Use of cyclic voltammetry, photochemiluminescence, and spectrophotometric methods for the measurement of the antioxidant capacity of buckwheat sprouts

This paper describes the use of cyclic voltammetry, photochemiluminescence (PCL), and spectrophotometric methods (TEAC and FCR reducing capacity) for the measurement of the antioxidant capacity of dark- and light-grown buckwheat sprouts. Moreover, the flavonoid profile of studied material is provided. Methanol extracts (80%) from ungerminated buckwheat grain and 6 and 8 DAS (days after seeding) sprouts were used. The 8 day germination period was sufficient to obtain good quality sprouts with completely removed pericarps. The ungerminated buckwheat grain contained only rutin, whereas in sprouts produced in dark or light, a high level of isoorientin, orientin, vitexin, rutin, and isovitexin was found. The flavonoid content in sprouts produced under light was almost 2 times higher than those of sprouts produced in the dark. The antioxidant capacity of light-grown sprouts was higher than that of dark-grown ones. The results from voltammetric experiments obtained for buckwheat seeds and 6 and 8 DAS sprouts harvested under dark or light conditions highly correlated with those obtained by PCL antioxidant capacity of water-soluble substances (PCL ACW) (r = 0.99), PCL antioxidant capacity of lipid-soluble substances (r = 0.99), TEAC (r = 0.99), and FCR reducing capacity (r = 0.99). The use of cyclic voltammetry, PCL ACW, and TEAC was fully applicable for the evaluation of the antioxidant capacity of buckwheat sprouts.     

Effect of processing on the flavonoid content in buckwheat (Fagopyrum esculentum M?ench) grain.

Six flavonoids have been isolated and identified in buckwheat grain. These are rutin, orientin, vitexin, quercetin, isovitexin, and isoorientin. Rutin and isovitexin are the only flavonoid components of buckwheat seeds while hulls contain all six identified compounds. The total flavonoid concentration in the seeds was 18.8 and in the hulls 74 mg/100 g of dry matter. Dehulling the grain by using different temperature regimes resulted in drastic reductions of the total flavonoid concentration in the grain (by 75% of the control) and smaller but significant (15-20%) reduction in the hulls.     

Trace element water improves the antioxidant activity of buckwheat (Fagopyrum esculentum Moench) sprouts

Buckwheat (Fagopyrum esculentum Moench) was grown in trace element water (TEW) (100, 200, 300, 400, and 500 ppm) and deionized water (DIW) to evaluate whether the beneficial effects of trace elements on the antioxidant activity could be accomplished with the supplement of TEW. At 300 ppm, TEW significantly increased the Cu, Zn, Mn, and Fe contents in buckwheat sprout but not the Se content. However, the levels of rutin, isoorientin, vitexin, and isovitexin did not differ between buckwheat sprouts grown in TEW and DIW. The ethanolic extract from buckwheat sprout grown in 300 ppm of TEW showed higher ferrous ion chelating activity and inhibitory activity toward lipid peroxidation than that grown in DIW. The extract in the TEW group also enhanced intracellular superoxide dismutase activity and lowered reactive oxygen species and superoxide anion in the human Hep G2 cell. It was concluded that TEW could increase the antioxidant activities of buckwheat sprouts.     

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.     

喷洒微酸性电解水对荞麦芽菜生长的影响

针对荞麦(F.tatarium)芽菜生产过程中微生物污染造成的烂种、烂苗和食品安全问题,该文利用微酸性电解水在荞麦芽菜浸种和发芽过程中进行喷洒,考察微酸性电解水对荞麦种子发芽特性、种子表面微生物和芽菜品质的影响。试验结果表明:荞麦种子浸种宜采用有效氯浓度(available chlorine concentration,ACC)为40mg/L、pH值为5.0的微酸性电解水,发芽过程喷洒ACC为50mg/L的微酸性电解水能有效控制荞麦芽菜表面细菌和真菌数量,增加芽菜的苗高以及还原糖、芦丁含量,对芽菜产量和干质量无影响。     

Exudation of allelopathic substances in buckwheat (Fagopyrum esculentum Moench)

Root exudates of the common buckwheat, especially phenolic compounds, were studied. Their contents, both in the soil during the growing season and in agar medium during germination, were determined by HPLC and GC-MS. The allelopathic activity of the soil from a buckwheat stand was evaluated, as well. Palmitic acid, squalene, epicatechin, vitexin, a gallic acid derivative, and a quercetin derivative were the main compounds of the agar medium. In the soil, palmitic acid methyl ester, vanillic acid, rutin, a gallic acid derivative, and a 4-hydroxyacetophenone derivative were identified. The effects of vitexin, squalene, epicatechin, 4-hydroxyacetophenone, and vanillic and gallic acids were tested on eight plant species. Inhibitive effects were observed in the cases of 4-hydroxyacetophenone and vanillic and gallic acids. Comparisons of the identified compounds and inhibitive effects of soil extracts indicated that palmitic acid and the gallic acid derivative probably have an important function in the allelopathic root response of buckwheat.     

Unfermented rooibos tea: quantitative characterization of flavonoids by HPLC-UV and determination of the total antioxidant activity

Unfermented rooibos originates from the leaves and the stems of the indigenous South African plant, Aspalathus linearis, and it has been reported to have a higher content of flavonoids compared to that of fermented rooibos. The HPLC/UV method developed in our laboratory for the analysis of the fermented rooibos was applied to the quantitative characterization of the major flavonoids present in the unfermented rooibos. Main compounds determined were aspalathin (49.92 +/- 0.80 mg/g), isoorientin (3.57 +/- 0.18 mg/g), orientin (2.336 +/- 0. 049 mg/g), and rutin (1.69 +/- 0.14 mg/g), followed in order by isovitexin, vitexin, isoquercitrin and hyperoside, quercetin, luteolin and chrysoeryol. The identity of detected flavonoids was confirmed by comparing their retention times and UV spectra with those of corresponding standards. The total antioxidant activity (TAA) of the tea infusions was measured by the ABTS*+ radical cation decolorization assay. The TAA of unfermented rooibos (0.8 Trolox meq/g) resulted 2-fold higher than that of the fermented rooibos. When compared with different water infusions of Camellia sinensis (green and black tea), this TAA value was about 50% lower.     

Predicting Buckwheat Flavonoids Bioavailability in Different Food Matrices Under In Vitro Simulated Human Digestion

The digestibility and bioaccessibility of major flavonoids (rutin, quercetin, and isoquercitrin) in various buckwheat food matrices were evaluated as a function of the rutin levels using an in vitro simulated digestion model. Food matrices were unprocessed samples (buckwheat flour [BF], flavonoid extract [FE], rutin‐enhanced flavonoid extract [REFE], and pure rutin) and processed samples (cakes with BF, FE, and REFE and a rutin‐spiked cake). FE showed the highest digestibility out of all the unprocessed samples (FE > REFE > BF > rutin), whereas BF exhibited the highest bioaccessibility (BF > FE > REFE = rutin). Moreover, the processed samples improved their flavonoid bioaccessibility upon baking. Thus, unprocessed FE is a good source for highly bioavailable flavonoids; moreover, baking exerts a positive effect on flavonoid digestibility and bioaccessibility. Because flavonoids can be further fermented by microorganisms in the large intestine into various metabolites, determining the digestibility and bioaccessibility of various flavonoids is useful for predicting flavonoid bioavailability in buckwheat and buckwheat‐based food products.     

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.     

Effect of buckwheat extract on the antioxidant activity of lipid in mouse brain and its structural change during in vitro human digestion

This study was conducted to investigate the effects of buckwheat ( Fagopyrum esculentum Moench cv. Yangjul No. 2) extract on the antioxidant activity of lipids in mouse brain and the structural change during in vitro human digestion. Buckwheat was collected from a wild farm and extracted with water. The buckwheat extracts were then passed through an in vitro human digestion model that simulated the composition of the mouth, stomach, and small intestine juice. The results confirmed that the main phenolics of buckwheat extract were rutin, quercitrin, and quercetin. The rutin content increased with digestion of the buckwheat (from 48.82 to 96.34 μg/g) and rutin standard samples (from 92.76 to 556.56 μg/g). Antioxidant activity was more strongly influenced by in vitro human digestion of both buckwheat and rutin standard. After digestion by the small intestine, the antioxidant activity values were dramatically increased (from 5.06 to 87.82%), whereas the antioxidant activity was not influenced by digestion in the stomach for both buckwheat extract and rutin standard. Inhibition of lipid oxidation of buckwheat in mouse brain lipids increased after digestion in the stomach for both buckwheat extract and the rutin standard. The major finding of this study was that in vitro human digestion may be an important modulator of the antioxidant capacity of buckwheat and that this may be because in vitro human digestion increased the antioxidative activity via an increase in antioxidants such as rutin and quercetin.     

Variation in phenolic content and antioxidant activity of fermented rooibos herbal tea infusions: role of production season and quality grade

Data are required to calculate the dietary exposure to rooibos herbal tea flavonoids and phenolic acids. Representative content values for the principal phenolic compounds and total antioxidant capacity of fermented rooibos infusion, taking into account variation caused by production seasons (2009, 2010, and 2011) and quality grades (A, B, C, and D), were determined for samples (n = 114) from different geographical areas and producers. The major phenolic constituents were isoorientin and orientin (>10 mg/L), with quercetin-3-O-robinobioside, phenylpyruvic acid glucoside, and aspalathin present at >5 mg/L. Isovitexin, vitexin, and hyperoside were present at <3 mg/L. Rutin, ferulic acid, and isoquercitrin were present at <2 mg/L. Nothofagin was present at <1 mg/L. Only traces of luteolin-7-O-glucoside and the aglycones quercetin, luteolin, and chrysoeriol were present. Substantial variation was observed in the individual content values of the phenolic compounds and total antioxidant capacity within production seasons and quality grades.     

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.     

茶园施肥模式对茶叶黄酮类及糖苷类代谢物含量的影响

  【目的】  研究施肥模式对茶叶生化成分含量和黄酮类及糖苷类代谢物累积的影响,为实现茶叶优质高产提供科学依据。  【方法】  田间试验设在四川茶树种植典型区域—雅安,试验连续进行了3年,设置不施肥 (CK)、常量化肥 (N 585 kg/hm2,TF)、有机肥替代25%化肥氮 (OF)、减施25%氮肥 (SF) 4个处理。分析了不同处理茶叶主要品质成分含量,并基于非靶向代谢组学检测结果分析了黄酮类及糖苷类代谢物累积差异。  【结果】  与CK处理相比,TF、OF、SF处理茶氨酸、游离氨基酸含量分别提高了7.22%～13.40%、23.15%～25.50%,可溶性糖和茶多酚含量分别降低了8.17%～13.86%、6.08%～11.49%,表儿茶素、表没食子儿茶素、芸香苷等10种代谢物累积水平显著下降。与TF处理相比,SF处理茶氨酸、咖啡碱和水浸出物含量分别降低了5.45%、5.97%、8.91%,茶多酚、可溶性糖、游离氨基酸含量和酚氨比无显著差异;芍药素-3-O-葡萄糖苷、肉桂萜醇C1-19-葡萄糖苷、查耳酮樱花素等7种代谢物累积水平显著降低,川陈皮素、牡荆素-4'-O-α-L-吡喃鼠李糖苷、(S)-橙花醇-3-O-[a-L-鼠李糖吡喃醇-(1->2)-β-D-吡喃葡萄糖苷]和Ranupenin3-rutinoside等4种代谢物累积水平显著增加。与TF处理相比,OF处理茶叶咖啡碱含量降低了5.73%;茶氨酸、游离氨基酸、可溶性糖、茶多酚、水浸出物含量差异不显著;10种黄酮苷、2种黄酮、6种萜苷、5种甾体皂甙和2种脂酰苷共25种代谢物累积水平显著增加,以川陈皮素、飞燕草素-3-(6-对香豆酰基半乳糖苷)增幅较大,分别为2280.20%、1355.11%。4个处理相比,OF处理下有15种黄酮类及糖苷类代谢物累积水平显著高于CK、TF和SF处理。  【结论】  有机肥替代25%化肥氮降低了茶叶咖啡碱含量,提高了川陈皮素、飞燕草素、槲皮素、芹黄素、山奈酚、牡荆素等多种黄酮类代谢物和去氢大豆皂甙I、珠子参苷R2、孕甾烷甙等多种糖苷组分累积水平。减施25%氮肥降低了茶氨酸、咖啡碱、水浸出物含量和糖苷类代谢物 (如杨梅酮-3-O-洋槐糖苷、芍药素-3-O-葡萄糖苷、肉桂萜醇C1-19-葡萄糖苷等)累积水平,对茶多酚、可溶性糖、游离氨基酸含量影响较小。有机肥替代部分化肥氮有利于茶叶黄酮类及糖苷类物质累积,单纯减少氮肥投入会降低茶叶品质。     

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。     

电解水对荞麦芽酚酸含量及抗氧化能力的影响

为了能够更好地将发芽荞麦用作制备功能食品的原料,本研究采用理化指标不同的电解水制备荞麦芽,考察荞麦的发芽率及芽长、总酚含量、DPPH和ABTS自由基清除能力,以及铁离子还原力在发芽期间的动态变化,并分析各处理组发芽第7天荞麦芽中游离酚和结合酚种类和含量。结果表明,在7 d观测期内,电解水有利于荞麦发芽及生长。不同评价方法测定荞麦芽抗氧化能力的结果显示,pH值11.13电解水处理组发芽1~3 d荞麦芽总酚含量及抗氧化水平均显著高于对照组(P<0.05),而pH值3.21、pH值5.02和pH值9.02电解水处理组发芽5~7 d荞麦芽总酚含量及抗氧化水平均显著高于对照组(P<0.05)。各处理组荞麦芽中主要的游离酚均为绿原酸,且电解水处理组荞麦芽游离绿原酸含量均显著高于对照组(P<0.05)。自来水处理组荞麦芽主要结合酚为香豆酸,而电解水处理组荞麦芽主要结合酚为咖啡酸。本研究结果为电解水应用于功能性荞麦芽的制备提供了理论依据。     

Edible neotropical blueberries: antioxidant and compositional fingerprint analysis

Edible blueberry species are well recognized for their potential health benefits. Ericaceae fruits including the North American highbush blueberry (Vaccinium corymbosum L.) and five less common edible blueberry relatives from the New World tropics, Anthopterus wardii Ball, Cavendishia grandifolia Hoerold, Macleania coccoloboides A. C. Smith, Sphyrospermum buxifolium Poeppig & Endlicher, and Sphyrospermum cordifolium Benth, were investigated for their antioxidant properties and phenolic profiles. The neotropical berries C. grandifolia and A. wardii exhibited significantly higher 1,1-diphenyl-2-picrylhydrazyl (DPPH(?)) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonate) (ABTS(??+)) free radical scavenging and iron chelation activities than V. corymbosum. Total phenolic content and high-performance liquid chromatography with photodiode array detection (HPLC-PDA) compositional fingerprint analyses were also carried out. Significant correlations were observed among total phenolic contents, DPPH(?) and ABTS(??+) scavenging, and iron chelation activities. By use of HPLC-PDA, the phenolic constituents in the berries were identified as chlorogenic acid, p-coumaric acid, hyperoside, quercetin-3-O-glucoside, isoorientin, isovitexin, orientin and vitexin. Principal component analysis reduced the dimensions of antioxidant and total phenolic data to two components, which accounted for 95% of total variation among the six fruits. Each fruit species formed its own cluster, and therefore the antioxidant profile of each species was shown to be distinct.     

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.     

Effect of nutrient solution concentration on photosynthesis,growth, and content of the active substances of passionfruit 1

The objective of this study was to investigate the effects of exposure of passionfruit plants (Passiflora edulis Sims. var edulis) to five different electrical conductivity (EC) levels (1.2, 2.3, 4.5, 6.8, and 9.0 mS/cm) of the nutrient solution on net gas exchange (Pn), content of chlorophyll, the biologically active substances, vitexin and orientin, total biomass production, and morphological traits using plants grown in a greenhouse. Important traits, such as plant height, number of internodes, and leaf number per plant were significantly influenced and showed a linear relation to the increased EC levels of the solution. The highest biomass production, chlorophyll concentration, net carbon dioxide (CO₂) assimilation and leaf area were obtained at an EC of 6.8 mS/cm. At an EC level of 9.0 mS/cm, the growth of the plants was depressed and photosynthetic rate sharply declined. The highest photosynthetic rate (8.45 μmol/m²/s) was measured at an EC of 6.8 mS/cm and a photosynthetic photon flux of 1600 μmol/m²/s. The trend of photosynthesis was closely related to those of chlorophyll content, transpiration and mesophyll conductance. There was significant regression relating the shoot yield to increasing EC of the nutrient solution. The leaf chlorophyll content increased from 0.70 mg/g (75 mg/m²) at an EC of 0 to 1.74 mg/g at an EC of 6.8 mS/cm. The leaf water potential sharply declined with the increasing EC levels. The specific leaf area was high at EC of 2.3,4.5, and 6.8 mS/cm. The mean ratio of the root to the shoots decreased as EC was increased up to 6.8 mS/cm. Orientin content was the highest (168 mg %) at an EC of 9.0 mS/cm, having a linear effect, while the lowest vitexin content (36 mg %) was achieved at an EC of 6.8 mS/cm. The results suggest a positive role of increasing nutrient solution EC for orientin accumulation than to vitexin.     

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.