Exposure estimates to Fusarium mycotoxins through cereals intake
Introduction
Mycotoxins are secondary metabolites produced by a wide variety of fungal species leading nutritional losses and representing a significant hazard to the food chain (Mankeviciene et al., 2011). In fact, the Food and Agricultural Organization (FAO) has estimated that one-quarter of the world’s food crop is contaminated, to some extent, with mycotoxins (JECFA, 2001). The molds ability to produce mycotoxins is greatly influenced by environmental factors, being the most important temperature, relative humidity, insect damage, drought and inadequate storage conditions (Miraglia et al., 2009, Prandini et al., 2009). The Fusarium toxins such as the trichothecenes and zearalenone, occur mainly in cereals grown in more moderate climates. While these contaminants may be of lower toxicity than aflatoxins, ochratoxin A, etc., their occurrence in food commodities that are eaten more widely by consumers means that levels must also be rigorously controlled in food and feed. Thus, the occurrence of mycotoxins in agricultural commodities has been recognized as a potential hazard for the human and animal health.
Among the thousands of species of fungi, only about 100 belonging to genera Fusarium Aspergillus, Penicillium, Alternaria, Claviceps, etc. are known to produce mycotoxins. Trichothecenes and zearalonene constitute one of the largest groups of mycotoxins produced by Fusarium in cereal grains. Most Fusarium species produce either type A or type B trichothecenes. Type A trichothecenes includes HT-2 and T-2 toxins, diacetoxyscirpenol (DAS) and neosolaniol (NEO). Type B trichothecenes is mainly represented by deoxynivalenol (DON) and byits derivates 3-acetyl-deoxynivalenol (3-ADON) and 15-acetyl-deoxynivalenol (15-ADON). Other type B trichothecenes are fusarenon-X (FUS-X) and nivalenol (NIV). Immunosuppressive and immunomodulatory effects were described as trichothecenes toxicological effect. Zearalenone (ZON) has high oestrogenic activity and can be an important etiologic agent of intoxication in young children (Zinedine et al., 2007, Metzler et al., 2010). Patulin (PAT) is produced by different species of Penicillium, Aspergillus, and Byssochlamys and it is related to neurotoxic, immunotoxic and gastrointestinal effects in animals.
The human risk to mycotoxins exposure is related to consumption of plant-based foods like cereal grains or animal origin as kidney, liver, milk and eggs (Erdogan, 2004, Atanda et al., 2007). Cereal foods are important in infants and young children diet and they are among the first solid food eaten. Wheat, rice and maize are the most consumed cereals worldwide; the latest data reported by FAO estimated an annual global mean consumption of 66, 53.3 and 17.1 kg/per capita for wheat, rice and maize, respectively. Additionally, the Spanish population is one of the largest consumers of wheat in the world, with an average consumption of 86.10 kg/per capita (FAO, 2009).
Mycotoxins are relatively stable to cooking and processing, thus food preparation procedures do not guarantee their removing. Monitoring studies are required in order to obtain reliable information about the real exposure of human population to mycotoxins. In this sense, in the European Union, efforts to assess human exposure to mycotoxins have been undertaken within SCOOP (Co-operation on Questions relating to Food) projects. The latest data were reported in the SCOOP task 3.2.10 and compiles about 35 000 results received about occurrence of thricothecenes in various food and food raw materials from 12 countries (Schothorst and van Egmond, 2004).
Nowadays, increased efforts have been carried out in order to improve analytical methods not only for the detection of very low concentrations of mycotoxins in foodstuffs but also for the simultaneous determination of different types of mycotoxins (Shephard, 2008, Koeppen et al., 2010, Meneely et al., 2011, Capriotti et al., 2012). In this way, the European Union Reference Laboratory (EU-RL) for mycotoxins aims to facilitate the implementation of European legislation related to monitoring of mycotoxins in food of plant origin and animal feed. The Joint Research Centre Institute for Reference Materials and Measurements and the EU-RL for mycotoxins develop analytical approaches and validate them in-house (JRC Technical Notes, 2011). In the same way, the Association of Official Analytical Chemist (AOAC Official Method 985.18, 2002, AOAC Official Method 986.18, 2002 International) reported official methods for mycotoxins determination, for instance Official Method 986.18 for deoxynivalenol determination in wheat or Official Method 985.18 for zearalenone determination in corn.
Taking into account the methods of analysis for the official control of the levels of contaminants in foodstuffs, an analytical strategy to determine ten mycotoxins using QuEChERS-based extraction and gas chromatography-tandem mass spectrometry (GC–MS/MS) with triple quadrupole (QqQ) mass analyzer have been recently optimized and validated in our laboratory in cereal products and bee pollen (Rodríguez-Carrasco et al., 2012, Rodríguez-Carrasco et al., 2013).
To ensure food safety, the European Commission (EC) has set maximum limits for some mycotoxins in foodstuffs (EC No. 1881, 2006, EC No. 1126, 2007) (Table 1). Maximum levels of DON permitted in Europe are 750 μg kg−1 in pasta, cereals intended for human consumption and cereal flour and 200 μg kg−1 in processed cereal-based foods and baby foods for infants and young children. The Joint FAO/WHO Expert Committee on Food Additives (JEFCA) established provisional maximum tolerable daily intakes (PMTDI) for DON and its acetylated derivates (3-ADON and 15-ADON) of 1 μg kg−1 bw, whereas for NIV and the sum of T-2 and HT-2 toxins the SCF proposed a temporary tolerable daily intake (t-TDI) of 0.7 and 0.06 μg kg−1 bw, respectively (SCF, 2002). Recently, the Scientific Committee on Food (SCF) concluded that a full TDI of 0.1 μg kg−1 bw for the sum of T-2 toxin and HT-2 toxin can be now established based on recent data (SCF, 2011a).
EC set also a maximum level of ZON of 75 μg kg−1 in cereals intended for direct human consumption and cereal flour, 100 μg kg−1 in maize intended for direct human consumption and maize flour and 20 μg kg−1 in processed cereal-based foods and baby foods for infants and young children. In 2000 the SCF established a t-TDI of 0.2 μg kg−1 bw for ZON. However, in 2011 the SFC concluded that a TDI of 0.25 μg kg−1 can be established based on recent data in the most sensitive animal species (SCF, 2011b). Concerning PAT, Codex Alimentarius have recommended PAT concentration of <50 μg kg−1 for apple products intended for human consumption and the European Union (EU) established a limit of 10 μg kg−1 for patulin in baby food and infant formulae. On the other hand a PMTDI of 0.4 μg kg−1 bw was established by JEFCA (JECFA, 1995).
The occurrence of PAT, DON, 3-ADON, DAS, NIV, FUS-X, NEO, T-2, HT-2 and ZON was evaluated in this study in cereal-based products at first stage for screening purposes followed by a risk assessment approach for these toxins in infants, children and adults.
Section snippets
Sampling
One hundred and fifty-nine cereal-based samples purchased in 2012 from different shops located in Valencia (Spain) were evaluated. The samples were classified as wheat-based (n = 119), rice-based (n = 23), and maize-based (n = 17). Additionally, they were categorized as store brands (n = 79) and name brands (n = 80). A subdivision of the collected samples depending on agricultural practice was also performed. Thereby, 70 out of 79 store brand samples were classified as conventional while the remaining
Presence of mycotoxins in samples
The analytical method was in-house validated with respect to selectivity, linearity, LOD, LOQ, precision and accuracy according to SANCO 12495/2011 document (SANCO, 2011). Matrix-matched calibrations were used for quantitation. Linear range studied was from LOQ to 100 times LOQ. A good linearity (R2 > 0.990) within the tested range was obtained in all food matrices evaluated. For the recovery analysis, one wheat, rice and maize-based sample, previously analyzed to assure the absence of studied
Conclusions
The occurrence of PAT, DON, 3-ADON, DAS, NIV, FUS-X, NEO, T-2, HT-2 and ZON has been evaluated in 159 cereal-based samples commonly consumed in Spain mainly by infants and children. Our findings show that the levels of contamination of mycotoxins in cereal-derived products were lower than the permitted levels by EU for safe consumption. As regards occurrence of mycotoxins, no significant differences were found neither in the type of brand nor in the agricultural practice employed. However, a
Acknowledgements
This work was supported by the Spanish Ministry of Science and Innovation (AGL2010-17024/ALI). Y. Rodríguez-Carrasco thanks the F.P.U. Grant (No. AP2010-2940) provided by the Ministry of Education.
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