**3.3. The reduction of T-2 and HT-2 toxin**

In general, different cereal treatments implemented by the food industry are known to decrease mycotoxin concentrations, but mostly do not eliminate these toxins completely. These food processing operations include sorting, trimming, cleaning, cooking, baking, frying, roasting, flaking and extrusion, and have variable effects on the level of contamination. In their recently published study, Schmidt et al. [39] stated that in comparison to other mycotoxins, thermal degradation of T-2 and HT-2 has not been the subject of many studies. In the last decades, some research on the effects of thermal degradation has mainly been performed on oats, known to be the cereal most contaminated with these mycotoxins [34, 39–41]. Scudamore [20] concluded that final processing, such as boiling, fermentation, baking, frying, and extrusion, has no impact on T-2 and HT-2 contamination. A greater extent of thermal degradation of T-2 as compared to HT-2 has been established, as well [34, 39, 41].

Nevertheless, the efficiency of T-2 and HT-2 toxin reduction using thermal processing techniques is still under-established, mostly because of the fairly small data pool on the subjectmatter provided insofar, obtained under various, mutually different thermal degradation conditions, which, in turn, yields inconsistent study outcomes and study conclusions. In light of the foregoing, in order to establish the degree of thermal degradation and reduction of T-2 and HT-2 toxin in naturally contaminated cereals, this study made use of three thermal processing methods, that is to say, cooking, roasting and extrusion, each of them running at three different temperatures for different lengths of time.

Cooking is the preparation of a meal using heat [42]. Several studies have reported about the effect of cooking on the reduction of *Fusarium* mycotoxins in contaminated cereals [34, 43, 44]. **Figure 2.** A typical LC-MS/MS-MRM chromatogram of the contaminated maize sample subsequently subjected to thermal reduction processing (128 μg/kg of T-2 and 256 μg/kg of HT-2); TIC; HT-2 MRM 447.2 → 285.1; HT-2 MRM

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447.2 → 345.1; T-2 MRM 489.2 → 245.1; and T-2 MRM 489.2 → 387.1.

When the individual concentrations of T-2 and HT-2 toxin obtained by the LC-MS/MS method were summed up, it was established that the latter sum slightly differs from, and is mainly lower than, the summary concentrations of these mycotoxins determined using the ELISA method. This can be explained by the cross-reactivity and a lower specificity of the ELISA method. It is known that the ELISA represents an easy-to-use purification technique with a lesser need for an extensive clean-up, but it may suffer from undesired cross-reactivity with

**Figure 2** presents the LC-MS/MS-MRM chromatogram of the most contaminated sample (maize), in which the concentration of T-2 toxin of 128 μg/kg and the concentration of HT-2 toxin of 256 μg/kg was determined (summary T-2/HT-2 toxins concentration, 384 μg/kg). Together with the most contaminated oat and the most contaminated triticale sample, this

Based on the comparison of the mean T-2 and HT-2 toxin concentrations, the shares of T-2:HT-2 were established to be in the range from 1:1.7 in wheat and barley to 1:2.4 in maize, with the mean share value of 1:1.9 in all investigated cereal samples. The determined share values are comparable to those stated by other literature sources, which show that HT-2 is present in cereals and their products in levels higher than those of T-2 toxin, representing

In general, different cereal treatments implemented by the food industry are known to decrease mycotoxin concentrations, but mostly do not eliminate these toxins completely. These food processing operations include sorting, trimming, cleaning, cooking, baking, frying, roasting, flaking and extrusion, and have variable effects on the level of contamination. In their recently published study, Schmidt et al. [39] stated that in comparison to other mycotoxins, thermal degradation of T-2 and HT-2 has not been the subject of many studies. In the last decades, some research on the effects of thermal degradation has mainly been performed on oats, known to be the cereal most contaminated with these mycotoxins [34, 39–41]. Scudamore [20] concluded that final processing, such as boiling, fermentation, baking, frying, and extrusion, has no impact on T-2 and HT-2 contamination. A greater extent of thermal degradation

Nevertheless, the efficiency of T-2 and HT-2 toxin reduction using thermal processing techniques is still under-established, mostly because of the fairly small data pool on the subjectmatter provided insofar, obtained under various, mutually different thermal degradation conditions, which, in turn, yields inconsistent study outcomes and study conclusions. In light of the foregoing, in order to establish the degree of thermal degradation and reduction of T-2 and HT-2 toxin in naturally contaminated cereals, this study made use of three thermal processing methods, that is to say, cooking, roasting and extrusion, each of them running at three

Cooking is the preparation of a meal using heat [42]. Several studies have reported about the effect of cooking on the reduction of *Fusarium* mycotoxins in contaminated cereals [34, 43, 44].

approximately two-thirds (1:2) of the summary T-2/HT-2 concentrations [16].

of T-2 as compared to HT-2 has been established, as well [34, 39, 41].

different temperatures for different lengths of time.

other trichothecenes that give rise to metric uncertainty [16].

52 Fusarium - Plant Diseases, Pathogen Diversity, Genetic Diversity, Resistance and Molecular Markers

sample was further subjected to thermal processing.

**3.3. The reduction of T-2 and HT-2 toxin**

**Figure 2.** A typical LC-MS/MS-MRM chromatogram of the contaminated maize sample subsequently subjected to thermal reduction processing (128 μg/kg of T-2 and 256 μg/kg of HT-2); TIC; HT-2 MRM 447.2 → 285.1; HT-2 MRM 447.2 → 345.1; T-2 MRM 489.2 → 245.1; and T-2 MRM 489.2 → 387.1.

The results descriptive of T-2 and HT-2 reduction achieved by various cooking times (10:30 min) are presented in **Table 6**.

of T-2 toxin in fortified samples of up to 76%. Schwake-Anduschus et al. [34] stated that T-2/ HT-2 toxin levels are relatively stable during short-time cooking. This was also confirmed by this study, as it resulted in a very small reduction of both mycotoxins despite the prolonged

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

Roasting is a cooking method that uses dry heat in form of an open flame, oven, or other heat sources. Roasting can enhance flavour through caramelisation and Maillard browning of the food surface [42]. The results of T-2 and HT-2 reduction via roasting, obtained in this study at three different temperatures (180–220°C), are presented in **Table 7**. **Figure 3** presents a typical LC-MS/MS-MRM chromatogram of a contaminated maize sample obtained after roasting at the temperature of 220°C during 30 min, in which a significant reduction of both T-2 toxin

**180°C 200°C 220°C**

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**Cerealsa T-2/HT-2 after roasting Temperature of roastingb**

Maize T-2/HT-2 (μg/kg) 270 225 188

Triticale T-2/HT-2 (μg/kg) 190 173 151

Oat T-2/HT-2 (μg/kg) 101 84.3 68.9

Concentrations in cereals before reduction: 384 μg/kg (128 μg/kg of T-2 and 256 μg/kg of HT-2) in maize, 267 μg/kg (107 μg/kg of T-2 and 160 μg/kg of HT-2) in oat and 151 μg/kg (47.0 μg/kg of T-2 and 104 μg/kg of HT-2) in triticale.

T-2 and HT-2 toxin concentrations are presented as the mean value of three replicates; R: reduction; ND: not detected;

**Table 7.** Reduction of T-2/HT-2 toxins achieved by roasting of contaminated cereals at different temperatures.

Roasting was carried out for 30 minutes at the default temperatures.

T-2/HT-2 R (%) 29.7 41.4 51.0 T-2 (μg/kg) 80.1 65.3 50.3 T-2 R (%) 37.4 49.0 60.7 HT-2 (μg/kg) 190 160 138 HT-2 R (%) 25.8 37.5 46.1

T-2/HT-2 R (%) 28.8 35.2 43.4 T-2 (μg/kg) 56.2 43.3 38.2 T-2 R (%) 47.5 59.5 64.3 HT-2 (μg/kg) 138 130 113 HT-2 R (%) 13.8 18.8 29.4

T-2/HT-2 R (%) 33.1 44.2 54.4 T-2 (μg/kg) 28.1 22.7 ND T-2 R (%) 40.2 51.7 CR HT-2 (μg/kg) 72.9 61.6 59.3 HT-2 R (%) 29.9 40.8 43.0

cooking time (30 min).

a

b

CR: completely reduced.

(60.7%) and HT-2 toxin (46.1%) can be seen.

The maximal reduction of T-2 toxin was observed in oat (14.7%) cooked for 30 min, whereas the greatest HT-2 (7.5%) and summary T-2/HT-2 concentration reduction (9.3%) were obtained in oat cooked for 20 min. A slightly greater reduction was observed for T-2 toxin (9.3%) in comparison to the HT-2 toxin, which was mostly unreduced in all three types of cereals despite cooking. A prolonged cooking time (30 min) achieved no significantly greater reduction of summary T-2/HT-2 concentrations or individual toxin levels. The results show that the reduction of T-2/HT-2 summary concentrations achieved by cooking can be considered negligible (R < 10%), suggesting that cooking as a thermal processing method does not represent a valuable tool when it comes to the decontamination of cereals contaminated with T-2 and HT-2. After cooking of noodles, Kamimura et al. [43] determined the residual rate


a Concentrations in cereals before cooking: 384 μg/kg (128 μg/kg of T-2 and 256 μg/kg of HT-2) in maize, 267 μg/kg (107 μg/kg of T-2 and 160 μg/kg of HT-2) in oat and 151 μg/kg (47.0 μg/kg of T-2 and 104 μg/kg of HT-2) in triticale. T-2 and HT-2 toxin concentrations are presented as the mean value of three replicates; R: reduction; NR: not reduced.

**Table 6.** Reduction of T-2/HT-2 toxins achieved by various cooking times.

of T-2 toxin in fortified samples of up to 76%. Schwake-Anduschus et al. [34] stated that T-2/ HT-2 toxin levels are relatively stable during short-time cooking. This was also confirmed by this study, as it resulted in a very small reduction of both mycotoxins despite the prolonged cooking time (30 min).

The results descriptive of T-2 and HT-2 reduction achieved by various cooking times (10:30 min)

54 Fusarium - Plant Diseases, Pathogen Diversity, Genetic Diversity, Resistance and Molecular Markers

The maximal reduction of T-2 toxin was observed in oat (14.7%) cooked for 30 min, whereas the greatest HT-2 (7.5%) and summary T-2/HT-2 concentration reduction (9.3%) were obtained in oat cooked for 20 min. A slightly greater reduction was observed for T-2 toxin (9.3%) in comparison to the HT-2 toxin, which was mostly unreduced in all three types of cereals despite cooking. A prolonged cooking time (30 min) achieved no significantly greater reduction of summary T-2/HT-2 concentrations or individual toxin levels. The results show that the reduction of T-2/HT-2 summary concentrations achieved by cooking can be considered negligible (R < 10%), suggesting that cooking as a thermal processing method does not represent a valuable tool when it comes to the decontamination of cereals contaminated with T-2 and HT-2. After cooking of noodles, Kamimura et al. [43] determined the residual rate

> T-2/HT-2 (μg/kg) 375 366 372 T-2/HT-2 R (%) 2.3 4.7 3.1 T-2 (μg/kg) 117 123 110 T-2 R (%) 8.6 3.9 14.1 HT-2 (μg/kg) 258 243 270 HT-2 R (%) NR 5.1 NR

> T-2/HT-2 (μg/kg) 252 271 262 T-2/HT-2 R (%) 5.6 NR 1.9 T-2 (μg/kg) 104 99.5 94.8 T-2 R (%) 2.8 7.0 11.4 HT-2 (μg/kg) 172 151 167 HT-2 R (%) NR 5.6 NR

> T-2/HT-2 (μg/kg) 152 137 144 T-2/HT-2 R (%) NR 9.3 4.6 T-2 (μg/kg) 43.2 40.8 40.1 T-2 R (%) 8.1 13.2 14.7 HT-2 (μg/kg) 108 96.2 104 HT-2 R (%) NR 7.5 NR

Concentrations in cereals before cooking: 384 μg/kg (128 μg/kg of T-2 and 256 μg/kg of HT-2) in maize, 267 μg/kg (107 μg/kg of T-2 and 160 μg/kg of HT-2) in oat and 151 μg/kg (47.0 μg/kg of T-2 and 104 μg/kg of HT-2) in triticale. T-2 and HT-2 toxin concentrations are presented as the mean value of three replicates; R: reduction; NR: not reduced.

**Table 6.** Reduction of T-2/HT-2 toxins achieved by various cooking times.

**10 min 20 min 30 min**

**Cerealsa T-2/HT-2 after cooking Cooking time (96°C)**

are presented in **Table 6**.

Maize

Triticale

Oat

a

Roasting is a cooking method that uses dry heat in form of an open flame, oven, or other heat sources. Roasting can enhance flavour through caramelisation and Maillard browning of the food surface [42]. The results of T-2 and HT-2 reduction via roasting, obtained in this study at three different temperatures (180–220°C), are presented in **Table 7**. **Figure 3** presents a typical LC-MS/MS-MRM chromatogram of a contaminated maize sample obtained after roasting at the temperature of 220°C during 30 min, in which a significant reduction of both T-2 toxin (60.7%) and HT-2 toxin (46.1%) can be seen.


a Concentrations in cereals before reduction: 384 μg/kg (128 μg/kg of T-2 and 256 μg/kg of HT-2) in maize, 267 μg/kg (107 μg/kg of T-2 and 160 μg/kg of HT-2) in oat and 151 μg/kg (47.0 μg/kg of T-2 and 104 μg/kg of HT-2) in triticale. b Roasting was carried out for 30 minutes at the default temperatures.

T-2 and HT-2 toxin concentrations are presented as the mean value of three replicates; R: reduction; ND: not detected; CR: completely reduced.

**Table 7.** Reduction of T-2/HT-2 toxins achieved by roasting of contaminated cereals at different temperatures.

Published data on the influence of roasting on the reduction of mycotoxins are mostly linked to aflatoxins, ochratoxins or some *Fusarium* mycotoxins other than T-2/HT-2 [45–48] and refer to baking [34, 49]. In this study, the reduction of summary T-2/HT-2 toxin levels, achieved with roasting at 180°C, ranged from 28.8 to 33.1%; at 200°C, the achieved reduction ranged from 35.2 to 44.2%, while at 220°C, a 43.4 to 54.4% reduction was witnessed. A complete T-2 toxin reduction was achieved when roasting oats; after roasting at 220°C, no detectable concentration of this mycotoxin was determined. The results show that in comparison to the HT-2 toxin, a significantly higher (nearly 2-fold) reduction of T-2 was achieved, which is in line with an earlier observation that thermal processing reduces T-2 concentrations to a greater extent as compared to those of HT-2 [39]. Roasting can be considered as an effective thermal processing method suitable for cereal decontamination, as it resulted in a roughly 40% T-2/

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Extrusion is used in the production of cereal products such as breakfast foods, snacks and animal feedstuffs, and represents one of the fastest-growing food-processing operations in the recent years due to its many advantages. Extrusion cooking of cereals combines pumping, kneading, mixing, shearing, cooking and forming, all in one processing session. As several operations are carried out simultaneously, they interact with each other [39]. Cereals are passed through an extruder under pressure, undergo mechanical shearing stresses at elevated temperature, and rapidly expand when forced through the outlet die [50]. Apart from its main goal in terms of improving the product quality, extrusion may also significantly improve the product safety because of its potential to reduce mycotoxin levels in cereals [51]. The effectiveness of mycotoxin reduction also depends on the presence of minor ingredients or additives. Scudamore et al. [20] explained that during extrusion, contaminants are subjected to both high temperatures and chemical reactions mediated by free radical mechanisms so that they might be susceptible to

some degree of breakdown, the effects on mycotoxins thereby generally being variable.

maize sample at the defined temperatures of 135-190-190°C.

the ELISA method showed higher T-2/HT-2 summary concentrations.

The reduction of T-2/HT-2 toxins achieved by extrusion cooking of contaminated cereals in this study using three different regimes of extrusion is shown in **Table 8**. **Figure 4** presents a typical LC-MS/MS-MRM chromatogram obtained after extrusion cooking of a contaminated

By virtue of extrusion cooking under three temperature regimes and with the same moisture content (25%), an almost complete reduction of T-2 and HT-2 toxin was achieved. The results show a similar effect of extrusion independent of the type of cereal and the applied temperature regime, based on which this method can be considered as the most effective and most valuable when it comes to the reduction of mycotoxins. As the LC-MS/MS method failed to determine any of the two mycotoxins in any of the extruded samples, the summary T-2/HT-2 concentrations were analysed using the ELISA method. The results showed T-2/HT-2 concentrations slightly higher than the method's LOQs, except for the oat subjected to the extrusion temperature regime of 135-190-190°C, in which the presence of the above toxins was not detected at all. The reduction of T-2/HT-2 achieved under the 135-150-150°C extrusion regime ranged from 73.0% in oats to 92.5% in maize. However, it should be taken into account that the presence of individual mycotoxins was not confirmed by the LC-MS/MS method and that

HT-2 toxins reduction in contaminated samples.

**Figure 3.** A typical LC-MS/MS-MRM chromatogram of a contaminated maize sample after roasting at the temperature of 220°C for 30 min (50.3 μg/kg of T-2 and 138 μg/kg of HT-2); TIC; HT-2 MRM 447.2 → 285.1; HT-2 MRM 447.2 → 345.1; T-2 MRM 489.2 → 245.1; and T-2 MRM 489.2 → 387.1.

Published data on the influence of roasting on the reduction of mycotoxins are mostly linked to aflatoxins, ochratoxins or some *Fusarium* mycotoxins other than T-2/HT-2 [45–48] and refer to baking [34, 49]. In this study, the reduction of summary T-2/HT-2 toxin levels, achieved with roasting at 180°C, ranged from 28.8 to 33.1%; at 200°C, the achieved reduction ranged from 35.2 to 44.2%, while at 220°C, a 43.4 to 54.4% reduction was witnessed. A complete T-2 toxin reduction was achieved when roasting oats; after roasting at 220°C, no detectable concentration of this mycotoxin was determined. The results show that in comparison to the HT-2 toxin, a significantly higher (nearly 2-fold) reduction of T-2 was achieved, which is in line with an earlier observation that thermal processing reduces T-2 concentrations to a greater extent as compared to those of HT-2 [39]. Roasting can be considered as an effective thermal processing method suitable for cereal decontamination, as it resulted in a roughly 40% T-2/ HT-2 toxins reduction in contaminated samples.

Extrusion is used in the production of cereal products such as breakfast foods, snacks and animal feedstuffs, and represents one of the fastest-growing food-processing operations in the recent years due to its many advantages. Extrusion cooking of cereals combines pumping, kneading, mixing, shearing, cooking and forming, all in one processing session. As several operations are carried out simultaneously, they interact with each other [39]. Cereals are passed through an extruder under pressure, undergo mechanical shearing stresses at elevated temperature, and rapidly expand when forced through the outlet die [50]. Apart from its main goal in terms of improving the product quality, extrusion may also significantly improve the product safety because of its potential to reduce mycotoxin levels in cereals [51]. The effectiveness of mycotoxin reduction also depends on the presence of minor ingredients or additives. Scudamore et al. [20] explained that during extrusion, contaminants are subjected to both high temperatures and chemical reactions mediated by free radical mechanisms so that they might be susceptible to some degree of breakdown, the effects on mycotoxins thereby generally being variable.

The reduction of T-2/HT-2 toxins achieved by extrusion cooking of contaminated cereals in this study using three different regimes of extrusion is shown in **Table 8**. **Figure 4** presents a typical LC-MS/MS-MRM chromatogram obtained after extrusion cooking of a contaminated maize sample at the defined temperatures of 135-190-190°C.

By virtue of extrusion cooking under three temperature regimes and with the same moisture content (25%), an almost complete reduction of T-2 and HT-2 toxin was achieved. The results show a similar effect of extrusion independent of the type of cereal and the applied temperature regime, based on which this method can be considered as the most effective and most valuable when it comes to the reduction of mycotoxins. As the LC-MS/MS method failed to determine any of the two mycotoxins in any of the extruded samples, the summary T-2/HT-2 concentrations were analysed using the ELISA method. The results showed T-2/HT-2 concentrations slightly higher than the method's LOQs, except for the oat subjected to the extrusion temperature regime of 135-190-190°C, in which the presence of the above toxins was not detected at all. The reduction of T-2/HT-2 achieved under the 135-150-150°C extrusion regime ranged from 73.0% in oats to 92.5% in maize. However, it should be taken into account that the presence of individual mycotoxins was not confirmed by the LC-MS/MS method and that the ELISA method showed higher T-2/HT-2 summary concentrations.

**Figure 3.** A typical LC-MS/MS-MRM chromatogram of a contaminated maize sample after roasting at the temperature of 220°C for 30 min (50.3 μg/kg of T-2 and 138 μg/kg of HT-2); TIC; HT-2 MRM 447.2 → 285.1; HT-2 MRM 447.2 → 345.1;

56 Fusarium - Plant Diseases, Pathogen Diversity, Genetic Diversity, Resistance and Molecular Markers

T-2 MRM 489.2 → 245.1; and T-2 MRM 489.2 → 387.1.



a Concentrations in cereals before reduction: 384 μg/kg (128 μg/kg of T-2 and 256 μg/kg of HT-2) in maize, 267 μg/kg (107 μg/kg of T-2 and 160 μg/kg of HT-2) in oat and 151 μg/kg (47 μg/kg of T-2 and 104 μg/kg of HT-2) in triticale. b Moisture in extruded samples ranges from 11.0 to 12.0%.

T-2 and HT-2 toxin concentrations are presented as the mean value of three replicates; R: reduction; ND: not detected; CR: completely reduced.

**Table 8.** Reduction of T-2/HT-2 toxins achieved by extrusion cooking of contaminated cereals at different temperatures.

When comparing the degradation rates of T-2 against those of HT-2 toxin, it was revealed that T-2 shows a higher mitigation in the extrusion cooking process [39, 41]. Some investigations showed that T-2 and HT-2 degradation during extrusion are not influenced by the heating temperature to the same extent and that other variables present during processing are responsible for a more complex degradation pattern. This observation can be linked to the results of this study, in which a significant influence of the extrusion temperature regime was not determined. Schmidt et al. [39] stated that due to the strong interference of various parameters during extrusion, it is not possible to attribute toxin degradation to just one of them. Among the factors of influence, the water content plays an important role in

**Figure 4.** A typical LC-MS/MS-MRM chromatogram of a contaminated maize sample after extrusion at 135-190-190°C (T-2 and HT-2 toxin not detected); TIC; HT-2 MRM 447.2 → 285.1; HT-2 MRM 447.2 → 345.1; T-2 MRM 489.2 → 245.1;

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and T-2 MRM 489.2 → 387.1.

**Figure 4.** A typical LC-MS/MS-MRM chromatogram of a contaminated maize sample after extrusion at 135-190-190°C (T-2 and HT-2 toxin not detected); TIC; HT-2 MRM 447.2 → 285.1; HT-2 MRM 447.2 → 345.1; T-2 MRM 489.2 → 245.1; and T-2 MRM 489.2 → 387.1.

When comparing the degradation rates of T-2 against those of HT-2 toxin, it was revealed that T-2 shows a higher mitigation in the extrusion cooking process [39, 41]. Some investigations showed that T-2 and HT-2 degradation during extrusion are not influenced by the heating temperature to the same extent and that other variables present during processing are responsible for a more complex degradation pattern. This observation can be linked to the results of this study, in which a significant influence of the extrusion temperature regime was not determined. Schmidt et al. [39] stated that due to the strong interference of various parameters during extrusion, it is not possible to attribute toxin degradation to just one of them. Among the factors of influence, the water content plays an important role in

Concentrations in cereals before reduction: 384 μg/kg (128 μg/kg of T-2 and 256 μg/kg of HT-2) in maize, 267 μg/kg (107 μg/kg of T-2 and 160 μg/kg of HT-2) in oat and 151 μg/kg (47 μg/kg of T-2 and 104 μg/kg of HT-2) in triticale.

T-2 and HT-2 toxin concentrations are presented as the mean value of three replicates; R: reduction; ND: not detected;

**Table 8.** Reduction of T-2/HT-2 toxins achieved by extrusion cooking of contaminated cereals at different temperatures.

**Cerealsa T-2/HT-2 after extrusion Regime of extrusionb**

Maize T-2/HT-2 (μg/kg) 28.9 49.7 32.2

58 Fusarium - Plant Diseases, Pathogen Diversity, Genetic Diversity, Resistance and Molecular Markers

Triticale T-2/HT-2 (μg/kg) 37.7 45.9 41.3

Oat T-2/HT-2 (μg/kg) 40.7 38.1 ND

a

b

CR: completely reduced.

Moisture in extruded samples ranges from 11.0 to 12.0%.

T-2/HT-2 R (%) 92.5 87.1 91.6 T-2 (μg/kg) ND ND ND T-2 R (%) CR CR CR HT-2 (μg/kg) ND ND ND HT-2 R (%) CR CR CR

T-2/HT-2 R (%) 85.9 82.8 84.5 T-2 (μg/kg) ND ND ND T-2 R (%) CR CR CR HT-2 (μg/kg) ND ND ND HT-2 R (%) CR CR CR

T-2/HT-2 R (%) 73.0 74.8 CR T-2 (μg/kg) ND ND ND T-2 R (%) CR CR CR HT-2 (μg/kg) ND ND ND HT-2 R (%) CR CR CR

**135-150-150°C 135-170-170°C 135-190-190°C**

extrusion cooking, because it is essential for starch gelatinization and strongly affects fluid viscosity and the expansion ratio. Extrusion cooking was shown to decrease the mycotoxin content at rates depending on the moisture level, screw centrifugation, extruder geometry, die temperature, die size, screw speed and additives [51], while the extrusion temperature was found to be a minor factor of influence. As opposed to that, high moisture levels and high shear rates substantially contribute to the toxin degradation [39, 52]. The authors elaborated that since the fate of T-2 and HT-2 and the formation of so far unknown degradation products or bound forms remains unclear, it cannot be concluded that extrusion cooking of contaminated oats is accompanied by a detoxification process. Scudamore et al. [52] pointed out that the inconsistency of the results presented in the literature may be a consequence of failure to control or report all conditions under which the extrusion process was taking place. For example, chemical breakdown taking place during an extrusion process is related to the duration of the process, so that the loss of mycotoxin will depend on the residence time of the material in the extruder. Differences in parameters implemented during extrusion cooking carried out in this study may also explain an almost complete reduction of T-2 and HT-2 toxin achieved, as opposed to other studies quoted above, in which only partial or smaller toxin reduction has been witnessed when using this thermal processing method.

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