Gelatinization Properties of Starches from Three Successive Generations of Six Exotic Corn Lines Grown in Two Locations

The objectives of this research were to evaluate the intra‐ and interpopulation variability in gelatinization properties of starches from exotic corn lines and their derivatives when grown 1) during two successive years in the same location; and 2) in both temperate and tropical environments. Six novel exotic corn lines (two 100% exotic and four 25% exotic derived from a breeding cross developed by crossing an exotic hybrid with Corn Belt lines) were selected for this research because their starches have significantly different (and potentially useful) thermal properties from those found in starch from normal Corn Belt corn. The S_n (n = 3 for 25% exotic lines and n = 1 for 100% exotic lines) generations of the six exotic lines were self‐pollinated and grown in the winter nursery in Puerto Rico. Two successive generations (S_n+1 and S_n+2) of lines selected for low onset of gelatinization temperature were self‐pollinated and grown in the same environment near Ames, IA. To evaluate the effect of environment, the S_n+2 generation also was self‐pollinated and grown in the winter nursery in Puerto Rico. Thermal properties of starches from 10 single kernels from each line were analyzed by using differential scanning calorimetry (DSC) at a ratio of 4 mg of dry starch to 8 mg of distilled water. After subsequent generations, the differences in DSC gelatinization properties between selected kernels within each progeny line narrowed, suggesting increased homogeneity of starch structural properties within each line. Unusual thermal properties were fixed in some progeny lines. Environmental factors also affected the thermal properties of starch and a significant interaction between environment and genotype was observed. These results suggest that introgression of adapted germplasm with useful genes from exotic corn would increase the available genetic variability for starch functionality and allow the development of hybrids with important value‐added traits.     

Thermal Properties of Starch from 62 Exotic Maize (Zea mays L.) Lines Grown in Two Locations

The variability in thermal properties among 62 S₃ lines derived from a high-yielding exotic corn (Zea mays) population, Antigua 1 (PI 484990), was evaluated by differential scanning calorimetry (DSC). The S₃ lines were grown in Puerto Rico (1990–1991) and Georgia (1994). Separate single-kernel starch extractions for five kernels (five replicates) from each line grown in each location were performed, and the starch was analyzed. The DSC values reported included gelatinization onset (T_oG), range (R_G), enthalpy (ΔH_G), and peak height index (PHI) and retrogradation onset (T_oR), range (R_R), enthalpy (ΔH_R), and percent retrogradation (%R) (an indication of the stability of gelatinized starch after storing at 4°C for 7 days). Significant differences (P < 0.05) were found among the 62 lines of Antigua 1 for T_oG, R_G, and PHI and highly significant differences (P < 0.01) were found for ΔH_G. The starches from plants grown in Georgia (1994) had significantly (P < 0.05) greater T_oG, ΔH_G, and PHI but a significantly lower R_G than those from Puerto Rico (1990–1991). These data suggest that the starch from plants grown in Georgia (1994) might have a greater degree of crystallinity than that from Puerto Rico (1990–1991). None of the retrogradation values were significantly different among starches of the 62 lines of Antigua 1 and the starches from plants grown in the two locations.     

Thermal Properties of Starch from Selected Maize (Zea mays L.) Mutants During Development

The changes in thermal properties of maize starches during five stages of kernel maturity, (12, 18, 24, 30, and 36 days after pollination [DAP]), from three mutant genotypes, amylose extender (ae), sugary-2 (su₂), and waxy (wx) in an OH43 background, and the OH43 genotype were studied using differential scanning calorimetry (DSC). Within a genotype, DSC values of starches at 24, 30, and 36 DAP were similar to each other and often were significantly different (P < 0.05) from the values at 12 DAP, indicating possible differences in the fine structure of starch during endosperm development. For su₂ starches, the gelatinization onset temperature (T_oG) significantly decreased after 12 DAP and remained low throughout the study. The gelatinization range (R_G) had a similar pattern. For wx starches, T_oG at 18 DAP was significantly lower than at 12 DAP but tended to increase after 18 DAP. The R_G increased significantly after 12 DAP and significantly decreased after 30 DAP. Thus, thermal properties of starches during early development were different from those of their mature counterparts, and differences among the mutant genotypes and the normal starch originated from the earliest endosperm development stage studied (12 DAP).     

Comparison of Flour Starch Properties in Half‐Sib Families of Opaque‐2 Maize (Zea mays L.) from Argentina

Since the discovery of the o2 mutation in maize, many studies have reported the characterization of the protein quality of opaque‐2 genotypes. However, few have reported the properties of their starch. The objective of this study was to characterize flour starch properties of 12 half‐sib families of opaque‐2 maize from Argentina. Chemical composition and thermal and pasting properties of whole grain flour were determined. Nonopaque genotypes were used as a control. Starch content of opaque‐2 genotypes did not show significant differences compared with nonopaque genotypes, yet amylose content was significantly lower. A high variability in pasting and thermal properties was observed in genotypes. Opaque samples showed a significantly higher peak viscosity and a lower pasting temperature compared with nonopaque samples, probably owing to larger and less compact starch granules in the floury endosperm. The higher the gelatinization enthalpy of opaque‐2 genotypes was, the lower the amylose content in relation to nonopaque varieties. Two retrogradation endotherms were observed in DSC analysis: one corresponding to amylopectin crystallization and the other to melting of amylose‐lipid complex. Both enthalpies were considered total starch retrogradation (ΔH _RT). A wide range of variation was obtained in ΔH _RT in opaque‐2 genotypes, but no significant differences between opaque and nonopaque genotypes were observed. The differences in starch properties found in this study would make it possible to identify opaque‐2 families with particular characteristics for the development of starchy food items adapted to specific processing traits.     

Retrogradation of Maize Starch After Thermal Treatment Within and Above the Gelatinization Temperature Range

Studies of starch retrogradation have not considered the initial thermal treatment. In this article, we explore the effect of heating to temperatures within and above the gelatinization range on maize starch retrogradation. In the first experiment, 30% suspensions of waxy (wx) starch were initially heated to final temperatures ranging from 54 to 72°C and held for 20 min. On reheating in the differential scanning calorimeter immediately after cooling, the residual gelatinization endotherm peak temperature increased, the endotherm narrowed, and enthalpy decreased. Samples stored for seven days at 4°C showed additional amylopectin retrogradation endotherms. Retrogradation increased dramatically as initial holding temperature increased from 60 to 72°C. In a second experiment, wx starch was initially heated to final temperatures from 54 to 180°C and rapidly cooled, followed by immediate reheating or storage at 4°C. Maximum amylopectin retrogradation enthalpy after storage was observed for initial heating to 82°C. Above 82°C, retrogradation enthalpy decreased as initial heating temperature increased. A similar effect for ae wx starch was observed, except that retrogradation occurred more rapidly than for wx starch. These experiments show that heating to various temperatures above the range of gelatinization may profoundly affect amylopectin retrogradation, perhaps due to varying extents of residual molecular order in starch materials that are commonly presumed to be fully gelatinized. This article shows that studies of starch retrogradation should take into account the thermal history of the samples even for temperatures above the gelatinization temperature range.     

Relationship of Gelatinization and Recrystallization of Cross‐Linked Rice to Glass Transition Temperature

Nonwaxy rice starch was cross‐linked with sodium trimetaphosphate and sodium tripolyphosphate to obtain different degrees of cross‐linking (9.2, 26.2, and 29.2%). The objective was to investigate the influence of cross‐linking on thermal transitions of rice starch. Starch suspensions (67% moisture) were heated at 2°C/min using differential scanning calorimetry (DSC) to follow melting transition of amylopectin. Biphasic transitions were observed at ≈60–95°C in all samples. Melting endotherms of amylopectin shifted to a higher temperature (≤5°C) with an increasing degree of cross‐linking, while there was no dramatic change in enthalpy. Recrystallization during aging for 0–15 days was significantly suppressed by cross‐linking. The delayed gelatinization and retrogradation in crosslinked starch were evident due to restricted swelling and reduced hydration in starch granules. Glass transition temperature (T_g) measured from the derivative curve of heat flow was ‐3 to ‐4°C. No significant change in T_g was observed over the storage time studied.     

Properties of Starches from Near‐Isogenic Wheat Lines with Different Wx Protein Deficiencies

To determine the effect of amylose content on the starch properties, the amylose content, pasting properties, swelling power, enzymatic digestibility, and thermal properties of partial and perfect waxy types along with their wild‐type parent were analyzed. As expected, amylose content decreases differently in response to the loss of each Wx gene, showing the least response to Wx‐A1a. Most of the characteristics, except the thermal properties of the amylose‐lipid complex in differential scanning calorimetry (DSC), differed significantly among the tested types. Furthermore, the breakdown, setback, and pasting temperatures from the Rapid Visco Analyser (RVA) and the enzymatic digestibility, swelling power, peak temperature, and enthalpy of starch gelatinization from DSC showed a correlation with the amylose content. The relationships between the peak viscosity from the RVA and the onset temperature of starch gelatinization determined by DSC with amylose content of the tested materials were not clear. Waxy starch, which has no amylose, showed a contrasting behavior in starch gelatinization compared with nonwaxy starches. Among the nonwaxy starches, lower setback, lower pasting temperature, higher enzyme digestibility, higher peak temperature, higher enthalpy of starch gelatinization, and higher swelling were generally associated with low amylose starches.     

Gelatinization and Retrogradation Kinetics of High‐Fiber Wheat Flour Blends: A Calorimetric Approach

The impact of dietary fiber (DF) mixtures on dough thermal properties needs to be investigated when designing high‐fiber wheat bread. Effects of flour replacement at different levels (6–34%) by soluble (inuline [FN]), partially soluble (sugar beet [FX], pea cell wall [SW]), and insoluble (pea hull [EX]) DF on wheat dough thermal profiles have been investigated by simulating baking, cooling, and storage in differential scanning calorimetry (DSC) pans. In general, DF incorporation into water‐flour systems delayed endothermic transition temperatures for both gelatinization and retrogradation phenomena except for the peak temperature (T_p) of retrogradation. With some exception, the pattern of the enthalpy of amylopectin retrogradation was lower and slower (lower constant of proportion, k) over 10 days of storage in gelatinized hydrated flour‐fiber blends when compared with control without DF. FX, a partially soluble fiber, provided major effects on gelatinization (T_p decrease and ΔH increase) and retrogradation kinetics (the Avrami exponent, n, increase). Single presence of EX allowed a significant reduction in the Avrami exponent n leading to slower kinetics for amylopectin retrogradation when included in the blends.     

Identification of QTL Controlling Thermal Properties of Maize Starch

Starch has many uses and some of these uses would be facilitated by altering its thermal properties. Genetic manipulation of starch thermal properties will be facilitated by a better understanding of the genetic control of starch gelatinization. We used differential scanning calorimetry to characterize the gelatinization parameters of maize (Zea mays L.) kernel starch prepared from two populations of recombinant inbred lines, an intermated B73xMo17 population (IBM) and an F_6:7 Mo17xH99 population. The traits examined were the onset and peak temperatures of gelatinization and the enthalpy of gelatinization. These traits were measured for both native starch and for gelatinized starch allowed to recrystallize, a process called retrogradation. Substantial variation in these traits was found in spite of the narrow genetic base of the populations. We identified several quantitative trait loci (QTL) controlling traits of interest in each population. In the IBM population, a significant QTL for the peak temperature of gelatinization of retrograded starch co‐localized to a molecular marker in the Wx1 gene, which encodes a granule bound starch synthase. The major QTL identified in this study explain, on average, ≈15% of the variation for a given trait, underscoring the complexity of the genetic control of starch functional properties.     

Starch Characteristics of the Rice Mutant du2‐2 Taichung 65 Highly Affected by Environmental Temperatures During Seed Development

The effects of environmental temperature (21 vs. 28°C) during rice seed development on the starch characteristics (apparent amylose content, amylopectin chain length distribution, and gelatinization properties) of nonwaxy Taichung 65 (T65), waxy Taichung (T65wx), du2‐2 mutated low‐amylose strain Taichung (76‐3/T65), and Koshihikari were studied. Amylose contents increased with decreasing environmental temperatures. Analysis of the amylopectin chain length distribution showed that the relative amounts of long chains with degree of polymerization (DP) > 25 in all starches decreased if maturation occurred at 21°C. Gelatinization onset, peak, and conclusion temperatures and enthalpies decreased with decreasing environmental temperatures. Of all starches studied, the du2‐2 mutated low‐amylose Taichung (76‐3/T65) was most affected by maturation temperatures. These results indicate that the du2‐2 mutated low‐amylose Taichung (76‐3/T65) may be a useful strain in understanding biochemical and genetic starch biosynthesis response to slight changes in temperature.     

Wet‐Milling Characteristics of 10 Lines from Germplasm Enhancement of Maize Project Compared with Five Corn Belt Lines

The use of corn (Zea mays L.) hybrids with high grain yield and starch extractability has steadily increased in the processing industry. In light of widespread corn seed industry participation in the Germplasm Enhancement of Maize Project (GEM), which seeks to enhance exotic germplasm, future hybrids may contain more exotic sources in genetic backgrounds. It is necessary to establish and monitor physical, compositional, and milling characteristics of the new exotic breeding materials to determine the processing value. The present study was conducted to determine the wet‐milling characteristics of a set of GEM lines compared with typical Corn Belt lines. Ten GEM lines introgressed with exotic materials from Argentina, Chile, Cuba, Florida, and Uruguay and previously identified as having different starch yields, three commercial inbred lines, and two public inbred lines (B73 and Mo17) were analyzed using both near‐infrared transmittance (NIT) and a 100‐g wet‐milling procedure. There were statistical differences (P < 0.05) in the yield of wet‐milled fractions (starch, fiber, gluten, and germ). The GEM lines AR16035:S19‐227‐1‐B and CUBA117:S1520‐562‐1‐B had similar or better starch yield and starch recovery than B73 and the other adapted inbred lines, indicating that they may be useful in improving the proportion of extractable starch present in kernels of hybrids. Residual protein levels in the starch and gluten fractions were 0.26–0.32% and 38–45%, respectively. The starch yield of GEM lines from wet milling correlated positively with starch content from NIT and was negatively correlated with protein content of the corn kernels. Oil content in the germ varied from 50 to 60%. Our results indicate that incorporating GEM lines in a breeding program can maintain or even improve wet‐milling characteristics of Corn Belt materials if lines with appropriate traits are used.     

Molecular and Gelatinization Properties of Rice Starches from IR24 and Sinandomeng Cultivars

The differences in pasting properties involving gelatinization and retrogradation of rice starches from IR24 and Sinandomeng cultivars during heating‐cooling processes were investigated using a Rapid Visco Analyser (RVA)and a dynamic rheometer. The results were discussed in relation to the molecular structure, actual amylose content (AC), and concentration of the starches. Generally, both starches possessed a comparable AC (≈11 wt%), amylose average chain length (CL), iodine absorption properties, and dynamic rheological parameters on heating to 95°C at 10 wt% and on cooling to 10°C at higher concentrations. In contrast to Sinandomeng, IR24 amylose had a greater proportion of high molecular weight species and number‐average degree of polymerization (DP_n). IR24 amylopectin possessed a lower DP_n and greater CL, exterior CL (ECL), and interior CL (ICL). Comparing the results of RVA analysis and dynamic rheology, the gelatinization properties and higher retrogradation tendencies of IR24 starch can be related to the structural properties and depend on starch concentration. In addition, the exponent n of starch concentration for storage moduli at 25°C (G′₂₅ ∝ Cⁿ) increased linearly with increasing AC.     

Variation Among Physical,Compositional, and Wet‐Milling Characteristics of the F1 Generation of Corn Hybrids of Introgressed Exotic and Adapted Inbred Lines

Hybrids with high grain yield and higher starch, protein, or oil content are available to corn growers; however, they result from crossing adapted Corn Belt inbred lines that rarely include exotic germplasm. This study was conducted to determine whether Corn Belt lines introgressed with exotic materials from Argentina, Chile, Uruguay, Cuba, and Florida have appropriate wet‐milling characteristics in their hybrids. Ten lines from the Germplasm Enhancement of Maize (GEM) project with different starch contents were crossed to three adapted inbred lines used as testers. The B73×Mo17 hybrid was used as a control. The F₁ generation of these 30 experimental hybrids was analyzed using both near‐infrared transmittance (NIT) technology and a 100‐g modified wet‐milling procedure, and measuring test and 1,000‐kernel weight. There was great variation among physical, compositional, and wet‐milling characteristics of the experimental hybrids, suggesting that exotic germplasm can be used to improve wet‐milling characteristics of Corn Belt hybrids.     

Quality Characteristics of Waxy Hexaploid Wheat (Triticum aestivum L.): Properties of Starch Gelatinization and Retrogradation

The viscoelastic properties and molecular structure of the starch isolated from waxy (amylose-free) hexaploid wheat (WHW) (Triticum aestivum L.) were examined. WHW starch generally had lower gelatinization onset temperature, peak viscosity, and setback than the starch isolated from normal hexaploid wheat (NHW). Differential scanning calorimetry (DSC) showed that WHW starch had higher transition temperatures (T_o, T_p, and T_c) and enthalpy (ΔH) than NHW starch. However, when compared on the basis of amylopectin (AP) content, ΔH of WHW starch was almost statistically identical to that of its parental varieties. Typical A-type X-ray diffraction patterns were observed for the starches of WHW and its parental varieties. Somewhat higher crystallinity was indicated for WHW starch. WHW starch was also characterized by having greater retrogradation resistance. The high-performance size-exclusion chromatography (HPSEC) of amylopectin showed that each amylopectin yielded two fractions after debranching. Although WHW amylopectin had somewhat long B chains, little difference was observed in the ratio of Fr.III/ Fr.II between WHW and its parental varieties.     

Quantitative Genetic Basis of Gelatinization Temperature of Rice

Gelatinization temperature of rice starch plays an important role in grain quality. We mapped eight loci responsible for the gelatinization parameters of rice: onset temperature (T_o), peak temperature (T_p), conclusion temperature (T_c) and enthalpy (ΔH) using a population of 242 recombinant inbred lines (RIL). Eight quantitative trait loci (QTL) were mapped onto the chromosome regions where starch‐synthesis related genes are located. The Wx gene (granule‐bound starch synthase [GBSS]), and the region of chromosome 11 played major roles in gelatinization trait determination; and other loci were starch‐branching enzyme (SBE) genes that modify the two major loci by epistasis. High‐level interactions were also detected indicating a trans mode of action of the genes. The genetic basis for these parameters was consistent with the relationship between the thermal properties and the structure of the starch granule. This information can be helpful for the improvement of starch‐producing crops, including the improvement of rice grain quality, both for food and industrial uses.     

Structural and Functional Characteristics of Selected Soft Wheat Starches

Starches from eight soft wheat samples (two parent lines and six offspring) were isolated; relationships between their structures and properties were examined. Branch chain‐length distributions of amylopectins were determined by using high‐performance anion exchange chromatography equipped with an amyloglucosidase reactor and a pulsed amperometric detector (HPAEC‐ENZ‐PAD). Results showed that the average chain length of the eight samples varied at DP 25.6–26.9. Starch samples of lines 02, 60, 63, 95, and 114 consisted of amylopectins with more long chains (DP ≥ 37) and longer average chain length (DP 26.2–26.9) than that of other samples. These starch samples of longer branch chain length displayed higher gelatinization temperatures (55.3–56.5°C) than that of other samples (54.4–54.9°C) and higher peak viscosity (110–131 RVU) and lower pasting temperature (86.3–87.6°C) than others (83–100 RVU and 88.2–88.9°C, respectively). The M_w of amylopectins, determined by using high‐performance size exclusion chromatography equipped with multiangle laser‐light scattering and refractive index detectors (HPSEC‐MALLS‐RI), were similar for all samples (6.17 × 10⁸ to 6.97 × 10⁸). There were no significant differences in amylose and phosphorus contents between samples. These results indicated that physical properties of wheat starch were affected by the branch‐chain length of amylopectin.     

Exotic Corn Lines with Increased Resistant Starch and Impact on Starch Thermal Characteristics

Ten parent corn lines, including four mutants (dull sugary2, amylose‐extender sugary2, amylose‐extender dull, and an amylose‐extender with introgressed Guatemalen germplasm [GUAT ae]) and six lines with introgressed exotic germplasm backgrounds, were crossed with each other to create 20 progeny crosses to increase resistant starch (RS) as a dietary fiber in corn starch and to provide materials for thermal evaluation. The resistant starch 2 (RS2) values from the 10 parent lines were 18.3–52.2% and the values from the 20 progeny crosses were 16.6–34.0%. The %RS2 of parents was not additive in the offspring but greater RS2 in parents was correlated to greater RS2 in the progeny crosses (r = 0.63). Differential scanning calorimetry (DSC) measured starch thermal characteristics, revealing positive correlations of peak gelatinization temperature and change in enthalpy with %RS2 (r = 0.65 and r = 0.67, P ≤ 0.05); however, % retrogradation (a measure of RS3) and retrogradation parameters did not correlate with %RS2. The %RS2 and onset temperature increased with the addition of the ae gene, likely because RS delays gelatinization.     

Molecular Characterization of Corn Starch Using an Aqueous HPSEC‐MALLS‐RI System Under Various Dissolution and Analytical Conditions

Molecular characteristics based on absolute weight‐average molecular weight (M_w) and z‐average radius of gyration (R_g) of normal corn starch were analyzed by high‐performance size‐exclusion chromatography (HPSEC) attached to multiangle laser‐light scattering (MALLS) and refractive index (RI) detectors under different starch dissolution and analytical conditions. Autoclaving (121°C, 20 min) or microwave heating (35 sec) provided better HPSEC recovery and higher M_w for starch molecules than simple dissolution in hot water. The M_w for the autoclaved corn amylopectin and amylose fractions separated with a TSK G5,000 column at 60°C were 201 × 10⁶ and 3.3 × 10⁶, respectively. The specific volume for gyration (SV_g) calculated from M_w and R_g could be used for the comparison of molecular compactness which was inversely related to the degree of branching. The SV_g values of amylopectin and amylose fractions in the chromatogram (TSK G5,000, autoclaved for 20 min) were 0.092 and 0.529, respectively. But a portion (20–30%) of large amylopectin molecules did not pass the injection membrane filter (3.0 μm) and the SEC column, resulting in incomplete recovery. The unfiltered portion varied according to the dissolution treatment. Homogenization (7,000 rpm, 5 or 10 min) of the starch solution improved the recovery of the amylopectin fraction, but significantly increased the M_w of the amylose fraction (17 × 10⁶). Sonication for 5 min degraded starch molecules. For accurate analysis of a native starch using an aqueous SEC, the starch should be fully dissolved with proper treatment such as autoclaving or microwaving, and the column should be improved for full recovery of large amylopectin molecules.     

Characterization of Thermal Traits of Starches from Argentinian Maize Inbreds: Genotypic and Crop Year Variability

Thermal properties are among the most important end‐use characteristics of starch from maize (Zea mays L.). Knowledge of the contribution of genotype and environment to the total variance for starch thermal properties is needed to aid in defining a testing strategy for selecting maize with desirable thermal starch properties. Thus, the objectives of this study were 1) to characterize the thermal properties of starches from a group of recently developed Argentine maize inbred lines, and 2) to assess the variability in starch properties attributable to genetic and crop year effects. Twelve inbred lines developed by the National Institute of Agricultural Technology (INTA) in Argentina derived from a wide array of germplasm sources were evaluated. Gelatinization and retrogradation properties were measured by differential scanning calorimetry. Enthalpy means for gelatinization were below means reported in the literature, suggesting possible energy savings when using these starches. The ratio between change in enthalpy for retrogradation and gelatinization was above the mean reported in the literature, suggesting a starch that may be useful as a dietary fiber. Significant environmental effects caused by crop year were detected. Some inbred lines, with smaller observed ranges and standard deviations across environments, may be more stable for some properties.     

Thermal Properties of Corn Starch Extraction Intermediates by Differential Scanning Calorimetry

Thermal properties of corn starch extraction intermediates from four types of corn were studied using differential scanning calorimetry. Starch at four different stages of extraction, including a standard single-kernel starch isolation procedure and three starch extraction intermediates, was isolated from mature corn kernels of B73 and Oh43 inbreds and the mutants of waxy (wx) and amylose extender (ae) in an Oh43 background. Differences in thermal properties and moisture and protein contents of starch from the extraction stages were statistically analyzed. Most thermal properties (gelatinization and retrogradation onset temperatures, gelatinization and retrogradation ranges, gelatinization and retrogradation peak temperatures, gelatinization and retrogradation enthalpies, peak height index, and percentage of retrogradation) of starches extracted at stage 3 intermediate (a procedure that did not include a final washing step) were similar to those of starch extracted by the standard single-kernel isolation procedure. Values for gelatinization peak temperature, gelatinization enthalpy, and peak height index were different between the standard and the stage 3 intermediate. The values obtained from starches extracted at stage 3, however, were consistent and predictable, suggesting that this extraction intermediate might be used in screening programs in which many starch samples are evaluated. By using the stage 3 extraction, samples could be evaluated in three rather than four days and the procedure saved ≈0.5 hr of labor time. The other two starch extraction intermediates, which excluded filtering and washing or filtering, washing, and steeping, produced starch with thermal properties generally significantly different from starch extracted by the standard single-kernel isolation procedure.