92 ISSN 1120-1770 online, DOI 10.15586/ijfs.v33iSP1.2065 P U B L I C A T I O N S CODON Italian Journal of Food Science, 2021; 33 (SP1): 92–102 et al., 2021; Heshmati et al., 2019). They pose carcino- genic, mutagenic, immunosuppressive, and teratogenic consequences. The most common AFs are aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1), and G2 (AFG2) (Heshmati et al., 2017). AFB1 is the most carcinogenic type for humans and animals (Mokhtarian et al., 2020). The International Agency for Research on Cancer (IARC) classified AFB1 as a group 1 carcinogen (Elzupir et al., 2010). Aflatoxins production may occur during harvesting, transportation, storage, or on the farm. AFs are very stable chemical compounds resistant to heat and food processes (Cui et al., 2020). They can contaminate var- ious foods, including cereal, dairy products, oilseeds, P U B L I C A T I O N S CODON The prevalence and risk assessment of aflatoxin in sesame-based products Ali Heshmati1, Mina Khorshidi1*, Amin Mousavi Khaneghah2* 1Department of Nutrition and Food Safety, School of Medicine, Nutrition Health Research Center, Hamadan University of Medical Sciences, Hamadan, Iran; 2Department of Food Science and Nutrition, Faculty of Food Engineering, University of Campinas (UNICAMP), São Paulo, Brazil *Corresponding Author: Mina Khorshidi, Department of Nutrition and Food Safety, School of Medicine, Nutrition Health Research Center, Hamadan University of Medical Sciences, Hamadan, Iran. Email: mkhorshidi2992@gmail.com; Mousavi Khaneghah, Department of Food Science and Nutrition, Faculty of Food Engineering, University of Campinas (UNICAMP), São Paulo, Brazil. Email: mousavi@unicamp.br Received: 9 May 2021; Accepted: 4 June 2021; Published: 5 July 2021 © 2021 Codon Publications OPEN ACCESS PAPER Abstract The contamination of aflatoxins (AFs) in 120 samples of sesame seeds, tahini, and tahini halva collected from Iran’s market were evaluated. The exposed risk due to ingestion of aflatoxin B1 (AFB1) via their consumption was estimated with the aid of the Monte Carlo simulation (MCS). The highest prevalence of AF (55%) was associated with sesame seed samples, followed by tahini (45%) and tahini halva (32.5%). The AFB1 concentration in sesame seeds, tahini, and tahini halva was in the ranges of 0.21–12.35, 0.23–5.81, and 0.27–3.56 μg/kg, respectively. The concentration of the total aflatoxin (TAF) in 7 (17.5%), 8 (20%), and 2 (5%) samples of sesame seeds, tahini, and tahini halva, respectively, was below the limit of European regulations (4 µg/kg), while the levels of AFB1 in 10 (25%), 7 (17.5%), and 6 (15%) samples of sesame seeds, tahini, and tahini halva, respectively, were higher than the European regulations (2 µg/kg). As the percentile 50 and 95 of margin of exposure (MOE) with AFB1 for sesame seed, tahini, and tahini halva was more than 10,000, it could conclude the intake of aflatoxin through the con- sumption of mentioned products did pose a not remarkable cancer risk for adults. Keywords: mycotoxin; contamination; risk assessment; traditional products; sesame based Introduction Today, humans pay considerable attention to food safety and contaminants (Milicevic et al., 2021; Rapa et al., 2021; Xinyu et al., 2020). Mycotoxins produced by fungi are among the most important food contami- nants and have a negative impact on public health, food safety, and the national economy of many countries, especially developing countries (Batrinou et al., 2020; Grumi et al., 2020). The most critical factors in food contamination with mycotoxins are moisture, intrin- sic properties and nutrients, long shelf life and pH, and high-water activity (Wang et al., 2018). Aflatoxins (AFs) are produced by Aspergillus fungi, especially A. flavus and A. parasitics and rarely by A. nomius (De Souza mailto:mkhorshidi2992@gmail.com mailto:mousavi@unicamp.br Italian Journal of Food Science, 2021; 33 (SP1) 93 Prevalence and risk assessment of aflatoxin in sesame-based products prevalence of AFs in sesame seeds and their products was demonstrated by previous studies (Anthony et  al., 2014; Apeh et al., 2016; Asadi et al., 2011; Esan et al., 2020; Fapohunda et al., 2018; Hosseininia et al., 2014; Kollia et al., 2016; Li et al., 2009; Reddy et al., 2011; Sabry et al., 2016; Sebaei et al., 2020; Sirhan et al., 2014; Tabata, 2007; Torlak and Akan, 2013; Var et al., 2007). No studies have been performed to evaluate mycotoxin contamination in tahini and tahini halva samples avail- able in Iran’s market. Therefore, the current study aimed to determine the level of AFs in sesame seed, tahini, and tahini halva, and halva consumed in western Iran. Furthermore, the exposed risk due to ingestion of AFB1 via their consumption was estimated with the aid of the Monte Carlo simulation (MCS). Material and Methods Materials Acetonitrile, HNO3 (65%), phosphate buffer solution (PBS), methanol, Potassium bromide (KBr), were pur- chased from Merck (Darmstadt, Germany). The AF stan- dards were purchased from Sigma-Aldrich (St. Louis, MO, USA) Sample collection Samples (n = 120; 40 fresh sesame seed, 40 tahini, and 40 tahini halva) were collected from the local market in Hamadan province, Iran, from April 2020 until August 2020. Samples were stored in a refrigerator (4°C) until analysis. Chemical properties Moisture content (%) of sesame seed, tahini, and tahini halva was determined by drying in an oven at 100°C. The protein and fat contents of samples were determined by the Kjeldahl method and Soxhlet extraction, respectively (Zebib et al., 2015). Extracted fat acidity was measured by titration by sodium hydroxide (0.01°N) in the presence of phenolphthalein as an indicator. Extract and cleanup of AF The method applied for the extraction and cleanup of AF was similar to the previous one with slight modifications (Torlak and Akan, 2013). Samples were wholly powdered and mixed. Fifty grams of ground sample was weighed and transferred into a 250 mL flask. Then, 150 mL of a mixture of water and methanol (30:70; v/v) was added and placed spices, and nuts (Javanmardi et al., 2020; Khaneghah et al., 2018; Mozaffari Nejad et al., 2020). Due to the high toxicity of AFs, exposure to these contam- inations could threaten human health. The Committee on Food Additives of Joint FAO/WHO (JECFA) recom- mends that the presence of mycotoxins in meals be mini- mized to reduce the potential risk (Di Sanzo et al., 2018). Therefore, in order to control AFs in food products, some regulations were established in many countries; the mea- surement and monitoring of AFs in food products are crucial (Sebaei et al., 2020). With the scientific name of Sesamum indicum L., sesame seed belongs to the Pedaliaceae family and is one of the oldest and most momentous oilseeds globally (Lee et al., 2020). It contains 58%–44% oil, 25%–25% protein, 13.5% carbohydrates, and 5% ash (Kollia et al., 2016). It is also a significant source of dietary fiber and micronutrients such as minerals, including calcium, phosphate, iron, potassium, vitamins such as E, thiamine, and niacin, lig- nans tocopherols, and phytosterols (Elleuch et al., 2011; Yao et al., 2021). Sesame seeds have antioxidant, anti-in- flammatory, anti-fungal, anti-viral, and natural anti- bacterial effects (Dravie et al., 2020). Sesame seeds are consumed in different forms in the world (Namiki, 2007). It is widely used in the Iranian food industry as an ingre- dient in confectionery products, bread, and pastries. Therefore, ensuring the mycotoxicological quality of sesame is very important (Asadi et al., 2011; Eghbaljoo- Gharehgheshlaghi et al., 2020; Elleuch et al., 2011; Kollia et al., 2016). Tahini is made by grinding and roasting sesame seeds known in the Middle East (Gholami et al., 2020; Sebaei et al., 2020). Tahini halva, also known as halva, helva, halawi, halawh, is produced by mixing tahini with sugar, citric or tartaric acid, and Saponaria officinalis root extract (Öğütcü et al., 2017; Osaili et al., 2018a; Var et  al., 2007). Consumption of tahini halva has increased due to its excellent nutritional value and health proper- ties in other countries, including the United States and European countries, Iran, Turkey, Saudi Arabia, Iraq, Greece, Jordan, Bulgaria, and Bosnia and Herzegovina (Anilakumar et al., 2010). Tahini halva consists of about 45% tahini, 45%–55% sugar, 2% ash, and 2.5% moisture and is consumed with bread in breakfast and dinner (Osaili et al., 2018b). When sesame seeds were stored improperly, they can be contaminated with mycotoxins, especially AFs (Anthony et al., 2014). Therefore, the EU sets limits for AFB1 and TAF in sesame and its products as 2 and 4 μg/kg, respec- tively. In contrast, the limit of these mycotoxins, accord- ing to the Iranian Institute of Standards and Industrial Research (ISIRI), was 5 and 15 μg /kg (INSO, 2020). The 94 Italian Journal of Food Science, 2021; 33 (SP1) Heshmati A et al. on a shaker and stirred for 10 min. Then, it was filtered through filter paper of Whatman No. 2. Twenty milliliters of filtered solutions were collected and transferred into a 100 mL flask, and 40 mL of distilled water was added and mixed for 5 min and filtered. For the cleanup of AFs, 15 mL of filtrate was transferred through the immunoaffinity column at a flow rate of 2–3 drops/s. Further, the column was washed three times with 10 mL distilled water at the same flow rate. The elution of AFs was performed by ace- tonitrile (1 mL). The eluate was gathered in a vial, and its volume was reached 2 mL with acetonitrile. Then, 100 µL of eluted AFs was injected into a high-performance liquid chromatography (HPLC) instrument. Analysis of AFs The quantification of AFs in sesame products was car- ried out by HPLC (Knauer-Germany), equipped with a Smartline Pump, fluorescence detector, reverse phased C18 (150 mm × 4.6 mm i.d and 5 μm particle size). In the fluorescence detector, an excitation wavelength of 333 nm and emission wavelength of 430 nm was applied for AFs determination. The mobile phase was water/methanol/ acetonitrile (6:102:94, v/v/v) and contained 100 mg KBr and 100 μL HNO3. Then, it is diverted into HPLC in the isocratic method with a flow rate of 0.8 mL/min. The col- umn temperature of HPLC was maintained at 40°C. The validation of AFs analysis method The linearity, accuracy, repeatability, limit of detection (LOD), and limit of quantification (LOQ) were deter- mined (Heshmati et al., 2017). Risk assessment For risk assessment of AFB1 intake through sesames seed, tahini, and tahini halva, the probabilistic approach was considered and estimated daily intake (EDI). The follow- ing equations are used to calculate the margin of expo- sure (MOE): EDI (ng/kg bw/day) = the concentration of AFB1 × average daily consumption (kg)/average body weight (kg) Per capita consumption of sesame in Iran is 3 g/day (Eghbaljoo-Gharehgheshlaghi et al., 2020). In this study, 70 kg is considered as the mean body weight for an adult in Iran. MOE = benchmark dose lower confidence limit 10% (BMDL10)/EDI where, BMDL10 is the lowest dose that is 95% certain to cause no more than 10% cancer prevalence. The EFSA Panel on Contaminants in the Food Chain (CONTAM) suggested 400 ng/kg BW/day for BMDL10 reference value (Chain et al., 2020). A MOE of 10,000 or larger has lit- tle concern for public health, while a MOE of less than 10,000 shows a potential danger for consumers (Heshmati et al., 2017). EDI and MOE were estimated by MCS using Crystal ball software (version 11.1.2.3 Oracle). Statistical analysis For statistical analysis, the SPSS software version 20 (IBM, PASW Statistics, USA) was applied. The mean and standard deviation of AFs levels in different samples were determined. One sample T-test was used to compare the mean of AFB1 and TAF with the allowable limit. The sig- nificant level was considered P < 0.05. One-way ANOVA and Tukey’s test were applied to determine the significant difference of moisture, fat, protein content, and extracted fat acidity levels of these samples. Result and Discussion The chemical properties of samples The moisture, fat, protein, and extracted fat acidity amount of sesame seed, tahini, and tahini halva samples are shown in Table 1. Sesame seed had a higher moisture, fat, and protein value than tahini and tahini halva samples. Method validation The LOD of AFs for sesame seeds, tahini, and tahini Halva samples ranged from 0.04 to 0.08, 0.05 to 0.09, and 0.07 to 0.08 µg/kg, respectively while LOQ for them ranged from 0.13 to 0.25, 0.18 to 0.31, and 0.21 to 0.27 µg/kg, respectively (Table 2). Moreover, the recovery range for AFs of sesame seeds, tahini, and tahini halva samples was 82.15–96.23, 77.36–95.63, and 83.65–98.65, respectively (Table 3). The determination coefficients (R2 > 0.992) of the Table 1. Chemical proprieties of sesame and related products. Parameters Sesame seed Tahini Tahini halva Moisture (%) 6.87 ± 0.38a 1.15 ± 0.07c 2.5 ± 0.06b Fat (%) 52.31 ± 2.89a 49.13 ± 2.12b 28.45 ± 2.32c Protein (%) 22.34 ± 1.87a 20.67 ± 1.76b 9.87 ± 0.75c Extracted fat acidity (%, in oleic acid) 0.82 ± 0.09a 0.90 ± 0.10a 0.76 ± 0.07a The different superscript letters within each row indicated significant differences (P < 0.05). Italian Journal of Food Science, 2021; 33 (SP1) 95 Prevalence and risk assessment of aflatoxin in sesame-based products Table 3. Recovery of aflatoxin from sesame and related products. Aflatoxin type Spiked level (µg/kg) Recovery ± RSD (%) Sesame seed Tahini Tahini halva AFB 1 0.5 82.15 ± 4.51 84.56 ± 10.36 87.65 ± 12.74 2 87.63 ± 8.56 88.32 ± 13.08 90.23 ± 15.32 5 85.25 ± 3.65 87.32 ± 8.98 85.32 ± 14.56 AFB 2 0.5 90.23 ± 10.23 79.65 ± 4.56 94.36 ± 12.02 1.5 92.35 ± 12.32 82.36 ± 7.25 98.65 ± 4.08 3 95.63 ± 15.36 77.36 ± 14.97 94.82 ± 6.23 AFG 1 0.5 89.69 ± 12.31 83.65 ± 4.56 88.63 ± 4.51 1.5 92.34 ± 8.96 86.53 ± 45.23 87.56 ± 14.23 3 94.23 ± 14.85 82.03 ± 12.78 83.65 ± 14.32 AFG 2 0.5 90.23 ± 11.57 90.23 ± 15.63 89.32 ± 4.36 1.5 96.23 ± 8.69 95.63 ± 10.56 90.56 ± 8.69 3 94.23 ± 10.23 92.53 ± 8.02 95.36 ± 10.26 AF: aflatoxin; RSD: Relative standard deviation Table 2. Validated parameters for aflatoxin analysis in sesame and related products. Aflatoxin type Equation of calibration curve Range of linearity (ng/mL) R2 Sesame seed Tahini Tahini halva LOD LOQ LOD LOQ LOD LOQ AFB 1 Y = 13562.23X + 456.03 0.15–25 0.998 0.06 0.21 0.07 0.23 0.08 0.27 AFB 2 Y =10623.15X – 4856.45 0.25–20 0.997 0.08 0.25 0.09 0.31 0.07 0.22 AFG 1 Y = 20354.67X + 120.09 0.16–20 0.996 0.06 0.22 0.05 0.18 0.08 0.26 AFG 2 Y = 14600.18X – 809.53 0.12–25 0.992 0.04 0.13 0.06 0.19 0.07 0.21 LOD and LOQ in µg/kg. AF: aflatoxin; LOD: limit of detection; LOQ: limit of quantification. regression equations showed acceptable linearity, and good recovery was obtained for spike samples and was similar to values reported in previous studies (Heshmati et al., 2017). The findings obtained during method validation were con- formed with accepted criteria (AOAC International, 2002). The prevalence of AFs in sesame seeds In this study, 40 samples of sesame seeds were analyzed for AFs. Sesame seeds had the highest prevalence (55%) of total AFs among the three analyzed samples. The detec- tion rates of AFs were higher in sesame seed samples than in the tahini and tahini halva. AFB1, AFB2, AFG1, and AFG2 were detected in 19 (47.5%), 5 (12.5%), 6 (15%), and 5 (12.5%) of 40 sesame seeds samples, respectively (Table  4). AFB1 was the most abundant AF, and its level varied from 0.21 to 12.35 μg/kg. In addition, 10 and 6 sam- ples contained AFB1 more than the accepted limit accord- ing to European (2 µg/kg) and Iranian standard (5 µg/kg), respectively, and the total AFs content of samples was lower than the permitted level in Iran (15 mg/kg). There are several reports of AF contamination in ses- ame seeds (Table 5). The different levels of total AFs and AFB1 have been reported in previous similar stud- ies. For example, Anthony et al. (2014) reported that 8 (26.67%) out of 30 sesame samples studied in Nigeria were contaminated with AFB1 at levels above the limit of European regulations (Anthony et al., 2014). Esan et al. (2020) surveyed the contamination of total AFs in ses- ame seeds of Nigeria. They demonstrated that the pos- itive samples contaminated with total AFs ranged from 0.29 to 88.5  µg/kg (Esan et al., 2020). In another study, Kollia et al. (2016) investigated 30 samples of sesame seeds from the Greek market. They observed that the amount of AFB1 in eight samples exceeded the limit of European regulations (Kollia et al., 2016). In an investi- gation, among 28 samples of sesame products from Egypt by Sabry et al. (2016), a higher prevalence of AFB1 and AFG1 than other mycotoxins in the range of 60%–100% and 33.33%–100%, respectively, were reported. The mean range of AFB1 and AFG1 in different provinces was 18.63–66.79 and 14.88–51.47 g/kg, respectively (Sabry et al., 2016). 96 Italian Journal of Food Science, 2021; 33 (SP1) Heshmati A et al. (2020) in Egypt, mean AFB1 in 16 samples of branded tahini and 101 samples of local tahini reported 0.10 ± 0.24 μg/kg and 13 ± 19.3 μg/kg, respectively (Sebaei et al., 2020). Li et al. (2009) reported that 19% and 32% of 100 of the sesame paste (Tahini) samples studied in China contained AFB1 at levels higher than the Chinese regula- tion (5 μg/kg) and European regulations (Li et al., 2009). In addition, Torlak and Akan (2013) studied AFs contam- ination in 104 samples of tahini from the Anatolia region of Turkey. The mean of AFB1 and total AFs of samples was 0.93 ± 0.62 μg/kg and 1.17 ± 0.55 μg, respectively, which both were lower than the Turkish standard (AFB1: 5 μg/kg and TAF: 10 μg/kg) and our study. These authors indicated that the roasting process applied in tahini pro- duction does not eliminate AFs (Torlak and Akan, 2013). For the traditional production of tahini, sesame is roasted for 2 h at a temperature of 100°C to 150°C (Torlak and Akan, 2013). AFs are resistant to heat and are difficult to eliminate at insufficient temperatures. In general, AFs are eliminated at 237°C to 306°C. In foods heated, import- ant parameters determining the reduction of AF include Table 4. The contamination status of aflatoxin of sesame and related products. Aflatoxin type Contamination status Sesame seed Tahini Tahini halva AFB 1 Contamination level (µg/kg) No. of contaminated samples 19 (47.5) 14 (35) 11 (27.5) Mean ± SD (µg/kg) 1.67 ± 0.45 0.85 ± 0.24 0.55 ± 0.17 0.15–2 9 (22.5) 7 (17.5) 5 (12.5) 2–5 4 (10) 4 (10) 6 (15) >5 6 (15) 3 (7.5) 0 Range (µg/kg) 0.21–12.35 0.23–5.81 0.27–3.56 AFB 2 No. of contaminated samples 5 (12.5) 3 (7.5) 4 (7.5) Mean ± SD (µg/kg) 0.13 ± 0.06 0.1 ± 0.06 0.07 ± 0.03 Range (µg/kg) 0.25–1.62 0.31–1.75 0.22–0.92 AFG 1 No. of contaminated samples 6 (15) 4 (10) 2 (5) Mean ± SD (µg/kg) 0.10 ± 0.05 0.07 ± 0.04 0.04 ± 0.03 Range (µg/kg) 0.22–1.41 0.18–1.32 0.26–1.02 AFG 2 No. of contaminated samples 5 (12.5) 5 (12.5) 4 (10) Mean ± SD (µg/kg) 0.05 ± 0.02 0.08 ± 0.04 0.06 ± 0.03 Range (µg/kg) 0.13–0.85 0.19–1.02 0.21–0.74 TAF Contamination level (µg/kg) No. of contaminated samples 22 (55) 18 (45) 13 (32.5) Mean ± SD (µg/kg) 1.95 ± 0.48 1.10 ± 0.30 0.72 ± 0.22 >4 15 (37.5) 14 (35) 11 (27.5) 4–15 7 (17.5) 8 (20) 2 (5) >15 0 0 0 AF: aflatoxin. The prevalence of AFs in tahini A lower rate of AF contamination was observed in tahini than that in sesame seed. Eighteen (45%) of Tahini sam- ples contained total AFs. The highest prevalence of AFs in tahini halva was AFB1 (35%), followed by AFG2 (12.5%), AFG1 (10%), and AFB2 (7.5%). It was observed that the AFB1 value of seven samples was higher than the limit of European regulations (2 µg /kg) while only three samples had AFB1 contamination higher than the recommended standard of Iran (5 µg/kg). Also, 18 samples contained total AF with an average value of 1.10 ± 0.30 µg/kg, among which the concentration of eight samples was higher than the limit of European regulations (4 µg/kg). It seems that washing and peeling of sesame seeds before tahini prepa- ration could reduce the amount of AFs. Before tahini preparation, sesame seeds were roasted. The previous studies indicated that roasting operation could cause AFs degradation (Emadi et al., 2021; Yazdanpanah et al., 2005). The prevalence of AFs in tahini was reported in other studies. For example, in a study performed by Sebaei et al. Italian Journal of Food Science, 2021; 33 (SP1) 97 Prevalence and risk assessment of aflatoxin in sesame-based products related to the dilution effect of other products, including sugar, emulsifier, in the formulation. Few studies have been performed to evaluate AFs in tahini halva. Var et al. (2007) investigated AFB1 contamination in 34 samples of helva in Turkey. No AFB1 was found in helva samples. It seemed the discrepancies in AFs contamination preva- lence in different studies could be due to differences in sampling geographical areas and storage conditions and the AFs measuring method. Risk assessment of AFB1 intake Findings of percentile 50 (as the median of the popula- tion) of EDI of AFB1 calculated using the MCS approach showed the lowest (0.02 ng/kg bw/day) and highest (0.05 ng/kg bw/day) of EDI in sesame seed and tahini halva, respectively (Figure 1). The percentile 95 of EDI of AFB1 through sesame seed, tahini, and tahini halva was 0.09, 0.05, and 0.03 ng/kg bw/day, respectively. As shown in Figure 2, the percentile 95 of MOE with AFB1 through sesame seed, tahini, and tahini halva was calculated as 25,485, 50,092, and 77,114, respectively. Because data Table 5. The contamination status of aflatoxin of sesame and related products. Country Sample type No of samples Positive n (%) Method Mycotoxin Range (µg/kg) Mean ± SD (μg/kg) Reference Iran Sesame seeds 269 50% HPLC TAF 1.43 ± 4.38 Hosseininia et al. 2014 Iran Sesame seeds 269 50% HPLC AFB 1 1.25 ± 3.7 Hosseininia et al. 2014 Egyptian Tahini (Brand) 16 HPLC AFB 1 0.10 ± 0.2 Sebaei et al. 2020 Egyptian Tahini (Local) 101 HPLC AFB 1 13 ± 19.3 Sebaei et al. 2020 China Sesame paste 100 37 (37%) LC TAF 0.54–56.89 6.75 Li et al. 2009 China Sesame paste 100 37 (37%) LC AFB 1 0.39–20.45 4.31 Li et al. 2009 Iran Sesame seeds 182 33 (18.1%) LC AFB 1 1.62 ± 1.32 Asadi et al. 2011 Malaysian Sesame seeds 8 7 (87.5%) ELISA AFB 1 0.5–1.82 0.9 Reddy et al. 2011 Nigeria Sesame seeds 60 12 LCMS/MS TAF 0.29–88.5 16.9 Esan et al. 2020 Nigeria Sesame seeds 60 12 LCMS/MS AFB 1 0.29–79.3 14.8 Esan et al. 2020 Nigeria Sesame seeds 30 26.66 HPLC AFB 1 14.71–140.90 69.72 ± 41.68 Anthony et al. 2014 Turkey Tahini 104 14.42 HPLC AFB 1 0.93 ± 0.62 Torlak and Akan 2013 Turkey Tahini 104 15.38 HPLC TAF 1.17 ± 0.55 Torlak and Akan 2013 Turkey Tahini Helva 34 0 TLC AFB 1 <1 <1 Var et al. 2007 Egyptian Sesame seeds 28 88.89 HPLC AFB 1 33.66 ± 1.35 Sabry et al. 2016 FCT, Abuja, Nigeria Sesame seeds 24 13 LCMS/MS AFB 1 3.6 Fapohunda et al. 2018 Nigeria Sesame seeds 46 23 (50%) TLC AFB 1 0.79–37.25 Apeh et al. 2016 Japan Sesame seeds 47 5 HPTLC AFB 1 0.6–2.4 Tabata 2007 Jordan Sesame seeds 46 2 HPLC TAF 100–1280 Sirhan et al. 2014 Greek Sesame seeds 30 77.60% HPLC AFB 1 2.0 ng AFB1 g-1 Kollia et al. 2016 moisture content, heating temperature, and nutrient media. It has been reported that roasting dried wheat at 150°C for 30 min, Green coffee beans at 150°C to 180°C for 10 to 15 min, and Pistachio nuts at 150°C for 30 min will reduce AF levels to 50%, 42.2%–55/9%, and 63%, respectively (Pankaj et al., 2018). The prevalence of AFs in tahini halva The results showed that tahini halva samples contained four types of AF, i.e., AFB1, AFB2, AFG2, and AFG2, with mean values of 0.55 ± 0.17, 0.07 ± 0.03, 0.04 ± 0.03, and 0.06 ± 0.03 μg/kg, respectively (Table 4). Except for AFB2, the detection rate of AFB1, AFG2, and AFG2 was lower in tahini halva samples than in the sesame seeds and tahini samples. The average total AFs of tahini halva samples was 0.72 ± 0.22 µg/kg. According to Iranian standards, the concentration of total AFs and AFB1 was not higher than the allowable limit. In contrast, the total AFs level of two samples was higher than the limit of European regu- lations (4 μg/kg). Also, in six samples, the concentration of AFB1 was higher than the limit of European regulations (2 µg/kg). The lower level of AFs in tahini halva could be 98 Italian Journal of Food Science, 2021; 33 (SP1) Heshmati A et al. Sesame seed 1.00 EDI 0.90 0.80 0.70 0.60 C um ul at iv e pr ob ab ili ty 0.50 0.40 0.30 0.20 0.10 0.00 0.00 0.01 0.02 0.03 0.04 0.05 ng/kg bw/day 0.06 0.07 0.08 0.09 0.10 0.11 Mean = 0.05 95% = 0.09 Tahini 1.00 EDI 0.90 0.80 0.70 0.60 C um ul at iv e pr ob ab ili ty 0.50 0.40 0.30 0.20 0.10 0.00 0.00 0.01 0.02 ng/kg bw/day 0.03 0.04 0.05 0.06 Mean = 0.03 95% = 0.05 Tahini halva 1.00 EDI 0.90 0.80 0.70 0.60 C um ul at iv e pr ob ab ili ty 0.50 0.40 0.30 0.20 0.10 0.00 0.00 0.01 0.02 ng/kg bw/day 0.03 0.04 Mean = 0.02 95% = 0.03 Figure 1. Cumulative probability plot of EDI of AFB1 through sesame seed, tahini, and tahini halva consumption. Italian Journal of Food Science, 2021; 33 (SP1) 99 Prevalence and risk assessment of aflatoxin in sesame-based products Sesame seed 1.00 MOE 0.90 0.80 0.70 0.60 C um ul at iv e pr ob ab ili ty 0.50 0.40 0.30 0.20 0.10 0.00 4000 8000 12000 16000 20000 24000 28000 32000 36000 40000 44000 48000 Mean = 11121 95% = 25485 Tahini 1.00 MOE 0.90 0.80 0.70 0.60 C um ul at iv e pr ob ab ili ty 0.50 0.40 0.30 0.20 0.10 0.00 10000 20000 30000 40000 50000 60000 70000 80000 90000 100000 110000 Mean = 21713 95% = 50092 Tahini halva 1.00 MOE 0.90 0.80 0.70 0.60 C um ul at iv e pr ob ab ili ty 0.50 0.40 0.30 0.20 0.10 0.00 20000 40000 60000 80000 100000 120000 140000 Mean = 33716 95% = 77114 Figure 2. Cumulative probability plot of MOE of AFB 1 through sesame seed, tahini, and tahini halva consumption. 100 Italian Journal of Food Science, 2021; 33 (SP1) Heshmati A et al. regarding the percentile 50 and 95 of MOE was more than 10,000 (Heshmati et al., 2017), it could be concluded that AFB1 intake through sesame seed, tahini, and tahini halva consumption has no remarkable cancer risk for adults. Conclusions The results of this study indicated the high prevalence of AFs in sesame seed (55%), tahini (45%), and tahini halva (32.5%) samples. In addition, the levels of TAF in 7 (17.5%), 8 (20%), and 2 (5%) samples of sesame seeds, tahini, and tahini halva exceeded the limit of European regulations (4 µg/kg), respectively. Furthermore, 10 (25%), 7 (17.5%), and 6 (15%) samples of sesame seeds, tahini, and tahini halva contained AFB1 more than the limit of European regulations (2 µg/kg), respectively. However, risk assessment indicated that the intake of AF through the consumption of mentioned products had no remarkable cancer risk for adults. Acknowledgments The ethical and scientific committee of Hamadan University of Medical Science approved this study (ethical code: IR.UMSHA.REC.1400.177). Conflict of interest The authors declare that they have no conflict of interest. References Anilakumar, K.R., Pal, A., Khanum, F. and Bawa, A.S., 2010. Nutritional, medicinal and industrial uses of sesame (Sesamum indicum L.) seeds–an overview. Agriculturae Conspectus Scientificus 75(4): 159–168. Anthony, M.H., Ojochenemi, A.D., Yemi, A.H.R., Tahir, N., Okechukwu, O.J., Saidu, M.A., et al. 2014. Determination of aflatoxins in sesame, rice, millet and acha from Nigeria using HPLC. Chemical Science Transactions 3(4): 1516–1524. https:// doi.org/10.7598/cst2014.749 AOAC International, 2002. AOAC Guidelines for single laboratory validation of chemical methods for dietary supplements and botanicals. AOAC International, Gaithersburg, MD, USA. Apeh, D., Ochai, D., Adejumo, A., Muhammad, H., Saidu, A., Atehnkeng, J., et al. 2016. Mycotoxicological concerns with sor- ghum, millet and sesame in Northern Nigeria. Analytical and Bioanalytical Chemistry 7: 336. https://doi.org/10.4172/2155- 9872.1000336 Asadi, M., Beheshti, H.R. and Feizy, J., 2011. A survey of aflatoxins in sesame in Iran. Mycotoxin Research 27(4): 259. https://doi. org/10.1007/s12550-011-0102-y Batrinou, A., Houhoula, D. and Papageorgiou, E., 2020. Rapid detec- tion of mycotoxins on foods and beverages with enzyme linked immunosorbent assay. Quality Assurance and Safety of Crops & Foods 12(1): 40–49. https://doi.org/10.15586/QAS2019.654 Chain, E.P.O.C.I.T.F., Schrenk, D., Bignami, M., Bodin, L., Chipman,  J.K., Del Mazo, J., et al. 2020. Risk assessment of aflatoxins in food. EFSA Journal18(3): e06040. https://doi. org/10.2903/j.efsa.2020.6040 Cui, P., Yan, H., Granato, D., Ho, C.-T., Ye, Z., Wang, Y., et al. 2020. Quantitative analysis and dietary risk assessment of aflatoxins in Chinese post-fermented dark tea. Food and Chemical Toxicology 146: 111830. https://doi.org/10.1016/j.fct.2020.111830 De Souza, C., Khaneghah, A.M. and Oliveira, C.A.F., 2021. The occur- rence of aflatoxin M1 in industrial and traditional fermented milk: a systematic review study. Italian Journal of Food Science 33(SP1): 12–23. https://doi.org/10.15586/ijfs.v33iSP1.1982 Di Sanzo, R., Carabetta, S., Campone, L., Bonavita, S., Iaria, D., Fuda, S., et al. 2018. Assessment of mycotoxins co-occurrence in Italian dried figs and in dried figs-based products. Journal of Food Safety 38(6): e12536. https://doi.org/10.1111/jfs.12536 Dravie, E.E., Kortei, N.K., Essuman, E.K., Tettey, C.O., Boakye, A.A. and Hunkpe, G., 2020. Antioxidant, phytochemical and physi- cochemical properties of sesame seed (Sesamum indicum L). Scientific African 8: e00349. https://doi.org/10.1016/j.sciaf.2020. e00349 Eghbaljoo-Gharehgheshlaghi, H., Shariatifar, N., Arab, A., Alizadeh- Sani, M., Sani, I.K., Asdagh, A., et al. 2020. The concentration and probabilistic health risk assessment of trace metals in three type of sesame seeds using ICP-OES in Iran. International Journal of Environmental Analytical Chemistry 1–15. https:// doi.org/10.1080/03067319.2020.1804896 Elleuch, M., Bedigian, D. and Zitoun, A., 2011. Sesame (Sesamum indicum L.) seeds in food, nutrition, and health. In: Victor R. Preedy, Ronald Ross Watson, Vinood B. Patel (Eds.), Nuts and seeds in health and disease prevention. Elsevier, Academic Press, London, pp. 1029–1036. https://doi.org/10.1016/ B978-0-12-375688-6.10122-7 Elzupir, A.O., Suliman, M.A., Ibrahim, I.A., Fadul, M.H. and Elhussein, A.M., 2010. Aflatoxins levels in vegetable oils in Khartoum State, Sudan. Mycotoxin Research. 26(2): 69–73. https://doi.org/10.1007/s12550-010-0041-z Emadi, A., Jayedi, A., Mirmohammadkhani, M. and Abdolshahi, A., 2021. Aflatoxin reduction in nuts by roasting, irradiation and fumigation: a systematic review and meta-analysis. Critical Reviews in Food Science and Nutrition 1–11. https://doi.org/10. 1080/10408398.2021.1881436 Esan, A.O., Fapohunda, S.O., Ezekiel, C.N., Sulyok, M. and Krska, R., 2020. Distribution of fungi and their toxic metabolites in melon and sesame seeds marketed in two major producing states in Nigeria. Mycotoxin Research 36(4): 361–369. https:// doi.org/10.1007/s12550-020-00400-0 Fapohunda, S.O., Anjorin, T.S., Sulyok, M. and Krska, R., 2018. Profile of major and emerging mycotoxins in sesame and soy- bean grains in the Federal Capital Territory, Abuja, Nigeria European Journal of Biology Research 8(3): 121–130. https://doi. org/10.5281/zenodo.1307184 https://doi.org/10.7598/cst2014.749� https://doi.org/10.7598/cst2014.749� https://doi.org/10.4172/2155-9872.1000336� https://doi.org/10.4172/2155-9872.1000336� https://doi.org/10.1007/s12550-011-0102-y� https://doi.org/10.1007/s12550-011-0102-y� https://doi.org/10.15586/QAS2019.654� https://doi.org/10.2903/j.efsa.2020.6040� https://doi.org/10.2903/j.efsa.2020.6040� https://doi.org/10.1016/j.fct.2020.111830� https://doi.org/10.15586/ijfs.v33iSP1.1982� https://doi.org/10.1111/jfs.12536� https://doi.org/10.1016/j.sciaf.2020.e00349� https://doi.org/10.1016/j.sciaf.2020.e00349� https://doi.org/10.1080/03067319.2020.1804896� https://doi.org/10.1080/03067319.2020.1804896� https://doi.org/10.1016/B978-0-12-375688-6.10122-7� https://doi.org/10.1016/B978-0-12-375688-6.10122-7� https://doi.org/10.1007/s12550-010-0041-z� https://doi.org/10.1080/10408398.2021.1881436� https://doi.org/10.1080/10408398.2021.1881436� https://doi.org/10.1007/s12550-020-00400-0� https://doi.org/10.1007/s12550-020-00400-0� https://doi.org/10.5281/zenodo.1307184� https://doi.org/10.5281/zenodo.1307184� Italian Journal of Food Science, 2021; 33 (SP1) 101 Prevalence and risk assessment of aflatoxin in sesame-based products Gholami, M., Davan, S.T., Gholami, M., Bolandparvaz, S., Gholami, M., Chamanpara, P., et al. 2020. Effects of topical ses- ame oil extracted from Tahini (Ardeh) on pain severity in trauma patients: A randomized double-blinded placebo- controlled clin- ical trial. Bulletin of Emergency and Trauma 8(3): 179. http:// dx.doi.org/10.30476/BEAT.2020.82561 Grumi, M., Kunova, A., Isotti, M., Barbiroli, A. and Pasquali, M., 2020. Occurrence of deoxynivalenol in beers commercialised in Italy. Italian Journal of Food Science 32(3): 712–720. https://doi. org/10.14674/IJFS-1826 Heshmati, A., Ghadimi, S., Ranjbar, A. and Khaneghah, A.M., 2019. Changes in aflatoxins content during processing of pekmez as a traditional product of grape. LWT 103: 178–185. https://doi. org/10.1016/j.lwt.2019.01.001 Heshmati, A., Zohrevand, T., Khaneghah, A.M., Nejad, A.S.M. and Sant’ana, A.S., 2017. Co-occurrence of aflatoxins and ochra- toxin A in dried fruits in Iran: dietary exposure risk assess- ment. Food and Chemical Toxicology 106: 202–208. https://doi. org/10.1016/j.fct.2017.05.046 Hosseininia, A.R., Vahabzadeh, M., Rashedinia, M., Riahi-Zanjani, B. and Karimi, G., 2014. A survey of aflatoxins in sesame seeds imported into Khorasan Province, Iran. Mycotoxin Research 30(1): 43–46. https://doi.org/10.1007/s12550-013-0186-7 INSO, 2020. Iranian National Standardization Organization. Food and feed – maximum tolerated level of mycotoxins. No: 5925. First Revision. Iranian National Standardization Organization., Karaj, Iran. Javanmardi, F., Khodaei, D., Sheidaei, Z., Bashiry, M., Nayebzadeh, K., Vasseghian, Y., et al. 2020. Decontamination of aflatoxins in edi- ble oils: A comprehensive review. Food Reviews International 1–17. https://doi.org/10.1080/87559129.2020.1812635 Khaneghah, A.M., Fakhri, Y. and Sant’ana, A.S., 2018. Impact of unit operations during processing of cereal-based products on the levels of deoxynivalenol, total aflatoxin, ochratoxin A, and zearalenone: a systematic review and meta-analysis. Food Chemistry 268: 611– 624. https://doi.org/10.1016/j.foodchem.2018.06.072 Kollia, E., Tsourouflis, K. and Markaki, P., 2016. Aflatoxin B1 in ses- ame seeds and sesame products from the Greek market. Food Additives & Contaminants Part B 9(3): 217–222. https://doi.org/ 10.1080/19393210.2016.1179349 Lee, B.M., Lee, E.M., Kang, D.J., Seo, J.-A., Choi, H.-K., Kim, Y.-S., et  al. 2020. Discovery study of integrative metabolic profiles of sesame seeds cultivated in different countries. LWT 129: 109454. https://doi.org/10.1016/j.lwt.2020.109454 Li, F.-Q., Li, Y.-W., Wang, Y.-R. and Luo, X.-Y., 2009. Natural occur- rence of aflatoxins in Chinese peanut butter and sesame paste. Journal of Agricultural and Food Chemistry 57(9): 3519–3524. https://doi.org/10.1021/jf804055n Milicevic, B., Tomović, V., Danilović, B. and Savić, D., 2021. The influence of starter cultures on the lactic acid bacteria microbi- ota of Petrovac sausage. Italian Journal of Food Science 33(2): 24–34. https://doi.org/10.15586/ijfs.v33i2.1918 Mokhtarian, M., Tavakolipour, H., Bagheri, F., Oliveira, C.A.F., Corassin, C.H. and Khaneghah, A.M., 2020. Aflatoxin B1 in the Iranian pistachio nut and decontamination methods: a systematic review. Quality Assurance and Safety of Crops & Foods 12(4): 15–25. https://doi.org/10.15586/qas.v12i4.784 Mozaffari Nejad, A.S., Heshmati, A. and Ghiasvand, T., 2020. The occurrence and risk assessment of aflatoxin M1 in cheeses sam- ples from Hamadan, Iran. Iranian Journal of Pharmaceutical Research 19(4): 44–50. https://doi.org/10.22037/ijpr.2020. 112399.13754 Namiki, M., 2007. Nutraceutical functions of sesame: a review. Critical Reviews in Food Science and Nutrition 47(7): 651–673. https://doi.org/10.1080/10408390600919114 Öğütcü, M., Arifoğlu, N. and Yılmaz, E., 2017. Restriction of oil migration in tahini halva via organogelation. European Journal of Lipid Science and Technology 119(9): 1600189. https://doi. org/10.1002/ejlt.201600189 Osaili, T.M., Al-Nabulsi, A.A. and Aljaafreh, T.F., 2018a. Use of gamma radiation for inactivating Salmonella spp., Escherichia coli O157: H7 and Listeria monocytogenes in tahini halva. International Journal of Food Microbiology 278: 20–25. https:// doi.org/10.1016/j.ijfoodmicro.2018.04.029 Osaili, T.M., Al-Nabulsi, A.A., Aljaafreh, T.F. and Olaimat, A.N., 2018b. Use of gamma radiation to inactivate stressed Salmonella spp., Escherichia coli O157: H7 and Listeria monocytogenes in tahini halva. LWT 98: 438–443. https://doi.org/10.1016/j. lwt.2018.09.017 Pankaj, S., Shi, H. and Keener, K.M., 2018. A review of novel phys- ical and chemical decontamination technologies for aflatoxin in food. Trends in Food Science & Technology 71: 73–83. https:// doi.org/10.1016/j.tifs.2017.11.007 Rapa, M., Ciano, S., Gobbi, L., Ruggieri, R. and Vinci, G., 2021. Quality and safety evaluation of new tomato cultivars. Italian Journal of Food Science 33(2): 35–45. https://doi.org/10.15586/ ijfs.v33i2.1921 Reddy, K.R., Farhana, N.I. and Salleh, B., 2011. Occurrence of Aspergillus spp. and aflatoxin B1 in Malaysian foods used for human consumption. Journal of Food Science 76(4): T99–T104. https://doi.org/10.1111/j.1750-3841.2011.02133 Sabry, B.A., Hathout, A.S., Nooh, A., Aly, S.E. and Shehata, M.G., 2016. The prevalence of aflatoxin and Aspergillus parasiticus in Egyptian sesame seeds. International Journal of Chemtech Research 9(11): 308–319. Sebaei, A.S., Refai, H.M., Elbadry, H.T. and Armeya, S.M., 2020. First risk assessment report of aflatoxins in Egyptian tahini. Journal of Food Composition and Analysis 92: 103550. https:// doi.org/10.1016/j.jfca.2020.103550 Sirhan, A.Y., Tan, G.H., Al-Shunnaq, A., Abdulra’uf, L. and Wong,  R.C., 2014. QuEChERS-HPLC method for aflatoxin detection of domestic and imported food in Jordan. Journal of Liquid Chromatography & Related Technologies 37(3): 321– 342. https://doi.org/10.1080/10826076.2012.745138 Tabata, S., 2007. Mycotoxin contamination in foods and foodstuffs in Japan Aflatoxins. JSM Mycotoxins 2006(Suppl 4): 123–129. https://doi.org/10.2520/myco1975.2006.Suppl4_123 Torlak, E. and Akan, I., 2013. Aflatoxin contamination in tahini. Quality Assurance and Safety of Crops & Foods 5(3): 221–225. https://doi.org/10.3920/QAS2012.0131 http://dx.doi.org/10.30476/BEAT.2020.82561� http://dx.doi.org/10.30476/BEAT.2020.82561� https://doi.org/10.14674/IJFS-1826� https://doi.org/10.14674/IJFS-1826� https://doi.org/10.1016/j.lwt.2019.01.001� https://doi.org/10.1016/j.lwt.2019.01.001� https://doi.org/10.1016/j.fct.2017.05.046� https://doi.org/10.1016/j.fct.2017.05.046� https://doi.org/10.1007/s12550-013-0186-7� https://doi.org/10.1080/87559129.2020.1812635� https://doi.org/10.1016/j.foodchem.2018.06.072� https://doi.org/10.1080/19393210.2016.1179349� https://doi.org/10.1080/19393210.2016.1179349� https://doi.org/10.1016/j.lwt.2020.109454� https://doi.org/10.1021/jf804055n� https://doi.org/10.15586/ijfs.v33i2.1918� https://doi.org/10.15586/qas.v12i4.784� https://doi.org/10.22037/ijpr.2020.112399.13754� https://doi.org/10.22037/ijpr.2020.112399.13754� https://doi.org/10.1080/10408390600919114� https://doi.org/10.1002/ejlt.201600189� https://doi.org/10.1002/ejlt.201600189� https://doi.org/10.1016/j.ijfoodmicro.2018.04.029� https://doi.org/10.1016/j.ijfoodmicro.2018.04.029� https://doi.org/10.1016/j.lwt.2018.09.017� https://doi.org/10.1016/j.lwt.2018.09.017� https://doi.org/10.1016/j.tifs.2017.11.007� https://doi.org/10.1016/j.tifs.2017.11.007� https://doi.org/10.15586/ijfs.v33i2.1921� https://doi.org/10.15586/ijfs.v33i2.1921� https://doi.org/10.1111/j.1750-3841.2011.02133� https://doi.org/10.1016/j.jfca.2020.103550� https://doi.org/10.1016/j.jfca.2020.103550� https://doi.org/10.1080/10826076.2012.745138� https://doi.org/10.2520/myco1975.2006.Suppl4_123� https://doi.org/10.3920/QAS2012.0131� 102 Italian Journal of Food Science, 2021; 33 (SP1) Heshmati A et al. Var, I., Kabak, B. and Gök, F., 2007. Survey of aflatoxin B1 in helva, a traditional Turkish food, by TLC. Food Control 18(1): 59–62. https://doi.org/10.1016/j.foodcont.2005.08.008 Wang, Y.-J., Nie, J.-Y., Zhen, Y., Li, Z.-X., Cheng, Y. and Farooq, S., 2018. Multi-mycotoxin exposure and risk assessments for Chinese consumption of nuts and dried fruits. Journal of Integrative Agriculture 17(7): 1676–1690. https://doi. org/10.1016/S2095-3119(18)61966-5 Xinyu, Z., Yi, W., Xue, H., Zixi, Y., Weiqiong, Y. and Zhaolin, L., 2020. Characterization of volatile compounds in five blueberry varieties using purge and trap coupled to gas chromatogra- phy-mass spectrometry. Italian Journal of Food Science 32(2): 482–497. https://doi.org/10.14674/IJFS-1675 Yao, Y.-G., Wang, W.-Y., Chen, L.-Y., Liu, H.-M., Yan, R.-Z., Li, S., et al. 2021. Structural changes of cellulosic polysaccharides in sesame hull during roasting at various temperatures. Quality Assurance and Safety of Crops & Foods 13(2): 13–24. https:// doi.org/10.15586/qas.v13i2.876 Yazdanpanah, H., Mohammadi, T., Abouhossain, G. and Cheraghali,  A.M., 2005. Effect of roasting on degradation of aflatoxins in contaminated pistachio nuts. Food and Chemical Toxicology 43(7): 1135–1139. https://doi.org/10.1016/j.fct. 2005.03.004 Zebib, H., Bultosa, G. and Abera, S., 2015. Physico-chemical prop- erties of sesame (Sesamum indicum L.) varieties grown in Northern Area, Ethiopia. Agricuture Science 6(02): 238–246. https://doi.org/10.4236/as.2015.62024 https://doi.org/10.1016/j.foodcont.2005.08.008� https://doi.org/10.1016/S2095-3119(18)61966-5� https://doi.org/10.1016/S2095-3119(18)61966-5� https://doi.org/10.14674/IJFS-1675� https://doi.org/10.15586/qas.v13i2.876� https://doi.org/10.15586/qas.v13i2.876� https://doi.org/10.1016/j.fct.2005.03.004� https://doi.org/10.1016/j.fct.2005.03.004� https://doi.org/10.4236/as.2015.62024� _Hlk71474928