Mixograph Responses of Gluten and Gluten‐Fortified Flour for Gluten Produced by Cold‐Ethanol or Water Displacement of Starch from Wheat Flour

The total protein of gluten obtained by the cold‐ethanol displacement of starch from developed wheat flour dough matches that made by water displacement, but functional properties revealed by mixing are altered. This report characterizes mixing properties in a 10‐g mixograph for cold‐ethanol‐processed wheat gluten concentrates (CE‐gluten) and those for the water‐process concentrates (W‐gluten). Gluten concentrates were produced at a laboratory scale using batter‐like technology: development with water as a batter, dispersion with the displacement fluid, and screening. The displacing fluid was water for W‐gluten and cold ethanol (≥70% vol, ‐12°C) for CE‐gluten. Both gluten types were freeze‐dried at ‐10°C and then milled. Mixograms were obtained for 1) straight gluten concentrates hydrated to absorptions of 123–234%, or 2) gluten blended with a low protein (9.2% protein) soft wheat flour to obtain up to 16.2% total protein. The mixograms for gluten or gluten‐fortified flour were qualitatively and quantitatively distinguishable. We found differences in the mixogram parameters that would lead to the conclusion of greater stability and strength for CE‐gluten than for W‐Gluten. Differences between the mixograms for these gluten types could be markedly exaggerated by increasing the amount of water to the 167–234% range. Mixograms for evaluation of gluten have not been previously reported in this hydration range. Mixograms for fortification suggest that less CE‐gluten than W‐gluten would be required for the same effect.     

Effect of Processing on Functional Properties of Wheat Gluten Prepared by Cold‐Ethanol Displacement of Starch

Functional properties of gluten prepared from wheat flour are altered by separation and drying. Gluten was separated and concentrated by batterlike laboratory methods: development with water, dispersion of the batter with the displacing fluid, and screening to collect the gluten. Two displacing fluids were applied, water or cold ethanol (70% vol or greater, ‐13°C). Both the water‐displaced gluten (W‐gluten) and ethanol‐displaced‐ gluten (CE‐gluten) were freeze‐dried at ‐20°C as a reference. Samples were dried at temperatures up to 100°C using a laboratory, fluidized‐bed drier. Tests of functionality included 1) mixing in a mixograph, 2) mixing in a farinograph, and 3) the baked gluten ball test. Dough‐mixing functionality was assessed for Moro flour (9.2% protein) that was fortified up to 16% total protein with dried gluten. In the mixograph, CE‐gluten (70°C) produced improved dough performance but W‐gluten (70°C) degraded dough performance in proportion to the amount added in fortification. In the microfarinograph, there was a desirable and protein‐proportional increase in stability time for CE‐gluten (70°C) but no effect on stability for W‐gluten (70°C). Baking was evaluated using the baked gluten ball test and the percentage increase in the baked ball volume relative to the unbaked gluten volume (PIBV). PIBV values were as high as 1,310% for freeze‐dried CE‐gluten and as low as 620% for W‐gluten dried at 70°C. PIBV for CE‐gluten was reduced to 77% of the freeze‐dried control by fluid‐bed drying at 70°C. Exposure of CE‐gluten to 100°C air gave a PIBV that was 59% of the reference, but this expansion was still greater than that of W‐gluten dried at 70°C. The highest values of PIBV occurred at the same mixing times as the peak mixograph resistance.     

Proteins Extracted by Water or Aqueous Ethanol During Refining of Developed Wheat Dough to Vital Wheat Gluten and Crude Starch as Determined by Capillary‐Zone Electrophoresis (CZE)

Fluids applied to large‐sale, technical separation of wheat starch and protein also extract soluble proteins. The degree and rate of extraction and the specific components extracted depend on the flour, the flour hydration and development, the starch‐displacing fluid composition, the temperature, and the mechanical processing method. This study sought to identify major extracted protein groups using high‐performance capillary zone electrophoresis (CZE) applied directly to fluids obtained during laboratory‐scale technical separations. A dough‐ball or compression separation method was applied using a Glutomatic system and a batter or dispersion method was applied using a a McDuffie mixer and Pharmasep vibratory separator. Process fluids were water at 22°C to model commercial practice and 70 vol% ethanol in water at ‐13°C to model the cold ethanol process being developed here. Data were referenced to use of 70 vol% ethanol in water at 22°C in the Glutomatic compression method. The dough processed by each method was developed by mixing to a separable state. When flooded with excess water, this dough immediately released starch and water‐soluble or albumin proteins. When flooded with excess cold aqueous ethanol, neither the albumin nor gliadin proteins appeared in significant amounts until the bulk of the starch had been displaced, regardless of the mechanical method. Even with extraction and manipulation well beyond that necessary for starch displacement, the net amount of gliadin proteins dissolved was only ≈10% of that available from wet developed dough using 70 vol% ethanol at 22°C. There was more gliadin protein in the fluids at earlier stages of processing when the batter dispersion method was applied using cold ethanol. The most common soluble proteins revealed in the electrophoresis patterns for the batter compression method using cold aqueous ethanol were initially albumins and later γ‐gliadins. Albumins not appearing as soluble in cold 70 vol% ethanol were found in the insoluble crude starch, suggesting their precipitation in the dough fluids during the change from free water to cold aqueous ethanol. These results establish that some protein is dissolved during starch displacement by cold aqueous ethanol, but that the amounts may be limited by control of the mechanical working of the dough in the presence of the displacing fluids.     

Optimizing Extraction of Zein and Glutelin‐Rich Fraction During Sequential Extraction Processing of Corn

This study was conducted to improve yields and qualities of corn protein co‐products produced by the sequential extraction process (SEP), a process using ethanol to fractionate corn in producing fuel ethanol. A two‐stage extraction protocol was evaluated to recover zein and subsequently recover a glutelin‐rich fraction (GRF). After the simultaneous oil‐extraction and ethanol‐drying step of SEP, zein was extracted from the anhydrous‐ethanol‐defatted, flaked corn by using 70% (v/v) ethanol at 60°C for 1.5 hr in a shaking water bath. Zein was recovered by ultrafiltering and then drying in a vacuum‐oven. Zein yield was 65% of the available zein in the flaked corn. SDS‐PAGE band patterns of the recovered zein closely resembled that of commercial zein. After zein extraction, the GRF was extracted using 45% ethanol and 55% 0.1M NaOH at 55°C for 2 hr. The extract was concentrated by ultrafiltration and then freeze‐dried. GRF yield was ≈65% of the available protein. Freeze‐dried GRF contained 90% crude protein (db), which classified the protein as a protein isolate. As with the protein concentrate from the original SEP, the GRF isolate was highly soluble in water at pH ≥ 7, had good emulsifying and foaming properties, formed stable emulsions, and was heat‐stable.     

Significance of Wheat Flour Particle Size on Sponge Cake Baking Quality

We evaluated the effect and magnitude of flour particle size on sponge cake (SC) baking quality. Two different sets of wheat flours, including flours of reduced particle size obtained by regrinding and flour fractions of different particle size separated by sieving, were tested for batter properties and SC baking quality. The proportion of small particles (<55 μm) of flour was increased by 11.6–26.9% by regrinding. Despite the increased sodium carbonate solvent retention capacity, which was probably a result of the increased starch damage and particle size reduction, reground flour exhibited little change in density and viscosity of flour‐water batter and produced SC of improved volume by 0.8–15.0%. The volume of SC baked from flour fractions of small (<55 μm), intermediate (55–88 μm), and large (>88 μm) particles of soft and club wheat was in the range of 1,353–1,450, 1,040–1,195, and 955–1,130 mL, respectively. Even with comparable or higher protein content, flour fractions of intermediate particle size produced larger volume of SC than flour fractions of large particle size. The flour fractions of small particle size in soft white and club wheat exhibited lower flour‐water batter density (102.6–105.9 g/100 mL) than did those of large and intermediate particle fractions (105.2–108.2 g/100 mL). The viscosity of flour‐water batter was lowest in flour fractions of small particle size, higher in intermediate particles, and highest in large particles. Flour particle size exerted a considerable influence on batter density and viscosity and subsequently on SC volume and crumb structure. Fine particle size of flour overpowered the negative effects of elevated starch damage, water absorption, and protein content in SC baking.     

Wheat Proteins Extracted from Flour and Batter with Aqueous Ethanol at Subambient Temperatures

Contact of wheat flour with aqueous ethanol may enrich protein by starch displacement or deplete protein by extraction depending on 1) extraction conditions and 2) the form of the substrate. Extraction at subambient temperatures has not been described for specific gliadins for either dry flour with the protein in native configurations or for wet, developed batter or dough. This limits the ability to interpret technologies such as the cold-ethanol method. Here, we describe specific albumin and gliadin composition of flour extracts by capillary zone electrophoresis CZE in 0–100% (v/v) ethanol from –12 to 22°C. Extraction was reduced for albumin and gliadin protein as the temperature was reduced and the concentration range for extraction narrowed. Extraction dropped precipitously between 0 and –7°C for both albumins and gliadins. Electrophoretically defined gliadins extracted in constant proportion at 22°C and 30–80%(v/v) ethanol, but at lower temperature, the α-gliadins were enriched and β-gliadins depleted in the 30–55% (v/v) range. For extracts from wheat flour batter, depletion of α and β and enrichment of γ relative to the dry flour contact suggested that the electrophoretically slow migrating γ- and ω-proteins are less well incorporated to the dough matrix than electrophoretically fast migrating α and β types.     

Tracking Arabinoxylans Through the Preparation of Pancakes

Arabinoxylans (AX) are well known to have a wide‐ranging influence on wheat (Triticum aestivum L.) end‐use quality and are associated with health benefits. There is little information on the effects of processing on AX properties in high‐water‐content batter‐based products and on the associations between AX properties and end‐use quality in such products. The objective of this study was to track total and water‐extractable AX (TAX and WEAX, respectively) contents and determine changes in AX characteristics throughout the baking process of pancakes, a batter‐based wheat product. The TAX and WEAX contents along with the arabinose‐to‐xylose (A/X) ratio were quantified in refined flour and wholemeal as well as batter and pancakes from two soft and three hard wheat varieties. ANOVA F values indicated that the variation in TAX content was influenced most by sample type differences (flour versus batter versus pancakes), whereas varietal differences were responsible for the greatest differences in WEAX. In separate analyses on refined and wholemeal flours, the highest F values were for variety WEAX, largely attributed to the higher WEAX content of the three hard varieties. WEAX levels generally increased slightly from flour to batter to pancakes in refined flour. The WEAX content in flour, batter, and pancakes of both refined flour and wholemeal was highly correlated with pancake volume. These observations suggest moderate changes in wheat AX characteristics during processing and a positive association of WEAX levels with end‐product volume in a batter‐based product.     

Selective enzyme-mediated extraction of capsaicinoids and caratenoids from chili guajillo puya (Capsicum annum L.) using ethanol as solvent

The selective extraction of capsaicinoids and carotenoids from chili guajillo "puya" flour was studied. When ethanol was used as solvent, 80% of capsaicinoids and 73% of carotenoids were extracted, representing an interesting alternative for the substitution of hexane in industrial processes. Additionally, when the flour was pretreated with enzymes that break the cell wall and then dried, extraction in ethanol increased to 11 and 7% for carotenoid and capsaicinoid, respectively. A selective two-stage extraction process after the treatment with enzymes is proposed. The first step uses 30% (v/v) ethanol and releases up to 60% of the initial capsaicinoids, and the second extraction step with industrial ethanol permits the recovery of 83% of carotenoids present in the flour.     

Changes in Secondary Protein Structures During Mixing Development of High Absorption (90%) Flour and Water Mixtures

Wheat flour and water mixtures at 90% absorption (dry flour basis) prepared at various mixing times were examined using Fourier transform infrared (FT‐IR) reflectance spectroscopy. Spectra were obtained using a horizontal attenuated total reflection (ATR) trough plate. The apparent amount of protein and starch on the surface of the dough varied with mixing time but this was likely due to the polyphasic nature of the substrate and the changing particle distributions as the batter matrix was developed. Deconvolution of the Amide I band revealed contributions from alpha helical, β‐turn, β‐strand, β‐sheet, and random conformations. The ratio of β‐sheet to nonsheet conformations reached its greatest value about the same time that the mixture was most effectively separated by a laboratory‐scale, cold‐ethanol‐based method but before the peak consistency measured by a microfarinograph.     

Pericarp Fiber Separation from Corn Flour Using Sieving and Air Classification

In the dry‐grind process, starch in ground corn (flour) is converted to ethanol, and the remaining corn components (protein, fat, fiber, and ash) form a coproduct called distillers dried grains with solubles (DDGS). Fiber separation from corn flour would produce fiber as an additional coproduct that could be used as combustion fuel, cattle feed, and as feedstock for producing valuable products such as “cellulosic” ethanol, corn fiber gum, oligosaccharides, phytosterols, and polyols. Fiber is not fermented in the dry‐grind corn process. Its separation before fermentation would increase ethanol productivity in the fermenter. Recently, we showed that the elusieve process, a combination of sieving and elutriation (air flow), was effective in fiber separation from DDGS. In this study, we evaluated the elusieve process for separating pericarp fiber from corn flour. Corn flour remaining after fiber separation was termed “enhanced corn flour”. Of the total weight of corn flour, 3.8% was obtained as fiber and 96.2% was obtained as enhanced corn flour. Neutral detergent fiber (NDF) of corn flour, fiber, and enhanced corn flour (dry basis) were 9.0, 61.5, and 5.7%, respectively. Starch content of corn flour, fiber, and enhanced corn flour (dry basis) were 68.8, 23.5, and 71.3%, respectively. Final ethanol concentration from enhanced corn flour (14.12% v/v) was marginally higher than corn flour (13.72% v/v). No difference in ethanol yields from corn flour and enhanced corn flour was observed. The combination of sieving and air classification can be used to separate pericarp fiber from corn flour. The economics of fiber separation from corn flour using the elusieve process would be governed by the production of valuable products from fiber and the revenues generated from the valuable products.     

Oil uptake properties of fried batters from rice flour

Batters were prepared, using rice flour as the main component, and analyzed for their oil uptake properties during frying. Rice flour resisted oil absorption better but was less effective as a thickening agent than wheat flour. Of the rice components, increased amylose in the amylopectin/amylose ratio of the starch decreased the batter oil uptake, whereas increased protein content had the opposite effect. Various additives were introduced and investigated for their ability to develop viscosity and other desirable characteristics for the batter. As additives to the rice flour batters, phosphorylated starch and gelatinized rice flour enhanced both the thickening and oil-reducing capacities of the batter. Compared with values for batters from wheat flour, the percent batter oil uptake in the fried crust for the modified rice flour batters was decreased by up to 62%, and the percent total oil uptake for the whole coated drumstick was reduced by up to 59%.     

Exploring Relationships Between Pancake Quality and Grain and Flour Functionality in Soft Wheats

The objective of this study was to observe the influence of differences in genotype (variety) and protein concentration on batter flow and pancake making performance of a collection of soft white winter wheats. Wheats were chosen to express contrasting absorption characteristics and oxidative gelation potentials. Pancakes were processed with two formulations, one (“old”) with egg, soy, and dairy and one (“new”) without. Pancake performance was compared with grain, milling, flour, solvent retention capacity (SRC), pasting, and oxidative gelation characteristics of the flours. Kernel texture, break flour yield, carbonate SRC, and lactic acid SRC were not significantly associated with pancake performance for either formulation. ANOVA showed that flour protein concentration had a dominant effect on pancake batter flow and dimensions. Flour protein concentration affected pancakes more than flour protein quality (lactic acid SRC). Water and sucrose SRCs and Rapid Visco Analyzer pasting temperature were negatively correlated with pancake batter flow and dimensions. Pasting temperature was significantly and positively correlated with flour protein, suggesting that correlations with pancake properties might be simply a cross‐correlation with protein concentration. Notably, and in contrast to our hypothesis, oxidative gelation potential had no relationship with pancake processing or quality.     

Polymer Conformation Structure of Wheat Proteins and Gluten Subfractions Revealed by ATR‐FTIR

The polymer conformation structure of gluten extracted from a Polish wheat cultivar, Korweta, and gluten subfractions obtained from 2 U.K. breadmaking and biscuit flour cultivars, Hereward and Riband, was investigated using attenuated total reflectance Fourier transform infrared spectroscopy (ATR‐FTIR). The results showed the conformation of proteins varied between flour, hydrated flour, and hydrated gluten. The β‐sheet structure increased progressively from flour to hydrated flour and to hydrated gluten. In hydrated gluten protein fractions comprising gliadin, soluble glutenin, and gel protein, β‐sheet structure increased progressively from soluble gliadin and glutenin to gluten and gel protein; β‐sheet content was also greater in the gel protein from the breadmaking flour Hereward than the biscuit flour Riband.     

Impact of xylanases with different substrate selectivity on gluten-starch separation of wheat flour

The influence on wheat flour gluten-starch separation of a xylanase from Aspergillus aculeatus (XAA) with hydrolysis selectivity toward water extractable arabinoxylan (WE-AX) and that is not inhibited by wheat flour xylanase inhibitors was compared to that of a xylanase from Bacillus subtilis (XBS) with hydrolysis selectivity toward water unextractable arabinoxylan (WU-AX) and that is inhibited by such inhibitors. XAA improved gluten agglomeration through degradation of WE-AX and concomitant reduction in viscosity, which in the laboratory scale batter procedure with a set of vibrating sieves (400, 250, and 125 microm), increased protein recoveries on the 400 microm sieve. In contrast, XBS had a negative effect as it decreased gluten protein recovery on this sieve, probably as a result of the viscosity increase that accompanied WU-AX solubilization. Hence, it was active even if most likely a considerable part of its activity was prevented by xylanase inhibitors. A combination of XAA and XBS at a low dosage yielded a distribution of gluten proteins on the different sieves comparable to that of the control. At a high combined dosage, the gluten agglomeration was better than that with XAA alone, indicating that both WE-AX and WU-AX have a negative impact on gluten agglomeration. Finally, experiments with endoxylanase addition at different moments during the separation process suggest that the status of the arabinoxylan population during dough mixing is far less critical for its impact on gluten agglomeration than that during the batter phase.     

Fermented Sorghum as a Functional Ingredient in Composite Breads

Whole sorghum flour was fermented (a five‐day natural lactic acid fermentation) and dried under forced draught at 60°C, and evaluated for its effect on sorghum and wheat composite bread quality. In comparison with unfermented sorghum flour, fermentation decreased the flour pH from 6.2 to 3.4, decreased total starch and water‐soluble proteins, and increased enzyme‐susceptible starch, total protein, and the in vitro protein digestibility (IVPD). Fermentation and drying did not decrease the pasting temperature of sorghum flour, but slightly increased its peak and final viscosity. In comparison with composite bread dough containing unfermented sorghum flour, fermented and dried sorghum flour decreased the pH of the dough from 5.8 to 4.9, increased bread volume by ≈4%, improved crumb structure, and slightly decreased crumb firmness. IVPD of the composite bread was also improved. Mixing wet fermented sorghum flour directly with wheat flour (sourdough‐type process) further increased loaf volume and weight and reduced crumb firmness, and simplified the breadmaking process. It appears that the low pH of fermented sorghum flour inactivated amylases and increased the viscosity of sorghum flour, thus improving the gas‐holding capacity of sorghum and wheat composite dough. Fermentation of sorghum flour, particularly in a sourdough breadmaking process, appears to have considerable potential for increasing sorghum utilization in bread.     

Sequential Acid,Alkaline, and Enzymatic Modifications of Chickpea and Lentil Flours Impacted Batter Physical Properties

The effect of sequential acid, alkaline, and enzymatic treatment of chickpea and lentil flours on batter rheological properties was investigated. Substitution of wheat with disrupted chickpea and lentil flours significantly (P < 0.05) increased water‐holding capacity from 66.8% in wheat flour to more than 70.0% based on the disruption treatment, indicating an improved adhesion of coated batter. Flow behavior index of batter treatments of partially replaced wheat flour with various ratios of disrupted chickpea and lentil flours ranged from 0.88 to 1.36 and was significantly (P < 0.05) lower than the flour (i.e., 2.15) and nondisrupted control (i.e., 1.28–1.38 for chickpea and 1.22–1.28 for lentil) flours. Consistency coefficients of disrupted chickpea and lentil flours were significantly (P < 0.05) greater when replacing wheat control, indicating a best fit for the shear‐thickening model. Flour disruption decreased the treatment's pasting properties, except the setback, providing support for the significant role of proteins in dictating the pasting characteristics of batter flour treatments. Results of this study suggested a potential use for treated chickpea and lentil flours in enhancing batter rheological properties including adhesion and water‐holding capacity.     

Investigation of Soft Wheat Flour Quality Factors Associated with Sponge Cake Sensory Tenderness

Relationships among soft wheat quality parameters relating to sponge cake volume and sensory tenderness were investigated. Sixteen soft wheats from the 2008–2009 crop and 11 from the 2009–2010 crop, including Japanese soft wheat cultivars, advanced breeders' lines, and western white wheat imported from the United States, were milled and evaluated for protein content, sucrose solvent retention capacity value, specific surface area, flour pasting properties, batter pasting viscosity, sodium dodecyl sulfate sedimentation (SDSS) volume, farinograph properties, specific cake volume, and sensory tenderness score to investigate their relationships. Batter pasting viscosity was measured with a Rapid Visco Analyzer (RVA) at 2 min after reaching 90°C in heating a mixture with equal weights of flour, sucrose, and water. RVA minimum viscosity of flour suspension in water was the most influencing factor and positively correlated to specific cake volume, and RVA batter pasting viscosity and SDSS volume were negatively correlated. Meanwhile, protein content and SDSS volume were strongly negatively correlated with sensory tenderness score. Stepwise multiple regression analysis selected protein content and specific cake volume as independent variables to predict sensory tenderness score; however, SDSS volume and farinograph properties relating to protein strength were not selected. Protein content affected sponge cake tenderness independently of specific cake volume, which was related to differences in cake density.     

Effect of Morphology of Mechanically Developed Wheat Flour and Water on Starch from Gluten Separation Using Cold Ethanol Displacement

The mechanical development of wheat flour and water creates micro and macro structures in dough or batter that critically influence the ability to separate starch from protein by fluid displacement. This study sought to identify specific structural and rheological features and to relate these to separation as indexed by the separation factor. Structural features, especially protein and starch distributions, were examined using visible light microscopy applied to dough samples that had been exposed to a protein dye. Flour and water samples were developed in a Brabender microfarinograph at conditions (water content and time of development) generally suitable for use of the USDA Western Regional Research Center, cold‐ethanol fluid‐displacement method. No truly homogenous structures were observed. However, distinct segregation of protein and starch were apparent at all conditions. Structural features correlated qualitatively with the success of separation indexed by the overall separation factor (α_p/s) for the separation process. Highly segregated states characterized by large protein bands, clustered starch, and large open spaces were obtained with intermediate development (25 ± 5 min) and were most readily separated (α_p/s = 118 ± 7). Segregated states with relatively thin protein bands (≤10 μm dia) in complex networks entrapping starch were obtained after additional development (≥45 min) and were less completely separable (α_p/s = 32 ± 2). Segregated states with irregularly organized protein in the form of clumps and bands were obtained with minimal development and were partially separable (α_p/s = 65 ± 4). Consistency indicated on the microfarinograph increases monotonically throughout and beyond the period of maximum separability. However, elasticity changes and a high rate of increase in consistency evident in the microfarinogram may reflect changes in the structure that also reduce separability. The study demonstrated the use of the ethanol method to isolate development from displacement phenomena forindependent study.     

Effect of Modified Oat Starch and Protein on Batter Properties and Quality of Cake

Starch and protein separated from oat were chemically modified using cross‐linking and acetylation protocols for starch, and deamidation and succinylation for protein isolate. Cross‐linking decreased swelling power of starch, whereas syneresis increased, but cross‐linking does not have a significant effect on gelatinization temperature. Acetylation increased swelling power, but gelatinization temperature and syneresis diminished. Deamidation and succinylation increased nitrogen solubility index, emulsion activity, foaming capacity, and water and oil binding capacity. Emulsion stability did not change with deamidation and it diminished with succinylation, while foaming stability decreased with both treatments. Acetylated starch and two types of modified proteins were substituted for 5, 10, 15, and 20% of oat flour to bake cake samples and then physical properties of the cakes were measured. Acetylated starch increased batter viscosity, cake volume, and whiteness of cake crust. Increased level of deamidated protein produced cakes with lower batter viscosity, higher volume, and darker color (increase in redness). Application of higher levels of succinylated protein led to higher batter viscosity and lightness, and lower cake volume. Therefore, substitution of deamidated protein and acetylated starch can improve cake properties.     

Effect on Dough Functional Properties of Partial Fractionation,Redistribution, and In Situ Deposition of Wheat Flour Gluten Proteins Exposed to Water,Ethanol, and Aqueous Ethanol

The application of the cold‐ethanol laboratory fractionation method to the bulk separation of wheat starch and gluten is accompanied by incidental dissolution, removal, or redeposition of a small part of the functional gliadin protein. The new distribution resulting from process incidental redeposition of soluble components or by purposeful add‐back of soluble and leached components can lead to differences in functionality and more difficult recovery of native properties. To assess this issue, we exposed several wheat flour types to ethanol and water (50–90% v/v) solutions, water, and absolute ethanol at 22°C and –12°C. The exposure was mass conserving (leached components returned to substrate by evaporation of the solvent without separation of phases) or mass depleting (leached components not returned to substrate). The result of the mass‐conserving contact would be flour with altered protein distributions and intermolecular interactions. The result of the mass‐depleting contact would also include altered protein content. Furthermore, the mass‐conserving contact would model an industrial outcome for a cold‐ethanol process in which leached components would be added back from an alcohol solution. The leaching result was monitored by mixography of the flour, nitrogen analysis, and capillary zone electrophoresis of extracts. Although dough rheology was generally like that of the source flour, there were notable differences. The primary change for mass‐conserving contact was an increase in the time to peak resistance and a decrease in the rate of loss of dough resistance following peak resistance. These changes were in direct proportion to the amount of protein mobilized by the solvent. Leaching at 22°C, prevented dough formation for most aqueous ethanol concentrations and greatly reduced gliadin protein content. Minimal changes were noted for solvent contact at –12°C regardless of the ethanol concentration. The data suggested that 1) the conditions applied in cold‐ethanol enrichment of protein from wheat will generally preserve vital wheat gluten functionality, 2) functionality losses can be recovered by returning the solubilized fractions, and 3) the flour to which the gluten is added may require more mixing.