Application of near-infrared reflectance spectroscopy to the evaluation of rutin and D-chiro-Inositol contents in tartary buckwheat

Tartary buckwheat [Fagopyrum tataricum (L.) Gaench] is rich in rutin and D- chiro-inositol (DCI), which have beneficial effects in the treatment of hemorrhagic diseases and insulin-resistant diseases, respectively. The current methods of extraction and detection of rutin and DCI are complex and time-consuming; a simple way of analyzing these compounds in the native matrix would be desirable. In this work, near-infrared reflectance spectroscopy (NIRS) was applied to determine the contents of rutin and DCI in tartary buckwheat. The spectral data were compared with those determined from high-performance liquid chromatography (HPLC) methods. Models for predicting rutin and DCI contents in buckwheat were developed using a partial least-squares algorithm. Cross-validation procedures indicated good correlations between HPLC data and NIRS predictions (R2 = 0.76 for rutin and R2 = 0.86 for DCI). The rutin content ranged from 0.998 to 1.75%, while the DCI content covered 0.179-0.200%. The results showed that NIRS, a well-established and widely applied technique, could be applied to determine rutin and DCI in tartary buckwheat rapidly and nondestructively.     

D-chiro-inositol-enriched tartary buckwheat bran extract lowers the blood glucose level in KK-Ay mice

D-chiro-inositol (DCI) is an active compound in tartary buckwheat [Fagopyrum tataricum (L.) Gaench] with an insulin-like bioactivity. The present study was performed to (i) prepare DCI-enriched tartary buckwheat bran extract (TBBE), (ii) evaluate its acute toxicity in mice, and (iii) examine its blood glucose lowering activity in diabetic mice. It was found that steaming buckwheat bran in an autoclave at 1.6 MPa and 120 degrees C for 60 min could significantly enrich the DCI level in TBBE from 0.03 to 0.22% and further to 22% after passage of the TBBE through activated carbon and ion exchange resins. An acute toxicity test demonstrated that the LD 50 of TBBE was >20 g/kg of body weight in mice, suggesting that TBBE was in general nontoxic and safe in mice. Male KK-A(y) mice (type 2 diabetic) and C57BL/6 mice (the control) were used to investigate the antidiabetic activity of TBBE. In KK-A(y) mice, the blood glucose, plasma C-peptide, glucagon, total cholesterol, triglyceride, and blood urea nitrogen (BUN) levels were significantly higher than those in the C57BL/6 mice. In addition, KK-A(y) mice showed an obvious decrease in insulin immunoreactivity in the pancreas. The present study clearly demonstrated that oral administration of DCI-enriched TBBE could lower plasma glucose, C-peptide, glucagon, triglyceride, and BUN, improve glucose tolerance, and enhance insulin immunoreactivity in KK-A(y) mice.     

黑苦荞米黄酮提取工艺优化及其降血糖活性研究

为了获得合适的黑苦荞米黄酮提取工艺,该文采用单因素试验和响应面设计,选择乙醇体积分数、料液比、提取时间、提取温度4个因素,优化黑苦荞米黄酮提取工艺。试验结果表明,黑苦荞米黄酮的最佳提取条件是:乙醇体积分数54%,料液比1:24 g/m L,提取时间62 min,提取温度71℃。在此条件下,理论黄酮得率为2.21%,实际黄酮得率为2.20%,相对误差为0.45%。在此基础上进一步研究了黑苦荞米黄酮的α-淀粉酶抑制活性,结果表明7.5 mg/m L的黑苦荞米黄酮对α-淀粉酶活性的抑制率为54.05%,与二甲双胍(5 mg/m L)效果相当;此外,与空白对照相比,50μg/m L黑苦荞米黄酮能显著(P0.05)提高肝脏细胞Hep G2的葡萄糖消耗量(48.73%),并促进肝脏细胞糖原的合成。研究结果表明,黑苦荞米黄酮具有较好的辅助降血糖功效。     

Pharmacokinetic profile of total quercetin after single oral dose of tartary buckwheat extracts in rats

As an important edible and medicinal material, tartary buckwheat ( Fagopyrum tataricum Gaertn.) is commonly used as a kind of food or drug in eastern Asian countries. To investigate the pharmacokinetic profile of total quercetin after a single oral dose of tartary buckwheat extract in rats, a sensitive, simple, and accurate HPLC-UV method was developed to determine total quercertin following plasma enzyme hydrolysis with glucuronidase/sulfatase. Eighteen male Wistar rats were randomly assigned into three groups, which were given three different doses of tartary buckwheat extract. The pharmacokinetic results showed that the area under the plasma concentration-time curve (AUC) of total quercetin after single oral doses of tartary buckwheat extract presented a linear relationship. The peak plasma concentrations (C(max)) of total quercetin afte plasma enzyme hydrolysis with glucuronidase/sulfatase were 0.55 ± 0.26, 1.10 ± 0.53, and 2.05 ± 0.26 μg/mL; the peak times (T(max)) were 2.33 ± 1.94, 2.75 ± 3.67, and 2.50 ± 1.82 h; the areas under the curves (AUC(0-36h)) were 5.29 ± 1.35, 10.02 ± 4.43, and 22.51 ± 3.05 μg·mL(-1)·h(-1) for three doses of tartary buckwheat extract (60, 120, and 240 mg/kg). The present study has provided a basic pharmacokinetic profile of total quercetin after a single oral dose of tartary buckwheat extract in rats.     

Physicochemical Properties of Common and Tartary Buckwheat Starch

Physicochemical properties of starch of three common (Fagopyrum esculentum) and three tartary (F. tataricum) buckwheat varieties from Shanxi Province, China, were compared. Starch color, especially b*, differed greatly between tartary (7.99–9.57) and common (1.97–2.42) buckwheat, indicating that removal of yellow pigments from tartary buckwheat flour may be problematic during starch isolation. Starch swelling volume in water of reference wheat starch (2.8% solids and 92.5°C) was 20.1 mL; for the three common buckwheat starches it was 27.4–28.0 mL; and for the three tartary buckwheat starches it was 26.5–30.8 mL. Peak gelatinization temperature (T_p) in water was 63.7°C for wheat starch, 66.3–68.8°C for common buckwheat and 68.8–70.8°C for tartary buckwheat. T_p of all samples was similarly delayed (by 4.0–4.8°C) by 1% NaCl. Enthalpy of gelatinization (ΔH) was higher for all six buckwheat starches than it was for wheat starch. However, one common buckwheat sample had significantly lower ΔH than the others. Starch pasting profiles, measured by a Rapid Visco-Analyzer, were characteristic and similar for all six buckwheat starches, and very different from the reference wheat starch. A comparison of pasting characteristics of common and tartary buckwheat starches to wheat starch indicated similar peak viscosity, higher hot paste viscosity, higher cool paste viscosity, smaller effect of NaCl on peak viscosity, and higher resistance to shear thinning. Texture profile analysis of starch gels showed significantly greater hardness for all buckwheat samples when compared to wheat starch.     

Tartary buckwheat (Fagopyrum tataricum Gaertn.) as a source of dietary rutin and quercitrin

Two samples of tartary buckwheat (Fagopyrum tataricum Gaertn.) from China and one from Luxembourg were studied by high-performance liquid chromatography (HPLC) to reveal the possibilities of growing tartary buckwheat herb as a possible source of rutin, quercetin, and quercitrin. The content of rutin was determined as up to 3% dry weight (DW) in tartary buckwheat herb. Quercitrin values were in the range of 0.01-0.05% DW. Only traces of quercetin were detected in just some of the samples. Tartary buckwheat seeds contained more rutin (about 0.8-1.7% DW) than common buckwheat seeds (0.01% DW). Rutin and quercetin content in seeds depends on variety and growing conditions. Tartary buckwheat seeds contained traces of quercitrin and quercetin, which were not found in common buckwheat seeds.     

硒对苦荞硒、总黄酮和芦丁含量、分布与累积的影响

采用盆栽试验探讨土壤施硒对苦荞硒、总黄酮和芦丁含量、分布与累积的影响。结果看出,在土壤施硒0.5～2.0 mg/kg范围,苦荞根在苗期（40 d）大量吸收并累积硒,全生育期各器官硒含量极显著提高;在生长中后期(60～80 d) 硒的累积最快,累积量最多。硒在苦荞各器官中的分布为:在40 d时,根＞叶＞茎;60 d时Se0.0处理为叶＞根＞花＞茎,施硒各处理则为花＞根＞叶＞茎;80 d时Se0.0处理以花＞根＞叶＞茎＞子粒,Se0.5处理以花＞叶＞子粒＞茎＞根,施硒≥1.0 mg/kg的处理则为花＞叶＞茎＞根＞子粒。土壤施硒≤1.0 mg/kg促进苦荞生长,提高地上部各器官干重和植株总干重以及各器官总黄酮和芦丁含量与累积量,不改变总黄酮和芦丁的器官分布,增加苦荞中后期对总黄酮的累积;以Se0.5处理效应最佳,各差异达显著水平。过量的硒（Se1.5～2.0 mg/kg）显著抑制苦荞生长,降低各器官干重、总黄酮和芦丁含量与累积,不利于硒在子粒中富集和总黄酮在子粒中分布。表明在低硒土壤上栽培苦荞,土壤施硒以不超过1.0 mg/kg为宜,既能最大限度的提高苦荞各器官硒、总黄酮和芦丁含量和累积量,又可降低施用硒肥的成本和减少硒肥对环境的影响。     

大孔树脂精制苦荞总黄酮工艺条件的优化研究

为了研究DM-2型大孔树脂精制苦荞总黄酮的优化工艺条件，以树脂对苦荞总黄酮的吸附率、解吸率为评价指标，采用响应曲面法(response surface methodology，RSM)建立树脂法精制苦荞总黄酮的二次多项回归模型方程，对精制苦荞总黄酮吸附解吸工艺条件进行优化。结果表明吸附最佳工艺条件为吸附速率2 mL/min，pH值2，上样液浓度2 mg/mL。解吸最佳工艺条件：乙醇浓度71.05%，pH值8.76，解吸速率1 mL/min。该方法简单可行，精制效果好，适合于工业化生产。     

Antioxidant activity of tartary (Fagopyrum tataricum (L.) Gaertn.) and common (Fagopyrum esculentum moench) buckwheat sprouts

This study compared the differences of two types of buckwheat sprouts, namely, common buckwheat ( Fagopyrum esculentum Moench) and tartary buckwheat ( Fagopyrum tataricum (L.) Gaertn.), in general composition, functional components, and antioxidant capacity. The ethanol extracts of tartary buckwheat sprouts (TBS) had higher reducing power, free radical scavenging activity, and superoxide anion scavenging activity than those of common buckwheat sprouts (CBS). As for chelating effects on ferrous ions, CBS had higher values than TBS. Rutin was the major flavonoid found in these two types of buckwheat sprouts, and TBS was 5 fold higher in rutin than CBS. The antioxidant effects of buckwheat sprouts on human hepatoma HepG2 cells revealed that both of TBS and CBS could decrease the production of intracellular peroxide and remove the intracellular superoxide anions in HepG2 cells, but TBS reduced the cellular oxidative stress more effectively than CBS, possibly because of its higher rutin (and quercetin) content.     

Social and environmental influences on tartary buckwheat (<Emphasis Type="Italic">Fagopyrum tataricum</Emphasis> Gaertn.) varietal diversity in Yunnan,China

Effective conservation strategies aimed to protect crop genetic resources require multiple sources of information. We used a combination of AFLP genotyping and farmer surveys to understand the extent, distribution and management of tartary buckwheat (Fagopyrum tataricum Gaertn.) diversity in its center of origin in Yunnan Province, China. We found genetic evidence of gene flow in tartary buckwheat throughout the study area, with small but statistically significant regional and village-level components. We also found genetic differentiation by seed color. Although most farmers reported exchanging seed in localized kinship networks, our results imply homogenizing gene flow is occurring. Yi ethnic farmers tend to plant more buckwheat than non-Yi farmers, and we found that in some communities, Yi farmers serve as seed sources for farmers of other ethnicities. Different tartary buckwheat varieties did not have different end uses; rather farmers maintained varietal diversity in order to protect crop yield and quality. Individual farmers’ seed exchange practices reflect their ideas about components of seed quality, as well as priorities in protecting buckwheat yield. From the standpoint of genetic resources conservation, the presence of a culturally rich farmer exchange network and hierarchical structuring of tartary buckwheat genetic diversity demonstrates the importance of maintaining an interlinked community of tartary buckwheat farmers in Yunnan.     

超高压处理改善苦荞淀粉理化性质及益生菌群落

为研究超高压(ultra-high pressure,HHP)处理对苦荞淀粉理化性质及功能性的影响,该文以6个不同压力处理后的苦荞淀粉为材料,利用扫描电镜、X射线衍射仪等分析了其淀粉颗粒大小、晶体结构、溶解度、透光率、冻融稳定性等特性,并通过建立体外模型模拟人体消化以及肠道发酵过程,利用气相色谱质谱联用技术和平板计数法,对小鼠粪便发酵液短链脂肪酸(short chain fatty acid、SCFAs)含量及主要菌群进行了测定。试验结果表明:超高压处理后苦荞淀粉颗粒出现凹陷与粘连状态,并逐渐失去原有形态,但其结晶类型仍为典型的A型,其结晶度在200 MPa时达到最大值41.8%;溶解度和膨胀度均随着压力的增加呈先减小后增大的趋势,并且其透光率会下降,其中在200 MPa时,溶解度和膨胀度都达到了最低0.83%和171%,而冻融稳定性会有一定的改善。此外,超高压处理后苦荞淀粉使肠道中双歧杆菌、乳酸杆菌的数量显著增加(P0.05),而大肠杆菌、肠球菌的生长受到显著抑制(P0.05)。同时,肠道内的pH值显著下降(P0.05),SCFAs中乙酸、丙酸、丁酸显著增加(P0.05),综上所述,超高压处理后苦荞可作为一种良好的天然改善肠道菌群的食物来源。     

Fagopyritols, D-chiro-inositol, and other soluble carbohydrates in buckwheat seed milling fractions

Fagopyritols are mono-, di-, and trigalactosyl derivatives of D-chiro-inositol that accumulate in seeds of common buckwheat (Fagopyrum esculentum Moench) and may be important for seed maturation and as a dietary supplement. Fagopyritols and other soluble carbohydrates were assayed in mature groats and 11 milling fractions of common buckwheat seed. Because fagopyritols are in embryo and aleurone tissues, differences in fagopyritol concentrations reflect varying proportions of these tissues in each milling fraction. Bran milling fractions contained 6.4 g of total soluble carbohydrates per 100 g of dry weight, 55% of which was sucrose and 40% fagopyritols. Flour milling fractions had reduced fagopyritol concentration [0.7 g/100 g of dry weight total fagopyritols in the dark (Supreme) flour and 0.3 g/100 g in the light (Fancy) flours]. Fagopyritol B1 was 70% of total fagopyritols in all milling fractions. Fagopyritols were 40% of total soluble carbohydrates in groats of two cultivars of common buckwheat but 21% in groats of tartary buckwheat [Fagopyrum tataricum (L.) Gaertn.], probably a reflection of environment and genetics. A rhamnoglucoside present in tartary buckwheat was not detected in common buckwheat.     

超微粉碎对苦荞麸功能特性的影响

为研究超微粉碎处理对苦养麸中功能成分的溶出、清除有毒有害物及抗氧化功能特性的影响。将苦养麸皮原粉进行超微粉碎处理得到苦养麸微粉,比较不同粒度的苦养微粉与麸皮原粉中总黄酮提取率、在体外对胆酸钠、Pb^2＋、Cd^2＋、Hg^2＋的吸附力以及对NO2^-、2,2-联苯-1-三砂基苯肼（DPPH）的清除力。结果表明,与苦养麸皮原粉相比,苦养麸微粉（平均粒径为20．621μm）的总黄酮提取率增加0．36个百分点;对胆酸钠的吸附量降低5．60mg／g;对Pb^2＋、Cd^2＋、Hg^2＋的吸附量增加0．184、0．840、1．341mg／g;对NO2的清除率增加24．25个百分点;对DPPH的清除率增加53．56个百分点。超微粉碎处理适合苦养麸皮的深加工;得到苦养麸微粉具有优良的营养、功能特性,是一种极具开发潜力的功能食品原料。     

Analyses of the seed protein contents on the cultivated and wild buckwheat <Emphasis Type="Italic">Fagopyrum esculentum</Emphasis> resources

The seed protein contents of 179 accessions belonging to ten species of genus Fagopyrum were determined by means of the Kjeldahl method. The results indicated that there are significant differences of seed protein content among different accessions within the same species. The average protein content of F. esculentum is 12.94%, ranging from 8.81–18.71%, and the protein content of Sibano is the highest, up to 18.71%, Gantian 1 the lowest with 8.81%. The average protein content of tartary buckwheat is 12.17%, with the range from 7.82–18.94%, and the protein content of cultivated tartary buckwheat Qianku 2 is the highest (18.59%), and that of Ganku 1 (7.82%) the lowest. Moreover, there are significant differences of seed protein content among different buckwheat species and between the different buckwheat types (between common buckwheat and tartary buckwheat, between the diploid and the tetraploid, between the cultivated and the wild, and between the big-achene group and the small-achene group). The average seed protein content of F. giganteum is the highest in all tested species, up to 17.81% with the range of 13.91–21.27%, and that of F. callianthum the lowest, only 8.31%. This study was financed by the Natural Science Foundation of China (30270852, 30471116), Program for New Century Excellent Talents in University (NCET-2004-0913), Guizhou Key International Cooperation Project (Qianke Hewai G Zi 2005#400108), Guizhou Oversea Talent Project (Qianren Xiangmu Zizhu Hetong 2004#02), and Key Project of Students in Guizhou Normal University (GNU-SP-2005-SJXY-A01)     

水分胁迫下新造地施磷深度对苦荞生长及根系分布的影响

  【目的】  根系构型影响作物的抗旱能力,研究磷肥施用深度调节苦荞根系分布的可行性,为贫瘠干旱地区苦荞的生长提供科学养分管理措施。  【方法】  以‘黑丰1号’苦荞 (Fagopyrum tataricum L.) 为试验材料,进行根管土柱 (直径25 cm、高50 cm) 栽培试验,设置田间持水量65%～75% (W1)、45%～55% (W2) 和35%～45% (W3) 3种土壤水分条件,磷肥施用深度分别设置距离地表10 cm (P10)、20 cm (P20)、30 cm (P30) 以及3层均匀施用 (P-all) 4种方式,共有12个处理。在苦荞幼苗三叶一心期进行处理,生长22天后取样,测定根系构型,并记录生物量。  【结果】  干旱胁迫抑制了苦荞植株生长、干物质量的积累以及根系发育,其中W3水分条件抑制作用最为明显,导致苦荞株高、茎粗和叶面积较W1水分条件分别下降17.20%、18.03%和23.17%;根长、根表面积和根体积分别下降16.97%、20.39%和17.39%;地上部干物质量和根系干物质量分别下降39.16%、28.60%。干旱胁迫促进根系下扎,增加深层土壤中的根系分布。与W1水分条件相比,W2、W3水分条件下0—10和10—20 cm土层平均根长分别下降30.18%和27.55%、41.83%和41.02%,根系干物质量分别下降36.62%和33.61%、49.72%和48.11%;而20—30和30—45 cm土层中的苦荞平均根长则分别增加33.53%和42.52%、31.74%和50.95%,根系干物质量分别增加13.70%和26.84%、5.85%和28.64%。深层施磷促进施磷层土壤根系生长,与P-all处理相比,P10处理10—20 cm土层根长平均增加18.96%,P20处理20—30 cm土层平均增加32.39%,P30处理30—45 cm土层平均增加28.73%,根系干物质量依次分别增加26.62%、30.74%和24.65%。方差分析结果表明,各水分处理条件下,0—10和10—20 cm土层根系干物质量均表现为P10处理显著高于其他施磷处理,且其他处理间差异也达显著水平;而20—30、30—45 cm土层根系干物质量则表现为P20、P30施磷处理显著高于其他处理。  【结论】  水分和施磷深度对苦荞苗期植株生长以及根系分布均有显著影响。在干旱胁迫下,增加磷肥的施用深度可促进苦荞根系在20—45 cm深土壤中的分布,显著扩大根系对土壤养分和水分的获取空间,并最终促进苦荞的生长。本试验条件下,采样仅限于苦荞苗期,在水分胁迫条件下磷肥以10 cm的施肥深度效果最佳。     

Molecular cloning, recombinant expression, and immunological characterization of a novel allergen from tartary buckwheat

Buckwheat is generally regarded as a nutritionally rich food source. However, earlier studies prove that it also causes allergies to subjects. Allergenic proteins with a strong IgE-binding activity have been identified in common buckwheat (CB) and a 24 kDa allergen (rTBa) in tartary buckwheat (TB). The objective of this research was to clone and express a novel allergen in tartary buckwheat and to evaluate its structure and immunological activity. The 1773 bp full-length cDNA was amplified and cloned from the total RNA of TB by polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE) methods. Its nucleotide sequence had high similarity with legume-like 13S storage protein mRNA in CB. The deduced amino acid sequence included a putative signal peptide and 18 fragments as its epitope sites. The predicted full-length TB allergen sequence was found to have two domains, and the recombinant protein reacted with sera from patients with positive IgE binding to buckwheat and had a lower binding ability than the recombinant TBa and recombinant TBb (C- and N-terminal amino acid sequence of TBt codes for protein). This fact suggests that full-length TB allergen may hydrolyze to two domains in vivo, decreasing the IgE-binding ability.     

Identification of anthocyanins in the sprouts of buckwheat

The anthocyanin profiles and varieties/breeding line differences of anthocyanin concentrations in common/tartary buckwheat sprouts have been studied. Four anthocyanins, cyanidin 3-O-glucoside, cyanidin 3-O-rutinoside, cyanidin 3-O-galactoside, and cyanidin 3-O-galactopyranosyl-rhamnoside, were isolated from the sprouts of common buckwheat, were separated using high-performance liquid chromatography (HPLC), and were identified using reversed-phase liquid chromatography (LC)/electrospray ionization-mass spectrometry (ESI-MS)/MS techniques. In tartary buckwheat sprouts, two anthocyanins (cyanidin 3-O-glucoside and cyanidin 3-O-rutinoside) were identified. Among 19 common/tartary buckwheat varieties/breeding lines, Hokkai T10 contained the highest amounts of anthocyanins. Cyanidin 3-O-glucoside and cyanidin 3-O-rutinoside concentrations in 6-10 days after seeding sprouts of Hokkai T10 ranged from 0.16 to 0.20 mg/g dry wt and from 5.55 to 6.57 mg/g dry wt, respectively. In addition, dark-grown sprouts of Hokkai T10 accumulated 0.091 and 2.77 mg/g dry wt of cyanidin 3-O-glucoside and cyanidin 3-O-rutinoside whereas other varieties/breeding lines accumulated trace amounts of anthocyanins. Given its anthocyanin-rich red cotyledons, Hokkai T10 is a promising line for use as "Moyashi" type sprouts and is strongly recommended as a new functional food, rich in dietary anthocyanins.     

Determination of 1-deoxynojirimycin in mulberry leaves using hydrophilic interaction chromatography with evaporative light scattering detection

A simple and rapid method for determining 1-deoxynojirimycin (DNJ), a potent glucosidase inihibitor present in mulberry leaves (Morus alba and Morus bombysis), by high-performance liquid chromatography coupled to an evaporative light scattering detector (ELSD) has been developed. DNJ was separated from an extract of mulberry leaves on a TSKgel Amide-80 column, which is a representative column for hydrophilic interaction chromatography. During postcolumn detection, DNJ was detected by ELSD and concurrently identified by mass spectrometry. The detection limit was 100 ng. This method is sufficiently sensitive for determining DNJ in mulberry leaves and other related products.     

Differential expression of anthocyanin biosynthetic genes and anthocyanin accumulation in tartary buckwheat cultivars 'Hokkai t8' and 'Hokkai t10'

Six genes involved in anthocyanin biosynthesis in tartary buckwheat have been cloned, namely, FtC4H, Ft4CL, FtCHI, FtF3H, FtF3'H, and FtANS, which encode cinnamate 4-hydroxylase (C4H), 4-coumarate:CoA ligase (4CL), chalcone isomerase (CHI), flavones 3-hydroxylase (F3H), flavonoid 3'-hydroxylase (F3'H), and anthocyanidin synthase (ANS), respectively. Then, these cDNAs were used, along with previously isolated clones for phenylalanine ammonia-lyase (PAL) and chalcone synthase (CHS), to compare gene expression in different organs, flowering stages, and maturing seeds of tartary buckwheat cultivars 'Hokkai T8' and 'Hokkai T10'. Quantitative real-time polymerase chain reaction analysis showed that these anthocyanin biosynthetic genes were most highly expressed in the stems and roots of Hokkai T10. The FtANS gene was more highly expressed than other genes during flowering and maturing seeds. In addition, the anthocyanin concentration was higher in 'Hokkai T10' than in 'Hokkai T8'; however, naringenin chalcone, a flavonoid, was absent from 'Hokkai T10' seedlings based on fluorescence microscopy.     

Determination of rutin, catechin, epicatechin, and epicatechin gallate in buckwheat Fagopyrum esculentum Moench by micro-high-performance liquid chromatography with electrochemical detection

A simple and sensitive method has been developed for determining rutin, catechin, epicatechin, and epicatechin gallate in buckwheat (Fagopyrum esculentum Moench) flour and seeds by micro-high-performance liquid chromatography with electrochemical detection. Chromatography was performed using an octadecylsilica column, acetonitrile-water-formic acid (13:87:1, v/v/v) as a mobile phase, and an applied potential at +0.5 V vs Ag/AgCl. We found that Japanese buckwheat flour contains rutin (12.7 mg/100 g), catechin (3.30 mg/100 g), epicatechin (20.5 mg/100 g), and epicatechin gallate (1.27 mg/100 g). The relative standard deviations for rutin, catechin, epicatechin, and epicatechin gallate peak heights were less than 0.86% (n = 5). The detection limit of rutin was 0.86 ng/mL. Moreover, the present method was applied to the distribution analysis of these compounds in buckwheat seed. The embryo proper and cotyledons of a mature buckwheat seed contained rutin with the highest concentration as compared to other parts. This method is useful in determining rutin, catechin, epicatechin, and epicatechin gallate in buckwheat with a small amount of sample for quality control in the food industry.