Initial Phase Solubilization of Normal Corn Starch by Methyl Sulfoxide (DMSO): Evidence from Scanning Electron Microscopy and Size Exclusion Chromatography

Initial structure and solubility changes of normal corn and starch granules, dispersed in varying methyl sulfoxide (DMSO) concentrations were investigated by scanning electron microscopy (SEM) and high-performance size exclusion chromatography (HPSEC). Larger granules from starches held for 6 or 15min and air dried showed increased surface etching with increasing DMSO concentration (0–70%): smaller granules showed surface peeling at 70% DMSO. At 90% DMSO, larger granules fractures into smaller pieces, while smaller granules showed full surface erosion. At 100% DMSO, large chunks eroded off both large and small granules. With 100% DMSO, a sheet-like layer formed over a mass of ruptured granules during drying, suggesting that in the absence of water, DMSO dissolves part of the granule, forming a viscous solution which may limit further DMSO granule dispersion. Granules processed with critical-point drying showed additional evidence of attack. HPSEC polymer solubility increased with DMSO concentration (0–90%). At 100% DMSO, however, there was no measured solubility. Similar solubilities for starches held at 6 or 15min (0–90% DMSO) suggest that DMSO may require an external force such as heat to facilitate polymer solubility. More amylopectin of a lower molecular weight was solubilized at 6min, while more amylose was solubilized at 15min. This suggests that granule surfaces were mostly composed of amylopectin under development, while amylose was mostly confined to amorphous regions inside the granule.     

Gelatinization and Solubility Properties of Commercial Oat Starch

Gelatinization and solubility characteristics of starch contribute to unique functionality in foods. Corn and oat starch viscoamylographs (35g db) showed peak viscosities of 400 and 390B.U., respectively. Oat starch had a more rapid (89.7 vs 85.6°C) and higher set back (790 vs. 740B.U.) than corn. Data on soluble components of cooled (85°C) starch pastes, as collected and analyzed by high-performance size exclusion chromatography (HPSEC), suggest that amylopectin plays a significant (P<0.05) role in oat paste set back; for corn starch, amylose is the dominant factor. Solubilities and apparent molecular weights (MW) of oat starch heated (65-120°C) under shear and subsequent sonication (0-40s) in water or 90% methyl sulfoxide (DMSO) were also determined by HPSEC. An intermediate MW fraction was eluted on the HPSEC chromatograms only when oat starch was heated in water (100-120°C/30min) or DMSO and sonicated, suggesting that this fraction may not be inherent in the native granule. in water, polymer solubility and peak MW increased with temperature (65-120°C), whereas in DMSO, solubility decreased with heating (65-100°C), while peak MW remained basically constant. Aqueous (aq) leaching at 75°C solubilized more corn amylose than amylopectin, but amylopectin and amylose co-leached from oat starch granules. Aq leaching, at 20°C above their DSC peak ends (85 and 95°C for oat and corn, resepctively), showed more amylopectin leached from oat starch granules whereas more amylose was leached from corn starch granules.     

Molecular Fractionation of Starch in Nycodenz‐Dimethyl Sulfoxide by Density‐Gradient Ultracentrifugation

Starches from various origins (normal corn, waxy corn, high‐amylose corn, normal rice, waxy rice, potato and tapioca) were fractionated by density‐gradient ultracentrifugation at 124,000×g, using a gradient of Nycodenz (0–40%, w/w) in 90% dimethyl sulfoxide (DMSO). The fractions of different densities were collected from a centrifuge tube, and then analyzed by the phenol‐sulfuric acid method and iodine‐binding test. Amylose and amylopectin were clearly separated by the density‐gradient centrifugation within 6 h of centrifugation in the total carbohydrate vs. density profile. The amylose content in the starches, measured from the centrifugation for 6 h, agreed with the values reported in the literature, and the content of intermediate fraction (5–13%) was also measured. Amylopectin quickly reached a density region (1.22–1.23 g/mL) with a sharp and stable peak, whereas amylose showed a band in a broad density range, moving toward higher density as the centrifugation continued. The buoyant density of amylose became similar to that of amylopectin after extensive centrifugation. Because the sedimentation rate of amylose was much lower than that of amylopectin, both starch chains are effectively fractionated in the gradient medium, and the iodine‐binding property of the fractionated starches provides valuable information on the chain conformation of starch.     

Effect of cooking time and ingredients on the performance of different starches in white sauces

The effect of white sauce ingredients and increased cooking time at 90 °C on the degree of gelatinization of corn, waxy corn, rice, potato and modified waxy corn starches was studied. The changes in pasting properties, linear viscoelastic properties, and microstructure were determined. In all the native starches in water, a longer cooking time at 90 °C caused greater starch granule swelling and more leaching of solubilized starch polymers into the intergranular space. These effects were more noticeable in the waxy corn and potato starches. The potato starch was the most affected, with complete disruption of the starch granules after 300 s at 90 °C. The microstructural changes which transformed a system characterized by starch granules dispersed in a continuous phase (amylose/amylopectin matrix) into a system with an increase in the continuous phase and a decrease in starch granules were associated with a decrease in system viscoelasticity. The elastic moduli were higher in the sauce than in the starch in water system. However, with the exception of potato starch, the white sauce showed lower viscoelasticity than the starch in water system. The white sauce ingredients decreased the effect of cooking time on the starch gelatinization process, particularly in potato starch.     

Amylopectin structure of heat–moisture treated starches

The effects of heat–moisture treatment (HMT; moisture content of 25%, at 100°C for 24 h) on starch chain distribution and unit chain distribution of amylopectin in normal rice, waxy rice, normal corn, waxy corn, normal potato, and waxy potato starches were investigated. After HMT, starch chain distribution (amylose and amylopectin responses) of waxy corn and potato starches were identical to those of untreated starches, whereas the chromatographic response of waxy rice starch showed a slight decrease, but with a slight increase in peak tailing. This result indicated that HMT had no (or very limited) effect on the degradation of amylopectins. Analysis of unit chain distribution of amylopectins revealed that waxy characteristics affected the molecular structure of amylopectin in untreated starches, i.e., the CL of normal‐type starches was greater than that of waxy‐type starches. After HMT, the CL and unit chain distribution of all starches were no different than those of untreated starches. The results implied that changes in the physico‐chemical properties of HMT starches would be due to other phenomena rather than the degradation of amylopectin molecular structure. However, the thermal degradation of amylopectin molecules of waxy starches could occur by HMT at higher treatment temperatures (120 and 140°C).     

酸解对不同链/支比含量玉米淀粉特性的影响

以4种不同链/支比含量的玉米淀粉为原料,酸解处理不同时间,以酸解玉米淀粉的形貌特性、冻融稳定性、膨胀度、溶解度、晶体性质为指标衡量不同酸解时间对玉米淀粉结构性质的影响。结果表明:4种玉米淀粉酸水解程度的顺序为:蜡质玉米普通玉米淀粉G50G80。酸解后,同品种的4种玉米淀粉的析水率随着酸解天数的增加而增加;溶解度增加,膨胀度降低。酸解并未改变淀粉的晶型,随着酸解时间的延长,蜡质玉米淀粉和普通玉米的相对结晶度先增大后保持不变,G50和G80的相对结晶度随着酸解时间的增加而增大。表明酸解对低直链淀粉(蜡质玉米淀粉和普通玉米淀粉)的结构、性能影响最大。     

Structural,thermal and viscoelastic characteristics of starches separated from normal,sugary and waxy maize

Starches separated from five types of maize (two normal, one sugary and two waxy) were investigated for physicochemical, thermal, amylopectin structure and viscoelastic properties. Kisan and Paras were normal maize while Parbhat and LM-6 were waxy maize type. Apparent amylose content of normal and sugary maize was 29.5–32.6 and 41.0%, respectively. Swelling power of normal, sugary and waxy maize starches was 11.6–15.2, 7.8 and 30.2–39.2 (g/g), respectively. X-ray diffraction of maize starches indicated typical A-pattern. Maize starch showed a single broad peak at 2θ=23.2° and a dual peak 2θ=17°–18.1, respectively. Waxy maize starches showed the presence of greater crystallinity than other starches while sugary maize starch showed the presence of lower crystallinity and a large amount of amylose–lipid complex. Intrinsic viscosity [η] of starches in 90% DMSO at 25 °C was 79.7–119.5 ml g⁻¹ for normal, 70.5 ml g⁻¹ for sugary and 107.2–118.1 ml g⁻¹ for waxy starches. Branch chain–length distribution of amylopectin revealed that the apparent amylose, long side chain- and short side chain-amylopectin proportion ranged between 0.0–41%, 13.4–31.5% and 41.5–66.8%, respectively, among the various maize starches. Maize sugary showed the highest apparent amylose content and the least amount of short- and long-side chains of amylopectin. LM-6 and Parbhat showed higher proportion of both long- and short-chain amylopectin as compared to other starches. Distribution of α-1, 4-chains of amylopectin (short-/long-chain) ranged between 2.1 and 3.4, the least for LM-6 and the highest for Paras starch. The transition temperatures (T_o–T_c) ranged between 60.5 and 76.1 °C for sugary, 63.5–76.3 °C for normal and 64.4–81.3 °C for waxy maize starch. The enthalpy of gelatinization (ΔH_gel) of sugary, normal and waxy maize starches was 2.47, 3.7–4.75 and 4.15–5.4 J/g, respectively. Normal and sugary maize starches showed higher G′ and G″ than waxy type starches. The change in the moduli during cooling and reheating of pastes cooked at different temperatures revealed low disintegration of granular structure in starch with higher amylose and amylose–lipid complex as well as low crystallinity. The changes in moduli during 10 h at 10 °C revealed highest retrogradation in maize sugary followed by Paras and Kisan starch.     

Influence of moisture content on the degradation of waxy and normal corn starches acid-treated in methanol

Waxy and normal corn starches with different moisture contents, 5.1-16.9% and 4.8-15.9%, respectively, were prepared and treated in methanol containing 0.36% HCl at 45 °C for 1 h. Recovery of all the treated starches was found to be above 90%. Peak viscosity, gelatinization temperature and enthalpy change of gelatinization of waxy and normal corn starches decreased after treatment and this decrement was found to be more in treated starches having lower initial moisture content. The weight-average degree of polymerization and chain length (CL) of waxy and normal corn starches decreased upon acid-methanol treatment. The decrement ratio of molecular weight of modified starches was found to be negatively correlated with the initial moisture content of the starches. The decrement ratio of normal corn starch was higher than waxy corn starch with similar moisture content of starch. The content and CL of long chain fraction of amylopectin for waxy corn starch slightly decreased after treatment, while no obvious trend was found among starches with different moisture contents. CL of amylose for acid-methanol-treated normal corn starch decreased and this change was found to be higher in starches with lower initial moisture contents. Results demonstrated that the initial moisture content of starch granules strongly influenced the functional properties and degradation of starch treated by acid in methanol.     

Analysis of Cereal Starches by High-Performance Size Exclusion Chromatography

Starch has unique physicochemical characteristics among carbohydrates. Two important characteristics of starch are amylose/amylopectin ratio and molecular weight. The objective of this study was to develop a simple, one-step, and accurate method for the determination of amylose/amylopectin ratio and weight-averaged molecular weights of starch in whole meal cereal samples. This method would allow for the simultaneous measurement of these important properties without the time-consuming step of large-scale starch extraction. Starch from spring wheat, barley, oat, rye, buckwheat, rice, durum wheat, and corn was precipitated with ethanol and solubilized with potassium hydroxide and urea before analysis by a high-performance size exclusion chromatography (HPSEC) system with a refractive index detector. To verify peak identity, fractions were collected and then gas chromatography analysis of soluble starch and beta-glucan assays was performed. All HPSEC fractions contained only glucose and no beta-glucan was detected, indicating that there was no contamination from non-starch polysaccharides. Weight-averaged molecular weights were calculated using a series of gel permeation chromatography grade dextrans. The method described in this study can be used to determine amylose/amylopectin ratio and weight-averaged molecular weight of cereal starches using as low as 25 mg whole meal samples without large-scale starch extraction.     

Rheology of Okenia hypogaea Starch Dispersions in Aqueous Solution of DMSO

The aim of this work was to study the rheological behavior of Okenia hypogea starch dispersions (OSD) in aqueous solution (90/10, v/v) of dimethyl sulfoxide (DMSO). Okenia starch dispersions with 3 and 7 % (w/v) total solids were prepared at 20 °C and rheological tests were undertaken at 20, 40 and 60 °C, using a shear rate controlled rotational viscometer. The data fitted better the power law than the Casson model. Flow curves and flow behavior index indicated shear‐thinning behavior. All rheological parameters were affected by the solids concentration and the measurement temperature. As regards amylose and amylopectin content okenia starch is grouped among “ordinary” (non‐waxy) starches. Overall, OSD behaved similarly to potato starch dispersions and corn starch dispersions used as controls.     

Effect of Aqueous Ethanol Cationization on Functional Properties of Normal and Waxy Starches

Cationic starch ethers of normal and waxy corn, normal and waxy barley and normal pea starch were prepared by an aqueous alcoholic process for evaluation of their functional properties as compared to the native starch controls. The native starches exhibited a wide range in average granule size (10–21 μm diameter), amylose content (0–34%) and swelling power (13–31). Cationization to degrees of substitution (DS) of 0.030–0.035 with 3-chloro-2-hydroxypropyltrimethylammonium chloride resulted in marked increases in swelling power of all starches, with little corresponding increases in starch solubility. Cationization also decreased the onset of endothermic transitions and pasting temperatures quite substantially, and promoted the development of sharp peak viscosities in the amylographs of all normal and waxy starches, including that of pea starch. Final cold viscosities of the cationic starches exhibited positive setbacks, and the cooked starch gels, after storage for 7 days at 4°C and −15°C, showed no syneresis. All cationic starches except for waxy corn were more susceptible to α-amylase hydrolysis than native control starches. The general improvement in functional properties, especially in the waxy corn, waxy barley and pea starches, due to the aqueous alcoholic-alkaline cationization process would greatly enhance their industrial applications.     

Effect of annealing on the semicrystalline structure of normal and waxy corn starches

The influence of amylose and amylopectin on structural reorganization occurred during annealing was studied for normal and waxy corn starches. Annealing caused an increase in crystallinity in the waxy corn starch, whereas the number of pores on the granule surface, observed by SEM, increased especially for normal corn starch. Amylose and amylopectin chains of the annealed normal corn starch were degraded to greater extension during enzymatic hydrolysis than those of the native starch. On contrary, the annealing caused a protective effect on waxy corn starch amylopectin toward the enzymatic reaction suggesting that this treatment promoted a better interaction between amylopectin chains of waxy corn starch. The amylose molecules of normal corn starch may have impaired the mobility of amylopectin molecules and restricted the reorganization of the crystalline structure during the annealing. The major increase in pores number on the granule surface of annealed normal corn starch, resulted of the endogenous amylase action during annealing, could facilitate the exogenous enzymes’ role in the degradation of the starch granules’ amorphous area.     

Rheological Properties of Mixed Gels using Waxy and Non‐waxy Wheat Starch

Mixed starches with an amylose content of 5, 10, 18, 20, 23, and 25% were prepared by blending starches isolated from waxy and non‐waxy wheat at different ratios. The dynamic viscoelasticity of mixed 30% and 40% starch gels was measured using a rheometer with parallel plate geometry. The change in storage shear modulus (G′) over time at 5 °C was measured, and the rate constant of G′ development was estimated. As the proportion of waxy starch in the mixture increased, starch gels showed lower G′ and higher frequency dependence during 48 h storage at 5 °C. Since the amylopectin of waxy starch granules was solubilized more easily in hot water than that of non‐waxy starch granules, mixed starch containing more waxy starch was more highly solubilized and formed weaker gels. G′ of 30% and 40% starch gels increased steadily during 48 h. 30% starch gel of waxy, non‐waxy and mixed starches showed a slow increase in G′. For 40% starch gels, mixed starch containing more waxy starch showed rapidly developed G′ and had a higher rate constant of starch retrogradation. Waxy starch greatly influenced the rheological properties of mixed starch gels and its proportion in the mixture played a major role in starch gel properties.     

Amylose Content and Chain Profile of Amylopectin from Normal,High Amylose and Waxy Barleys

The amylose content and the chain profile of amylopectin from normal, waxy and high amylose barley starches were determined after enzymatic debranching and gel permeation chromatography and the degree of branching of the amylopectin was analysed by 1H-n.m.r. spectroscopy. The normal barley starch contained around 30%, the high amylose around 40% and the waxy starch 9% amylose. The amylopectin of the high amylose starches had longer chains than those of the normal or waxy starches, especially in the molecular weight interval 5,400-8,000, but less of those below 2,400 in molecular weight. The chain length of amylopectin from high amylose barley was on average 5 units longer than those of normal or waxy barleys.     

The Fine Structure of Corn Starches of Various Amylose-Percentage: Waxy,Normal and Amylomaize

The fine structure of high-amylose corn starches has been studied after dispersion of the starch and fractionation into their components amylose and amylopectin. The resulting amylopectin fraction reported in the literature possesses anomalous properties with regard to the waxy and normal amylopectin. However, the experimental results obtained by different authors for determining the structure lead to controversal explanations. Therefore, using an enzymic method, which permits the direct examination of the constitutive chains of the starch, the fine structure of the amylopectin of an amylomaize starch (64% amylose) has been investigated and compared with those of waxy and normal starches. The pullulanase – debranched chains are fractionated by gel permeation and their linearity are checked under the action of β-amylase. The inner chains of the amylopectin fraction are studied after debranching of the β-limit dextrins. The results show the identity between amylopectins from waxy and normal starches. The amylopectin fraction of amylomaize has its own structure with longer inner chains than those of waxy-maize amylopectin.     

Development and Physicochemical Characterization of New Resistant Citrate Starch from Different Corn Starches

Resistant starch has drawn broad interest for both potential health benefits and functional properties. In this study, a technology was developed to increase resistant starch content of corn starch using esterification with citric acid at elevated temperature. Waxy corn, normal corn and high‐amylose corn starches were used as model starches. Citric acid (40% of starch dry weight) was reacted with corn starch at different temperatures (120–150°C) for different reaction times (3–9 h). The effect of reaction conditions on resistant starch content in the citrate corn starch was investigated. When conducting the reaction at 140°C for 7 h, the highest resistant starch content was found in waxy corn citrate starch (87.5%) with the highest degree of substitution (DS, 0.16) of all starches. High‐amylose corn starch had 86.4% resistant starch content and 0.14 DS, and normal corn starch had 78.8% resistant starch and 0.12 DS. The physicochemical properties of these citrate starches were characterized using various analytical techniques. In the presence of excess water upon heating, citrate starch made from waxy corn starch had no peak in the DSC thermogram, and small peaks were found for normal corn starch (0.4 J/g) and Hylon VII starch (3.0 J/g) in the thermograms. This indicates that citrate substitution changes granule properties. There are no retrogradation peaks in the thermograms when starch was reheated after 2 weeks storage at 5°C. All the citrate starches showed no peaks in RVA pasting curves, indicating citrate substitution changes the pasting properties of corn starch as well. Moreover, citrate starch from waxy corn is more thermally stable than the other citrate starches.     

Starch Related Changes in Stored Soft Sorghum Porridge

Soft sorghum (Sorghum bicolor (L.) Moench) unfermented porridge was prepared by cooking pearled sorghum flour (6% solids in water) in a microwave oven. Fermented porridge was prepared by mixing with Lactobacillus delbruekii for 12 hr at 48°C. followed by cookine. Porridge was aged (0–12 hr) at 25°C or 7.5%. Starch polymer solibility was determined by high performance size exclusion chromatography (HPSEC) and rheology followed with a rotational viscometer. During aging, solubilized and gelled amylose and amylopectin retrogradation rates were higher in porridge stored at 7.5 than at 25°C. Amylose polymer solubility decreased with storage, while solubilized amylopectin remained in solution. Fermented porridges were lighter in color and their initial consistency indexes increased faster. HPSEC of soluble polymers was more sensitive than enzymatic starch digestion.     

Comparison between Size Exclusion Chromatography and Micro‐Batch Analyses of Corn Starches in DMSO using Light Scattering Detector

The molecular structure of corn starches different in amylose content (waxy, normal, and high‐amylose) was analyzed in 90% dimethyl sulfoxide (DMSO) solution by refractive index (RI) and multi‐angle laser light scattering (MALLS) detectors. The starch sample solutions were measured either by medium‐pressure size exclusion chromatography (MPSEC) or by the micro‐batch mode. For waxy corn starch, the average molar mass (M_w) and radius of gyration (R_g) values were similar in both methods. However, for normal and high‐amylose corn starches, M_w measured by the micro‐batch mode was 2–4 times greater than that by the chromatographic method, although R_g values obtained from both methods were not very different. The M_w difference was the greater the higher the amylose content of starch.     

Production and characterization of digestion‐resistant starch by the reaction of Neisseria polysaccharea amylosucrase

Recombinant amylosucrase (200 U/mL) from Neisseria polysaccharea was used to produce digestion‐resistant starch (RS) using 1–3% (w/v) corn starches and 0.1–0.5 M sucrose incubated at 35°C for 24 h. Characterization of the obtained enzyme‐modified starches was investigated. Results show that the yields of the enzyme‐modified starches were inversely proportional to the original amylose contents of corn starches. After enzymatic reaction, insoluble RS contents increased by 22.3 and 20.7% from 6.9% of waxy and 7.7% of normal corn starches, respectively, using 3.0% starch as acceptor and 0.3 M sucrose as donor, while amylomaize VII showed the lowest increase (8.5%) in RS content. The crystalline polymorph of these enzyme‐modified starches resulted in the B‐type immediately after enzymatic reaction. The enzyme‐modified starches displayed higher melting peak temperatures (85.6–100.6°C) compared to their native starch counterparts (70.1–78.4°C). After enzymatic reaction, pasting temperature increased in waxy (71.9 → 77.6°C) and normal corn starches (75.3 → 80.6°C), and the peak viscosity of waxy corn starches increased from 264 to 349 RVU, whereas that of normal corn starches decreased from 235 to 66 RVU.     

Microstructural and physicochemical properties of heat-moisture treated waxy and normal starches

Starches from normal rice (21.72% amylose), waxy rice (1.64% amylose), normal corn (25.19% amylose), waxy corn (2.06% amylose), normal potato (28.97% amylose) and waxy potato (3.92% amylose) were heat-treated at 100 °C for 16 h at a moisture content of 25%. The effect of heat-moisture treatment (HMT) on morphology, structure, and physicochemical properties of those starches was investigated. The HMT did not change the size, shape, and surface characteristics of corn and potato starch granules, while surface change/partial gelatinization was found on the granules of rice starches. The X-ray diffraction pattern of normal and waxy potato starches was shifted from B- to C-type by HMT. The crystallinity of the starch samples, except waxy potato starch decreased on HMT. The viscosity profiles changed significantly with HMT. The treated starches, except the waxy potato starch, had higher pasting temperature and lower viscosity. The differences in viscosity values before and after HMT were more pronounced in normal starches than in waxy starches, whereas changes in the pasting temperature showed the reverse (waxy > normal). Shifts of the gelatinization temperature to higher values and gelatinization enthalpy to lower values as well as biphasic endotherms were found in treated starches. HMT increased enzyme digestibility of treated starches (except waxy corn starch); i.e., rapidly and slowly digestible starches increased, but resistant starch decreased. Although there was no absolute consistency on the data obtained from the three pairs of waxy and normal starches, in most cases the effects of HMT on normal starches were more pronounced than the corresponding waxy starches.