.2 |
| Grilled chicken | Chicken (boneless and skinless) | 2.85 ± 0.9 |
It has been demonstrated that fire sources influence PAHs formation. The highest level of PAHs is generated during charcoal grilling, followed by gas-flame grilling and electric grilling3,6. In a previous study, fish grilled on an electric broiler produced fewer PAHs than fish grilled on gas34. Akkaya et al. (2024) found that the total PAHs levels in meat doner cooked on electric, open gas, wood, and charcoal were 18.41, 20.47, 28.54, and 34.56 µg/kg, respectively. Also, the total PAH levels in chicken doner cooked on electric, open gas, wood, and charcoal were 23.85, 26.07, 35.87, and 42.53 µg/kg, respectively. The highest BaP level in the doner kebab samples was formed during charcoal cooking, and electric cooking generated lower PAHs in samples10. Nor Hasyimah et al. (2018) investigated the formation of PAHs in gas-grilled beef satay prepared at five different temperatures (150, 200, 250, 300, and 350 °C) using HPLC-FLD. They reported that PAH levels were minimum at 150 °C, and PAH levels increased with increasing temperature18. In this area, research has emphasized that direct contact with the flame or grill should be avoided during the cooking of the meat at high temperatures20,35.
Health risk assessment
The TEQ, EDI, and HQ values of PAHs in grilled meat for Iraqi adults and children are presented in Table 5. The TEQ values of Acy, Ace, Flu, Flt, Pyr, BaA, Chr, BbF, and BaP in grilled beef samples were 8.48E-7, 7.13E-7, 1.49E-6, 1.96E-6, 1.52E-6, 6.95E-5, 4.88E-6, 4.76E-5, and 9.83E-4 mg/kg, respectively. The TEQ values of Nap, Acy, Ace, Flu, Flt, Pyr, BaA, Chr, BbF, BaP, DBA, and BghiP in kebab samples were 2.85E-7, 1.11E-6, 8.16E-7, 1.79E-6, 1.54E-6, 3.12E-6, 9.06E-5, 1.61E-5, 5.97E-5, 1.59E-3, 5.08E-4, and 7.19E-6, mg/kg, respectively. The TEQ values of Acy, Ace, Flu, Flt, Pyr, Chr, BbF, BkF, BaP, and DBA in the grilled chicken sample were 8.86E-7, 5.53E-7, 1.37E-6, 2.60E-6, 1.44E-6, 5.20E-6, 6.53E-5, 5.73E-5, 7.65E-4, and 3.78E-4 mg/kg, respectively. The total TEQ values of PAHs in the sample were as follows: kebab = 2.28E-3 mg/kg > grilled chicken = 1.28E-3 mg/kg > grilled beef = 1.11E-3 mg/kg. In another study, the TEQ values of total PAH in meat doner, chicken doner, meatball, grilled chicken, and grilled fish were 328.46, 570.9, 1,746.48, 310.46, and 1,116.42 ng/kg, respectively14. Jiang et al. (2018) found that the TEQ value of BaP in 52 grilled and fried meat products was 360 ng/kg6.
Table 5.
TEQ, EDI, and HQ values of PAHs in grilled meat samples for Iraqi adults and children.
| PAHs | Grilled beef | Kebab | Grilled chicken | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| TEQ | Adults | Children | TEQ | Adults | Children | TEQ | Adults | Children | |||||||
| EDI | HQ | EDI | HQ | EDI | HQ | EDI | HQ | EDI | HQ | EDI | HQ | ||||
| Nap | ND* | ND | ND | ND | ND | 2.85E-7 | 6.68E-8 | 3.32E-6 | 3.12E-7 | 1.55E-5 | ND | ND | ND | ND | ND |
| Acy | 8.48E-7 | 1.84E-7 | NT** | 8.59E-7 | NT | 1.11E-6 | 2.60E-7 | NT | 1.21E-6 | NT | 8.86E-7 | 2.49E-7 | NT | 1.16E-6 | NT |
| Ace | 7.13E-7 | 1.55E-7 | 2.58E-6 | 7.22E-7 | 1.20E-5 | 8.16E-7 | 1.91E-7 | 3.17E-6 | 8.93E-7 | 1.48E-5 | 5.53E-7 | 1.56E-7 | 2.59E-6 | 7.26E-7 | 1.21E-5 |
| Flu | 1.49E-6 | 3.23E-7 | 8.07E-6 | 1.51E-6 | 3.77E-5 | 1.79E-6 | 4.19E-7 | 1.05E-5 | 1.95E-6 | 4.89E-5 | 1.37E-6 | 3.86E-7 | 9.65E-6 | 1.80E-6 | 4.50E-5 |
| Phe | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND |
| Ant | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND |
| Flt | 1.96E-6 | 4.25E-7 | 1.06E-5 | 1.98E-6 | 4.96E-5 | 1.54E-6 | 3.61E-7 | 9.02E-6 | 1.68E-6 | 4.21E-5 | 2.60E-6 | 7.31E-7 | 1.83E-5 | 3.41E-6 | 8.53E-5 |
| Pyr | 1.52E-6 | 3.29E-7 | 1.10E-5 | 1.54E-6 | 5.12E-5 | 3.12E-6 | 7.31E-7 | 2.44E-5 | 3.41E-6 | 1.14E-4 | 1.44E-6 | 4.04E-7 | 1.35E-5 | 1.88E-6 | 6.28E-5 |
| BaA | 6.95E-5 | 1.51E-7 | NT | 7.04E-7 | NT | 9.06E-5 | 2.12E-7 | NT | 9.91E-7 | NT | ND | ND | ND | ND | ND |
| Chr | 4.88E-6 | 1.06E-7 | NT | 4.95E-7 | NT | 1.61E-5 | 3.76E-7 | NT | 1.75E-6 | NT | 5.20E-6 | 1.46E-7 | NT | 6.84E-7 | NT |
| BbF | 4.76E-5 | 1.03E-7 | NT | 4.82E-7 | NT | 5.97E-5 | 1.40E-7 | NT | 6.53E-7 | NT | 6.53E-5 | 1.84E-7 | NT | 8.58E-7 | NT |
| BkF | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | 5.73E-5 | 1.61E-7 | NT | 7.53E-7 | NT |
| BaP | 9.83E-4 | 2.14E-7 | 7.12E-4 | 9.96E-7 | 3.32E-3 | 1.59E-3 | 3.72E-7 | 1.24E-3 | 1.73E-6 | 5.78E-3 | 7.65E-4 | 2.15E-7 | 7.18E-4 | 1.01E-6 | 3.35E-3 |
| Ind | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND | ND |
| DBA | ND | ND | ND | ND | ND | 5.08E-4 | 1.19E-7 | NT | 5.56E-7 | NT | 3.78E-4 | 1.06E-7 | NT | 4.96E-7 | NT |
| BghiP | ND | ND | ND | ND | ND | 7.19E-6 | 1.68E-7 | NT | 7.86E-7 | NT | ND | ND | ND | ND | ND |
* ND (not detected), ** NT (not tested, RfD is unavailable).
The 95th percentile EDI values of PAHs in grilled beef ranged from 1.03E-7 to 4.25E-7 mg/kg bw/day for adults, while they ranged from 4.82E-7 to 1.98E-6 mg/kg bw/day for children. The range of 95th percentile EDI values of PAHs kebab for adults was 6.68E-8 to 7.31E-7 mg/kg bw/day, and for children, it was 3.12E-7 to 3.41E-6 mg/kg bw/day. The 95th percentile EDI values of PAHs in grilled chicken varied from 1.06E-7 to 7.31E-7 mg/kg bw/day for adults and varied from 4.96E-7 to 3.41E-6 mg/kg bw/day for children. The 95th percentile EDI values of Σ4PAH for adults in grilled beef, kebab, and grilled chicken were 1.94E-7, 3.67E-7, and 2.07E-7 mg/kg bw/day, respectively, while for children were 9.07E-7, 1.71E-6, and 9.67E-7 mg/kg bw/day, respectively. The 95th percentile EDI values of Σ8PAH for adults in grilled beef, kebab, and grilled chicken were 1.94E-7, 4.03E-7, and 1.97E-7 mg/kg bw/day, respectively, and for children were 9.07E-7, 1.88E-6, and 9.20E-7 mg/kg bw/day, respectively. In the study by Jiang et al. (2018), the mean EDI values of Σ15PAH, Σ8PAH, and Σ4PAH in grilled meat products were 120, 4.99, and 3.96 ng/kg bw/day, respectively6. Another study indicates that the EDI values of Σ4PAH, Σ8PAH, and Σ16PAH in grilled chicken were 1.79, 3.09, and 4.12 ng/kg bw/day, respectively14. Alomirah et al. (2011) found that the EDI values of BaP in the meat kebab for Kuwaiti adults and children were 5.62 and 5.92 ng/day3.
The 95th percentile HQ values of Ace, Flu, Flt, Pyr, and BaP in grilled beef for children were 1.20E-5, 3.77E-5, 4.96E-5, 5.12E-5, and 3.32E-3, respectively, while these values for adults were 2.58E-6, 8.07E-6, 1.06E-5, 1.10E-5, and 7.12E-4, respectively. The 95th percentile HQ values of Nap, Ace, Flu, Flt, Pyr, and BaP in kebab for children were 1.55E-5, 1.48E-5, 4.89E-5, 4.21E-5, 1.14E-4, and 5.78E-3, respectively, and for adults, these were 3.32E-6, 3.17E-6, 1.05E-5, 9.02E-6, 2.44E-5, and 1.24E-3, respectively. The 95th percentile HQ values of Ace, Flu, Flt, Pyr, and BaP in grilled chicken for children were 1.21E-5, 4.50E-5, 8.53E-5, 6.28E-5, and 3.35E-3, respectively, whereas for adults were 2.59E-6, 9.65E-6, 1.83E-5, 1.35E-5, and 7.18E-4, respectively. The HQ values of all PAH compounds were less than one, so non-carcinogenic health risk is not expected. In another study, the 95th percentile HQ values of BaP and Pyr, Nap, and Ace in burger samples were 0.0296, 0.0159, 0.0078, and 0.0036, respectively24.
Table 6 provides EDI and MoE values of BaP, Σ4PAH, and Σ8PAH in grilled meat samples for Iraqi adults and children. The MoE values of BaP in grilled beef, kebab, and grilled chicken for adults were 327,837, 188,362, and 324,978, respectively, and for children, these values were 70,251, 40,363, and 69,640, respectively. The MoE values of Σ4PAH for adults in grilled beef, kebab, and grilled chicken were 1,748,680, 925,687, and 1,460,549, respectively, while these values for children were 374,717, 198,361, and 351,546, respectively. The MoE values of Σ8PAH in grilled beef, kebab, and grilled chicken for adults were 2,520,155, 1,195,285, and 2,468,904, respectively, whereas for children these values were 540,033, 256,133, and 529,051, respectively. All MoE values calculated in the present study were above 10,000, so they are acceptable. In the study by Sahin et al. (2020), the MoE values of BaP and Σ4PAH in grilled fish were 388,888 and 425,00014. Another study in Egypt showed that the MoE value of BaP in grilled meat was 290,4536.
Table 6.
EDI and MoE values of BaP, Σ4PAH, and Σ8PAH in grilled meat samples for Iraqi adults and children.
| Risk | Age groups | Grilled beef | Kebab | Grilled chicken | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| BaP | Σ4PAH | Σ8PAH | BaP | Σ4PAH | Σ8PAH | BaP | Σ4PAH | Σ8PAH | ||||
| EDI | Adults | 2.14E-7 | 1.94E-7 | 1.94E-7 | 3.72E-7 | 3.67E-7 | 4.03E-7 | 2.15E-7 | 2.07E-7 | 1.97E-7 | ||
| Children | 9.96E-7 | 9.07E-7 | 9.07E-7 | 1.73E-6 | 1.71E-6 | 1.88E-6 | 1.01E-6 | 9.67E-7 | 9.20E-7 | |||
| MoE | Adults | 327,837 | 1,748,680 | 2,520,155 | 188,362 | 925,687 | 1,195,285 | 324,978 | 1,460,549 | 2,468,904 | ||
| Children | 70,251 | 374,717 | 540,033 | 40,363 | 198,361 | 256,133 | 69,640 | 351,546 | 529,051 | |||
The 95th percentile ILCR values of BaP in grilled meat samples are indicated in Fig. 2. The 95th percentile ILCR values of BaP for children and adults in grilled beef samples were 7.27E-6 and 1.56E-6, in kebab samples were 1.27E-5 and 2.71E-6, and in grilled chicken samples were 7.34E-6 and 1.57E-6, respectively. The ILCR values of BaP for both age groups were acceptable, and carcinogenic health risks resulting from BaP do not exist. In another study, the ILCR values were 2.69E-6 and 3.75E-6 for children and adults, respectively6. The 95th percentile ILCR values of BaP in the juje kebab and kebab koobideh estimated by Khalili et al. (2023) were 7.1E-07 and 6.06E-07, respectively11.
Fig. 2.
95th percentile ILCR values of BaP in grilled meat samples for Iraqi adults and children.
Conclusion
For the first time in Iraq, this study investigated PAHs in three types of grilled meat sold in the Sulaymaniyah, Kurdistan region. The Σ16PAH levels found in samples were as follows: kebab = 11.09 µg/kg > grilled chicken = 7.68 µg/kg > grilled beef = 7.18 µg/kg. The BaP and Σ4PAH concentrations in the grilled meats were below the standard limits. The results showed that kebab had the highest amounts of PAHs, which could be due to its high fat content. It is recommended to reduce the fat content of kebab, as well as use lean meat for grilled meat and avoid charcoal flare-ups. Additionally, it is recommended that industry collaborations focus on PAH-reduction strategies. The risk associated with PAHs in grilled meat was evaluated by HQ, ILCR, and MoE. In all cases, HQ, ILCR, and MoE values of PAHs for Iraqi children and adults were acceptable. However, cumulative PAH exposure from throughout the entire food basket may jeopardize human health. In general, our results suggest that grilled meat intake does not pose a health risk to consumers. In spite of this, it is recommended that PAHs be examined in other types of processed meat in Iraq.
Supplementary Information
Acknowledgements
This work is conducted in cooperation with the food safety and hygiene division in the School of Public Health at Tehran University of Medical Sciences.
Author contributions
S.M.A. Supervision, Conceptualization, Methodology, Project administration, Writing – review & editing. A.E.F. Methodology, Project administration, Formal analysis, Data curation, Writing – review & editing. P.S. Methodology, Data curation, Writing – review & editing. N.J. Investigation, Writing – original draft. Z.R. Investigation, Writing – original draft. R.A. Supervision, Conceptualization, Project administration, Writing – review & editing.
Funding
Not applicable.
Data availability
The datasets generated and/or analysed during the current study are available from the corresponding author on reasonable request.
Declarations
Competing interests
The authors declare no competing interests.
Footnotes
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
The online version contains supplementary material available at 10.1038/s41598-025-16484-6.
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Associated Data
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Supplementary Materials
Data Availability Statement
The datasets generated and/or analysed during the current study are available from the corresponding author on reasonable request.