Dietary exposure to aflatoxins, ochratoxin A and ...

FCT 8058

No. of Pages 9, Model 5G

11 August 2014 Food and Chemical Toxicology xxx (2014) xxx–xxx 1

Contents lists available at ScienceDirect

Food and Chemical Toxicology journal homepage: www.elsevier.com/locate/foodchemtox 5 6

Dietary exposure to aflatoxins, ochratoxin A and deoxynivalenol from a total diet study in an adult urban Lebanese population

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a

Lebanese Agricultural Research Institute, Lebanon Laboratoire d’Evaluation du Risque Chimique pour le Consommateur (LERCCo), Université de Bretagne Occidentale (UEB-UBO), UFR Sciences et Techniques, France c Department of Nutrition and Food Science, Faculty of Agricultural and Food Sciences, American University of Beirut, Lebanon d ARVALIS – Institut du Végétal, Pôle Analyses et Méthodes, Station Expérimentale, 91720 Boigneville, France e Université Paris-Sud, Inserm UMR-S 996, 92296 Châtenay-Malabry, France b

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F. Raad a,b, L. Nasreddine c,⇑, C. Hilan a, M. Bartosik d, D. Parent-Massin b,e

a r t i c l e Q3

i n f o

Article history: Received 31 December 2013 Accepted 23 July 2014 Available online xxxx Keywords: Dietary exposure Total diet study Mycotoxins Adults Risk characterization Lebanon

a b s t r a c t Exposure to mycotoxins may be associated with carcinogenic, immunosuppressant and estrogenic effects. In the Middle-East, studies investigating food contamination and dietary exposure to mycotoxins are particularly scarce. This study aims at evaluating the dietary exposure of an adult Lebanese urban population to four mycotoxins (AFB1, AFM1, OTA, DON) classified as priority food contaminants by the WHO. Dietary exposure assessment was performed by means of the total diet study approach. Average and excessive consumer exposure estimates (p95) were calculated and compared with appropriate toxicological reference values (TRVs). Average dietary exposure levels to OTA and DON represented 29.9% and 156.8% of the respective TRVs, with the p95 exposure estimates approaching or exceeding the TRVs for these mycotoxins (95.1% and 355.8%, respectively). Based on the mean dietary exposure level to AFB1, cancer risk was estimated at 0.0527–0.0545 cases/100,000 persons/year, while mean exposure to AFM1 was associated with a population risk of 0.0018–0.0027 cases/100,000 persons/year. The study’s findings place Lebanon among countries that are highly exposed to mycotoxins through the diet and call for larger-scale studies aiming at providing a comprehensive assessment of the dietary exposure of the Lebanese population to mycotoxins as well as to other food contaminants. Ó 2014 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).

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1. Introduction

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Human exposure to toxic chemicals may be associated with a wide array of human health disorders such as reproductive disorders and birth defects, immune system suppression, kidney and liver dysfunction, mental health problems and promoting some types of cancer (WHO, 2005). One of the main routes of human exposure to contaminants is the diet, with most foods potentially containing natural or synthetic chemicals that could present a toxic hazard for the consumer. Food contamination by mycotoxins has been recognized as a public health threat (JECFA, 2002). Mycotoxins have been included as priority food contaminants by the Global Environment Monitoring System/Food Contamination Monitoring Assessment Programme (GEMS/Food) of the WHO (2002). Mycotoxins are usually thermostable, tend to persist during the transformation

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⇑ Corresponding author. Address: Department of Nutrition and Food Science, Faculty of Agricultural and Food Sciences, American University of Beirut, P.O. Box 11-0236, Riad El Solh 1107 2020 Beirut, Lebanon. Tel.: +961 1350000. E-mail address: [email protected] (L. Nasreddine).

and processing of contaminated plants and are usually not eliminated during cooking and sterilization (Turner et al., 2009; Sirot et al., 2013). Given their capacity to bind to plasma proteins, mycotoxins can persist in organisms if the exposure is repeated or chronic (Galtier et al., 2006). Due to their diverse chemical structures and biosynthetic origins, mycotoxins have a myriad of biological effects with some being classified as carcinogenic, immunosuppressant or estrogenic, thus potentially causing severe metabolic disorders in humans (Richard et al., 2003; AFSSA, 2009). Aflatoxin B1 (AFB1), Aflatoxin M1 (AFM1) as well as Ochratoxin A (OTA) and Deoxynivalenol (DON) are considered potent toxic mycotoxins (Diaz, 2005). They occur in staple foods and highly consumed commodities such as cereals, milk and dairy products, dried fruits and legumes, coffee, wine and beer (Weidenborner, 2008). Aflatoxin B1 (AFB1) is considered as the strongest naturally occurring genotoxic carcinogen and chronic exposure to AFB1 was reported to increase the risk of liver cancer especially when it is associated with hepatitis B or C (Kew, 2003). AFB1 has been classified as ‘‘carcinogenic to humans’’ (group 1) by the International Agency for Research on

http://dx.doi.org/10.1016/j.fct.2014.07.034 0278-6915/Ó 2014 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).

Please cite this article in press as: Raad, F., et al. Dietary exposure to aflatoxins, ochratoxin A and deoxynivalenol from a total diet study in an adult urban Lebanese population. Food Chem. Toxicol. (2014), http://dx.doi.org/10.1016/j.fct.2014.07.034

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Cancer (WHO and IARC, 1993). The carcinogenicity of AFM1, the hydroxylated metabolite of AFB1 found in the milk of animals exposed to AFB1, is considered as 10 times lower than that of AFB1. AFM1 has been classified as ‘‘possibly carcinogenic to humans’’ (group 2B) by IARC (WHO and IARC, 1993). Chronic exposure to OTA is associated with a renal pathological condition referred to as Balkan Endemic Nephropathy (BEN), in addition to its association with immunotoxic and neurotoxic effects (EFSA, 2006). Chronic exposure to DON is associated with delayed growth as well as immunotoxic and hematotoxic effects (EFSA, 2007; AFSSA, 2009; Sirot et al., 2013). Mycotoxins have been ranked as ‘‘the most important chronic dietary risk factor, higher than synthetic contaminants, plant toxins, food additives, or pesticide residues’’ (Kuiper-Goodman, 1998; Bennet and Klich, 2003). Protection of diets from these contaminants should therefore be considered as one of the essential and priority public health functions in any country. This is the main purpose of dietary exposure assessment, an integral component of the risk assessment process (Verger, 2013). Dietary exposure assessment consists of associating food consumption data with concentration/contamination data and typically includes the application of statistical adjustment factors that allow conclusions about the amount of a substance being consumed on a ‘usual’ basis or over a lifetime (FAO/WHO, 2006). Data on the dietary exposure of different populations to mycotoxins are limited. In most of the JECFA reports, data on food contamination with mycotoxins has been often described as inadequate for developed countries and non- existent for most developing ones (JECFA, 1999, 2001, 2002). In the Middle-East, studies investigating food contamination and dietary exposure to mycotoxins are particularly scarce. In Lebanon, and in a study conducted by Soubra et al. (2009), the occurrence of total aflatoxins, OTA and DON in some foodstuffs available on the Lebanese market was investigated. The study highlighted high levels of total aflatoxins, OTA and DON in some food samples, namely nuts, biscuits and bread (Soubra et al., 2009) and called for routine monitoring of the levels of mycotoxin contamination in foods that are highly consumed by the Lebanese population and the need for dietary exposure investigations in the country. Based on the recommendations of several international bodies (WHO, 2005, 2006; EFSA, FAO and WHO, 2011), the Total Diet Study (TDS) has been the most widely applied method in studies aiming at evaluating the dietary exposure to mycotoxins. The TDS approach has the advantage of yielding refined exposure data since it consists of analyzing a representative ‘‘market basket’’ of foods, reflective of the population’s dietary habits’’ (WHO, 1985, 2005; Nasreddine et al., 2010). It differs from other food surveillance programmes since it focuses on chemicals in the diet as a whole rather than on individual food items and it analyzes foods that are processed as for normal consumption, thus taking into consideration the impact of home cooking and preparation techniques on the levels of chemicals in food items (Betsy et al., 2012). The TDS approach was adopted by studies conducted in the United States, the Netherlands, the Czech Republic, Belgium, China, the United Kingdom, Spain, France, Denmark, Finland and Q4 Germany to assess the dietary exposure to different mycotoxins (Cuadrado et al., 1995; Ruprich, 1998; Henry et al., 1998; Egan et al., 2002; Sommerfeld, 2004; Leblanc et al., 2005; Sirot et al., 2013). The objective of the present study is to assess, by the TDS approach, the dietary exposure of an adult urban Lebanese population to AFB1, AFM1, OTA and DON, which are classified as priority food contaminants by the GEMS/Food programme of the WHO (2002). The types and quantities of foods that make up the average ‘total diet’ are based on the results of a previous food consumption study targeting the adult urban population living in Beirut, the capital of Lebanon (Nasreddine et al., 2006). Consumer exposure esti-

mates are compared with the appropriate toxicological reference values and with data provided from other countries.

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2. Materials and methods

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2.1. Food selection and collection of food samples

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The first step in the undertaken TDS consisted of establishing the list of foods to be analyzed. This list was based on food consumption data provided by an individual food consumption survey that was conducted on an adult urban population in Lebanon, the details of which have been published elsewhere (Nasreddine et al., 2006). The selection of food items to be analyzed in the present study was based on two criteria: first, the foods must be identified by GEMS/Food as potential sources of AFB1, AFM1, OTA or DON (WHO, 2002) and second, their level of consumption must exceed 1 g person 1 day 1 (Nasreddine et al., 2010). Accordingly, 47 food items were selected for the TDS (Table 1). Based on general guidelines provided by the WHO (WHO, 1985) and on sampling schemes described in the literature (Leblanc et al., 2005), a composite sampling approach has been applied in this study and consisted of purchasing the same item from five different sites or in five different brands/varieties and of combining the five items to represent a composite sample of the food product in question. Since market shares of the different brands are not established in Lebanon, the contribution of each sub-sample to total weight was equal to 20%. Even though the composite sampling scheme may dilute high contamination levels that may be found in one of the collected sub-samples, this scheme has the advantage of increasing the representativeness of food sampling (WHO, 1985; Nasreddine et al., 2010). Three complete sets of foods (market baskets) were collected at three months intervals and the sampling of foods was performed at the most popular retail markets in Beirut. Thus, 47 food items x five sub-samples x three market baskets = 705 food items were collected for this study. The quantities of purchased foods reflected that necessary for analysis of mycotoxins, as well as for the storage of reserve portions after allowing for amounts lost during preparation and cooking.

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2.2. Preparation of food samples

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A popular local cookbook was consulted for the preparation of cooked dishes (Kamal and Osman, 1995) which included chickpeas, lentils, fava beans, beans, pasta, burghol (parboiled wheat), rice and milk-based pudding. These food items were prepared and cooked in a manner similar to local cooking practices. In compliance with international WHO/GEMS/Food recommendations, foods were prepared using drinking water from the city where the foods were bought, i.e. Beirut for this study. Salt and spices were also added as in regular cooking practices. Suitable stainless steel cooking equipment was used for the preparation of food items. Other food items were not subjected to any cooking or preparation as these food items are typically purchased as ready to eat. For each of the three market baskets and 47 foods specified on the food list, the five sub-samples were combined (20% w/w) and blended to give a homogeneous sample representative of the food item in question. In accordance with good laboratory practices and with the internal standard operating procedures of LARI/Central lab, in compliance with the ISO/IEC 17025 norm (ISO/IEC 17025 2005), the equipment used for preparing and homogenizing the composite samples was thoroughly washed between each preparation (e.g. cleaning with a laboratory-grade detergent, rinsing thoroughly with hot tap water, rinsing or soaking with acid solution) to avoid the risk of cross-contamination. For each of the three market baskets, the 47 food items were aggregated into 15 groups of similar foods for the analysis of mycotoxins. The appropriate amount of each raw, prepared or cooked food item to be included in its composite food group was determined from the food consumption data (Table 1). Food items of each group were combined and blended using an ordinary domestic mixer to give a homogeneous sample. Thus, in total, 15 composite food groups per market basket x three market baskets = 33 composite food group samples were prepared for analysis in this study. Samples were stored at 18 °C prior to analysis. Based on available data from the literature and the evidence that mycotoxins can occur only in certain types of food products, it was not deemed necessary to analyze all types of food samples for all the four mycotoxins (WHO, 2002; Sirot et al., 2013). Analyses performed on each composite group are presented in Table 2.

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2.3. Analysis of food samples

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Food samples were analyzed in the Lebanese Agricultural Research Institute. All analyses were conducted by the same researcher who underwent extensive training in analytical techniques at ARVALIS – Institut du Végétal, Pôle Analyses et Méthodes. Arvalis-Institut du Végétal – is accredited by the French Committee for Accreditation (Cofrac) and fulfills the requirements of the standard NF EN ISO/CEI 17025:2005 and Cofrac rules of application for the activities of testing/calibration. One fourth of the samples were also sent to ARVALIS – Institut du Végétal – for inter-laboratory comparison of analytical results.

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F. Raad et al. / Food and Chemical Toxicology xxx (2014) xxx–xxx Table 1 Aggregation of the 47 food items into 15 composite food groups: weight of each item as consumed (g day 1) and percentage weight of each food item in its group. Composite food group

Daily intake (g day 1)

% Weight

136.8 6.2 3.2 146.2

93.6 4.2 2.2 100.0

2. Biscuits and croissants Biscuits Croissant Doughnuts Total

13.6 4.9 1.3 19.8

68.7 24.7 6.6 100.0

3. Cakes and pastries Cakes Traditional pastry (Knefah) Other traditional pastry Total

11.8 8.2 5.5 25.5

46.3 32.2 21.5 100.0

4. Pasta and other cereal products Pasta, cooked Burghol, Cooked Corn Burghol, raw Total

23.9 5.50 4.61 2.80 36.81

65.0 15.0 12.4 7.6 100.0

5. Pizza and pies Pies, type Manaeesh Pizza Other traditional pies Total

32.1 11.3 6.6 50.0

64.0 23.0 13.0 100.0

6. Rice and rice-based products Rice, cooked Total

50.1 50.1

100.0 100.0

7. Pulses Chickpeas Lentils Fava Beans Beans Fava bean-based (falafel) Total

12.7 9.8 9.1 5.3 2.7 39.6

32.1 24.7 22.9 13.4 6.9 100.0

8. Olive oil, sesame oil, and other oils Olive oil Sesame oil, type tahineh Oil and/or vegetable fat utilized in frying Total

10.3 3.1 4.8 18.1

56.5 16.8 26.6 100.0

1. Bread and toast Traditional bread Traditional crackers (Ka’ak) Toast Total

9. Nuts, seeds, olives, dried dates Nuts and seeds (peanuts, pistachios, almonds, roasted corn, etc.), Olives Dried dates Total

6.0

42.5

6.8 1.4 14.2

47.4 10.1 100.0

10. Cheese Fermented cheese Cheese Akkawi Cheese Halloum Cheese Bulgarian Braided cheese Other white cheese Cheese Kashkawal Swiss cheese Creamy packaged cheese Other imported cheeses Total

9.8 3.6 4.4 1.8 0.4 3.1 5.8 0.4 5.3 0.4 35.0

27.8 10.1 12.7 5.1 1.3 8.9 16.5 1.3 15.2 1.3 100.0

11. Milk and milk-based beverages Milk, liquid Milk, reconstituted from powder Total

29.9 69.8 99.7

30.0 70.0 100.0

12. Milk-based ice cream and pudding Pudding Milk-based ice cream Total

6.1 6.0 12.1

50.2 49.8 100.0

Table 1 (continued) Composite food group

Daily intake (g day 1)

13. Yogurt and yogurt-based products Yogurt Strained yogurt, type Lebneh Total 14. Caffeinated beverages Lebanese traditional coffee Coffee, type Nescafé Total 15. Alcoholic beverages Wine Beer Total

% Weight

68.3 27.8 96.1

71.1 28.9 100.0

128.9 84.7 213.6

60.3 39.7 100.0

1.6 30.4 32.0

5.1 94.9 100.0

Table 2 Mycotoxin analyses performed in different composite food groups. Composite food group

AFB1

AFM1

OTA

DON

Bread and toast Biscuits and croissants Cakes and pastries Pasta and other cereal products Pizza and pies Rice and rice based products Pulses Olive oil, sesame oil and other oils Nuts, seeds, olives and dried dates Alcoholic beverages Caffeinated beverages Cheese Milk and milk-based beverages Yogurt and yogurt-based products Milk based ice cream and pudding

X X X X X X X X X / / / / / /

/ / / / / / / / / / / X X X X

X X X X X X / / X X X / / / /

X X X X X X X / X X / / / / /

X: analyzed, /: not analyzed.

The standard ISO 16050, 2003 was used for the detection and determination of AFB1 in ‘‘cereals and cereal based food samples’’, ‘‘pulses’’ and ‘‘nuts, seeds, olives and dried dates’’. An in-house validated method based on AOAC 991.31 (1994) was performed for the detection and determination of AFB1 in ‘‘olive oil, sesame oil and other oils’’. Food samples (except for ‘‘Olive oil, sesame oil and other oils’’) were extracted by methanol/water mixture (70:30 v/v). For ‘‘Olive oil, sesame oil and other oils’’, liquid–liquid extraction with methanol had been applied. Extracts from the different food matrices were filtered through microfiber filter paper, diluted with water, and purified through an immunoaffinity column (AFLAPREP RBIOPHARM)). AFBl was eluted with methanol. The purified extract was injected to an isocratic reversed phase Liquid Chromatography followed by a derivatization system and fluorescence detector (Shimadzu, Japan). The mobile phase (0.45 lm filtered) consisted of de-ionized water:acetonitrile:methanol (60:20:20, v/v/v). The flow rate was 1 ml min 1, the column was at 45 °C and the injection volume was 50 ll. Fluorescence detection wavelengths were 360 nm for excitation and 435 nm for emission. The retention time under these conditions was approximately 11 min. The method reported by Dragacci and Grosso (2001) was used for the detection and determination of AFM1 in ‘‘Milk and milk-based beverages’’. The analytical technique consisted of a combined extraction and clean-up step of defatted liquid samples by using an immunoaffinity column (containing anti-AF antibodies). As such, samples of ‘‘Milk and milk-based beverages’’ were centrifuged at 2000 rpm to separate the fat and to discard the thin upper fat layer. The centrifugate was purified on immunoaffinity columns at a slow steady rate of 2–3 ml per min. Subsequently, the column was washed with distilled water at a steady flow rate. AFM1 was eluted with pure acetonitirile. The purified extract was injected to an isocratic reversed phase Liquid Chromatography followed by a fluorescence detector (Shimadzu, Japan). The mobile phase consisted of deionized water:acetonitrile:methanol (65:25:10, v/v/v) filtered through a 0.45 lm filter membrane, degased and used at a flow rate of 0.8 ml min 1. The column was at 45 °C and the injection volume was 100 ll. Excitation and emission wavelengths were 365 and 435 nm. The retention time under these conditions was approximately 3.8 min. For the analysis of AFM1 in ‘‘Cheese’’, ‘‘Yogurt and yogurt-based products’’ and ‘‘Milk-based ice cream and pudding’’, the method reported by Kamkar et al. (2008) was used. Briefly, the


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Table 3 Limits of quantification and limits of detection for the analyzed mycotoxins.

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Mycotoxin

Limit of detection (LOD, lg kg

AFB1 AFM1 OTA DON

0.01 0.01 0.05 62.50

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)

Limit of quantification (LOQ, lg kg 0.03 0.03 0.21 125.00

analytical technique consisted first of an extraction by using dichloromethane. The collected filtrates were evaporated to dryness and the residues suspended in methanol:water:n-hexane (30:50:20 vv/v). The toxin was eluted with water/acetonitrile mixture, evaporated to dryness in a rotary evaporator and re-dissolved in HPLC mobile phase. The purified extract was injected to an isocratic reversed phase Liquid Chromatography followed by a fluorescence detector (Shimadzu, Japan). The mobile phase consisted of deionized acetonitrile:methanol:water (20:20:60, v/v/v) filtered through a 0.45 lm filter membrane, degased and used at a flow rate of 1 ml min 1. The column was at 45 °C and the injection volume was 200 ll. Excitation and emission wavelengths were 360 and 440 nm. The retention time under these conditions was approximately 8 min. The standard NF EN 1432 – October 2003 (AFNOR, 2003) was used for the detection and determination of OTA in ‘‘cereals and cereal based food samples’’. An inhouse validated method based on the standard NF EN 14132 – October 2003 (AFNOR, 2003) was performed for the detection and determination of OTA in ‘‘Nuts, seeds, olives and dried dates’’. OTA was extracted from cereals and cereal-based food samples, by acetonitrile/water mixture (60:40 v/v). Extraction by methanol/ water mixture (80:20 v/v) was used for ‘‘Nuts, seeds, olives and dried dates’’. Filtrates were diluted with Phosphate Buffered Saline (PBS), loaded onto an immunoaffinity column (OCHRAPREP R-BIOPHARM) and washed with PBS. OTA was eluted with a methanol:glacial acetic acid solution (98:2, v/v). The eluate was then evaporated to dryness under N2 and the residue dissolved in injection solvent (methanol 30v/deionized water 70v/glacial acetic acid 1v). The purified extract was injected to an isocratic reversed phase Liquid Chromatography followed by a fluorescence detector (Shimadzu, Japan). The mobile phase (0.45 lm filtered) consisted of deionized water:glacial acetic acid (98:2, v/v) with 52% and 48% of acetonitrile. The flow rate was 1 ml min 1, the column was at 45 °C and the injection volume was 100 ll. Fluorescence detection wavelengths were 333 nm for excitation and 460 nm for emission. The retention time of OTA under these conditions was approximately 9.7 min. For ‘‘Caffeinated beverages’’, the method described by Taniwaki et al. (2003) was used. OTA was extracted by 3% sodium bicarbonate/methanol (50:50 v/v). Filtrates were diluted with PBS, loaded onto an immunoaffinity column (OCHRAPREP R-BIOPHARM) and washed with PBS. OTA was eluted with methanol. The purified extract was injected to an isocratic reversed phase Liquid Chromatography followed by a fluorescence detector (Shimadzu, Japan). The mobile phase (0.45 lm filtered) consisted of acetonitrile:sodium acetate with 0.5% acetic acid solution (42:58, v/v). The flow rate was 1 ml min 1, the column was at 45 °C and the injection volume was 100 ll. Fluorescence detection wavelengths were 333 nm for excitation and 470 nm for emission. The retention time of OTA under these conditions was approximately 9 min. For ‘‘Alcoholic beverages’’, the method described by Domijan and Peraica (2005) was used. ‘‘Alcoholic beverages’’ were directly purified on immunoaffinity columns (after being pH adjusted and buffered with PBS). OTA was eluted with a methanol:glacial acetic acid solution (98:2, v/v). The eluate was then evaporated to dryness under N2. Before analyzing on HPLC (Shimadzu, Japan), evaporated samples were dissolved in mobile phase. The mobile phase (0.45 lm filtered) consisted of methanol:deionized water:glacial acetic acid (70:30:20, v/v/v). The flow rate was 0.5 ml min 1, the column was at 45 °C and the injection volume was 50 ll. Fluorescence detection wavelengths were 336 nm for excitation and 464 nm for emission. The retention time of OTA under these conditions was approximately 8 min. The methods described by Ormutag and Beyog˘lu (2003, 2007) were used for the detection and determination of DON in ‘‘cereals and cereal-based food samples’’, ‘‘pulses’’, ‘‘nuts, seeds, olives and dried dates’’ and ‘‘Alcoholic beverages’’. DON was extracted from ‘‘cereals and cereal based food samples’’, ‘‘pulses’’ and ‘‘nuts, seeds, olives and dried dates’’ using acetonitrile/water mixture (21:4, v/v). The filtrate was loaded onto an immunoaffinity column (DONPREP R-BIOPHARM) and washed with acetonitrile–water (21:4, v/v), and eluted with acetonitrile–water (21:4, v/v). The eluate was collected and evaporated to dryness under N2 at 60 °C. Sample extract was dissolved in methanol/water solvent mixture (50:500 v/v). A UVPDA detector coupled with HPLC (Shimadzu, Japan) was used for DON quantification. Samples were injected into the HPLC system under the same conditions used for preparing calibration graphs. The UV PDA detector was set at 220 nm. The volume of sample injected was 100 ll. The mobile phase (0.45 lm filtered) was methanol/water (70:30, v/v) with flow rate of 0.7 ml min 1. The column temperature was at ambient. The retention time was approximately 8.9 min. For quality control, a number of Certified Reference Materials (CRMs) were used and included maize reference material (R-biopharm Rhone LTD P64/A230), peanut butter reference material (R-biopharm Rhone LTD P64/AP01), powder milk reference material (R-biopharm Rhone LTD P64/A230), coffee reference material (R-biopharm Rhone LTD P64/OR 11) in addition to wheat (BIPEA 41650). All CRMs

1

)

Spiking recovery (%)

Repeatability (RSDr, %)

Reproducibility (RSDR, %)

96 74 75 85

19.4 11.45 12.80 19.33

20.47 27.10 13.00 20.06

were used as provided without further grinding. CRMs were stored under the same conditions, extracted and eluted with the same protocol as the analyzed food samples. Each test run included spiked test samples and reference materials. All samples were analyzed in duplicates, which were extracted and measured in separate batches to eliminate any batch specific error. Average recovery percentages based on CRMs were of 75% for AFM1, 97% for OTA, 89% for DON and 90% for AFB1. Corrections based on recovery percentages were not performed. Performance criteria including average spiking recovery, repeatability (RSDr), reproducibility (RSDR), limits of detection (LOD) and limits of quantification (LOQ) for the analyzed mycotoxins are shown in Table 3.

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2.4. Calculation of dietary exposure

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The deterministic approach was applied for the calculation of the dietary exposure to mycotoxins. For OTA and DON, the percentage of censored data (results reported below LOD and/or LOQ) was less than 60%. Accordingly, for these 2 mycotoxins, and based on GEMS/Food guidelines for samples containing levels of elements below the LOQ, a value equal to half the LOQ was assigned and used for calculation purposes (GEMS/Food-Euro, 1995; EFSA, 2010), while for samples containing levels of elements below the LOD, a value equal to half the LOD was assigned and used for calculation purposes (GEMS/Food-Euro, 1995). However for AFB1 and AFM1, where the proportion of censored data exceeded 60%, two scenarios were adopted: (1) the lower bound (LB) approach by replacing the results below LOD by zero and results below LOQ by LOD; (2) the upper bound (UB) approach by replacing the results below LOD by LOD and results below LOQ by LOQ (GEMS/ Food-Euro, 1995; EFSA, 2010). To take into account the variability that exits in food consumption patterns and to provide data on the distribution of exposure levels within the studied population, the distribution of food intakes as provided by the individual dietary survey was combined with the average concentration for each contaminant under study. Accordingly, mean and 95 percentile exposure levels (p95) were computed and risk characterization was performed for average and high exposure levels. All calculations were carried out using the Statistical Analysis Package for Social Sciences, version 18.0 (SPSS Inc., Chicago, IL, USA). To compare dietary exposure levels with toxicological reference values which are expressed per kg body weight, an average body weight of 72.8 kg was used since it was the average weight of the participants in the food consumption survey (Nasreddine et al., 2006). The contributions of each food group to the average dietary exposure to each mycotoxin were also calculated.

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3. Results and discussion

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The analytical results showed that 33.4%, 25.0%, 66.7%, and 48.2% of the data obtained for AFB1, AFM1, OTA and DON, respectively, were above the LOQ. The estimated levels of mycotoxins in the different food groups, as determined from the three ‘market baskets’, are presented in Tables 4 and 5.

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3.1. AFB1

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The highest concentrations of AFB1 were observed in cerealbased products (‘‘Bread and toast’’, ‘‘Cakes and pastries’’) and ‘‘Nuts, seeds, olives and dried dates’’ while the lowest concentrations were found in ‘‘Pulses’’, ‘‘Pasta and other cereal products’’, ‘‘Rice and rice based products’’ as well as in ‘‘Olive oil, sesame oil and other oils’’ (Table 4). Based on AFB1 levels in the analyzed food samples, the average daily exposure to AFB1 was estimated to range between 0.046 (LB estimate) and 0.048 lg person 1 day 1 (UB estimate), while for excessive consumers, the 95th percentile exposure level was estimated to range between 0.102 (LB estimate) and 0.106 lg person 1 day 1 (UB estimate) (Table 6). Using an average body weight of 72.8 kg, the average level of exposure to AFB1 was found to range between 0.63 (LB estimate) and 0.66 ng kg 1 bw day 1 (UB estimate) and the p95 exposure level

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Q9

Table 4 Estimation of the mean concentrations (lg kg in Lebanon.

1

1

or mg kg

fresh weight) of AFB1, OTA and DON in 11 composite food groups of the total diet study of an adult urban population

Composite food group

AFB1 levels lower bound estimatea (lg kg 1)

AFB1 levels upper bound estimateb (lg kg 1)

Mean OTA levels (lg kg 1)

Mean DON levels (lg kg 1)

Bread and toast Biscuits and croissants Cakes and pastries Pasta and other cereal products Pizza and pies Rice and rice based products Pulses Olive oil, sesame oil and other oils Nuts, seeds, olives and dried dates Caffeinated beverages Alcoholic beverages

0.242 0.000 0.105 0.000

0.275 0.010 0.115 0.010

0.298 2.844 0.152 0.183

524.170 340.330 109.670 62.500c

0.043 0.000 0.000 0.000

0.048 0.010 0.010 0.010

0.223 0.680 N.A N.A

121.160 322.000 31.250d N.A

0.180

0.260

0.078

62.500c

N.A N.A

N.A N.A

0.508 1.472

NA 52.080

a,b For AFB1, the percentage of censored data exceeded 60%. Therefore, based on GEMS/Food recommendations, two estimates were provided. N.A.: Not analyzed. a Lower bound estimate: the undetected values were replaced by zero; the un-quantified values were replaced by LOD. b Upper bound estimate: the undetected values were replaced by LOD; the un-quantified values were replaced by LOQ. c Values were below the LOQ. Accordingly a value equal to half the LOQ was assigned to these food samples. d Values were below the LOD. Accordingly a value equal to half the LOD was assigned to these food samples.

Table 5 Estimation of the mean concentrations (lg kg

1

fresh weight)) of AFM1 in 4 composite food groups of the total diet study of an adult urban population in Lebanon.

Composite food group

AFM1 level (lg kg

Milk and milk-based beverages Cheese Yogurt and yogurt based products Milk-based ice cream and pudding

0.11 0.00 0.00 0.00

1

) lower bound estimatea

AFM1 level (lg kg

1

) upper bound estimateb

0.18 0.05 0.05 0.05

a,b

For AFM1, the percentage of censored data exceeded 60%. Therefore, based on GEMS/Food recommendations, two estimates were provided. Lower bound estimate: the undetected values were replaced by zero; the un-quantified values were replaced by LOD. b Upper bound estimate: the undetected values were replaced by LOD; the un-quantified values were replaced by LOQ. a

Table 6 Mean and p95 dietary exposure to AFB1 and AFM1 from the total diet study of an urban adult Lebanese population. Mycotoxin

Exposure (lg person

1

day

Mean LB AFB1 AFM1

b

0.046 0.016

1

)

Exposure (ng kg P95

b

b

1

bw day

Mean b

UB

LB

UB

0.048 0.023

0.102 0.040

0.106 0.058

LB

b

0.63 0.22

1 a

)

P95 UB

b

0.66 0.31

LBb

UBb

1.40 0.55

1.46 0.80

LB: Lower bound estimate: the undetected values were replaced by zero; the un-quantified values were replaced by LOD. UB: Upper bound estimate: the undetected values were replaced by LOD; the un-quantified values were replaced by LOQ. a bw: Body weight. b For AFB1 and AFM1, the percentage of censored data exceeded 60%. Therefore, based on GEMS/Food recommendations, two estimates were provided.

381 382 383 384 385 386 387 388 389 390 391 392 393 394 395 396

between 1.40 (LB estimate) and 1.46 ng kg 1 bw day 1 (UB estimate). Since aflatoxins are classified as carcinogenic and genotoxic contaminants, the ALARA approach (As Low As Reasonable Achievable) is recommended (JECFA, 2001). Based on JECFA and SCF reports (JECFA, 1999; SCF, 1994), even a very low exposure level to aflatoxins (1 ng kg 1 bw day 1) may induce liver cancer cases. Observational studies highlighted an interaction between hepatitis B and exposure to aflatoxins, suggesting that two separate aflatoxins’ potencies exist, one in populations in which chronic hepatitis infections are common and the second where such infections are rare (such as European countries) (Pitt, 2004). It was therefore suggested that, for non-European countries, an incidence of 0.083 cases of liver cancer per year per 100,000 persons would be attributed to the ingestion of 1 ng kg 1 bw day 1 of aflatoxins (JECFA, 1999). Lebanon and most Middle-Eastern countries are characterized by

a higher prevalence of hepatitis B as compared to countries in Western Europe (Soubra et al., 2009). In particular, the percentage of HbsAg carriers in the Middle-Eastern population is estimated to range between 5% and 15% (Toukan et al., 1990; Qirbi and Hall, 2001), while in Western Europe this rate is estimated to range between 0.5% and 2% (Van Damme et al., 2004). Therefore, according to the aforementioned potency value of aflatoxins in inducing liver cancers in non-European countries (0.083 cases/100,000 persons), the number of excess cancer cases in the Lebanese adult population would be estimated to range between 0.0527 and 0.0545 cases per 100,000 people per year based on the average level of exposure, while the p95 exposure levels would be associated with a population risk of 0.11–0.12 cancers per 100,000 people per year. It is important to note that liver cancer incidence rates in Lebanon have been following an increasing trend over time, with the incidence among men rising from 1.8 to 4 cases per 100,000 between


397 398 399 400 401 402 403 404 405 406 407 408 409 410 411 412

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2003 and 2008 and from 1.5 to 3.9 cases per 100,000 in women, during the same period of time (Shamseddine et al., 2014). Previous studies assessing the dietary exposure of the Lebanese population to AFB1 are scarce (Raad, 2005; Soubra, 2008). The average exposure values to AFB1, as documented in the present study (0.63–0.66 ng kg 1 bw day 1), are similar to those reported by Raad (2005) (0.6 ng kg 1 bw day 1), but lower than those reported by Soubra (2008) where the exposure estimates to AFB1 were found to range between 1.1 ng kg 1 bw day 1 and 4.04 ng kg 1 bw day 1. It is important to note that the higher values obtained by Soubra (2008) could be a reflection of methodological differences, whereby Soubra (2008) compiled existing contamination data from various available sources including the Lebanese Agricultural Research Institute (IRAL), the Toxicology Research Department of Saint Joseph University and the Agriculture Department of the American University, whereas, in the present study, all food samples were collected and analyzed using the same standard protocol described in the methodology section. Mean exposure levels to AFB1 as estimated in this study are compared with those reported from other countries. Accordingly, the average dietary exposure to AFB1 (0.63–0.66 ng kg 1 bw day 1) is higher than that reported from the TDS conducted in France (0.002–0.22 ng kg 1 bw day 1; Sirot et al., 2013), the United states (0.26 ng kg 1 bw day 1) (Henry et al., 1998), Australia (0.15 ng kg 1 bw day 1) (European commission, 1997) and Netherlands (0.1 ng kg 1 bw day 1) (Sizoo and van Egmond, 2005) but lower than that reported from Korea (1.19 ng kg 1 bw day 1) (Park et al., 2004) and Canada (1 ng kg 1 bw day 1) (Kuiper-GoodQ6 man et al., 1993). The mean exposure estimate obtained in the present study is comparable to that reported from Sweden (0.8 ng kg 1 bw day 1) (Thuvander et al., 2001) by the same general approach (TDS). It is important to note that, even though comparisons with data reported by other countries may provide useful benchmarking, such comparisons should be interpreted with caution since in many instances the studies may differ in the method and the model used to evaluate dietary exposure, the limits of detection/quantification of the analytical technique, the assumptions made in exposure assessment calculations for censored data, the degree of preparation of the foods included in the assessment etc. (Kroes et al., 2002; Nasreddine et al., 2010). This study helped in identifying the specific food groups that are mostly contributing to the dietary exposure to AFB1. This information is warranted since the contribution of a specific food item to daily exposure depends not only on the concentration of the contaminant in this particular food but also on the amount of that specific food typically consumed by the population. Consequently, food groups that represent the major source of a contaminant in one country may not be the same in another where the population has a different dietary pattern (Nasreddine et al., 2010). This can be illustrated by data provided by total diet studies conducted in different parts of the world. For instance, the main dietary source of AFB1 were eggs, nuts and oilseeds in the first TDS conducted in France (Leblanc et al., 2004) and chocolate in the second French TDS (Sirot et al., 2013), whereas rice was identified as the main contributor to AFB1 in Korea (98.96%) (Park et al., 2004). In the present study, and based on both the LB and UB estimates, the food groups that contributed the most to the dietary exposure to AFB1 were ‘‘Bread and toast’’ (79.4–82.2%), followed by ‘‘Nuts, seeds, olive and dried dates’’ (6.8–7.0%) (Table 8). The preponderance of ‘‘Bread and toasts’’ as the main contributor to AFB1 results not only from the fact that these food items had the highest level of contamination by AFB1, but also from the fact that they represent the major staple food for the Lebanese population. In fact, bread and cereals were found to provide, alone, 35% of the total energy intake of the average Lebanese urban adult (Nasreddine et al., 2006). It is important to mention that dried figs, which are specified by the

GEMS/Food comprehensive list as potential sources of AFB1, were excluded from the exposure assessment performed in this study. This exclusion resulted from the fact that food consumption data pertinent to dried figs or other dried fruits are not available in the country.

479

3.2. AFM1

484

The average concentration of AFM1 in ‘‘Milk and milk based beverages’’ varied between less than 0.05 lg kg 1 and a maximum of 0.18 lg kg 1 (Table 4). The mean daily exposure to AFM1 ranged between 0.016 (LB estimate) and 0.023 lg person 1 day 1 (UB estimate) whereas for excessive consumers (p95), the daily exposure levels ranged between 0.04 (LB estimate) and 0.058 lg person 1 day 1(UB estimate) (Table 6). AFM1 is considered a genotoxic and carcinogenic contaminant with its liver cancer induction potency being described as 10 times lower compared to AFB1 (JECFA, 2001). Accordingly, it is estimated that, for non-European countries, the ingestion of 1 ng kg 1 bw day 1 of aflatoxins would induce 0.0083 liver cancer cases per year per 100,000 persons (JECFA, 1999). Based on these assumptions, the mean dietary exposure to AFM1 as estimated in this study would be associated with an average population risk ranging between 0.0018 and 0.0027 additional cancer cases per 100,000 – people per year, while the p95 exposure levels would be associated with a population risk of 0.0046 to 0.0066 additional cancer cases per 100,000 people per year. The exposure estimate to AFM1 as obtained in this study (0.22– 0.31 ng kg 1 bw day 1) is similar to that reported from the Netherlands (0.19 ng kg 1 bw day 1; Sizoo and Van Egmond, 2005) and Brazil (0.188 ng kg 1 bw day 1; Shundo et al., 2009), while being higher than levels reported from France (0.03 ng kg 1 bw day 1; Sirot et al., 2013), United states (0.03 ng kg 1 bw day 1; JECFA, 2001), Ireland (0.0093 ng kg 1 bw day 1; Coffey et al., 2008), and Morocco (0.05 ng kg 1 bw day 1, Zinedine and Manes, 2009). The average exposure estimate to AFM1 as obtained in this study (16–23 ng person 1 day 1) is also higher than the level suggested by the WHO within the framework of GEMS/Food program for the Middle-Eastern diet (1.3 ng person 1 day 1) (JECFA, 2001), which may be due to discrepancies in the source of food contamination and food consumption data that were used for the generation of the exposure estimates. ‘‘Milk and milk-based beverages’’ appeared as the main contributors to the dietary exposure to AFM1 (68.9–100.0%), (Table 8), which is in agreement with previous reports from other countries including Brazil (Shundo et al., 2009) and France (Leblanc et al., 2005).

485

3.3. OTA

524

The food products presenting the highest concentrations of OTA were cereal-based products, particularly ‘‘Biscuits and Croissants’’ followed by ‘‘Alcoholic beverages’’ (Table 4). The lowest OTA concentrations were observed in ‘‘Nuts, seeds, olives and dried dates’’. The estimated average and excessive consumers’ exposure levels to OTA were 0.31 and 0.99 ng day 1, respectively. Mean and excessive consumers’ exposure levels to OTA represented respectively 29.9% and 95.1% of the PTWI set by JECFA (100 ng kg 1 week 1; JECFA, 2007), thus highlighting the risk for the excessive consumer to approach the toxicological reference value for this mycotoxin (Table 7). When compared to previous studies conducted in the country, the average exposure to OTA (4.28 ng kg 1 bw day 1) for the Lebanese urban adult population falls within the range reported by Soubra (2008) (2.2–5.5 ng kg 1 bw day 1 in 8–13 year old children and 2.1–4.1 ng kg 1 bw day 1 in 14–18 year old adolescents) while

525


480 481 482 483

486 487 488 489 490 491 492 493 494 495 496 497 498 499 500 501 502 503 504 505 506 507 508 509 510 511 512 513 514 515 516 517 518 519 520 521 522 523

526 527 528 529 530 531 532 533 534 535 536 537 538 539 540

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Table 7 Risk characterization: mean and P95 dietary exposure to OTA and DON and percent contribution to Toxicological Reference Values (TRVs) from the total diet study of an urban adult Lebanese population. Mycotoxin

OTA DON a b c d

Exposure per kg body weight per day

Dietary exposure expressed as %TRV SCF/EFSA/JECFA

Mean

Exposure per day P95

Mean

P95

Mean

P95

0.31 (lg day 1) 114.22 (lg day 1)

0.99 (lg day 1) 259.30 (lg day 1)

4.28 (ng kg 1.56 (lg kg

13.60 (ng kg 1 bw day 1) 3.55 (lg kg 1 bw day 1)

85.6a/25.0b/29.9c 156.8d

271.9a/79.5b/95.1c 355.8d

1

bw day 1) 1 bw day 1)

OTA PTDI according to SCF: 5 ng kg 1 day 1 (SCF, 1998). OTA PTWI according to EFSA: 120 ng kg 1 week 1 (EFSA, 2006, 2010). OTA PTWI according to JECFA: 100 ng kg 1 week 1 (JECFA, 2007). DON PMTDI according to SCF, EFSA and JECFA: 1 lg kg 1 bw day 1 (SCF, 2002; EFSA, 2007; JECFA, 2010).

Table 8 Estimation of the contribution (%) of the various food categories to the average exposure to mycotoxins in the total diet study of an adult urban population in Lebanon. Food group

Bread and toast Biscuits and croissants Cakes and pastries Pasta and other cereal products Pizza and pies Rice Pulses Olive oil, sesame oil and other oils Nuts and seeds, olives and dried dates Alcoholic beverages Caffeinated beverages Cheese Milk and milk based beverages Yogurt and yogurt based products Milk based ice cream and pudding

Contribution (%) to average exposure AFB1 LB

AFB1 UB

AFM1 LB

AFM1 UB

OTA

DON

82.2 0.0 5.9 0.0 4.9 0.0 0.0 0.0 7.0 NA NA NA NA NA NA

79.4 0.4 5.7 0.7 4.7 1.0 0.8 0.4 6.8 NA NA NA NA NA NA

NA NA NA NA NA NA NA NA NA NA NA 0.0 100.0 0.0 0.0

NA NA NA NA NA NA NA NA NA NA NA 7.6 68.9 20.8 2.6

14.0 18.1 1.2 1.9 3.6 10.9 NA NA 0.4 15.2 34.8 NA NA NA NA

67.1 5.9 2.4 1.8 5.3 14.1 1.1 NA 0.8 1.5 NA NA NA NA NA

NA: Not analyzed for the mycotoxin in question.

541 542 543 544 545 546 547 548 549 550 551 552 553 554 555 556 557 558 559 560 561 562 563 564 565 566 567 568 569 570 571 572

being lower than that reported by Raad (2005) for 15–25 year old adults (10 ng kg 1 bw day 1). The discrepancy with the results reported by Raad (2005) may, in part, be explained by differences in the analytical techniques used for the determination of OTA levels whereby contamination data reported by Raad (2005) were based on analysis by ELISA. The average dietary exposure to OTA as estimated in the present study (4.28 ng kg 1 bw day 1) is similar to values reported from Canada (4.2 ng kg 1 bw day 1), but higher than those reported from Ireland (0.000049 ng kg 1 bw day 1), Greece (0.15 ng kg 1 bw day 1), United Kingdom (0.53 ng kg 1 bw day 1), Portugal (0.81 ng kg 1 bw day 1), Norway (0.97 ng kg 1 bw day 1), Italy (1.13 ng kg 1 bw day 1), Spain (1.18 ng kg 1 bw day 1), Denmark (1.19 ng kg 1 bw day 1), Netherlands (1.59 ng kg 1 bw day 1), Sweden (1.42 ng kg 1 bw day 1), Finland (1.71 ng kg 1 bw day 1), Germany (1.82 ng kg 1 bw day 1) and France (0.28–1.92 ng kg 1 bw day 1) (Kuiper-Goodman et al., 1993; Cuadrado et al., 1995; Gilbert et al., 2001; Backer and Pieters, 2002; Miraglia and Brera, 2002; Sommerfeld, 2004; Sizoo and van Egmond, 2005; Coffey et al., 2008; Sirot et al., 2013). The results obtained in this study suggest that the Lebanese excessive consumer may approach the toxicological reference value of OTA. This exposure may be linked to cases of nephropathy widely reported in the country in both sexes of the population (Assaf, 2004). This is supported by studies investigating OTA levels in blood samples in the Lebanese population, whereby OTA was detected in 33% of tested plasma samples (n = 250) with an average contamination of 0.29 mg L 1 (Assaf, 2004). Exposure to OTA has been associated with an increased risk of kidney cancer (JECFA, 1999, 2001) and available data from the country suggest an increasing trend in the incidence of this type of cancer in the Lebanese population (Adib and Daniel, 2006).

In the present work, the main contributors to the dietary exposure to OTA were caffeinated beverages (34.8%), and cereal derivatives notably ‘‘Biscuits and Croissants’’ (18.1%) (Table 8). These food groups have also been reported by other countries including Denmark (coffee 15.9%) (Miraglia and Brera, 2002), Finland (coffee 22.8%) (Miraglia and Brera, 2002), France (cereals 38.1%) (Leblanc et al., 2005) and Germany (Bread 33.0%) (Sommerfeld, 2004) as being the major source of OTA dietary exposure. It is important to mention that OTA can be found in important quantities (10– 920 lg kg 1) in pigs’ kidneys and meat (Turgeon et al., 2009), which were excluded from the exposure assessment performed in this study. This exclusion of pork products should not have resulted in a significant underestimation of exposure estimates due to their very low level of consumption by the studied population (0.65 g day 1).

573

3.4. DON

588

The food products presenting the highest concentrations of DON were cereal-based products such as ‘‘Bread and toast’’, ‘‘Biscuits and croissants’’ in addition to ‘‘Rice’’ (Table 4). The lowest DON concentrations were observed in ‘‘Pulses’’ and ‘‘Nuts, seeds, olives and dried dates’’. The estimated levels of average and excessive consumers’ exposure (p95) to DON were 114.22 and 259.30 lg day 1, respectively (Table 7), representing 156.8% and 355.8% of the PMTDI, respectively (1 lg kg 1 bw day 1: JECFA, 2001). These findings highlight the risk of the average as well as the excessive consumer of exceeding the toxicological reference value for DON (Table 7). The average exposure level to DON as estimated in the present study (1.56 lg kg 1 bw day 1) is higher than that estimated by Raad (2005) (0.19 lg kg 1 bw day 1 in a sample of 432 subjects

589


574 575 576 577 578 579 580 581 582 583 584 585 586 587

590 591 592 593 594 595 596 597 598 599 600 601 602

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11 August 2014 8


627

aged 15–25 years) and that reported by Soubra (2008) (0.39 lg kg 1 bw day 1 in a sample of 285 Lebanese adolescents aged 15–18 years). Several factors, notably variability in food consumption levels, the age of the individuals included in the exposure assessment and the differences in the ways contamination data were generated may have led to the observed discrepancies in exposure estimates between the studies. The average dietary exposure to DON as estimated in the present study (1.56 lg kg 1 bw day 1) is comparable to that reported from the United states (1.50 lg kg 1 bw day 1), but higher than that reported from Denmark (0.17 lg kg 1 bw day 1), United Kingdom (0.17 lg kg 1 bw day 1), Belgium (0.25 lg kg 1 bw day 1), Germany, (0.27 lg kg 1 bw day 1), France (0.373–0.379 lg kg 1 bw day 1), Austria (0.3 lg kg 1 bw day 1), Netherlands 1 1 (0.34 lg kg bw day ), Portugal (0.37 lg kg 1 bw day 1), Canada (0.40 lg kg 1 bw day 1) and Ireland (0.20 lg kg 1 bw day 1) while being lower than that determined in Norway (2.6 lg kg 1 bw day 1) (Gareis et al., 2003; Leblanc et al., 2004; Sommerfeld, 2004; Sirot et al., 2013). In the present work, the main contributors to the dietary exposure to DON were ‘‘Bread and toast’’ (67.1%) (Table 8), a food group that has also been reported as the main sources of DON exposure by other countries including Germany (Bread, 77%) (Sommerfeld, 2004), the Netherlands (Bread, 52.30%) (Schothorst et al., 2005) and France (Cereals, 90%) (Leblanc et al., 2004).

628

4. Conclusion

629

The exposure assessment conducted in the present study by the TDS approach has provided the first estimate of the average dietary exposure of the adult Lebanese population to four mycotoxins. It showed that exposure to DON is high, exceeding the toxicological reference value for the average as well as the excessive consumer (156.8% and 355.8% of the PMTDI, respectively). Similarly, the excessive consumers’ exposure to OTA was shown to approach the TRV of this mycotoxin (95.1%), highlighting a relative risk for this population group. The average exposure levels to AFB1 and AFM1 were also found to exceed those reported by several developed and developing countries from various parts of the world. This study has some limitations as the population sample was only selected from the Beirut area. The choice of Beirut can be explained by the fact that it comprises 40% of the Lebanese population and is considered as the ‘‘melting pot’’ of the country. Furthermore, this study was confined, like many exposure assessments conducted around the world, to the adult fraction of the population. Thus, while not excluding the possibility that the daily exposure assessment performed in the present study may not be representative of the population as a whole, this study has provided a first estimate of the exposure of the adult consumer to four mycotoxins through the diet in Lebanon. The results highlight the necessity to reduce the consumer’s exposure to DON and OTA. Reducing the exposure of the consumer may be achieved by reducing the consumption and/or contamination levels of foods, while focusing on the main contributors to the exposure, as proposed by JECFA (2007), Sirot et al. (2013). There is therefore an urgent need for rigorous monitoring of the contamination levels of the local food supply to ensure that maximal limits in foods are not exceeded. The study’s findings call for further larger-scale studies aiming at providing a comprehensive assessment of the dietary exposure of the Lebanese population to mycotoxins as well as to other food contaminants. Such studies are crucial to catalyze the development and adoption of proper management strategies to ensure the safety of the food supply in the country.

603 604 605 606 607 608 609 610 611 612 613 614 615 616 617 618 619 620 621 622 623 624 625 626

630 631 632 633 634 635 636 637 638 639 640 641 642 643 644 645 646 647 648 649 650 651 652 653 654 655 656 657 658 659 660 661 662 663 664

Conflict of Interest

665

The authors declare that there are no conflicts of interest.

666

Transparency Document

667

The Transparency document associated with this article can be found in the online version.

668 670 669

5. Uncited reference

671

Richard (2007).

Q7

672

Acknowledgement

673

The study was supported by the Lebanese National Research Institute (LARI) and by the French Cooperation Program (French Embassy).

674

References

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