Physicochemical Properties of Zero Amylose Hull-less Barley Starch

Zero amylose starch isolated from hull-less barley (HB) showed a typical A-type diffraction pattern. The X-ray analysis suggested that granules of zero amylose (SB94794) and 5% amylose (CDC Candle) HB starches had lower crystallinity than did commercial waxy corn starch. Differential scanning calorimetry showed lower transition temperatures and endothermal enthalpies for the HB starches than for the waxy corn starch. The zero amylose HB starch showed a Brabender pasting curve similar to that of waxy corn starch, but with lower pasting and peak temperatures and a higher peak viscosity. Noteworthy characteristics of zero amylose HB starch were its low pasting temperature and high paste clarity and freezethaw stability, which make this starch useful for many food and industrial applications.     

Physical properties of octenyl succinic anhydride modified rice,wheat, and potato starches

The physical properties of octenyl succinic anhydride (OSA) starches prepared from rice, wheat, and potato starches were studied. Rice and wheat OSA starches had significantly higher peak viscosity (PV), hot paste viscosity (HPV), and cool paste viscosity (CPV), but potato OSA starch had only significantly higher CPV, relative to the native starch. The gel hardness was higher with lower degree of substitution (DS) but lower with higher DS OSA compared to native starch. The swelling volumes (SV) of rice and wheat OSA starches were significantly higher compared to native starch, but the SV of potato OSA starch was slightly lower at high DS. The gelatinization temperature (GT) of rice OSA starches was sharply lower at low DS; for wheat OSA starch it was slightly lower even at high DS, but potato OSA starches had higher GT than the native starch. The enthalpy of all the OSA starches decreased gradually with increased DS. This study showed that the magnitude of changes in physical properties of OSA-modified starches depends not only on their DS but also on the botanical origin of the native starches.     

Starch Pasting Properties and Amylose Content from 17 Waxy Barley Lines

Starch pasting properties and amylose content from 17 waxy barley lines (waxy gene originating from indigenous lines and an artificial mutant) were analyzed using rapid viscosity analysis (Rapid Visco Analyser [RVA]). Amylose contents varied from 0% (Shikoku‐hadaka 97) to 9.5% (Shikoku‐hadaka 96) compared with 30% for normal barley. Eight parameters were obtained from RVA profiles of these lines and correlation between each of these parameters and amylose content were evaluated. These parameters include pasting temperature (PT), peak viscosity (PV), temperature at PV, minimum viscosity (MV), final viscosity (FV), breakdown (BD), setback (SB), and time maintained at >80% PV (hot paste stability [HPS]). Significant correlations (0.64 and 0.61) were found between amylose content and FV and SB, respectively. High correlation (0.72) was found between amylose content and temperature at PV. HPS calculated from RVA profiles showed the highest correlation (0.79) to amylose content. Outer part of barley grains contained higher amounts of amylose than the inner part. There was a tendency that both PT and FV positively correlated to the amylose content of these parts.     

Sources of Variation for Starch Gelatinization,Pasting, and Gelation Properties in Wheat

The starch of wheat (Triticum aestivum L.) flour affects food product quality due to the temperature-dependent interactions of starch with water during gelatinization, pasting, and gelation. The objective of this study was to determine the fundamental basis of variation in gelatinization, pasting, and gelation of prime starch derived from seven different wheat cultivars: Kanto 107, which is a partial waxy mutant line, and six near-isogenic lines (NILs) differing in hardness. Complete pasting curves with extended 16-min hold at 93°C were obtained using the Rapid Visco Analyser (RVA). Apparent amylose content ranged from 17.5 to 23.5%; total amylose content ranged from 22.8 to 28.2%. Starches exhibited significant variation in onset of gelatinization. However, none of the parameters measured consistently correlated with onset or other RVA curve parameters that preceded peak paste viscosity. Peak paste viscosity varied from 190 to 323 RVA units (RVU). Higher peak, greater breakdown, lower final viscosity, negative setback, and less total setback were associated with lower apparent and total amylose contents. Each 1% reduction in apparent or total amylose content corresponded to an increase in peak viscosity of about 22 and 25 RVU, respectively, at 12% starch concentration. Of the seven U.S. cultivars, the lower amylose cultivars Penawawa and Klasic were missing the granule-bound starch synthase (GBSS; ADPglucose starch glycosyl transferase, EC 2.4.4.21) protein associated with the Waxy gene locus on chromosome 4A (Wx-B1 locus). Kanto 107 was confirmed as missing both the 7A and 4A waxy proteins (Wx-A1 and Wx-B1 loci). The hardness NIL also were shown to be null at the 4A locus. Apparent and total amylose contents of prime starch generally corresponded well to the number of GBSS proteins; although the hardness NIL tended to have somewhat higher amylose contents than did the other GBSS 4A nulls. We concluded that reduced quantity of starch amylose due to decreased GBSS profoundly affects starch gelatinization, pasting, and gelation properties.     

转基因抗虫水稻矮秆突变体的品质性状分析

对转cry1Ab基因抗虫水稻矮秆突变体的品质性状与原亲本秀水11进行了对比分析,发现在最高粘度、热浆粘度和冷胶粘度等RVA谱特征值上两者有显著差异,而在外观品质、碾磨品质、表观直链淀粉含量、胶稠度和淀粉粘滞特性的崩解值、消减值等淀粉食用品质方面基本相同,在蛋白、脂肪、灰分、矿物质和氨基酸等关键营养成分上也不存在显著差异,在蒸煮后的矮杆突变体米粒中也未检测到Cry1Ab杀虫蛋白。     

Role of Phosphorus in Viscosity,Gelatinization, and Retrogradation of Starch

The effect of starch crystallinity and phosphorus on starch gelatinization and retrogradation were studied using wide-angle X-ray powder diffraction, cross polarization/magic angle spinning (CP/MAS) ¹³C nuclear magnetic resonance (NMR) spectroscopy, ³¹P NMR spectroscopy, Rapid Visco Analyzer (RVA) and differential scanning calorimetry (DSC). Two starches differing significantly in peak viscosity (cv. Stephens, 283 BU; cv. Crew, 560 BU) were comparable in amylose content and starch crystallinity, while differing significantly in phospholipids content. Starch of lower peak viscosity had a higher phospholipids content and showed a slower rate of retrogradation. Starch from Stephens (0.098% phosphorus) had an enthalpy value of retrograded starch of 2.2 J/g after 14 days of storage, while starch from Crew (0.062% phosphorus) had an enthalpy value as high as 4.4 J/g. Defatting with a hot n-propanol and water (3:1) mixture caused substantial changes in peak viscosity. Peak viscosity for starch from Crew decreased by 75 RVU due to defatting, while starch from Stephens decreased by as much as 125 RVU. After defatting with the hot n-propanol water mixture, the rate and extent of starch retrogradation were comparable between the prime starches, which differed significantly in peak viscosity.     

Genetic variation in the physical properties of sweet potato starch.

Sweet potato starch, prepared from 44 genotypes adapted to Philippine conditions, showed wide variation and distinctly different pasting profiles in Rapid Visco-Analyzer (RVA) analysis at 11% and 7% starch concentration. At 11% starch concentration, the pasting profiles were type A, characterized by high to moderate peak with a major breakdown and low cold paste viscosity. At 7%, the pasting profile was generally type C, characterized by the absence of a distinct peak with none to very slight breakdown and high cold paste viscosity. However, differentiation among genotypes was better achieved from RVA pasting profiles at 11% starch concentration. Peak viscosity (PV) and hot paste viscosity (HPV) at 11% starch paste concentration had significant negative correlation with amylose content. PV, HPV, and setback ratio were significantly correlated to adhesiveness of the starch gel. Sweet potato starch generally had high swelling volume but low solubilities at 92.5 degrees C.     

辐照改良水稻淀粉特性的研究

以相似AAC的不同类型水稻 (籼稻、粳稻和杂交稻 )和不同AAC早籼稻品种为材料 ,研究了辐照对淀粉粘滞性、理化品质和淀粉颗粒的影响。研究表明 ,最高粘度 (PKV)、热浆粘度 (HPV)、冷胶粘度 (CPV)、消减值 (SBV)和回复值 (CSV)等 5个RVA谱特征参数随剂量增加均显著下降。辐照明显降低了低AAC品种、糯稻和中等AAC品种的AAC ,但对高AAC品种的AAC影响不显著。辐照对碱消值 (ASV)无影响 ,但最高粘度时间 (PKT)随剂量增加而下降。胶稠度尤其是高AAC的胶稠度 (GC)随剂量增加明显提高。胚乳内淀粉颗粒经辐照处理也有所变形。结果表明 ,辐照可提高稻米食用和蒸煮品质。     

Mechanism and enzymatic contribution to in vitro test method of digestion for maize starches differing in amylose content

To determine the rapidly digestible starch (RDS), slowly digestible starch (SDS), and resistant starch (RS) contents in a starch sample, the addition of amyloglucosidase is often used to convert hydrolyzates from α-amylase digestion to glucose. The objectives of this study were to investigate the exact role of amyloglucosidase in determining the digestibility of starch and to understand the mechanism of enzymatic actions on starch granules. Four maize starches differing in amylose content were examined: waxy maize (0.5% amylose), normal maize (≈27% amylose), and two high-amylose starches (≈57 and ≈71% amylose). Notably, without amyloglucosidase addition, the RS content increased from 4.3 to 74.3% for waxy maize starch, 29.7 to 76.5% for normal maize starch, 65.8 to 88.0% for starch with 57% amylose, and 68.2 to 90.4% for the starch with 71% amylose. In the method without α-amylase addition, less RS was produced than without added amyloglucosidase, except in maize at 71% amylose content. Scanning electron microscopy (SEM) revealed the digestive patterns of pinholes with α-amylase and burrowing with amyloglucosidase as well as the degree of digestion between samples. To understand the roles of amyloglucosidase and α-amylase in the in vitro test, multiple analytical techniques including gel permeation chromatography, SEM, synchrotron wide-angle X-ray diffraction, and small-angle X-ray scattering were used to determine the molecular and crystalline structure before and after digestion. Amyloglucosidase has a significant impact on the SDS and RS contents of granular maize starches.     

Physicochemical Properties of Rice Starch Modified by Hydrothermal Treatments

Rice starches of long grain and waxy cultivars were annealed (ANN) in excess water at 50°C for 4 hr. They were also modified under heat-moisture treatment (HMT) conditions at 110°C with various moisture contents (20, 30, and 40%) for 8 hr. The modified products were analyzed by rapid-viscosity analysis (RVA), differential scanning calorimetry (DSC), and X-ray diffraction (XRD). Generally, these hydrothermal treatments altered the pasting and gelling properties of rice starch, resulting in lower viscosity peak heights, lower setbacks, and greater swelling consistency. The modified starch showed increased gelatinization temperatures and narrower gelatinization temperature ranges on ANN or broader ones on HMT. The effects were more pronounced for HMT than for ANN. Also, the typical A-type XRD pattern for rice starch remained unchanged after ANN or HMT at low moisture contents, and the amorphous content increased after HMT at 40% moisture content.     

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.     

Pasting Characteristics of Maize Starch Heat‐Treated with Different Water‐Ethanol Mixtures

Pasting characteristics of maize starch heat‐treated with six different water‐to‐ethanol ratios (%wt base 0:100, 10:90, 20:80, 30:70, 40:60, 50:50) were investigated; treated starches were called EW 0, 10, 20, 30, 40, and 50, respectively. Endotherms in DSC analysis shifted to a higher temperature as the water content in water‐ethanol mixture increased. The removed amount of fatty acids was much higher in treatments for EW 10, 20, and 30. The RVA peak viscosity of EW 10 and 20 were highest among the treated starches and setbacks were more than twice that of untreated starch. The characteristic change in the RVA viscogram corresponded to the amount of leached amylose from the granule. EW 30 displays similar properties as conventional heat‐moisture‐treated starch, but maintained a higher viscosity of ≈300 RVU throughout the heating process. In treatment with water‐ethanol mixtures, heat‐moisture treatment and defatting effects generated new types of modified starches. EW 40 and 50 had no clear pasting peak on RVA, and showed a viscosity at low temperature similar to granular cold water gelling.     

Effect of Dry Heating on Physicochemical Properties of Pregelatinized Rice Starch

Japonica and indica rice starches (10% w/w) were pregelatinized in a boiling water bath for 5 or 10 min and subsequently heat‐treated in a dry state for 0, 1, 2, or 3 h at 130°C to examine the effects of dry heating on pasting viscosity, paste clarity, thermal properties, X‐ray diffraction pattern, and gel strength of pregelatinized starches. Heat treatment obviously changed the physicochemical properties of pregelatinized rice starch. The pregelatinized rice starches had higher peak viscosity and final viscosity than the corresponding native rice starches. Heat treatment of pregelatinized rice starch for 1 h increased the peak viscosity, but treatment for 2 or 3 h decreased the peak viscosity compared with the unheated pregelatinized rice starch. The indica rice starch exhibited more substantial changes in pasting viscosity than did japonica rice starch during heat treatment. The melting enthalpy of the endothermic peak occurred at 90–110°C, and the intensity of the X‐ray diffraction peak at 20° was increased by dry heating, possibly owing to the enhanced amylose‐lipid complexes. The dry heat treatment of pregelatinized starch caused an increase in paste clarity and a decrease in gel strength.     

Paste Properties of Modified Starches from Partial Waxy Wheats

Properties of modified starches from partial waxy wheats have not been examined. Protease digestion of cracked kernels of three hard winter wheats varying in amylose content led to 82–85% recovery of starch, whereas kneading of the flour-water doughs gave 75–83% recovery. All starches had a protein content of <0.3% and ash content of <0.01%. Granule size distributions showed that starch from Ike kernels contained 86% A-type granules with a peak size of ≈18μm, and Karl-92 starch contained 77% A-type granules with a peak size of ≈16μm. The A-type granules (82%) from Rio Blanco starch were intermediate in size. The amylose content of Karl-92 starch, determined by concanavalin-A precipitation of amylpectin, was 28%, which was 17% higher than that of Ike starch (23%). The amylose content of Rio Blanco starch was 26%. The lipid content of Karl-92 starch, determined as fatty acid methyl esters, also was 18% higher than that of Ike starch (601 vs. 488 mg/100 g of starch, respectively). Wheat starches were modified with hydroxypropyl (HP) groups to low (1.5–2.5%) and medium (≈4.0%) levels, and the HP starches were cross-linked with phosphoryl chloride at levels of 0.003–0.075%. Pasting curves (amylograms) showed that Ike starch substituted with a low level of HP and optimally cross-linked with 0.025% phosphoryl chloride (starch basis) had a greater paste consistency than low HP cross-linked Karl-92, and Rio Blanco starches. At 4% HP and optimum cross-linking (0.003% phosphoryl chloride), the paste consistencies of the modified starches were nearly the same. The clarity of unmodified Ike starch paste was higher than that of Karl-92 or Rio Blanco starch pastes, and the clarity of all three pastes decreased as cross-linking was increased. Unmodified Ike starch formed a stronger gel than unmodified Karl-92 and Rio Blanco starches, but gel properties largely converged as the starches were modified.     

Effects of Amylose,Corn Protein,and Corn Fiber Contents on Production of Ethanol from Starch‐Rich Media

The effects of amylose, protein, and fiber contents on ethanol yields were evaluated using artificially formulated media made from commercial corn starches with different contents of amylose, corn protein, and corn fiber, as well as media made from different cereal sources including corn, sorghum, and wheat with different amylose contents. Second‐order response‐surface regression models were used to study the effects and interactions of amylose, protein, and fiber contents on ethanol yield and conversion efficiency. The results showed that the amylose content of starches had a significant (P < 0.001) effect on ethanol conversion efficiency. No significant effect of protein content on ethanol production was observed. Fiber did not show a significant effect on ethanol fermentation either. Conversion efficiencies increased as the amylose content decreased, especially when the amylose content was >35%. The reduced quadratic model fits the conversion efficiency data better than the full quadratic model does. Fermentation tests on mashes made from corn, sorghum, and wheat samples with different amylose contents confirmed the adverse effect of amylose content on fermentation efficiency. High‐temperature cooking with agitation significantly increased the conversion efficiencies on mashes made from high‐amylose (35–70%) ground corn and starches. A cooking temperature of ≥160°C was needed on high‐amylose corn and starches to obtain a conversion efficiency equal to that of normal corn and starch.     

Bread Quality of Spelt Wheat and Its Starch

Flours from five spelt cultivars grown over three years were evaluated as to their breadbaking quality and isolated starch properties. The starch properties included amylose contents, gelatinization temperatures (differential scanning calorimetry), granule size distributions, and pasting properties. Milled flour showed highly variable protein content and was higher than hard winter wheat, with short dough‐mix times indicating weak gluten. High protein cultivars gave good crumb scores, some of which surpassed the HRW baking control. Loaf volume was correlated to protein and all spelt cultivars were at least 9–51% lower than the HRW control. Isolated starch properties revealed an increase in amylose in the spelt starches of 2–21% over the hard red winter wheat (HRW) control. Negative correlations were observed for the large A‐type granules to bread crumb score, amylose level, and final pasting viscosity for cultivars grown in year 1999 and to pasting temperature in 1998 samples. Positive correlations were found for the small B‐ and C‐type granules relative to crumb score, loaf volume, amylose, and RVA final pasting viscosity for cultivars grown in 1999, and to RVA pasting temperature for samples grown in 1998. The environmental impact on spelt properties seemed to have a greater effect than genetic control.     

Physicochemical and Pharmaceutical Properties of Carboxymethyl Rice Starches Modified from Native Starches with Different Amylose Content

Carboxymethyl rice starches (CMRS) were prepared from nine strains of native rice starches with amylose contents of 14.7–29.1%. The reaction was conducted at 50°C for 120 min using monochloroacetic acid as a reagent under alkaline conditions and 1-propanol as a solvent. After determining the degree of substitution (DS), the physicochemical properties including water solubility, pH, and viscosity of 1% (w/v) solution, scanning electron microscopy (SEM), and X-ray diffraction (XRD) analyses of the granules, as well as some pharmaceutical properties of CMRS powders and pastes were investigated. The DS range was 0.25–0.40. All CMRS dissolved in unheated water and formed viscous gel. A good positive correlation was observed between amylose content and DS (r = 0.9278) but not viscosity. SEM and XRD concurrently revealed significant physical alteration of CMRS granules compared with those of native starches, which reflected the changes in the properties of CMRS. At 3% (w/w), CMRS can function as tablet binder in the wet granulation of both water-soluble and water-insoluble diluents. The tablets compressed from these granules showed good hardness with fewer capping problems compared with those prepared using the pregelatinized native rice starch as a binder. In addition, most CMRS pastes formed clear films with varying film characteristics, depending upon the amylose content of the native starches. This type of modified rice starch can potentially be employed as a tablet binder and film-former for pharmaceutical dosage formulations.     

Amylose-lipid complexes as controlled lipid release agents during starch gelatinization and pasting

The effect of amylose-lipid (AM-L) complexes consisting of amylose populations with different peak degrees of polymerization (DP) and complexed with glyceryl monostearate (GMS) or docosanoic acid (C22) on the pasting properties of wheat and rice starches was evaluated with a rapid visco analyzer (RVA). AM-L complexes were formed by both (i) addition of lipids to amylose fractions with peak DP 20, 60, 400, or 950 at 60 degrees C or (ii) potato phosphorylase-catalyzed amylose synthesis in the presence of lipids. All AM-L complexes affected pasting properties in line with their dissociation characteristics. AM-L complexes therefore have potential as "controlled lipid release agents" with effects markedly different from those observable with emulsifier addition in starch pasting. More in particular, short chain AM-L complexes resulted in a starch pasting behavior comparable to that of cross-linked starch, as evidenced by reduced granule swelling, good viscosity stability in conditions of high temperature and shear, and a stable cold paste viscosity.     

优质小麦子粒淀粉组成与糊化特性对氮素水平的响应

在大田条件下,选用3个不同类型优质小麦品种: 豫麦47（强筋品种）、山农8355（中筋品种）和豫麦50（弱筋品种）,设置3个氮肥水平: 施N 0、15和30 g/m2,研究了小麦子粒淀粉的粒度分布、直支链淀粉组成、糊化特性及其对氮素水平的响应。结果表明,优质小麦子粒中淀粉粒的粒径分布范围为1～45 μm,其数目分布呈单峰或双峰曲线变化,体积和表面积分布均呈双峰曲线变化,峰谷位于10 μm处; 据此可将淀粉粒分为两种类型: A型大淀粉粒（10～45 μm）和B型小淀粉粒（1～10 μm）。优质小麦子粒淀粉粒组成存在显著的基因型差异。强筋品种豫麦47子粒中B型淀粉粒的比例较高,弱筋品种豫麦50子粒中A型淀粉粒的比例较高,中筋品种山农8355居中。施氮水平对优质小麦子粒中淀粉的粒度分布存在显著影响。在本试验条件下,随氮素水平的提高,强筋品种豫麦47子粒中A型淀粉粒的比例提高,而B型淀粉粒的比例下降; 增施氮肥后弱筋品种豫麦50和中筋品种山农8355子粒中B型淀粉粒的比例增大,而A型淀粉粒的比例降低,且前者变化的幅度较大。适量增施氮肥提高优质小麦子粒中的淀粉含量,氮肥用量进一步增大后,淀粉含量降低; 增施氮肥后优质小麦子粒中直链淀粉含量降低。增施氮肥对优质小麦子粒淀粉的糊化特性存在较大影响,且此影响的趋势因基因型和施氮量而异。其中强筋品种豫麦47表现为低谷粘度、最终粘度、反弹值、糊化温度和峰值时间提高,而高峰粘度和稀懈值降低; 当氮肥用量增大至30 g/m2时,糊化温度和峰值时间降低,而以粘度为单位的参数均提高。弱筋品种豫麦50表现为增施氮肥后,RVA参数呈下降趋势,与之相对应中筋品种山农8355的呈上升趋势。相关性分析表明,B型淀粉粒的数目、体积和表面积比例与高峰粘度和稀懈值存在显著正相关; 与低谷粘度、最终粘度和反弹值存在显著负相关。子粒中直链淀粉含量、支链淀粉含量和总淀粉含量与高峰粘度和稀懈值呈显著负相关,与低谷粘度、最终粘度、反弹值和峰值时间呈一定程度正相关; 直链淀粉相对含量与RVA特征参数之间的相关趋势与子粒中直链淀粉含量的趋势一致,但均未达显著水平。由此可以认为,氮肥通过调控小麦子粒中淀粉的直、支链组成和粒度分布而影响其糊化特性。     

Increase in Apparent Amylose Content and Change in Starch Pasting Properties at Cool Growth Temperatures in Mutant Wheat

Some mutant wheat lines with low‐amylose content were grown in a field and greenhouse (15 or 20°C) to compare apparent amylose content and starch pasting properties. The apparent amylose content of flour and starch increased and starch pasting parameters as measured by a Rapid Visco Analyser (RVA) changed in the greenhouse (at cool temperatures) during seed maturation. Densitometric analysis of the protein band separated by electrophoresis suggested that the increase in amylose content by cool temperature was related to the amount of Wx‐D1 protein. This data suggests that the Wx‐D1 gene was responsible for these changes. In wheat starch from Tanikei A6099 and Tanikei A6598 at 15°C, the value of final viscosity and total setback was higher than that from the field. In wheat starch from Tanikei A6599‐4 (waxy mutant with stable hot paste viscosity), the peak viscosity temperature was higher and time maintained >80% of the peak was shorter at 15°C than that from the field. Genetic analysis using doubled‐haploid (DH) lines from a combination of Tanikei A6599‐4 and Kanto 118 (low‐amylose line) showed that apparent amylose content increased and the starch pasting curve and properties changed in waxy progenies similar to Tanikei A6599‐4.