Extraction of Wheat Starch with Aqueous Sodium Hydroxide

Wheat starch was extracted with aqueous sodium hydroxide at 30–38% starch solids, pH 11.5–12.3, and 25–42°C for 0.17–24 hr. Stirring wheat starch at pH 12.3 and 25°C for 3 and 24 hr, then washing with water, neutralizing, and washing again, removed 70 and 90% phosphorus (P), respectively. Adding 16% sodium sulfate (dry starch basis) into the alkaline medium removed ≈80% of P at pH 12.0 and 25°C in 3 hr and >95% of P at pH 11.7 and 42°C in 3 hr. Sulfate ion was absorbed strongly by wheat starch in aqueous sodium hydroxide at pH 12.0, and sodium sulfate also increased the starch's uptake of hydroxide ion. Low-P wheat starch (>90% of P removed) retained the fatty acids in the untreated starch, but a fatty acid-amylose complex was not detectable by differential scanning colorimetry. The enthalpy of gelatinization of the low-P wheat starch almost matched that of prime starch, as did its X-ray diffraction pattern. Those data are consistent with saponification of the lysophospholipid in the amorphous phase of the starch to form fatty acid salts and glycerol-choline or glycerol-ethanolamine phosphodiesters that slowly diffused out of the granules. The low-P wheat starch was judged to have less “cereal” odor than the prime starch, and its pasting temperature at 9.3% starch solids was lowered by ≈10°C.     

Study of Internal Structure of Waxy Wheat Starch Granules by KI/I2 Solution

Wheat starch granules were separated from waxy wheat flour by the acetic acid (pH 3.5) fractionation technique. Waxy wheat starch samples were stained red‐brown using KI/I₂ solution. Subsequently, the concentration of the KI/I₂ solution was increased to >2.5% KI and 1.0% I₂. Then the appearance of the waxy wheat starch granules quickly became ghostlike. The internal structure of the ghost could be observed. This structure consisted of two parts with different colors: a red‐brown central part and a slightly pinkish surrounding part. Sonication of the ghost at 20 kHz for 20 sec separated the central part from the surrounding part. Each of the separated parts were subjected to gel‐filtration chromatography, and their chromatography profiles indicated only amylopectin and no amylose. The central and surrounding parts had molecular weights of 54.3 × 10⁵ and 23.8 × 10⁶ from Berry plots, respectively.     

Structure of Lysophosphatidylglyceryl-Remazolbrilliant Blue (LPG-RBB) from the Surface of Dyed Wheat Starch Granules

Wheat starch granule surface was covalently stained with Remazolbrilliant blue-R dye (RBB) and then extracted with 1% SDS containing 1% 2-mercaptoethanol (2-ME) at room temperature for 14.5 hr. The extracted blue-staining material (A₆₅₀) separated into two fractions. Low molecular weight (LMW) material was further purified by Sephadex G-75 size-exclusion chromatography and thin-layer chromatography. Infrared and nuclear magnetic resonance (¹H-NMR and ¹³C-NMR) spectroscopy indicated that the structure of the purified LMW material was 18-O-(6-lysophosphatidylglyceryl)-RBB.     

Molecular Structure and Organization of Starch Granules from Developing Wheat Endosperm

Wheat starches isolated from seeds harvested between 7 and 49 days after anthesis (DAA) were fractionated into large (>8 μm) and small (<8 μm) granules and studied for starch structure and architecture. Starch granules at 7 DAA possessed unimodal size distribution, whereas it was bimodal at later maturity stages. The apparent amylose fraction of starch granules at early maturity (7 and 14 DAA) consisted of intermediate‐type materials, whereas starch at later maturity stages (28 and 49 DAA) contained branched amylose. Wide‐angle X‐ray scattering (WAXS) revealed a well‐developed polymorphic structure already at 7 DAA. Although the presence of a small proportion of B‐type crystallites mixed with A‐type crystallites was observed in the X‐ray diffractogram of starches at early maturation (7 and 14 DAA), it was masked by the A‐type crystallites at later maturity stages. However, the large granules had a higher proportion of B‐type crystallites and lower relative crystallinity (RC) than their small‐granule counterpart. The iodine absorption properties of the starch granules demonstrated different levels of mobility of the starch polymers at different stages of maturity and the mobility of more glucan polymers in the large granule population compared with the small granules at the same maturity stage. Iodine did not change the characteristic A‐type crystalline pattern of starch, but it increased RC. Changes in peak width at half height based on WAXS data further suggested the possible interaction of iodine with amylopectin intercluster chain segments and branch chains in formation of inclusion complexes.     

Effect of Small and Large Wheat Starch Granules on Thermomechanical Behavior of Starch

The physicochemical properties of small‐ and large‐granule wheat starches were investigated to reveal whether gelatinization properties and rheological behavior differ between size classes of wheat starch. All samples contained 60% water (w/w, wb). The starch granule size and shape were examined by scanning electron microscopy in the separated A‐ and B‐type granule populations and in the whole wheat starch granule population. Differential scanning calorimetry (DSC) and electron spin resonance (ESR) analyses were performed in parallel with rheological measurements using dynamic mechanical thermal analysis (DMTA) to relate the viscoelastic changes to modifications in dynamic properties of aqueous solutions and structural disorganization of starch. The small (B‐type) granules had slightly higher gelatinization temperature and lower gelatinization enthalpy than did the large (A‐type) granules. Also, B‐type granules had higher enthalpy for the amylose‐lipid complex transition. Moreover, our results suggested that small granules have higher affinity for water at room temperature. It seems that there is a less ordered arrangement of the polysaccharide chains in the smaller granules when compared with the larger ones. These differences in functional properties of small and large granules suggested that the granule size distribution is an important parameter in the baking process.     

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.     

Separation and Characterization of A- and B-Type Starch Granules in Wheat Endosperm

Mature wheat (Triticum aestivum L.) endosperm contains two types of starch granules: large A-type and small B-type. Two methods, microsieving or centrifugal sedimentation through aqueous solutions of sucrose, maltose, or Percoll were used to separate A- and B-type starch granules. Microsieving could not completely separate the two types of starch granules, while centrifuging through maltose and sucrose solutions gave a homogenous population for B-type starch granules only. Centrifuging through two Percoll solutions (70 and 100%, v/v) produced purified populations of both the A- and B-type starch granules. Analysis of starch granule size distribution in the purified A- and B-type granule populations and in the whole-starch granule population obtained directly from wheat endosperm confirmed that the purified A- and B-type starch granule populations represented their counterparts in mature wheat endosperm. Centrifugations through two Percoll solutions were used to purify A- and B-type starch granule populations from six wheat cultivars. The amylose concentrations and gelatinization properties of these populations were analyzed. All of the A-type starch granules contained higher amylose concentrations and had higher gelatinization enthalpies than did B-type starch granules. Although A- and B-type starch granules started to gelatinize at a similar temperature, B-type starch granules had higher gelatinization peak and completion temperatures than did A-type starch granules     

Adsorption Kinetics of Blue 5G Dye from Aqueous Solution on Dead Floating Aquatic Macrophyte: Effect of pH, Temperature, and Pretreatment

The textile industry is known to generate large quantities of effluents contaminated with dyes that are not fixed to the fibers during the dyeing process. The available technologies to remove these dyes from the wastewater are expensive and ineffective. Within this context, low-cost, easy-maintenance technologies for the removal of dyes have been studied, such as adsorption on aquatic macrophytes. Thus, the macrophyte Salvinia sp., raw or pretreated with NaOH or H₃PO₄, was used as biosorbent of Blue 5G reactive dye. The study showed that pH and temperature affect the dye removal capacity. The analysis of the infrared spectrum (FTIR) showed that chemical treatment of the Salvinia sp. modified the biomass surface and affected dye adsorption capacity. The pseudo-second-order kinetic model satisfactorily described the experimental data for raw and NaOH-pretreated biomass, and the pseudo-first-order model was more appropriate to describe the experimental data obtained with H₃PO₄-pretreated biomass. The highest capacity of Blue 5G dye removal was obtained with raw biomass, at 333?K and pH?1.0, with 98.35?% adsorption.     

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.     

Differences in Starch Granule Composition and Structure Influence In Vitro Enzymatic Hydrolysis of Grain Meal and Extracted Starch in Two Classes of Canadian Wheat (Triticum aestivum L.)

Starch granule composition and amylopectin structure affect starch digestibility, an important factor influencing wheat grain utilization for human food consumption. Six bread wheat cultivars with four belonging to the Canada Western Red Spring (CWRS) and two Canada Prairie Spring Red (CPSR) market classes were analyzed for the relationship between their grain constituents and in vitro enzymatic hydrolysis of starch. CPSR cultivars had higher starch and amylose concentrations compared with CWRS cultivars, which had a higher protein concentration. Starch granule size distribution did not differ among the genotypes, except AC Foremost, which had significantly (P < 0.05) higher volume percent of B‐type starch granules (≈15%) and lower volume percent of A‐type starch granules (≈9%) compared with other cultivars. Fluorophore‐assisted capillary electrophoresis revealed a lower content of R‐IV (DP 15–18, ≈6%) and a higher content of R‐VII (DP 37–45, ≈7%) chains in the CPSR cultivars compared with the CWRS cultivars. Starch in vitro enzymatic hydrolysis showed that compared with CWRS cultivars, the two CPSR cultivars had reduced amounts of readily digestible starch and higher amounts of slowly digestible starch and resistant starch. Consequently, the two CPSR cultivars also showed lower hydrolysis indexes in grain meal as well as extracted starch. CPSR cultivars, with higher starch and amylose concentrations, as well as a higher content of long chains of amylopectin, showed a reduced starch in vitro enzymatic hydrolysis rate.     

Oil Binding of Heat‐Treated Waxy Wheat Starch Granules and Starch Granule Ghosts Produced with Concentrated KI and I2

Dry waxy wheat starch granules were heat‐treated at 120°C for 5 hr, and then shaken vigorously in a biphasic system of oil and water. Non‐heat‐ treated starch remained in the aqueous phase, whereas the heat‐treated starch granules showed a strong oil‐binding ability that was lost by trypsin treatment. This result showed that the starch granule surface protein changed from hydrophilic to hydrophobic due to the heat treatment. The presence of starch granule surface protein was ascertained by staining with fluorescamine and fluorescence microscopic observation. Heat‐treated waxy wheat starch granules were incubated with a 25% KI/10% I₂ (w/v) solution, which produced “ghosts” (exterior and interior) structures. The exteriors stained red‐brown, whereas the interiors stained black‐brown. Sonication (20 kHz for 255 sec) followed by centrifugation separated the structures, which were then shaken vigorously in an oil and water system. Only the exterior ghosts exhibited a remarkable emulsification property, which disappeared after trypsin treatment. The ghosts from unheated control granules did not show emulsification. The presence of protein in the exterior ghost fraction was further substantiated by fluorescamine treatment. No protein was detectable in the interior fraction with this dye. From these results, we suggest that the ghost fraction of the waxy wheat starch contained the starch granule surface protein that was made hydrophobic by heat treatment. Also, the nature of the induced emulsification property of the exterior fraction (ghosts) and the oil‐binding ability of the heat‐treated waxy wheat starch granules coincided. Both were due to the hydrophobic nature of the same starch granule surface protein, which showed that the ghosts were the swollen form of the outer region of the waxy starch granule.     

Effects of Environmental Temperature on Structure and Gelatinization Properties of Wheat Starch

The effects of environmental temperature on gelatinization properties and amylopectin structures of wheat endosperm starch were examined by isolating starches from four wheat cultivars matured in growth chambers at daytime temperatures of 15, 20, 25, or 30°C. Kernel weight and starch content per kernel were reduced by high maturation temperature. Amylose content showed no significant change at high maturation temperature in some cultivars; in other cultivars, there was a slight increase. Principal component analysis of data on relative peak areas of debranched amylopectin showed that amylopectin from wheat grown at a lower temperature had a greater proportion of shorter chains. Amylopectin branch chains were classified into three groups based on the correlation coefficients between the data of branch chain length distribution and principal component scores, degree of polymerization (DP) of 6–12, DP 13–34, and DP ≥ 35. The gelatinization temperature of starches increased markedly at a higher maturation temperature, with increases exceeding 10°C at high maturation temperatures. Gelatinization properties correlated significantly with amylopectin chain length distribution.     

Decolorization and Degradation of Reactive Dye in Aqueous Solution by Ozonation in a Semi-batch Bubble Column Reactor

The decolorization and degradation of anionic sulphonated azo dye (Reactive orange 16 (RO16)), which is suspected to be carcinogenic, were investigated using ozone. The decolorization process of the reactive dye was carried out by bubbling ozone at the bottom of a bubble column reactor containing the dye solution. The effect of pH, reaction time, dye concentration, ozone concentration, and decolorization time was studied. Also, degradation products and possible degradation mechanism were investigated. The results showed that ozonation was a highly effective way to remove color from wastewater. The color of a synthetic waste solution containing water-soluble reactive dye was reduced to 69.69 % under the basic condition (pH 12), with complete RO16 degradation occurring in 8 min. Ozone consumption continued for a further 16 min after which time most of the degradation reactions were complete. Kinetic studies showed that direct ozonation of the aqueous dyes represented a pseudo-first-order reaction with respect to the dye. The apparent rate constant increased with both the applied ozone dose and higher pH values and declined logarithmically with the initial dye concentration. Intermediates such as 6-acetylamino-3-aminonaphthalene-2-sulfonic acid, 2-(4-nitrosophenyl) sulfonylethyl hydrogen sulfate, and 6-acetamido-4-hydroxy-3-nitroso naphthalene-2-sulfonic acid were detected by gas chromatograph coupled with mass spectrometry in the absence of pH buffer, while nitrate and sulfate ions and formic, acetic, and oxalic acids were detected by ion chromatography.     

Effects of Varying Weight Ratios of Large and Small Wheat Starch Granules on Experimental Straight‐Dough Bread

One commercial bread wheat flour with medium strength (11.3% protein content, 14% mb) was fractionated into starch, gluten, and water solubles by hand‐washing. The starch fraction was separated further into large and small granules by repeated sedimentation. Large (10–40 μm diameter) and small (1–15 μm diameter) starch fractions were examined. Flour fractions were reconstituted to original levels in the flour using composites of varying weight percentages of starch granules: 0% small granules (100% large granules), 30, 60, and 100% (0% large granules). A modified straight‐dough method was used in an experimental baking test. Crumb grain and texture were significantly affected. The bread made from the reconstituted flour with 30% small granules and 70% large granules starch had the highest crumb grain score (4.0, subjective method), the highest peak fineness value (1,029), and the second‐highest elongation ratio (1.55). Inferior crumb grain scores and low fineness and elongation ratios were observed in breads made from flours with starch fractions with 100% small granules or 100% large granules. As the proportion of small granules increased in the reconstituted flour, it yielded bread with softer texture that was better maintained than the bread made from the reconstituted reference flour during storage.     

Effect of Surfactant Structure on the Pasting Properties of Wheat Flour and Starch Suspensions

Systematic studies were performed on the effect of the surfactant alkyl chain length (10–16 carbon atoms) and the head group charge/structure (anionic, cationic, nonionic) on the pasting properties of wheat flour and starch aqueous suspensions by means of a Rapid Visco Analyser (RVA). An excellent agreement was observed between the effect of surfactants on the onset temperature of the pasting process (PT) and the time to reach peak viscosity (t_peak) of wheat flour and wheat starch suspensions. Moreover, a correlation was found between the effect of different surfactants on these two parameters. With the exception of the cationic surfactants (alkyl trimethyl ammonium bromides), the effect of surfactants (alkyl sulfates, maltosides, monoglycerides, and sucrose esters) was found to be strongly dependent on the surfactant chain length. Shorter chain surfactants (C10–C12) induced an earlier pasting, while longer chain surfactants (C14–C16) had the opposite effect. The effect of surfactants on PT and t_peak of flour suspensions was enlarged when the surfactant concentration was increased from ≈1% to 15% (w/w) on a dry starch basis.     

Size Distribution and Properties of Wheat Starch Granules in Relation to Crumb Grain Score of Pup‐Loaf Bread

Twelve hard winter wheat flours with protein contents of 11.8–13.6% (14% mb) were selected to investigate starch properties associated with the crumb grain score of experimentally baked pup‐loaf bread. The 12 flours were classified in four groups depending on the crumb grain scores, which ranged from 1 (questionable‐unsatisfactory) to 4 (satisfactory). Flours in groups 1, 2, 3, and 4 produced breads with pup‐loaf volumes of 910–1,035, 1,000–1,005, 950–1,025, and 955–1,010 cm³, respectively. Starches were isolated by a dough handwashing method and purified by washing to give 75–79% combined yield (dry flour basis) of prime (62–71%) and tailing (7–16%) starches. The prime starch was fractionated further into large A‐granules and small B‐granules by repeated sedimentation in aqueous slurry. All starches were assayed for weight percentage of B‐granules, swelling power (92.5°C), amylose content, and granular size distribution by quantitative digital image analysis. A positive linear correlation was found between the crumb grain scores and the A‐granule sizes (r = 0.65, P < 0.05), and a polynomial relationship (R² = 0.45, P < 0.05) occurred between the score and the weight percentage of B‐granule starch. The best crumb grain score was obtained when a flour had a weight percentage of B‐granules of 19.8–22.5%, shown by varietal effects.     

Optimization of the Adsorption Conditions for the Decolorization and COD Reduction of Methylene Blue Aqueous Solution using Low-Cost Adsorbent

The performance of raw bagasse (RB), and tartaric acid-modified bagasse (TAMB) as adsorbents on decolorization and chemical oxygen demand (COD) reduction of methylene blue (MB) aqueous solution was studied. The effects of five factors namely: adsorbent dosage, pH, shaking speed, contact time, and temperature on decolorization and COD reduction were studied and optimized using central composite design (CCD). The results of the analysis show that all selected factors exhibit significant effect on decolorization and COD reduction. Maximum decolorization (78.16%) and COD reduction (77.95%) for RB was achieved at 0.82 g of adsorbent dosage, pH 9.4, 122 rpm of shaking speed, 44 min of contact time, and 55°C. For TAMB, maximum decolorization (99.05%) and COD reduction (98.45%) was achieved at 0.78 g adsorbent dosage, pH 9.4, shaking speed of 120 rpm, 34 min contact time, and 49°C. TAMB was found to be more effective than RB in decolorization and COD reduction of MB aqueous solution.     

Presence of a Minor Amylose Fraction in the Central Portion of Waxy Wheat Starch Granules and Its Contribution to Granular Stability

When wheat starch granules containing various amounts of amylose (2.1–25%) were stained with 25% KI/10% I₂ solution, the granules largely changed to ghost structures below ≈5.0% amylose. The ghost showed a typical double structure: a black‐brown central portion and a red‐brown surrounding portion. The proportion of the black‐brown central portion in the ghost was strongly correlated to the amylose content (%) in the starch, that is, the black‐brown central portion decreased with a decrease in amylose. This suggests that amylose is possibly present in the black‐brown central portion. Sonication (20 kHz) followed by centrifugation of the ghost separated the black‐brown central portion from the red‐brown surrounding portion, and the amylose content in each portion was determined. The results indicated that the amylose content in the black‐brown central portion was 6.9%, whereas in the surrounding portion, it was only 1.0%. Furthermore, the central and surrounding portions were subjected to Sepharose CL‐2B gel‐filtration column chromatography and the presence of amylose could only be observed in the black‐brown central portion.