**1. Introduction**

Worldwide, cereals are at risk from contamination with mycotoxins, i.e. secondary mould metabolites, both while still in field and during storage. *Fusarium* fungi produce several

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trichothecene mycotoxins having a common chemical structure and a similar mode of action [1]. Among them, T-2 toxin (3α-hydroxy-4β,15-diacetoxy-8α-(3-methylbutoxy)-12,13 epoxytrichothec-9-ene), and its major metabolite HT-2 toxin (3α,4β-dihydroxy-15-acetoxy-8α-(3-methylbutoxy)-12,13-epoxy-trichothecene), represent not only type A trichothecene mycotoxins produced mainly by the *Fusarium langsethiae*, but also the *Fusarium poae* and the *Fusarium sporotrichioides* [2, 3]. These toxins are produced at temperatures ranging from −2 to 35°C and with water activities above 0.88 [4, 5], and are frequently responsible for the contamination of different grains such as maize, oat, barley, wheat, rice and soya beans. Weather conditions, varieties and the sowing time are the most important factors influencing the T-2 and HT-2 toxin production [6].

be implemented in order to prevent contamination during cultivation and storage of cereals

The Incidence of T-2 and HT-2 Toxins in Cereals and Methods of their Reduction Practice by the Food Industry

http://dx.doi.org/10.5772/intechopen.71550

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Literature data on the impact of cereal processing on the levels of T-2 and HT-2 contamination are also very scarce. Data gathered insofar have suggested that toxins in reference, when milled, get to be relocated into various milling fractions, but are not eliminated [19]; in addition, they have been proven resistant to processing. Even more so, due to their hull binding, subsequent processing of cereals leads to significant rise in levels of these toxins in finished products [16]. Scudamore [20] concluded that final processing, such as boiling, fermentation, baking, frying, and extrusion, has no impact on the level of contamination with these mycotoxins. Some studies evidenced that processing of raw cereals greatly reduces T-2 and HT-2 levels in food products, but makes these toxins concentrate in high levels in by-products [6]. In industrial food processing settings, the processing time and temperature combination has been shown to be crucial for the reduction of mycotoxin content in a finished food product. While conventional food preparation at temperatures up to 100°C has a negligible effect on most mycotoxins, higher temperatures used with frying, roasting, toasting and extrusion have been shown to be capable of decreasing mycotoxin

The studies quoted under this chapter bring data on the incidence of T-2 and HT-2 toxin in different unprocessed cereals during a two-year period. They also demonstrate the influence of certain food processing methods, such as cooking, roasting and extrusion, performed under predefined conditions, on the levels of T-2 and HT-2 toxin in contaminated cereals. For this purpose, after application of the quantitative screening method termed the enzymelinked immunosorbent assay (ELISA), which enabled the determination of summary T-2 and HT-2 toxin concentrations, the concentrations of each mycotoxin in positive samples were determined using a confirmatory method in terms of liquid chromatography tandem-mass spectrometry (LC-MS/MS), also used in the investigations devoted to the possibilities of

During the period spanning from May 2015 to April 2017, a total of 285 samples of unprocessed cereals, in specific maize (n = 84), wheat (n = 56), oat (n = 72), barley (n = 44) and triticale (n = 29), were sampled from households situated in the Northern, Central and Eastern part of Croatia. All cereals were grown in the crop season 2015 and 2016 and had not undergone any physical or thermal treatment other than drying, cleaning and sorting prior to sampling. Sampling and sample preparation of unprocessed cereals were performed in full line with the Commission Regulation 401/2006 [22], laying down the methods of sampling to be exercised within the frame of the official control of mycotoxin levels in foodstuffs. The aggregate

and their final products.

contamination [21].

reduction of concentrations of these mycotoxins.

**2. Materials and methods**

**2.1. Sampling and sample preparation**

Type A trichothecenes, T-2 and HT-2 toxins included, are generally more toxic than type B trichothecenes (e.g. deoxynivalenol and nivalenol). Structure/activity-relationship studies revealed that 12,13-epoxide group and C9-C10-double bond are essential for their toxicity [7]. Toxicological studies show that T-2 toxin is a very potent cytotoxic and immunosuppressive agent, which can cause acute intoxication and chronic diseases in both humans and animals [6]. Given that T-2 toxin is metabolised into HT-2 toxin after ingestion, they are considered to be equally toxic [8]. The symptoms of acute T-2 intoxication of different mammalian species include skin necrosis, asthenia, lack of appetite, panting, vomiting, diarrhoea, anorexia, myocardial damage, lethargy, as well as haemorrhages and necrosis of the epithelium of the stomach and intestines, bone marrow, spleen, testis and ovary [1, 8–10]. The International Agency for Research on Cancer (IARC) classified T-2 toxin into Group 3 carcinogens because of the lack of data on its carcinogenicity in humans and only limited evidence on its carcinogenicity in experimental animals [11].

Data collected in a number of European countries have shown substantial variations in *Fusarium* mycotoxin levels across various cereal types, various countries and various investigated periods [12, 13]. Croatia, as a Central European country, falls into the group of countries in which contamination with *Fusarium* mycotoxins is a frequent occurrence [14, 15]. Since data on the occurrence of T-2 and HT-2 toxins in cereals and cereal by-products are still very limited, the European Commission recommended the member states to gather reliable data on year-to-year variations of these mycotoxins in order to be able to establish their maximal levels (MLs) in different food and feed in the near future [6, 16, 17]. In 2013, the European Commission gave recommendations (Commission Recommendation 2013/165/EU) regarding indicative levels of T-2 and HT-2 toxin in cereals and cereal-based products intended for food and feed. In this document, the Commission also recommended further investigations into the effects of food processing and agronomic factors on the presence of T-2 and HT-2 toxin and different factors favouring high level-contaminations with these toxins, so as to be able to identify measures to be taken to avoid or reduce the above [18]. In view of the evidenced toxicity of T-2 and HT-2 toxin, there exists the need for further collection of data on their presence in different cereals intended for food and feed production. In their recent study, Pleadin et al. [17] stated that further studies shall also be performed in Croatia in order to investigate into the conditions favouring T-2 and HT-2 production and to identify measures that are to be implemented in order to prevent contamination during cultivation and storage of cereals and their final products.

Literature data on the impact of cereal processing on the levels of T-2 and HT-2 contamination are also very scarce. Data gathered insofar have suggested that toxins in reference, when milled, get to be relocated into various milling fractions, but are not eliminated [19]; in addition, they have been proven resistant to processing. Even more so, due to their hull binding, subsequent processing of cereals leads to significant rise in levels of these toxins in finished products [16]. Scudamore [20] concluded that final processing, such as boiling, fermentation, baking, frying, and extrusion, has no impact on the level of contamination with these mycotoxins. Some studies evidenced that processing of raw cereals greatly reduces T-2 and HT-2 levels in food products, but makes these toxins concentrate in high levels in by-products [6]. In industrial food processing settings, the processing time and temperature combination has been shown to be crucial for the reduction of mycotoxin content in a finished food product. While conventional food preparation at temperatures up to 100°C has a negligible effect on most mycotoxins, higher temperatures used with frying, roasting, toasting and extrusion have been shown to be capable of decreasing mycotoxin contamination [21].

The studies quoted under this chapter bring data on the incidence of T-2 and HT-2 toxin in different unprocessed cereals during a two-year period. They also demonstrate the influence of certain food processing methods, such as cooking, roasting and extrusion, performed under predefined conditions, on the levels of T-2 and HT-2 toxin in contaminated cereals. For this purpose, after application of the quantitative screening method termed the enzymelinked immunosorbent assay (ELISA), which enabled the determination of summary T-2 and HT-2 toxin concentrations, the concentrations of each mycotoxin in positive samples were determined using a confirmatory method in terms of liquid chromatography tandem-mass spectrometry (LC-MS/MS), also used in the investigations devoted to the possibilities of reduction of concentrations of these mycotoxins.
