WELL-TOLERATED FLOUR COMPOSITION

Abstract

The present invention relates to the field of food production, in particular the provision of flour mixtures for the production of bakery products, pasta and bread, which are characterized by a reduced ATI content and can nevertheless be processed into doughs which meet the technical and rheological requirements of a wheat dough.

Claims

1. A flour mixture comprising one or more alpha-amylase/trypsin inhibitor (ATI)-reduced protein sources, one or more isolated ATI-reduced starch sources, one or more baking additives and, optionally one or more hydrocolloids, wherein the content of ATI 0.19 and/or the content of ATI 0.28 of the flour mixture is reduced by at least 40% compared to type 550 flour, wherein at least one protein source comprises cereal gluten protein, isolated wheat gluten, isolated gluten variants of spelt, rye, barley, emmer, oats, einkorn, or isolated gluten components or mixtures thereof and, wherein the one or more baking additives comprise sourdough, sourdough concentrate and/or dried sourdough.

2-15. (canceled)

16. The flour mixture according to claim 1, wherein the flour mixture comprises 5-25% by weight of gluten proteins.

17. The flour mixture according to claim 1, wherein the one or more starch sources comprise wheat starch, soft wheat starch or durum wheat starch.

18. The flour mixture according to claim 17, wherein the one or more starch sources is a mixture comprising wheat starch and corn starch, potato starch, tapioca starch, hydrolyzed starch, rye starch, oat starch, or barley starch or mixtures thereof.

19. The flour mixture according to claim 1, wherein the flour mixture comprises 50-95% by weight of starch.

20. The flour mixture according to claim 1, wherein the flour mixture comprises psyllium, guar gum, chia seed flour, linseed flour, xanthan gum, tragacanth, konjac, gum arabic, karaya, HPMC (hydroxy-propyl-methyl cellulose), or sunflower seed flour or mixtures thereof.

21. The flour mixture according to claim 1, wherein the flour mixture comprises up to 3% by weight of hydrocolloids.

22. The flour mixture according to claim 1, wherein an ATI extraction of the flour mixture has a bioactivity corresponding to an IL-8 release of 8 to 60 ng IL-8 per gram of the mixture and is detectable in an IL-8 specific ELISA assay.

23. The flour mixture according to claim 1, wherein the flour mixture further comprises leavening agents and yeast.

24. The flour mixture according to claim 23, wherein the flour mixture comprises Candida milleri, Candida humilis or Kazachstania milleri and/or mixtures thereof; and optionally L. pontis, L. sanfranciscensis or L. reuteri or mixtures thereof.

25. The flour mixture according to claim 23, further comprising fermentable oligo-, di-, monosaccharides or polyols or mixtures thereof.

26. A process for preparing the flour mixture according to claim 1, comprising: a. mixing the flour mixture of claim 1 with a sourdough or a sourdough starter comprising Candida milleri, Candida humilis, Kazachstania milleri, Kazachstania exigua, Debaryomyces hansenii, Dekkera bruxellensis, Kazachstania unispora, Kluyveromyces lactis, Torulaspora delbrueckii, T. pretoriensis, Wickerhamomyces anomalus, Pichia anomala, Hansenula anomala, Pichia kudriavzevii, Issatschenkia orientalis, or Candida krusei or mixtures thereof; and optionally L. pontis, L. sanfranciscensis or L. reuteri or mixtures thereof; and b. fermenting the product generated by step a.

27. A pasta comprising the flour mixture according to claim 1, water and/or milk, and optionally egg, egg white, egg yolk or ready-made egg, wherein the total ATI content is reduced by at least 40% compared to pasta produced from type 550 flour.

28. A bakery product comprising the flour mixture according to claim 1, wherein said bakery product comprises leavening agents, yeast, sugar, salt, bread spices, gluten-free cereals and/or pseudo-cereals as grains and/or ground and water, wherein the total ATI content is reduced by at least 40% compared to bakery products produced from type 550 flour.

Description

SHORT DESCRIPTION OF THE FIGURES

[0090] FIG. 1 shows the results of the differentiated ATI determination of the LC/MS+MS analysis. Double determination; white bar=commercial wheat flour of type 550; black bar=flour mixture according to invention, triple determination; peak areas are compared; however, all samples were treated in the same way and recorded in the same volume. For the ATIs 0.19, 0.28 and CM3 two peptides were used; of CM2 and CM16 only one peptide each. While the ATIs 0.29 and 0.28 are significantly reduced in Flour B, no difference can be seen for ATIs CM2, CM3 and CM16.

[0091] FIG. 2 shows the results of the farinograph measurement and thus the kneading behaviour of the different test mixtures 1-7. The farinograms of various test mixtures are shown; Experiment 1 (- wheat flour 550), Experiment 2 (--- without gluten), Experiment 3 (-- -- without psyllium), Experiment 4 (- - - 13% gluten 2.9% psyllium), Experiment 5 (13% gluten 0.8% psyllium), Experiment 6(-- - -- 6.8% gluten 3.2% psyllium), Experiment 7 (--- --- 13% gluten 0.8% psyllium 2% Germe).

[0092] FIG. 3 shows the influence of different wheat gluten on the kneading behaviour.

[0093] FIGS. 4A and B show the influence of different amounts of wheat gluten on the baking behavior. The visually recognizable differences show bread baked from doughs with a gluten content of (from left to right) approx. 4%, approx. 12% and approx. 19% wheat gluten.

[0094] FIG. 5 shows the influence of different amounts of starch on the kneading behaviour (farinograph measurement).

[0095] FIG. 6 shows the influence of different amounts of starch on baking behaviour (photo): crumb pictures of tin loaves with different starch contents based on ATI-reduced basic mix; from left to right: approx. 95%, approx. 83%, approx. 70% and approx. 50% starch.

[0096] FIGS. 7A and B show the results of baking trials with various baking additives (from left to right (from left to right): ascorbic acid, diastase malt and ascorbic acid, a amylase and ascorbic acid.

[0097] FIG. 8 shows the result of a tasting by 14 tasters. Here, rolls and breads made from commercial wheat flour were compared with rolls and breads made from the flour mixture according to the invention.

[0098] FIG. 9 shows the results of the ATI bioactivity analysis according to Junker et al (2012) on human monocytes.

EXAMPLES

Example 1: Influence of Different Compositions on the Dough Properties

[0099] Various mixing ratios of ATI-reduced wheat starch and ATI-reduced isolated wheat gluten were compiled and tested. The test doughs were kneaded in the farinograph until the maximum viscosity was reached. The doughs were round knitted with the roll press. The dough was then left to rest for 15 minutes. The doughs were then gently kneaded for a long time and formed into small strips of dough using the press. After a further rest period of 35 minutes, the measurement was carried out.

TABLE-US-00001 TABLE 1 Test formulations and production overview Formulation Ingredients (g) #1 #2 #3 #4 #5 #6 #7 Wheat flour (type 550) 300 0 0 0 0 0 0 (100%) Wheat starch 0 294 252 249 254 267 245 (96%) (85%) (83%) (85%) (89%) (82%) Wheat gluten A 0 0 39.6 39.1 39.9 20.3 38.5 (13%) (13%) (13%) (6.8%) (13%) Psyllium 0 8.9 0 8.9 2.4 9.5 2.3 (2.9%) (2.9%) (0.8%) (3.2%) (0.8%) Rapeseed oil 0 4.2 3.6 3.6 3.6 3.8 3.5 (1.4%) (1.2%) (1.2%) (1.2%) (1.3%) (1.2%) Sourdough 0 0 0 0 0 0 10.5 (Bcker Germe) (3.5%) Water 174 225 179 197 205 225 198 Kneading time 6 6.8 10 8 10 11.8 8.1 (min) max. viscosity 547 333 390 561 364 342 326 in the Farino- graph (FE)

[0100] FIG. 2 shows the results of the farinograph measurement and thus the kneading curves or kneading behaviour of the different test mixtures 1-7 in comparison with type 550 wheat flour.

[0101] For this purpose, the doughs were examined with the various test mixtures (see Table 1 for an overview of the recipes) in the farinograph at a kneading time of 20 min at 30 C. and a speed of 63 rpm.

[0102] Table 2 below also lists the elongation and elongation resistance. The tests show clear differences between all the compounds used. Test mix 1, represents a control and shows the extensibility and max. viscosity of normal wheat flour.

TABLE-US-00002 TABLE 2 Measurement parameters microextensogram Elongation max. elongation Trial (mm) resistance (mN) 1 33.43 164 2 7.75 126 3 11.74 131 4 10.96 317 5 9.04 114 6 7.59 152 7 8.81 86

Example 2: Influence of Different Wheat Gluten on the Kneading Behaviour

[0103] The influence of different adhesives and isolated adhesive proteins on the rheological properties of the dough was investigated.

[0104] The different adhesives were used in the following formulation (#8), the amounts were not varied.

TABLE-US-00003 TABLE 3 Formulation #8 Quantity (g) Wheat starch 245 Wheat gluten 38.5 Psyllium 2.3 Rapeseed oil 3.5 Germe 10.5 Water 198

[0105] For this purpose, gluten, i.e. isolated wheat gluten, was tested by various manufacturers. The results of the kneading curves are shown in FIG. 3.

[0106] Of particular interest was the result on hydrolysed gluten, where the kneading resistance is significantly reduced, as can be seen from the kneading curve. The other glues show a comparable kneading resistance, but the 2nd increase is at different times.

Example 3: Influence of Different Amounts of Wheat Gluten on Baking Behaviour

[0107] The influence of different doses of wheat gluten should be investigated to determine the tolerance ranges of wheat gluten quantities. The amount of wheat gluten used in the previous test series was approx. 13% and represents the standard. In comparison, mixtures with approx. 4% up to approx. 20% gluten (based on the total amount of ingredients without water) were tested.

TABLE-US-00004 TABLE 4 Test formulations Recipe: #9 Formulation #10: Formulation #11: Raw material with approx. with approx. with approx. in (g) 4% adhesive 12% adhesive 19% adhesive Wheat starch 1050 1050 1050 Wheat gluten 50 165 280 Psyllium 10 10 10 Germe 24.8 24.8 24.8 Rapeseed oil 12 12 12 Yeast 40 40 40 Salt 25 25 25 Sugar 15 15 15 TOTAL (g) 1226.8 1341.8 1456.8 Water (22 C.) 735 810 900 Kneading time 2 + 8 2 + 9 2 + 9 Dough rest 15 minutes 15 minutes 15 minutes Weighing in (g) 650 650 650 Fermentation time 40 min 40 min 40 min

[0108] The doughs with an adhesive content of less than 5% had almost no elastic properties. The doughs with an adhesive quantity of approx. 20% or more, on the other hand, had a rubbery consistency.

[0109] It was noticeable that the doughs had a very strong gluten smell. The ready-baked breads with a gluten dosage of approx. 20% had a very rubbery chewing behaviour, which is very untypical compared to a conventional wheat bread.

[0110] Overall it can be said that the doses of 5 and 20% show extremes and have a negative influence on the quality of the baked goods in different ways. Ideally, an adhesive quantity of 6%-19% should be used.

[0111] FIGS. 4A and 4B show the visually recognisable differences between breads baked from doughs with a gluten content of (from left to right) 4%, 12% and 19% wheat gluten.

Example 4: Use of Different Amounts of Starch and their Effect on Dough Properties and Baking Behaviour

[0112] In this test series, the influence of different starch contents in the basic formulation of the ATI-reduced mixture was investigated. This makes it possible to estimate the variability of the base mix for subsequent product variants.

[0113] In the test series, the dough properties were investigated using a farinograph. Baking trials were carried out to assess the influence of the different starch contents on the baking properties. The following starch contents were examined: 83% (hrs), 95%, 70% and 50%.

[0114] The following Table 5 shows the recipe and the measured parameters.

TABLE-US-00005 Quantity (g) Quantity (g) Quantity (g) Quantity (g) for approx. for approx- for approx- for approx- 83% starch imately imately imately Formulation (standard) 95% starch 70% starch 50% starch Wheat starch 249.2 285 210.0 150.0 Wheat gluten 39.2 3.3 78.3 138.3 Psyllium 3.0 3.0 3.0 3.0 Germe 5.3 5.3 5.3 5.3 Rapeseed oil 3.3 3.3 3.3 3.3 Water 195.8 195.8 195.8 195.8 (TA 165) Measurement parameters Speed (rpm) 63 Dough tem- 30 perature ( C.) Measuring 20 duration (min)

[0115] The Farinograph measurement (FIG. 5) shows that the use of different starch contents has a significant influence on the kneading properties of the doughs/masses, thus changing the overall curve characteristics. The similarity of the curves for wheat flour 550 and the test mass with 95% starch content is striking, but this provides little information about baking properties. The extent of the differences in processing and baking properties can be assessed on the basis of the baking results (FIG. 6) and the following description.

Baking Trial:

[0116] The following Table 6 shows the recipe and the production parameters of the baking tests, as well as their results.

TABLE-US-00006 Quantity (g) Quantity (g) Quantity (g) Quantity (g) for approx. for approx- for approx- for approx- 83% starch imately imately imately Formulation (standard) 95% starch 70% starch 50% starch Wheat starch 1050 1200 885 632 Wheat gluten 165 14 330 583 Hydrocolloid X 12.62 12.62 12.62 12.62 Germe 22.5 22.5 22.5 22.5 Rapeseed oil 14 14 14 14 Yeast 45 45 45 45 Salt 30 30 30 30 Sugar 15 15 15 15 Water (22 C.) 825 825 825 825 Production parameters Kneading (min) 2 slow + 9 fast Dough tem- 29 perature ( C.) Weighing in 750 (in loaf pan) (g) Gare (min) 45 Baking tem- 240 falling to 210 perature ( C.) Baking time 45 (min) Miscellaneous Cut the bread after cooking

[0117] Results:

TABLE-US-00007 Dough temperature Baking loss Volume Specific volume Sample ( C.) (%) (ml) (ml/g) Remarks 83% 29 16.1 1960 3.1 Starch 95% 26 18.6 1235 2.0 Dough: moist, sticky, not starch workable Bread: no bread quality 70% 36 18.2 2820 4.6 Dough: rubbery, strong starch heating of the dough, not workable Bread: very strong oven baking, crumbling crumb (still acceptable) 50% 37 19.6 4750 7.9 Dough: rubbery, strong starch heating of the dough, not workable Bread: very strong oven drift, balling and rubbery crumb

Example 5: Use of Different Hydrocolloids and their Effect on Dough Properties and Baking Behaviour

[0118] In this test series, the influence of different hydrocolloids on the base mix for the production of ATI-reduced baked goods was investigated. The results should ensure that the functionality of the base mix is not bound to a specific hydrocolloid. The dosages of all hydrocolloids were 1% (based on ingredients WITHOUT water). This represents a typical dosage of hydrocolloids in baked goods.

[0119] In order to assess the influence of the different hydrocolloids on the baking properties, baking tests were carried out. The following hydrocolloids were investigated: [0120] No hydrocolloid, missing hydrocolloid was replaced by wheat starch (Abbreviation: NONE) [0121] Psyllium, is used as standard with 0.8% in the basic recipe (abbreviation: Psy (standard)) [0122] Guar gum (3500 cps) (Abbreviation: Guar) [0123] HPMC (4000 cps) (Abbreviation: HPMC) [0124] Xanthan gum (1550 cps) (Abbreviation: Xan) [0125] Traganth (400 cps) (Abbreviation: Trag) [0126] Konjac (36000 cps) (Abbreviation: Konj) [0127] Gum Arabic (-cps) (Abbreviation: Ara) [0128] Karaya (-cps) (Abbreviation: Kara)

Baking Trial:

[0129] Table 7 shows the basic recipe and the production parameters of the baking trials:

TABLE-US-00008 Formulation ingredients Quantity (g) Wheat starch 1245.9 Wheat gluten A 195.8 Hydrocolloid X 15.0 Germe 26.7 Rapeseed oil 16.6 Yeast 45 Salt 30 Sugar 15 Water (22 C.) 979 Manufacturing Kneading (min) 2 slow + 9 fast parameters Dough temperature ( C.) 29 Weighing in (g) 750 (in loaf pan) Gare (min) 45 Baking temperature 240 falling to 210 ( C.) Baking time (min) 45 Miscellaneous Cut the bread after cooking

[0130] The following Table 8 shows the results of the baking tests using the various hydrocolloids or without hydrocolloid.

TABLE-US-00009 TABLE 8 Back Vol- Specific loss ume volume (%) (ml) (ml/g) Remarks NO 15.9 2213 3.5 dough: soft and sticky, difficult to cut after cooking, no stall Bread: sides no cracks Psy 15.7 1875 3.0 Dough: good working properties, long (hrs.) kneading time for dough formation Bread: low oven drift, slight cracks on the side Guar 16.7 2300 3.7 Dough: comparable with Psy (hrs) bread: no cracks on the side HPMC 18.9 2360 3.9 Dough: soft, moist and sticky, little standing, difficult to cut after cooking Bread: very good baking, slight cracks on the side (less than at Psy Std.) Xan 15.2 1805 2.8 Dough: doughs appear dry Bread: strong cracks on the side Trag 16.1 2230 3.5 Dough: soft and moist, no stall Bread slight cracks on the side Account 16.1 2180 3.5 Dough: firm and not sticky (compare with Psy Std.) Bread: very strong cracks on the side Macaw 17.2 2120 3.4 Dough: soft and moist, no stall, difficult to cut after cooking Bread: slight cracks on the side Kara 16.5 2115 3.4 Dough: compare with Psy Std. Bread: strong cracks on the side

Example 6: Use of Different Baking Additives and their Effect on Dough Properties and Baking Behaviour

[0131] The influence of different baking additives should be investigated. Formulation 10 from the previous trials was used as a basis.

TABLE-US-00010 TABLE 9 Formulation #11: Formulation #12: Formula #13: Raw material with ascorbic Diastase Malt and Amylase and in (g) acid (g) ascorbic acid (g) ascorbic acid (g) Wheat starch 1050 1050 1050 Wheat gluten 165 165 165 (Krner) Psyllium 10 10 10 Germe 24.8 24.8 24.8 Rapeseed oil 15 15 15 Yeast 40 40 40 Salt 25 25 25 Sugar 12 12 12 Ascorbic acid 0.1 0.1 0.1 Diastase Malt 38.55 Fungamyl 0.07 Water (22 C.) 810 810 810 Kneading time 2 + 9 2 + 9 2 + 9 Dough rest 15 minutes 15 minutes 15 minutes Weighing in 650 650 650 (g) Fermentation 40 min 40 min 40 min time Volume (ml) 1790 1800 1890

[0132] The doughs showed almost no differences in their properties.

[0133] In the baked goods, in the baking test, differences could be detected depending on the additives. Bread produced with diastatic malt showed a stronger browning, but no increase in volume compared to bread produced with ascorbic acid alone. The breads without ascorbic acid showed a comparable volume (not shown in the experiment).

[0134] Through the use of amylase, an increase in the pastry volume of approx. 100 ml could be achieved. The biscuits did not appear more strongly browned.

[0135] FIGS. 7A and 7B show the results of the baking tests (from left to right): ascorbic acid, diastase malt and ascorbic acid, a amylase and ascorbic acid.

Example 7: Standardization of an ATI-Reduced Formulation

[0136]

TABLE-US-00011 TABLE 10 Test formulation #14 Ingredient Quantity (g) % share in baking mix Wheat starch 1050 81.3 Wheat gluten A 165 12.8 Psyllium 37.5 2.9 Germe 22.5 1.7 Rapeseed oil 15 1.2 Yeast 60 Salt 37.5 Water 830 Dough temp. ( C.) 26 Kneading time (min) 2 + 7 Fermentation time 40 (min) Processing/Form Split roll

Dough Properties:

[0137] The dough yield was slightly increased with TA 164 compared to typical wheat doughs. The kneading time was slightly longer than with conventional wheat doughs. During processing, clear differences in the dough properties became apparent. The surface of the doughs appeared very dry and the doughs were more plastic than elastic, which can be attributed to the psyllium used.

[0138] The dough was processed as cut rolls, this processing was almost impossible. No acceptable conclusion could be reached with the dough pieces. This jumped up, which is due to the dry dough characteristics. During the cooking process there was only a very slight increase in volume.

Baking Properties:

[0139] The poor gas retention of the dough pieces during cooking resulted in baked goods with a very small volume. Despite the low volume, the cut of the rolls had a very pronounced effect. In addition, there was no typical browning of the baked goods during baking. The baked goods appeared greyish and showed an uneven leathery surface. The surface and crumb also appeared stippled, which can also be attributed to the psyllium.

[0140] For further experiments, the amount of psyllium was significantly reduced, as it has a significant influence on the dough and baking properties. The properties seem to be dominant in relation to the gluten properties.

TABLE-US-00012 TABLE 11 Test recipe #15 #16 #17 Quantity (g) Quantity (g) Quantity (g) Ingredient Share % er Share % er share % Wheat starch 1050 87.14 1050 83.1 1050 83.1 Wheat gluten 80 6.6 165 13.1 165 13.1 Psyllium 37.5 3.1 10 0.8 10 0.8 Germe 22.5 1.9 22.5 1.8 22.5 1.8 Rapeseed oil 15 1.2 15 1.2 15 1.2 Yeast 60 60 60 Salt 37.5 37.5 37.5 Water 930 880 850 Sugar 11 Dough temp. ( C.) 26 26 26 Kneading time (min) 2 + 8 2 + 8 2 + 9 Fermentation time 35 35 35 (min) Refurbishment/form Loaf pan and free Loaf pan and free Loaf pan and free

Experimental Formulation 15

[0141] The dough and baking properties were comparable with those of test recipe 14. This confirmed the impression that if the psyllium dosage is too high, the properties of this hydrocolloid predominate and no viscoelastic properties are produced.

Experimental Formulation 16

[0142] The dough properties were comparable to those of a wheat dough. The chosen dough yield of TA 169 was high, the doughs were very soft for processing. The doughs were comparable to conventional breads with regard to crumb structure and formation of crumbs. The atypical greyish crust colour was clearly noticeable.

Experimental Formulation 17

[0143] The dough yield (TA 167) and the kneading time were adjusted, resulting in dough properties that are typical of conventional wheat doughs. As a result of this and the use of sugar, the volume, texture, crumb structure and colour of the crust were improved to such an extent that there is almost no difference to conventional wheat bread.

Tasting

[0144] In taste tastings, it was determined how the rolls and breads produced with trial mix 17 (mixture according to the invention) and trial mix 1 (wheat flour) affected 14 testers in comparison. The optical impression (colouring and appearance), odour and aroma, taste as well as texture and haptics were evaluated.

[0145] The result (FIG. 8) shows that although the testers gave different evaluations and differences could be perceived, particularly with regard to taste, the overall impression is that the products made from standard flour or the ATI-reduced flour mixture are highly similar and comparable in terms of feel, texture, colour and appearance.

Example 8: ATI-BIOACTIVITY Determination for Test Formulations

[0146] The ATI bioactivity of test recipe 17 (flour mixture according to invention) and test recipe 1 (commercial wheat flour) was compared with the method Junker et al., 2012 (JEM, 2012) and e.g. described in WO2017075456.

[0147] For this purpose the bioactivity of ATIs is tested on human monocytes, e.g. the THP-1 cell line. Confluently grown THP-1 cells are stimulated with flour mixture extraction dissolved in neutral buffer. Depending on the amount of ATI, IL-8 is released from this cell line into the supernatant, which is then determined by a standardized ELISA according to the manufacturer's (ThermoFischer scientific) specifications.

[0148] Interestingly, clearly different cytokine releases were found in the supernatant of test cells, indicating different bioactivity of the contained ATIs.

[0149] FIG. 9 shows the result of the IL-8 specific ELISA assay, for which suitable test cells, namely ATI-reactive cellsaccording to WO2017075456 e.g. TLR4-expressing cells, preferably TLR4-expressing monocytesare brought into contact with ATI extracts (buffer extraction) and then the cytokine release, namely an IL-8 release, is measured in the supernatant by ELISA. FIG. 9 shows that the flour mixture according to the invention triggers a significantly lower (more than 90% lower) IL-8 release in the ATI bioactivity test compared to a commercial standard flour.

[0150] From these data, it can be clearly concluded that patients who are ATI-sensitive and who show various NCGS symptoms can expect a significantly better tolerance and digestibility of products made from the flour mixture according to the invention.

Example 9: Differentiated ATI Determination

[0151] For test recipe 17 (flour mixture according to inventionblack bar) and test recipe 1 (commercial standard flourwhite bar), a differentiated ATI determination was carried out using LC-MS/MS in accordance with Prandi et al. (Food Chemistry, 2013, 141-146).

[0152] For this purpose, peptides derived from enzymatic cleavage of the salt-soluble extracts were identified using the LC-MS/MS method. Lead peptides for ATI quantification are the ATI 0.19, 0.28 and CM3 with 2 cleavage peptides each, and the ATI CM2, and CM16 with one cleavage peptide. All measurements were performed at least in duplicate determinations. FIG. 1 shows the results.

[0153] As can be seen in FIG. 1, the flour mixture according to the invention has a much lower content of ATI 0.19 and ATI 0.28 in contrast to the standard flour, while the ATI CM3 content and the ATI CM2 and CM16 content remain unchanged. The Y-axis in FIG. 1 describes the area of the peaks determined by chromatography. It is generally known to the expert that the (peak) area is proportional to the concentration of the analyte (in our case the different ATI proteins).

[0154] The inventors were thus able to show that their experimental mixture 17, for which a reduced ATI bioactivity was found, surprisingly mainly reduced the content of ATI 0.19 and ATI 0.28, while the other ATs showed smaller differences.

Example 10: Sugar Determination According to AOAC Method (997.08)

[0155] For test recipe 17 (flour mixture according to the invention and test recipe 1 (commercial standard flour) a total sugar determination was carried out using the standard method AOAC method (997.08).

TABLE-US-00013 sugar Commercial Flour mixture according substance (g/100 g) wheat flour to the invention Glucose (g/100 g) <0.1 <0.1 Fructose <0.1 <0.1 Sucrose 0.3 <0.1 Fructooligosaccharides 1.0 0.1