紫薯中四种花色苷抗氧化活性的密度泛函理论研究

本研究旨在分析紫薯中四种不同花色苷的抗氧化机理。利用DPPH试验和脂肪过氧化抑制能力试验分析四种花色苷的体外抗氧化活性,进而通过密度泛函理论对花色苷及其简化分子模型的优化结构、键离能、电离势等进行分析,探讨抗氧化机理。结果显示,四种花色苷的OH-4'酚羟基的活性最高,键离能小于白藜芦醇的键离能,抗氧化活性的理论计算结果与体外抗氧化试验结果一致。研究结果表明,紫薯花色苷具有较好的抗氧化活性,并且DFT方法为花色苷抗氧化活性的开发提供了有力的理论基础。     

黑豆种皮花色苷酶法辅助提取工艺优化及其抗氧化活性分析

优化黑豆种皮花色苷复合酶法辅助提取工艺,并对其抗氧化活性进行评价。通过单因素试验和响应面法优化确定了黑豆种皮花色苷生物酶法辅助提取的最佳工艺为:复合酶(纤维素酶400 U/g,α-淀粉酶50 U/g),酶解温度50℃,液料比26∶1 mL/g,乙醇体积分数64%,酶解时间为59 min。在此条件下,提取花色苷的含量为2.019 mg/g。抗氧化试验研究表明,黑豆种皮花色苷的还原能力、对超氧阴离子自由基清除能力低于抗坏血酸,但对亚硝酸根离子和DPPH自由基、ABTS自由基的清除能力强于抗坏血酸。因此,生物酶法辅助提取是一种高效的黑豆种皮花色苷提取方法,且作为一种新型花色苷资源,黑豆种皮花色苷有着挖掘和应用价值。     

黑小豆种皮花色苷的提取及体外抗氧化活性研究

为了探索新的花色苷资源,以黑小豆种皮为原料,对其花色苷类色素的提取工艺进行了研究。通过单因素和L_9(3~4)正交试验,考察了乙醇浓度、料液比、温度和p H对粗提液中花色苷含量的影响。结果表明,最佳提取条件为:乙醇浓度60%、料液比1∶20(g∶m L)、温度50℃、p H=2.0。此条件下,黑小豆种皮粗提液中花色苷的含量最大(5.912 mg/g);黑小豆种皮花色苷粗提物得率为19.1%,纯度为3.06%;粗提物具有一定的总抗氧化能力和清除O~(-·)_2、·OH和DPPH自由基的能力。黑小豆种皮可作为一种新型花色苷资源加以利用。     

唐古特白刺果实花色苷体外清除自由基的活性

为探讨唐古特白刺(Nitraria tangutorun Bobr.)果实花色苷体外清除自由基活性,研究了其对超氧阴离子(O-2·)体系、羟基自由基(·OH)体系和二苯代苦味酰基自由基(DPPH·)体系的清除效果,并与抗坏血酸(vc)进行了比较.结果表明,该花色苷对O-2·、·OH、DPPH·均具有清除作用,且与浓度呈量效关系.对·OH和DPPH·清除效果相对较好,优于Vc.     

灯盏花乙素及其苷元结构与抗氧化活性关系研究

采用密度泛函理论B3LYP方法对灯盏花乙素及其苷元分子进行几何结构全优化,获得两个化合物的最低能量结构。从分子的几何结构、NBO电荷、酚羟基解离能BDE、绝热电离势IP和前线分子轨道等方面分析了两个化合物的结构与抗氧化活性之间的关系。计算结果表明,C6位的酚羟基为灯盏花乙素及其苷元的最大反应活性位点,灯盏花乙素的抗氧化活性弱于其苷元的抗氧化活性,计算结果与实验结果吻合。天然的糖苷类物质水解为苷元后,其抗氧化活性增强。此外,还考虑了溶剂效应,考察溶剂极性对抗氧化活性的影响。在所有环境中,抽氢反应机制是化合物自由基清除反应的最主要机制。     

超高压处理对蓝莓酚类物质生物可获得率及抗氧化性的影响

目的：探讨不同条件超高压处理对蓝莓酚类物质的生物可获得率以及抗氧化性的影响。方法：在不同压力下对蓝莓果实进行不同时间的超高压处理。分析测定不同处理条件下蓝莓中酚类物质的含量。通过测定1,1 二苯基 2 三硝基苯肼（DPPH）和2,2 联氮 双 3 乙基苯并噻唑啉 6 磺酸（ABTS）自由基清除能力评价超高压处理后蓝莓的抗氧化能力变化。利用超高效液相色谱 电喷雾串联四级杆质谱分析鉴定蓝莓中主要酚类物质及其含量变化情况。结果：不同压力与处理时间下,超高压处理均能提高蓝莓总酚、总黄酮和总花色苷的生物可获得率,其中在500MPa下处理5min效果最佳。DPPH和ABTS自由基清除能力实验结果表明,超高压处理可以显著提高蓝莓抗氧化性,在500 MPa处理5 min的蓝莓抗氧化能力最强。超高压前后共鉴定出9种酚类物质,其中7种为花色苷及其衍生物、2种类黄酮物质,这些物质中除了矮牵牛素外,其余物质在超高压处理后生物可获得率均显著增加。结论：超高压处理可以显著提高蓝莓中总酚、总黄酮及花色苷等酚类物质的生物可获得率,增强其清除自由基的能力,提高抗氧化性。     

紫山药色素的提取工艺及抗氧化性能研究

研究紫山药色素的最佳提取工艺及其抗氧化性能。在单因素试验的基础上,采用L9(34)正交试验法,以pH示差法测定花色苷得率为考察指标,优化了溶剂提取法提取紫山药色素的工艺参数。通过DPPH体系测定该色素清除自由基能力。试验结果表明:紫山药色素属于花色苷类物质,优化的紫山药色素提取条件为:提取温度60℃,提取时间80 min,料液比1∶30,提取溶剂为0.5%盐酸乙醇溶液,提取液花色苷含量可达2.075mg/鲜紫山药g。紫山药色素提取液清除DPPH自由基的IC50为98.14μg/mL。紫山药具有开发功能性色素的潜力。     

黑莓果渣中花色苷的提取工艺研究

通过提取方法优选,表明超声法对黑莓果渣中花色苷的提取效果最佳。以60%乙醇为超声法提取溶剂,通过单因素实验分别考察了料液比、超声时间、温度、功率、提取次数对提取黑莓果渣中花色苷的影响。综合单因素实验结果,通过响应面法筛选出最佳的工艺条件为:液料比14∶1(mL∶g),时间40 min,温度58℃,功率300 W,提取2次。通过提取物中花色苷含量与其清除DPPH.活性的相关性分析发现,黑莓果渣提取物清除DPPH.活性与其花色苷含量之间存在相关性,由此推断花色苷为黑莓果渣中主要的自由基清除物质。     

对九种天然产物清除自由基活性的理论评价

用量子化学方法计算了源自灯盏花、丹参、银杏和红花的九种天然产物--灯盏花乙素、焦袂康酸、3,5-二咖啡酰氧基奎宁酸、飞蓬酯B、丹酚酸B、丹参素、银杏双黄酮、银杏苦内酯B和羟基红花黄色素A的O-H键解离焓和电离势,并以此为理论指标评价了其清除自由基活性.结果发现,在非极性溶剂中3,5-二咖啡酰氧基奎宁酸、飞蓬酯B、丹酚酸B、丹参素等均具有较高的活性;而在极性溶剂中,相比其它八种化合物,灯盏花乙素显示出优良的清除自由基活性,提示这些化合物有望作为药物的有效成分治疗自由基引起的心、脑血管疾病.     

香樟果皮花色苷提取工艺优化及抗氧化研究

为了优化香樟果皮花色苷的最佳提取工艺条件,用乙醇作为提取溶剂,选取提取时间、提取温度、提取剂浓度、料液比、pH五因素,采用Box-Behnken建立三因素三水平模型,用响应面法优化各因素及其相互作用的最佳组合,用Design-Expert 8.0设计回归正交实验;同时用水杨酸比色法和DPPH法,测定了香樟果皮花色苷的抗氧化能力,并比较了不同放置时间对香樟果花色苷稳定性的影响。结果表明:香樟果花色苷最佳提取工艺的回归方程为Y=58.64+2.27A+12.78B+10.18C-14.01A2-11.00B2-7.56C2,R2=0.9796,模型拟合程度良好,在该试验范围内,模型能够准确反映花色苷的提取结果;最佳工艺参数分别为pH1.0、料液比1∶15、乙醇浓度78.59%、提取温度77.14℃、提取时间42.48min,在此条件下香樟果花色苷的得率为67.99mg·100g-1;在一定浓度范围内,香樟果皮花色苷清除羟自由基率和总抗氧化能力均与浓度成正线性相关,回归方程分别为y=0.3388x+13.485(R2=0.9856),y=0.0275x+0.0221,(R2=0.9966)。利用响应面法确定的最佳工艺条件合理,可用于香樟果皮花色苷的提取,同时香樟果花色苷具有良好的抗氧化活性,为天然色素的开发利用提供一定指导意义。     

Isolation and characterization of the fragrant cyclamen O-methyltransferase involved in flower coloration

Anthocyanin O-methyltransferase (OMT) is one of the key enzymes for anthocyanin modification and flower pigmentation. We previously bred a novel red-purple-flowered fragrant cyclamen (KMrp) from the purple-flowered fragrant cyclamen 'Kaori-no-mai' (KM) by ion-beam irradiation. Since the major anthocyanins in KMrp and KM petals were delphinidin 3,5-diglucoside and malvidin 3,5-diglucoside, respectively, inactivation of a methylation step in the anthocyanin biosynthetic pathway was indicated in KMrp. We isolated and compared OMT genes expressed in KM and KMrp petals. RT-PCR analysis revealed that CkmOMT2 was expressed in the petals of KM but not in KMrp. Three additional CkmOMTs with identical sequences were expressed in petals of both KM and KMrp. Genomic PCR analysis revealed that CkmOMT2 was not amplified from the KMrp genome, indicating that ion-beam irradiation caused a loss of the entire CkmOMT2 region in KMrp. In vitro enzyme assay demonstrated that CkmOMT2 catalyzes the 3' or 3',5' O-methylation of the B-ring of anthocyanin substrates. These results suggest that CkmOMT2 is functional for anthocyanin methylation, and defective expression of CkmOMT2 is responsible for changes in anthocyanin composition and flower coloration in KMrp.     

EVOLUTION IN THE GENUS RUELLIA (ACANTHACEAE): A DISCUSSION BASED ON FLORAL FLAVONOIDS

A clear dichotomy exists in the genus Ruellia, separating the blue from the red flowered species. Flavonoids differ in a qualitative rather than a quantitative way. Apigenin 7-glucuronide and malvidin 3,5-diglucoside are common to all the blue flowered species, whereas chalcononaringenin 2'-glucoside (isosalipurposide) and pelargonidin 3,5-diglucoside are shared by the red flowered ones. The blue flowered species are linked with the red via apigenin 7-glucuronide and 3,5-diglucosylation of their respective anthocyanins. Both groups are involved in flavonoid race formation. All examined species (and some populations within species) differ in flavonoid content. The patterns of variability displayed provide a basis upon which an evolutionary scheme is constructed. Genetic drift is hypothesized as the effector of race formation in the blue flowered group.     

Distribution of anthocyanins in aceraceae leaves

The distribution of anthocyanins in spring sprouted and/or autumn coloured leaves of Dipteronia sinensis and Acer (119 taxa) was studied.

Dipteronia contained four cyanidin glycosides: the 3-glucoside, 3-rutinoside, 3-galloylglucoside and 3,5-diglucoside. Acer contained five cyanidin glycosides: 3-glucoside, 3-rutinoside, 3-galloylglucoside, 3-galloylrutinoside and 3,5-diglucoside, two delphinidin glucosides: 3-glucoside and 3-rutinoside and three unidentified anthocyanins. Both Dipteronia and Acer contained the recently reported cyanidin 3-galloylglucoside. The anthocyanin constituents in spring leaves were more complex than those found in autumn coloured leaves: nine in spring and six in autumn. The presence/absence of the major anthocyanins in the spring sprouted leaves of 111 Acer taxa analysed were grouped into 17 distribution patterns. In the autumn the number of anthocyanin distribution patterns was found to be 11. In Acer, cyanidin glycosides were found in 20 sections and delphinidin glycosides in 17 out of the 21 sections analysed. Although the distribution of anthocyanins showed no clear relations among sections, delphinidin glycosides were mainly found in sections Macrantha, Goniocarpa and Saccharina. There were no differences in the pigment constituents in the species native to different countries, such as A. rubrum in North America and A. pycnanthum in Japan, both containing the same pigments: cyanidin 3-glucoside, 3-rutinoside, 3-galloylglucoside, 3-galloylrutinoside and 3,5-diglucoside.     

Anthocyanins in flowers of genus Rosa, sections Cinnamomeae (=Rosa), Chinenses, Gallicanae and some modern garden roses

Forty-four taxa of three sections (Cinnamomeae (=Rosa) 26, Chinenses 8 and Gallicanae 10) and eight modern garden roses in the genus Rosa were surveyed for their floral anthocyanins. Eleven anthocyanins: 3-glucosides and 3,5-diglucosides of cyanidin (Cy), pelargonidin (Pg) and peonidin (Pn), 3-rutinosides and 3-rho-coumaroylglucoside-5-glucosides of Cy and Pn, and Cy 3-sophoroside, were isolated from flowers of these taxa and identified by chemical and spectroscopic techniques. Four anthocyanins: Cy 3-rutinoside, Pn 3-rutinoside, Pn 3-rho-coumaroylglucoside-5-glucoside and Cy 3-sophoroside were found for the first time in Rosa flowers.Investigated sections of wild roses showed characteristic distribution of anthocyanins. Cy 3,5-diglucoside was the dominant anthocyanin detected in all three sections, but accumulation of Pn 3,5-diglucoside distinguished sections Cinnamomeae from other sections, and the occurrence of Cy 3-glucoside separates section Chinenses from others.Cy 3-sophoroside was detected in large amount in some taxa of section Cinnamomeae: e.g., R. moyesii and its related cultivars, and R. rugosa cv. Salmon Pink. The acylated Cy glycoside was found in all sections and also in some modern garden roses, while the acylated Pn glycoside was detected in the section Cinnamomeae, but not in sections Chinenses and Gallicanae. According to anthocyanin distribution patterns, eight groups were classified chemotaxonomically in genus Rosa.     

Chemotaxonomic and ecological implications of anthocyanins in Elythranthera

Elythranthera emarginata (Lindl.) A. S. George and E. brunonis (Endl.) A. S. George both contain cyanidin-3,5-diglucoside and delphinidin-3,5-diglucoside. Pigment concentrations in E. marginata are 2 · 5 times those of E. brunonis. Cyanidin/delphinidin ratios in both species are essentially the same. These findings are discussed in relation to the taxonomy, color, and the life cycle of Elythranthera.     

THE DIFFERENTIATION OF PIGMENTATION IN FLOWER PARTS. VIII. THE EFFECT OF INHIBITORS OF PROTEIN AND RNA SYNTHESIS ON DEVELOPMENTAL CHANGES OF ANTHOCYANINS IN CULTURED PETALS OF IMPATIENS BALSAMINA

If inhibitors of protein or RNA synthesis are administered to flower petals of the red genotype (HHHP^rP^r) of Impatiens balsamina at a very early stage of development, an alteration in the normal pattern of anthocyanin pigmentation results. Whereas control petals are mainly pigmented with acyl pelargonidin-3,5-diglucoside and pelargonidin-3,5-diglucoside, petals cultured in the presence of inhibitors are mainly pigmented with pelargonidin-3-monoglucoside. The complete absence of the more highly substituted forms of pelargonidin in treated petals suggests that the biochemical reactions required for the addition of glucosyl and hydroxycinnamoyl residues to pelargonidin-3-monoglucoside have been prevented. The ability to block the normal developmental pattern of pigmentation with these inhibitors suggests that de novo synthesis of active enzymes is required, and as indicated by the effectiveness of actinomycin D, specific RNA synthesis is a necessary prerequisite for the synthesis of the normal anthocyanin complement in this tissue. The ability of the white flowered genotype (llhhpp) to metabolize exogenously supplied pelargonidin-3-monoglucoside was found to be prevented by prior culture of immature petals in the presence of DL-ethionine. The data indicate that the enzymes required for this ability are not products of induction by the substrate but rather their presence is a normal feature of petal development. Treatment with inhibitors has failed to produce any inhibition in the formation of specific anthocyanins found in the flower petals of some other genotypes of I. balsamina.     

Streptococcal oxidation of malvin: The role peroxidogenic bacteria can play in decolorization

The possible use of microorganisms which yield substantial levels of hydrogen peroxide in treatments for decolorization of a natural pigment (malvidin-3,5-diglucoside) was studied. Streptococcus durans S-76 was able to oxidize about 38% malvin implicating biogenic hydrogen peroxide in a buffered solution. Malvin by-products and the coumarinic derivative have identical properties and high-performance liquid chromatographic patterns.     

Distribution pattern of anthocyanins in the polygonaceae

A survey of anthocyanins in the flowers and other organs of thirty-three species of three genera belonging to the Polygonaceae has been carried out. There are thirteen anthocyans. Cyanidin glycosides, especially the 3-glycoside (chrysanthemin), are present most commonly and peonidin glycosides including the arabinosylglucoside are found with low frequency. The distribution of malvidin 3,5-diglucoside (malvin) is confined to the species belonging to the sectionEchinocaulon of the genusPolygonum. It is noted that cyanidin itself occurs in the stems ofPolygonum perfoliatum andP. senticosum.     

The anthocyanin in blue flowers ofCentaurea cyanus

The anthocyanin in the blue cornflower (Centaurea cyanus) has been known for many years to be cyanidin 3,5-diglucoside, namely cyanin. However, in the course of this study, it became evident that the major anthocyanin in the blue cornflower is not cyanin but cyanidin 3-succinyl glucoside 5-glucoside. This anthocyanin has not been reported in the literature and is tentatively called “centaurocyanin”. Centaurocyanin is chromatographically identical with the anthocyanin contained in crystalline protocyanin, the blue pigment from the cornflower. thus, there seems no doubt that this anthocyanin, but not cyanin, forms the blue complex pigment protocyanin.     

Impact of specific environmental characteristics of the site of origin (shady,sunny) on anthocyanin and flavonol contents of replanted plants at common cyclamen (<Emphasis Type="Italic">Cyclamen purpurascens</Emphasis> Mill.)

The value of plant provenance (plant origin) is well-known phenomena in woody plants, but less is known in herbaceous plants (perennials). This study with common cyclamen (Cyclamen purpurascens Mill.) was conducted to reveal the importance of specific environmental site properties of plant origin for plant growth and plant quality in the next years. The plants were observed in years 2013 and 2014, more than 10 years after removing and replanting them from the original sites. Morphological characteristics of plants were evaluated by measuring the length and the width of plant rosettes, whereby plants originated from different sites did not show any significant differences. Additionally, the pigment composition, flavonol and anthocyanin content of plant leaves were evaluated. Plants removed from sunny sites showed significantly lower chlorophyll values (total chlorophyll, chlorophyll a) in the both observed years; lower carotenoid and total pigment values were measured only in year 2013. The prevailing anthocyanin in cyclamen leaves was malvidin-3,5-diglucoside with 57.28 µg l^?1 FW in the year 2013 and with 103.68 µg l^?1 FW in the year 2014. Plants originated from the sunny sites accumulated in 2013 significantly more malvidin-3,5-diglucoside in comparison with plants from shady sites of origin. The major substances from the flavonol group were quercetin-3-O-rutinoside and quercetin-dirhamnosyl-glucoside in both analysed years. The cyclamen leaves originated from sunny sites contained in 2013 significant more quercetin-dirhamnosyl-glucoside than cyclamen leaves from shady sites. The results of the study show that different stress parameters (irradiation and water supply in specific year) have a significant impact on the morphological and also internal parameters of cyclamen leaves.