Structural Modifications of Gluten Proteins in Strong and Weak Wheat Dough During Mixing

The network‐forming attributes of gluten have been investigated for decades, but no study has comprehensively addressed the differences in gluten network evolution between strong and weak wheat types (hard and soft wheat). This study monitored changes in SDS protein extractability, SDS‐accessible thiols, protein surface hydrophobicity, molecular weight distribution, and secondary structural features of proteins during mixing to bring out the molecular determinants of protein network formation in hard and soft wheat dough. Soft wheat flour and dough exhibited greater protein extractability and more accessible thiols than hard wheat flour and dough. The addition of the thiol‐blocking agent N‐ethylmaleimide (NEM) resulted in similar results for protein extractability and accessible thiols in hard and soft wheat samples. Soft wheat dough had greater protein surface hydrophobicity than hard wheat and exhibited a larger decrease in surface hydrophobicity in the presence of NEM. Formation of high‐molecular‐weight (HMW) protein in soft wheat dough was primarily because of formation of disulfides among low‐molecular‐weight (LMW) proteins, as indicated by the absence of changes in protein distribution when NEM was present, whereas in hard wheat dough the LMW fraction formed disulfide interaction with the HMW fraction. Fourier transform infrared spectroscopy indicated formation of β‐sheets in dough from either wheat type at peak mixing torque. Formation of β‐sheets in soft wheat dough appears to be driven by hydrophobic interactions, whereas disulfide linkages stabilize secondary structure elements in hard wheat dough.     

Structural Modification of Gluten Proteins in Strong and Weak Wheat Dough as Affected by Mixing Temperature

The effects of temperature (≥25°C) on dough rheological properties and gluten functionality have been investigated for decades, but no study has addressed the effect of low temperature (<30°C) on gluten network attributes in flours with strong and weak dough characteristics. This study monitored changes in protein extractability in the presence and absence of reducing agents, the contents of readily accessible and SDS‐accessible thiols, and the secondary structural features of proteins in doughs from commercial hard wheat flour (HWF) and soft wheat flour (SWF) mixed at 4, 15, and 30°C. SWF mixed at 4 and 15°C showed similar mixing properties as HWF mixed at 30°C (which is the standard temperature). The effect of mixing temperature is different at the molecular level between the two flours studied. Protein features of HWF did not change as mixing temperature decreased, with the only exception being an increase in SDS‐accessible thiols. Decreasing mixing temperature for SWF caused an increase in SDS protein solubility and SDS‐accessible thiols as well as an increase in β‐turn structures at the expense of β‐sheet structures. Thus, noncovalent interactions appear to drive protein network at low temperatures (4 and 15°C), whereas covalent interactions dominate at standard mixing temperature (30°C) in doughs from both flours.     

Wheat Gluten Polymer Structures: The Impact of Genotype,Environment, and Processing on Their Functionality in Various Applications

For a number of applications, gluten protein polymer structures are of the highest importance in determining end‐use properties. The present article focuses on gluten protein structures in the wheat grain, genotype‐ and environment‐related changes, protein structures in various applications, and their impact on quality. Protein structures in mature wheat grain or flour are strongly related to end‐use properties, although influenced by genetic and environment interactions. Nitrogen availability during wheat development and genetically determined plant development rhythm are the most important parameters determining the gluten protein polymer structure, although temperature during plant development interacts with the impact of the mentioned parameters. Glutenin subunits are the main proteins incorporated in the gluten protein polymer in extracted wheat flour. During dough mixing, gliadins are also incorporated through disulfide‐sulfhydryl exchange reactions. Gluten protein polymer size and complexity in the mature grain and changes during dough formation are important for breadmaking quality. When using the gluten proteins to produce plastics, additional proteins are incorporated in the polymer through disulfide‐sulfhydryl exchange, sulfhydryl oxidation, β‐eliminations with lanthionine formation, and isopeptide formation. In promising materials, the protein polymer structure is changed toward β‐sheet structures of both intermolecular and extended type and a hexagonal close‐packed structure is found. Increased understanding of gluten protein polymer structures is extremely important to improve functionality and end‐use quality of wheat‐ and gluten‐based products.     

Influence of High‐Temperature Drying on Structural and Textural Properties of Durum Wheat Pasta

Starch and protein are the main polymeric ingredients of pasta and they determine the structural and textural properties of cooked pasta. The present investigation sought better understanding of the impact of high‐temperature (HT) drying on the starch and the protein fraction, and their role in structure and texture of pasta. Durum wheat spaghetti was prepared in a pilot‐plant installation. The drying conditions were selected for the HT phase at 80 or 100°C applied at high, intermediate, or low product moisture content. Spaghetti dried at 55°C served as a reference sample. The color of dry pasta was measured and the changes in the starch and protein fractions were determined by protein solubility, light microscopy, confocal scanning laser microscopy (CSLM), cooking tests, and texture measurements. HT drying at 100°C and low product moisture promoted browning of pasta. At the molecular level, HT drying promoted protein denaturation. At the microscopic level, HT drying contributed to a better preservation of the protein network and reduced swelling of starch and disintegration of granules. At the macroscopic level, HT drying enhanced the firmness of cooked pasta and reduced surface stickiness. In general, the changes were more pronounced by increasing the drying temperature from 80 to 100°C and by shifting the HT phase from an early to a late stage of the drying process. The drying conditions are determinant for the phase morphology of protein and starch in cooked pasta which, in turn, govern the textural properties of pasta.     

A Fast,Simple, and Reliable Method to Predict Pasta Yellowness

Pasta yellowness depends on the semolina carotenoid content, carotenoid degradation by lipoxygenase (LOX), and pasta processing conditions. In breeding programs, early generation lines are selected for high grain yellow pigment content with the intent to improve pasta color. This approach has been successful in increasing the grain yellow pigment of Canadian durum wheat in the last few decades. In recent years, however, a weak relationship between pasta yellowness (b*) as measured by a Minolta spectrophotometer and semolina yellow pigment content (r = 0.19–0.52) was noted in the Canadian durum wheat lines. Thus, total semolina yellow pigment content cannot effectively predict the yellowness of its pasta product. Therefore, a fast and simple method was developed to predict pasta yellowness by measuring semolina dough sheet color at different time intervals after sheeting (0.5, 2.0, and 24 hr). Spaghettis were processed from the semolina samples at two drying temperature cycles (70 and 90°C). There were significant correlations between dough sheet b* values at all three times and spaghetti b* values at both drying temperatures (r = 0.87–0.94). Semolina dough sheet can be easily prepared in 15 min and requires only 30 g of material. Shortly after sheeting (30 min), dough sheet b* values can be used to predict pasta yellowness without producing the end product (involving mixing, extrusion, and drying). In this study, we also found that dough sheet b* values increased significantly with time over the sampling intervals after sheeting for those breeding lines with superior pasta color. DNA analysis revealed that all those lines lacked the Lpx‐B1.1 duplication.     

Tyrosine cross-links: molecular basis of gluten structure and function

The formation of the large protein structure known as "gluten" during dough-mixing and bread-making processes is extremely complex. It has been established that a specific subset of the proteins comprising gluten, the glutenin subunits, directly affects dough formation and breadmaking quality. Glutenin subunits have no definitive structural differences that can be directly correlated to their ability to form gluten and affect dough formation or breadmaking quality. Many protein structural studies, as well as mixing and baking studies, have postulated that disulfide bonds are present in the gluten structure and contribute to the process of dough formation through the process of disulfide-sulfhydryl exchange. Evidence presented here indicates that tyrosine bonds form in wheat doughs during the processes of mixing and baking, contributing to the structure of the gluten network. The relative contributions of tyrosine bonds and disulfide--sulfhydryl interchange are discussed.     

Impact of Bran Addition on Water Properties and Gluten Secondary Structure in Wheat Flour Doughs Studied by Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy

The aim of this work was to elucidate the underlying physical mechanism(s) by which bran influences whole grain dough properties by monitoring the state of water and gluten secondary structure in wheat flour and bran doughs containing 35–50% moisture and 0–10% added bran. The system was studied with attenuated total reflectance (ATR) FTIR spectroscopy. Comparison of the OH stretch band of water in flour dough with that in H₂O‐D₂O mixtures having the same water content revealed the formation of two distinct water populations in flour dough corresponding to IR absorption frequencies at 3,600 and 3,200 cm^–1. The band intensity at 3,200 cm^–1, which is related to water bound to the dough matrix, decreased and shifted to lower frequencies with increasing moisture content of the dough. Addition of bran to the dough caused redistribution of water in the flour and bran dough system, as evidenced by shifts in OH stretch frequency in the 3,200 cm^–1 region to higher frequencies and a reduction in monomeric water (free water). This water redistribution affected the secondary structure of gluten in the dough, as evidenced by changes in the second‐derivative ATR‐FTIR difference spectra in the amide I region. Bran addition caused an increase in β‐sheet content and a decrease in β‐turn (β‐spiral) content. However, this bran‐induced transconformational change in gluten was more significant in the 2137 flour dough than in Overley flour dough. This study revealed that when bran is added to flour dough, water redistribution among dough components promotes partial dehydration of gluten and collapse of β‐spirals into β‐sheet structures. This transconformational change may be the physical basis for the poor quality of bread containing added bran.     

Influence of Gluten Proteins and Drying Temperature on the Cooking Quality of Durum Wheat Pasta

It is well known that gluten plays a major role in determining cooking quality in durum wheat pasta. This work is an attempt to systematically elucidate the role of gluten quantity and nature in determining cooking quality as a function of the drying cycle used in the manufacturing process. Gluten and starch were fractionated from two durum wheat cultivars possessing good and poor gluten quality. Either of them were then added back to the original base semolina to alter its protein content and to produce two semolina series with identical protein contents. Semolinas were processed into pasta and dried following three drying programs (low, medium, and high temperature). Cooking quality was determined with sensorial, chemical, and instrumental methods. The results indicate that optimum cooking time is governed by gluten quality. The positive effect on cooking quality of increasing gluten contents and of the application of HT drying is evident in weak gluten samples, but it is not significant in the strong gluten samples.     

Protein Structural Features in Winter Wheat: Benchmarking Diversity in Ontario Hard and Soft Winter Wheat

A set of 32 winter wheat lines and varieties was selected to benchmark Ontario winter wheat as a first step toward improving quality. Protein secondary structure, total and accessible thiols, rheological properties, gluten aggregation kinetics, and network forming capabilities of different polymers were determined for each wheat line. Results revealed that there were statistically significant differences among the lines selected (P < 0.05). The differences between hard and soft wheat classes were not as large as would be expected, however, despite the range of quality parameters measured. Benchmarks revealed that several soft wheat lines outperformed hard wheat lines in standard breadmaking quality measures. Protein conformation changed significantly as the moisture content of the samples increased to mimic different model product systems: flour, dough, and batter. The conformation of the flour samples exhibited different patterns between hard and soft wheat classes, although these differences became narrower in the dough and batter states. Principal component analysis (PCA) factors included most quality parameters measured, with the notable exceptions of solvent retention capacity tests and total thiols. Protein conformation and accessible thiols were significant PCA factors that tended to override the rheological measures of quality they represented, suggesting that protein secondary structure and disulfide bonding patterns are fundamental aspects of rheological quality measures.     

Effect of Transglutaminase,Citrate Buffer,and Temperature on a Soft Wheat Flour Dough System

Transglutaminase (TGase) can improve the functional characteristics of proteins by introducing covalent bonds inter‐ or intrachains. Temperature and pH interfere with the protein structure and the catalytic activity of enzymes. Because these three factors can act synergistically, TGase, citrate buffer, and temperature were evaluated for their effects on the rheological and chemical changes in low‐protein wheat flour dough. Dough strength, measured by microextension test, significantly increased with increasing levels of TGase (8 U/g of protein), with changes in pH of the citrate buffer (pH 6.5), and by the effect of interaction between these factors. The same trend was observed in the size‐exclusion HPLC measurements, indicating that these two parameters have the effect of increasing gluten protein aggregation. Temperature had a significant effect on dough extension, measured by microextension test. The changes in secondary structure of gluten protein were investigated by FTIR second‐derivative spectra (amide I region, 1,600–1,700 cm⁻¹) and showed an increase in β‐sheet structures initiated by TGase, citrate buffer pH, and their interaction.     

Pasta made from durum wheat semolina fermented with selected lactobacilli as a tool for a potential decrease of the gluten intolerance

A pool of selected lactic acid bacteria was used to ferment durum wheat semolina under liquid conditions. After fermentation, the dough was freeze-dried, mixed with buckwheat flour at a ratio of 3:7, and used to produce the "fusilli" type Italian pasta. Pasta without prefermentation was used as the control. Ingredients and pastas were characterized for compositional analysis. As shown by two-dimensional electrophoresis, 92 of the 130 durum wheat gliadin spots were hydrolyzed almost totally during fermentation by lactic acid bacteria. Mass spectrometry matrix-assisted laser desorption/ionization time-of-flight and reversed phase high-performance liquid chromatography analyses confirmed the hydrolysis of gliadins. As shown by immunological analysis by R5-Western blot, the concentration of gluten decreased from 6280 ppm in the control pasta to 1045 ppm in the pasta fermented with lactic acid bacteria. Gliadins were extracted from fermented and nonfermented durum wheat dough semolina and used to produce a peptic-tryptic (PT) digest for in vitro agglutination tests on cells of human origin. The whole PT digests did not cause agglutination. Affinity chromatography on Sepharose-6-B mannan column separated the PT digests in three fractions. Fraction C showed agglutination activity. The minimal agglutinating activity of fraction C from the PT digest of fermented durum wheat semolina was ca. 80 times higher than that of durum wheat semolina. Pasta was subjected to sensory analysis: The scores for stickiness and firmness were slightly lower than those found for the pasta control. Odor and flavor did not differ between the two types of pasta. These results showed that a pasta biotechnology that uses a prefermentation of durum wheat semolina by selected lactic acid bacteria and tolerated buckwheat flour could be considered as a novel tool to potentially decrease gluten intolerance and the risk of gluten contamination in gluten-free products.     

Textural Images Analysis of Pasta Protein Networks to Determine Influence of Technological Processes

The structure of pasta is largely governed by the presence of a structured protein network. This work analyzed the protein network textures of various cooked pasta products through textural image analysis. Six different pasta types were investigated: reference pasta made from durum semolina; pasta enriched with gluten proteins from soft wheat flour at 10 and 20%; autoclaved pasta; soft wheat flour pasta; and pasta made from reconstituted flour fractions. Pasta samples were sectioned, and each crosssection consisted of three distinct zones (central, intermediate, and external) based on the state of swelling of starch granules for each pasta product. Digital images of the protein network in each zone were acquired using confocal laser scanning microscopy. Textural image analysis was then performed. Similarities and differences in protein network texture were assessed by principal component, stepwise discriminant, and variance analyses. With the exception of autoclaved pasta, protein network structure differed greatly with the position in the pasta. Furthermore the effect of technological treatments was greatly influenced by the position in pasta. The most significant differences in protein network structure were obtained with the autoclaved and 20% protein-enriched samples.     

Effects of Mixing Conditions on Pasta Dough Development and Biochemical Changes

Mixing of commercial durum wheat semolina with water was performed under different conditions in a Brabender micromixer equipped with pastamaking shafts. Semolina filling of the mixing chamber was 30.4–42.9% (v/v), shaft speed was 10–110 rpm, temperature was 10–40°C, and hydration level was 47–52.5% (db). The blend of water and semolina evolved from individualized hydrated particles (HP) to a dough product (DP) as a function of these conditions. Torque values (T) and the specific mechanical energies (SME) were recorded during mixing as a function of time. Terms from these curves were defined to characterize the mixing process: t_o (starting time of dough development), t_d (time to reach the maximum dough consistency), T_m (mean torque value after dough development), and SME_f (total energy applied to the dough during mixing). Transformation of HP into DP and the mixing temperature were the main parameters affecting t_o, t_d, T_m, and SME_f. Protein aggregate distribution was measured by size-exclusion HPLC, protein solubility in 0.01N acetic acid, free -SH content, soluble arabinoxylans, reducing sugars, ferulic acid, carotenoid content, and oxidase activities to characterize the biochemical changes that occurred during pasta dough formation. DP was characterized by lower amounts of insoluble glutenin aggregates, lower protein solubility in dilute acetic acid, lower free -SH content, ferulic acid, carotenoid content, and lower oxidoreductase activities as compared to HP. Once the dough was developed, the effects of mixing speed, temperature, or hydration level on the biochemical composition of the blend were null or low compared to the modifications that were observed when the blends changed from HP to DP. The t_o and SME_f were the most significant parameters in characterizing the pasta dough mixing process in relation to biochemical changes.     

Characteristics of Perennial Wheatgrass (Thinopyrum intermedium) and Refined Wheat Flour Blends: Impact on Rheological Properties

Intermediate wheatgrass (IWG) (Thinopyrum intermedium) is a perennial grass with desirable agronomic traits and positive effects on the environment. It has high fiber and protein contents, which increase the interest in using IWG for human consumption. In this study, IWG flour was blended with refined wheat at four IWG‐to‐wheat ratios (0:100, 50:50, 75:25, and 100:0). Samples were analyzed for proximate composition, microstructure features, pasting properties (Micro Visco‐Amylo‐Graph device), protein solubility, and total and accessible thiols. Gluten aggregation properties (GlutoPeak tester) and mixing profile (Farinograph‐AT device) were also evaluated. IWG flour enrichment increased the pasting temperature and decreased the peak viscosity of blended flours. IWG proteins exhibited higher solubility than wheat, with a high amount of accessible and total thiols. The GlutoPeak tester highlighted the ability of IWG proteins to aggregate and generate torque. Higher IWG flour enrichment resulted in faster gluten aggregation with lower peak torque, suggesting weakening of wheat gluten strength. Finally, the addition of IWG to refined wheat flour resulted in a decrease in dough development time and an increase in consistency, likely because of the higher levels of fiber in IWG. The 50% IWG flour enrichment represents a good compromise between nutritional improvement and maintenance of the pasting properties, protein characteristics, and gluten aggregation kinetics.     

Effect of Environment and Genotype on Durum Wheat Gluten Strength and Pasta Viscoelasticity

Data on the quality of durum wheat genotypes grown under eight environments (site-year combinations) were evaluated to determine the relative effects of genotype and environment on quality characteristics associated with gluten strength, protein content, and pasta texture. The 10 durum wheat genotypes assessed in this study represented a range of gluten strength types from the very strong U.S. desert durum genotype, Durex, to the medium strength Canadian genotype, Plenty. Considerable genetic variability was detected for all quality characteristics studied. Genotype-environment interaction was significant for all quality parameters evaluated, with the exception of mixograph development time. Genotypeenvironment interaction was most important in determining protein content and least important in determining gluten index, gluten viscoelasticity, and SDS sedimentation volume. The nature of the genotype-environment interaction was evaluated by determining the number of significant crossover (rank change) interactions. There was at least one significant crossover interaction between pairs of genotypes and environments for five of eight quality traits tested. Of 45 genotype pairs, eight and six showed significant crossover interactions for protein content and pasta disk viscoelasticity, respectively. Significant crossover interactions were at least partially due to the differential response of Canadian genotypes as compared with U.S. genotypes. With the exception of protein content and pasta disk viscoelasticity, our results suggest that among the selected sample of 10 genotypes, genotype-environment interactions were minor and due primarily to changes in magnitude rather than changes in rank.     

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.     

Description of Chemical Changes Implied During Bread Dough Mixing by FT‐ATR Mid‐Infrared Spectroscopy

The aim of the present study was to investigate the ability of mid‐infrared (MIR) spectroscopy to identify physicochemical changes in the French bread dough mixing process. An ATR FT‐MIR spectrometer at 4000–800 cm^–1 was used. The MIR spectra collections recorded during mixing were analyzed after standard normal variate using principal component analysis (PCA) and after second‐derivative treatment. The results were interpreted in terms of chemical changes involved in dough development and more particularly in terms of secondary structural protein changes (amide III). The loading spectrum associated with principal component 1 (PC1) allows three MIR wave number regions of variations (3500–3000, 1700–1200, and 1200–800 cm^–1) to be identified. The loading spectrum associated with PC1 describes an increase in the relative protein band intensities and a decrease in relative water and starch band intensities. The variation during bread dough mixing time of the different amide III bands identified after the second‐derivative show that α‐helical, β‐turn, and β‐sheet structures increase while random coil structure decreases, suggesting that the gluten structure is becoming a more ordered structure. The MIR mixing time identified as being the maximum scores value on the PC1 scores plots was associated with the time at which the dough apparent torque begin to collapse, suggesting that the MIR spectroscopy could monitor bread dough development.     

Relationship Between Physicochemical Properties of Wheat Flour,Wheat Protein Composition,and Textural Properties of Cooked Chinese White Salted Noodles

Physicochemical properties and protein composition of 39 selected wheat flour samples were evaluated and correlated with the textural properties of Chinese hard‐bite white salted noodles. Flour samples were analyzed for their protein and wet gluten contents, sedimentation volume, starch pasting properties, and dough mixing properties by farinograph and extensigraph. Molecular weight distribution of wheat flour proteins was determined with size‐exclusion (SE) HPLC, SDS‐PAGE, and acid‐PAGE. Textural properties of Chinese hard‐bite white salted noodles were determined through texture profile analysis (TPA). Hardness, springiness, gumminess, and chewiness of cooked noodles were found to be related to the dough mixing properties. Both protein content and protein composition were found to be related to TPA parameters of noodles. The amount of total flour protein was positively correlated to hardness, gumminess, and chewiness of noodles. The absolute amounts of different peak proteins obtained from SE‐HPLC data showed positive correlations with the hardness, gumminess, chewiness, and springiness of noodles. The proportions of these peak proteins were, however, not significantly related to texture parameters. The proportions of low‐molecular‐weight glutenins/gliadins and albumins/globulins, as observed from SDS‐PAGE, were correlated positively and negatively, respectively, to the hardness, gumminess, and chewiness of cooked noodles. Among the alcohol‐soluble proteins (from acid‐PAGE data), β‐gliadins showed strong correlations with the texture properties of cooked noodles. For the selected flour samples, the total protein content of flour had a stronger relationship with the noodle texture properties than did the relative proportion of different protein subgroups. Prediction equations were developed for TPA parameters of cooked noodles with SE‐HPLC and rapid visco analysis data of the 30 flour samples, and it was found that about 75% of the variability in noodle hardness, gumminess, and chewiness values could be explained by protein composition and flour pasting properties combined together. About 50% of the variations in cohesiveness and springiness were accounted for by these prediction equations.     

GlutoPeak: A Breeding Tool for Screening Dough Properties of Durum Wheat Semolina

A rapid shear‐based test using a GlutoPeak instrument was compared with tests commonly used by durum wheat breeders to assess the potential of this instrument to discriminate between samples. Thirty‐two durum wheat semolina samples were analyzed by mixograph, SDS sedimentation (SDSS), gluten index (GI), and GlutoPeak testing. A subset was also tested for pasta quality. GlutoPeak peak maximum time (PMT) was the best indicator of gluten strength and correlated well with the other tests except SDSS. Samples with higher levels of SDS‐unextractable glutenin (insoluble protein [IP]) had stronger dough and longer PMT, but the GlutoPeak test only correlated with pasta stickiness using a smaller set of samples. The range in mixogram profiles encountered in breeding material was related to the IP content, and the pasta made from the different types was of similar quality, differing more because of protein content rather than mixogram type. The GlutoPeak test is faster than GI and uses less sample, requires little technical skill, and is suitable for evaluating large numbers of breeder's lines. The GlutoPeak test is best suited to discriminating weak from strong dough samples and allows for testing with small samples, thus facilitating quality evaluations at early stages of a breeding program.     

Phytochemical Profile and Antiproliferative Activity of Dough and Bread Fractions Made from Refined and Whole Wheat Flours

Phytochemical profile (phenolic acids, carotenoids, and tocopherols) and antiproliferative properties of bread processing fractions, including the dough, crumb, and upper crust made from refined wheat and whole wheat flours were analyzed for two wheat cultivars. Ferulic acid, lutein, and α‐tocopherol were the predominant phenolic acid, carotenoid, and tocopherol, respectively, extracted from all fractions. The levels of all phytochemicals in whole wheat samples were over eightfold higher than their corresponding refined wheat samples. The concentrations of total phenolic acids (soluble and insoluble bound) were higher in the upper crust of refined (∼60–90%) and whole wheat (∼15–40%) breads than their corresponding dough fractions. However, the dough of whole wheat had higher levels of tocopherols and carotenoids compared with the crumb and upper crust, suggesting that phenolic acids were relatively stable during baking, whereas tocopherols (∼25–80%) and carotenoids (∼20–80%), were partially degraded. The antiproliferative activity of whole wheat bread extracts against HT‐29 cancer cells was weakly correlated with total phenolic acids but showed no correlations with total carotenoid and total tocopherol contents.