Structural Properties of Starch in Bread and Bread Model Systems: Influence of an Antistaling α‐Amylase

The influence of an antistaling α‐amylase on bread crumb and on wheat starch gels was investigated taking into account different levels of structural hierarchy. Bread was prepared by a conventional baking procedure. Starch gels were produced by heating a concentrated starch dispersion in closed molds. Bread and starch gels were characterized by compression tests, light microscopy (LM), differential scanning calorimetry, and X‐ray measurements. The α‐amylase enhanced the initial firmness of starch gels and reduced the firming rate of bread and starch gels on aging. LM revealed that amylose and amylopectin phase‐separated within the starch granules and that freshly baked control bread and starch gels showed weak birefringence which became more intense during aging. Amylase‐containing bread and starch gels exhibited strong birefringence in the amylose rich region of the granules directly after baking which did not significantly increase during aging. The enzyme hindered the retrogradation of amylopectin as detected by differential scanning calorimetry, whereas X‐ray diffraction indicated that the enzyme induced low levels of starch crystallinity which did not change during aging. It is hypothesized that the antistaling effect of the amylase is based on the capacity to partially degrade amylopectin and, by this, to hinder its recrystallization. On the other hand, the enzyme slightly degrades amylose by an endo‐mechanism which, in turn, promotes the rapid formation of a partly crystalline amylose network in fresh bread and hinders amylose rearrangements during aging.     

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.     

Model approach to starch functionality in bread making

We used modified wheat starches in gluten-starch flour models to study the role of starch in bread making. Incorporation of hydroxypropylated starch in the recipe reduced loaf volume and initial crumb firmness and increased crumb gas cell size. Firming rate and firmness after storage increased for loaves containing the least hydroxypropylated starch. Inclusion of cross-linked starch had little effect on loaf volume or crumb structure but increased crumb firmness. The firming rate was mostly similar to that of control samples. Presumably, the moment and extent of starch gelatinization and the concomitant water migration influence the structure formation during baking. Initial bread firmness seems determined by the rigidity of the gelatinized granules and leached amylose. Amylopectin retrogradation and strengthening of a long-range network by intensifying the inter- and intramolecular starch-starch and possibly also starch-gluten interactions (presumably because of water incorporation in retrograded amylopectin crystallites) play an important role in firming.     

Method for Determining the Rate and Extent of Accelerated Starch Retrogradation

A method to accelerate and quantitate retrogradation of starch pastes using a freeze-thaw cycle (FTC) process and turbidometric analysis has been developed. Using this method and differential scanning calorimetry (DSC), it was determined that the rate of retrogradation in 2.5% waxy maize pastes was inversely correlated to the rate of freezing, and that the thawing temperature affected perfection of the crystallites in retrograded amylopectin. DSC and X-ray diffraction were used to determine whether the crystallites formed during the FTC process were the same as those formed in starch pastes held isothermally at 4°C. Analysis of retrogradation of pastes of starches from various botanical sources indicated that the method reflects retrogradation in higher concentration pastes. Retrogradation rates were reduced by the addition of sodium dodecyl sulfate. Microstructures of freeze-thaw processed waxy maize and common corn starch pastes were examined.     

Physicochemical and structural properties of maize and potato starches as a function of granule size

Chemical composition, molecular structure and organization, and thermal and pasting properties of maize and potato starches fractionated on the basis of granule size were investigated to understand heterogeneity within granule populations. For both starches, lipid, protein, and mineral contents decreased and apparent amylose contents increased with granule size. Fully branched (whole) and debranched molecular size distributions in maize starch fractions were invariant with granule size. Higher amylose contents and amylopectin hydrodynamic sizes were found for larger potato starch granules, although debranched molecular size distributions did not vary. Larger granules had higher degrees of crystallinity and greater amounts of double and single helical structures. Systematic differences in pasting and thermal properties were observed with granule size. Results suggest that branch length distributions in both amylose and amylopectin fractions are under tighter biosynthetic control in potato starch than either molecular size or amylose/amylopectin ratio, whereas all three parameters are controlled during the biosynthesis of maize starch.     

Effect of Partial Gelatinization and Lipid Addition on α‐Amylolysis of Barley Starch Granules

The effect of partial gelatinization with and without lipid addition on the granular structure and on α‐amylolysis of large barley starch granules was studied. The extent of hydrolysis was monitored by measuring the amount of soluble carbohydrates and the amount of total and free amylose and lipids in the insoluble residue. Similarly to the α‐amylolysis of native large barley starch granules, lipid‐complexed amylose (LAM) appeared to be more resistant than free amylose and amylopectin. Partial gelatinization changed the hydrolysis pattern of large barley starch granules; the pinholes typical of α‐amylase‐treated large barley starch granules could not be seen. Lipid addition during partial gelatinization decreased the formation of soluble carbohydrates during α‐amylolysis. Also free amylose remained in the granule residues and mostly amylopectin hydrolyzed into soluble carbohydrates. These findings indicate that lysophospholipid (LPL) complexation with amylose occurred either during pretreatment or after hydrolysis, and free amylose was now part of otherwise complexed molecules instead of being separate molecules. Partial gelatinization caused the granules to swell somewhat less during heating 2% starch‐water suspensions up to 90°C, and lipid addition prevented the swelling completely. α‐Amylolysis changed the microstructure of heated suspensions. No typical twisting of the granules was seen, although the extent of swelling appeared to be similar to the reference starch. The granules with added LPL were partly fragmented after hydrolysis.     

Using Visible and Near‐Infrared Reflectance Spectroscopy and Differential Scanning Calorimetry to Study Starch,Protein, and Temperature Effects on Bread Staling

Starch, protein, and temperature effects on bread staling were investigated using visible and near‐infrared spectroscopy (NIRS) and differential scanning calorimetry (DSC). Bread staling was mainly due to amylopectin retrogradation. NIRS measured amylopectin retrogradation accurately in different batches. Three important wavelengths, 970 nm, 1,155 nm, and 1,395 nm, were associated with amylopectin retrogradation. NIRS followed moisture and starch structure changes when amylopectin retrograded. The amylose‐lipid complex changed little from one day after baking. The capability of NIRS to measure changes in the retrograded amylose‐lipid complex was limited. Two important wavelengths, 550 nm and 1,465 nm, were key for NIRS to successfully classify the starch‐starch (SS) and starch‐protein (SP) bread based on different colors and protein contents in SS and SP. Low temperature dramatically accelerated the amylopectin retrogradation process. Protein retarded bread staling, but not as much as temperature. The starch and protein interaction was less important than the starch retrogradation. Protein hindered the bread staling process mainly by diluting starch and retarding starch retrogradation.     

Physicochemical,Structural, and Proteomic Analysis of Starch Granules from Maize Landraces of Northwest Mexico

The variability in grain and starch characteristics and their relationship with the accumulation of starch granule associated proteins were investigated in five maize landraces of Northwest Mexico (Blando de Sonora, Chapalote, Elotero de Sinaloa, Reventador, and Tabloncillo). Significant differences were observed in grain hardness related traits, starch physicochemical properties, and structural properties. Blando de Sonora showed very soft grains, whereas the hardest grains were observed for Chapalote and Reventador. Starch granules isolated from landraces with hard grains contained more amylose and showed polygonal shapes, lower crystallinity and enthalpy of gelatinization, and greater retrogradation and proportion of long amylopectin chains. Proteomic analysis identified the enzymes granule‐bound starch synthase I (GBSSI), starch synthase I and IIa, starch branching enzyme IIb, sucrose synthase 1, and pyruvate phosphate dikinase 2 as granule‐associated proteins. The abundance of GBSSI correlated significantly with amylose content, consistent with the positive correlation observed between amylose and grain hardness. These results showed that the variability in the characteristics evaluated was mainly related to changes in the proportion of amylose in the starch granules, which were associated with differences in the expression of GBSSI. This information may be useful to define strategies for the exploitation and conservation of the landraces.     

Environmental factors that affect the ability of amylose to contribute to retrogradation in gels made from rice flour

Retrogradation in rice is a trait that describes the hardening of cooked rice after storage or cooling, and it has significant implications for many consumers of rice, since many people cook rice in the morning and consume it several hours later or the next day. Tools to select against retrogradation in breeding programs are yet to be described. Here, we aim to determine the effect on retrogradation of storage time and temperature and the role of starch, protein, and lipids using gels made from Koshihikari grown in either Australia or Japan. Immediately after cooking, cooling from 60 to 40 degrees C had a minimal effect on firmness, but cooling to 20 degrees C led to significantly firmer gels. Storing the gels at low temperatures did not have an additional effect on the firmness as compared with storing the gels at 20, 40, or 60 degrees C. The removal of proteins led to significantly softer gels at all storage treatments but did not affect the change in firmness on cooling. The removal of lipids increased the rate of retrogradation and the firmness of gels significantly for all treatments. Koshihikari grown in Japan retrograded much less than Koshihikari grown in Australia. The amount of amylose that could be washed from gels made from Australian flour was much greater than for gels made from Japanese flour. After storage, only low molecular weight amylose chains were released from the gel and only after rewarming them to 60 degrees C. Despite the fact that flours from both origins were 18% amylose, the amount of long amylose chains that were complexed with lipids was much greater for the Japanese rice, and the unavailability of the complexed long amylose chains explained the lack of retrogradation in the Japanese rice. Once the long chains were released from amylose-lipid complexes, the Japanese rice retrograded. Thus, the environmental factor affecting retrogradation in this variety is type or amount of lipids synthesized, and the degree of retrogradation was determined by the availability of long chains of amylose.     

Effect of alpha-amylases from different sources on the retrogradation and recrystallization of concentrated wheat starch gels: relationship to bread staling

Concentrated starch gels were supplemented with four alpha-amylases from different sources. The retrogradation and recrystallization of the gels were evaluated using differential scanning calorimetry (DSC) and X-ray crystallography. Correlations between the retrogradation data and the carbohydrate fractions extracted from these gels were determined. The thermostable (TBA) and intermediate temperature stability (ISBA) bacterial alpha-amylases were most effective in decreasing the rate of retrogradation of the starch in the gels. The cereal alpha-amylase at the high level (CAH) was also effective. Supplementation with the alpha-amylases increased the crystallinity of the gels. Gels supplemented with TBA or ISBA were most crystalline and retrograded to a lesser extent. The results indicated that DSC gives not only a measure of recrystallized amylopectin but also a measure of total order (recrystallized amylopectin and double-helical content). The maltooligosaccharides produced by the enzymes did not appear to be responsible for the reduced rates of retrogradation, but they appeared to be an expression of the degree of starch modification that was responsible for the inhibition of retrogradation. The crystallinity and retrogradation data were similar to results reported for bread and strongly suggest that bread staling is caused by the retrogradation of starch. The results also indicate that alpha-amylases decrease the rate and extent of retrogradation of starch gels by inhibiting the formation of double helices.     

Structure-retrogradation relationship of rice starch in purified starches and cooked rice grains: a statistical investigation

Amylose content and amylopectin chain length distribution, the two most commonly used structural parameters of starch, have significant effects on starch retrogradation. In the present work, starches were separated and purified from 18 rice cultivars. The amylopectin was purified from each starch. Amylopectin chain length distribution was analyzed by high-performance size-exclusion chromatography after debranched using isoamylase. The blue value was used to measure the amylose content before and after the defatting of starch. The amount of amylose associated with lipid was calculated. Pulsed nuclear magnetic resonance was used to follow the retrogradation of starch both in cooked rice grains and in the purified form. The Avrami equation was employed to describe the retrogradation kinetics of rice starch. To look into the relationship between the starch structure and retrogradation behavior, the structural parameters were correlated with retrogradation kinetics parameters using both Pearson and partial correlations. The results indicated the following: first, the retrogradation behavior of rice starch remains similar in both the purified form and cooked rice grains; second, the peak value of amylopectin short-chain length has a significant positive relationship with the amylopectin crystallization rate constant k; third, the amylose content after defatting has a significant positive relationship with the parameter k and a negative relationship with the Avrami exponent n; and fourth, the amount of amylose associated with lipid has a negative relationship with the parameter k.     

冻结-高压湿热解冻对多种淀粉中支链淀粉断裂的影响

冻结和高压湿热解冻是含淀粉面团在食品加工过程的必要工艺,在此过程中淀粉球破裂、部分支链淀粉断裂成为直链淀粉,给面团和食品的物理性状控制带来许多不可预见的结果。该文通过研究加热时间、加热温度、冷冻时间和解冻时间对糊化后的小麦、甘薯、马铃薯和玉米淀粉中直链淀粉含量的影响,间接反映支链淀粉断裂情况,结合冻结解冻前后淀粉分子量分布、链长分布、光学和电子显微镜图谱提出了冻结-解冻过程不同淀粉中支链淀粉可能断裂方式。结果表明,4种淀粉中的直链淀粉含量先升高后下降,直链淀粉含量在4种淀粉的中达到峰值的时间分别为48,48,48,72 h。光学显微照片观察表明,冻融处理会导致更多凝胶化淀粉球的破裂。对于小麦支链淀粉,冻融解冻过程支链淀粉中侧链长度为5、6、7个葡萄糖残基的侧链对应3种可能的断裂方式:2+2+1、2+2+2及2+2+2+1;对于甘薯支链淀粉,支链淀粉中侧链长度为10、11和13个葡萄糖残基的侧链对应3种可能的断裂方式:3+3+4、2+2+3+4及2+2+2+3+4;对于马铃薯支链淀粉,支链淀粉中侧链长度为5和6个葡萄糖残基的侧链对应3种可能的断裂方式:2+3、2+4、3+3;而玉米支链淀粉中,支链淀粉中侧链长度为7、8、9个葡萄糖残基的侧链对应3种可能的断裂方式:2+5,3+5,和3+3+3(其中1表示1个葡萄糖;2表示含2个葡萄糖的麦芽糖、3表示含3个葡萄糖的麦芽多糖、4表示含4个葡萄糖的麦芽多糖和5表示含5个葡萄糖的麦芽多糖)。该论文结果为培育具有冻融稳定性的淀粉种子提供一种全新的思路,即通过基因方法控制植物减少容易断裂淀粉侧链的合成。     

α-Amylolysis of Large Barley Starch Granules

The α-amylolysis of large (volume average 16 μm) barley starch granules was studied by measuring the amount of carbohydrates solubilizing during hydrolysis, and the changes in morphology and molecular structure of the granule residues by scanning electron microscopy, particlesize analysis, size-exclusion chromatography, X-ray diffraction, and differential scanning calorimetry. X-ray diffraction showed that, in the earlier stages of α-amylolysis, both amorphous and crystalline parts of the granules were equally solubilized. More extensive hydrolysis caused a gradual decrease in A-type crystallinity and degradation of the granular structure. Scanning electron microscopy revealed that hydrolysis proceeded through pinholes, and pitted and partially hollow granule residues were formed. The lipid-complexed amylose was less susceptible to α-amylolysis than free amylose and amylopectin. Lipid-complexed amylose started leaching out of the granule residues only after half of the starch had solubilized due to the α-amylase treatment. Even though scanning electron microscopy indicated that there were intact granules left throughout the hydrolysis, the results obtained suggested that α-amylolysis of large barley starch granules proceeded rather evenly among the granules.     

Increasing Slowly Digestible Starch Content of Normal and Waxy Maize Starches and Properties of Starch Products

Changes in the digestibility and the properties of the starch isolated from normal and waxy maize kernels after heat‐moisture treatment (HMT) followed by different temperature cycling (TC) or isothermal holding (IH) conditions were investigated. Moist maize kernels were heated at 80°C for 2 hr. The HMT maize kernels were subjected to various conditions designed to accelerate retrogradation of the starch within endosperm cells. Two methods were used to accelerate crystallization: TC with a low temperature of –24°C for 1 hr and a high temperature of 20, 30, or 50°C for 2, 4, or 24 hr for 1, 2, or 4 cycles, and IH at 4, 20, 30, or 50°C for 24 hr. The starch granules were then isolated from the treated kernels. The starch isolated from HMT normal maize kernels treated by TC using –24°C for 1 hr and 30°C for 2 hr for 2 cycles gave the greatest SDS content (24%) and starch yield (54%). The starch isolated from HMT waxy maize kernels treated by TC using –24°C for 1 hr and 30°C for 24 hr for 1 cycle had an SDS content of 19% and starch yield of 43%. The results suggest that TC after HMT changes the internal structure of maize starch granules in a way that results in the formation of SDS (and RS). They also suggest that thermal treatment of maize kernels is more effective in producing SDS than is the same treatment of isolated starch. All starch samples isolated from treated normal maize kernels exhibited lower peak viscosities, breakdown, and final viscosities and higher pasting temperatures than did the control (untreated normal maize starch). Although peak viscosities and breakdown of the starch isolated from treated waxy maize kernels were similar to those of the control (untreated waxy maize starch), their pasting temperatures were higher. The starch isolated from treated normal and waxy maize kernels with the highest SDS contents (described above) were further examined by DSC, X‐ray diffraction, and polarized light microscopy. Onset and peak temperatures of gelatinization of both samples were higher than those of the controls. Both retained the typical A‐type diffraction pattern of the parent starches. The relative crystallinity of the starch from the treated normal maize kernels was higher than that of the control, while the relative crystallinity of the starch from the treated waxy maize kernels was not significantly different from that of the control. Both treated starches exhibited birefringence, but the granule sizes of both starches, when placed in water, were slightly larger than those of the controls.     

Preparation and Properties of Starch Phosphates Using Waxy,Common, and High‐Amylose Corn Starches. I. Oven‐Heating Method

The structure and physicochemical properties of waxy, common, and high‐amylose corn starch phosphates prepared by oven heating were studied. Starch phosphates prepared by either slurry or dry‐mixing treatment before oven heating were also compared. The slurry treatment more efficiently incorporated phosphorus into starch relative to the dry‐mixing treatment under the reaction conditions studied. In general, the phosphorylated starch prepared by the slurry treatment exhibited a lower gelatinization temperature, a higher peak viscosity, a lesser degree of retrogradation, and improved freeze‐thaw stability compared with those prepared by the dry‐mixing treatment. Phosphorylation occurred probably in both amylose and amylopectin, and the amount and location of incorporated phosphate groups varied with starch types likely due to their different amylose and amylopectin contents. Waxy starch was more prone to phosphorylation, followed by common and high‐amylose starches, respectively.     

A Study of Maize Endosperm Hardness in Relation to Amylose Content and Susceptibility to Damage

The relative amounts of amylose and amylopectin in maize starch were determined in samples representing hard and soft endosperm. Although differences were small, amylose content differed significantly (P < 0.001 and P < 0.05) between the two types of endosperm, with hard endosperm containing a higher percentage of amylose. Scanning electron microscopy was used to determine that the surface appearance of starch granules from hard and soft endosperm differed. Starch granules from soft endosperm had randomly distributed pores on their surfaces, which had a rough appearance. Few pores were observed on granules from hard endosperm. A fairly common occurrence with starch granules from soft endosperm was the development of wrinkles or fissures upon prolonged exposure to the beam of the electron microscope. Thus, a correlation existed between endosperm hardness, amylose content, and susceptibility to wrinkling and fissures. The granules of the soft endosperm of maize, presumably less mature than the granules of the hard endosperm, have a lower amylose content (20.5 ± 1.9% vs. 23.0 ± 1.0%), exhibit more surface pores, and are more susceptible to wrinkling in an electron beam, compared with granules of the hard endosperm. Results suggested that the composition and internal architecture of the starch granule differ depending on the hardness of the endosperm from which it was obtained.     

Effects of Preparation Temperature on Gelation Properties and Molecular Structure of High‐Amylose Maize Starch

In this study, 3% aqueous high‐amylose maize starch (Hylon VII) dispersions were heated to temperatures of 140–165°C. The onset and rate of gel formation was observed using a small‐strain oscillation rheometer as a function of temperature from 90 to 25°C. The gel formation clearly began earlier in high‐amylose starch paste preheated at lower temperatures, but the rate of gelation was slower and the resulting gel was weaker in comparison with starch pastes preheated at higher temperatures. In addition, the structure of the final gels was studied using large deformation compression measurements. The most rigid gel structure on the basis of small and large deformation tests was obtained for high‐amylose starch gel preheated to 150–152°C, depending on the type of measurement. The rate of gelation was also fastest in that temperature range. High‐amylose gels heated to higher temperatures lost their rigidity. The molecular weight distribution of starch molecules was measured by size‐exclusion chromatography. Heating caused extensive degradation of amylopectin, which had a great effect on amylose gel formation and the final gel properties of high‐amylose maize starch. Micrographs of Hylon VII gels showed that phase separation of starch components visible in light microscopy occurred on heating to higher temperatures.     

Comparative study of the retrogradation of intermediate water content waxy maize, wheat, and potato starches

The retrogradation of extruded starches from three different botanical sources was studied in concentrated conditions (34 +/- 1% water) at 25 degrees C using differential scanning calorimetry (DSC) and isothermal calorimetry, Fourier transform infrared spectroscopy (FTIR), and wide-angle X-ray scattering. Potato starch showed the highest rate of retrogradation (approximately 0.17 h(-1)) followed by waxy maize (approximately 0.12 h(-1)), while the retrogradation of wheat starch was the slowest (approximately 0.05 h(-1)). In addition to the kinetics, the extent of molecular order in the retrograded samples was studied in detail in terms of "short-range" (helical) and "long-range" (crystalline) distance scales. The amylopectin crystallinity indices were essentially the same (approximately 47-51% amylopectin basis) for the three starches. However, significant differences were found in the enthalpy of melting measured by DSC after "full" retrogradation (potato, 11.6 +/- 0.7; waxy maize, 9.0 +/- 0.5; and wheat, 6.1 +/- 0.3 J/g of amylopectin). The degree of short-range molecular order in the retrograded state determined by FTIR was waxy maize > potato > wheat. The effect of amylopectin average chain length and the polymorphism of the crystalline phase were taken into account to explain the differences in the retrogradation enthalpies.     

Investigation of Digestibility In Vitro and Physicochemical Properties of A‐ and B‐Type Starch from Soft and Hard Wheat Flour

In this study, the functional properties of A‐ and B‐type wheat starch granules from two commercial wheat flours were investigated for digestibility in vitro, chemical composition (e.g., amylose, protein, and ash content), gelatinization, retrogradation, and pasting properties. The branch chain length and chain length distribution of these A‐ and B‐type wheat starch granules were also determined using high‐performance anion exchange chromatography (HPAEC). Wheat starches with different granular sizes not only had different degrees of enzymatic hydrolysis and thermal and pasting properties, but also different molecular characteristics. Different amylose content, protein content, and branch chain length of amylopectin in A‐ and B‐type wheat starch granules could also be the major factors besides granular size for different digestibility and other functional properties of starch. The data indicate that different wheat cultivars with different proportion of A‐ and B‐type granular starch could result in different digestibility in wheat products.     

Slowly digestible state of starch: mechanism of slow digestion property of gelatinized maize starch

The mechanism underlying the previously reported parabolic relationship between amylopectin fine structure, represented by the weight ratio of linear short chains [degree of polymerization (DP < 13) to long chains (DP >/= 13], and slowly digestible starch (SDS) content was investigated from the viewpoint of starch retrogradation and substrate susceptibility to enzyme hydrolysis. A maize mutant sample, termed "highest long-chain starch" (HLCS) representing group I samples with a higher proportion of long chains, showed a bell-shaped SDS pattern with retrogradation time, whereas insignificant changes in SDS were found for the sample termed "highest short-chain starch" (HSCS) representing group II samples with a higher proportion of short chains. This corresponded to results from X-ray powder diffraction and differential scanning calorimetry that showed a rapid increase of crystallinity and enthalpy for HLCS during retrogradation, but negligible changes for sample HSCS. Therefore, retrogradation was associated with SDS content for group I samples, but not for group II samples. Analysis of amylopectin fine structure, SDS content, retrogradation enthalpy, SDS material debranching profile, and hydrolysis pattern demonstrated, for group I samples, that linear branched chains of DP 9-30 of amylopectin may act as anchor points to slow the digestion of branced-chain fractions of DP > 30, which constitute the major slowly digestible portion, whereas for group II samples, it is the inherent molecular structure of amylopectin with a higher amount of branches and shorter chains that is not favorable for rapid enzyme digestion. The concept of a slowly digestible starch state (SDS state) that could be a chemical or physical entity is proposed to better describe the mechanistic underpinning of the slow digestion property of starches.