Changes in the Antioxidative Activity and the Content of Phenolics and Iridoids during Fermentation and Aging of Natural Fruit Meads.

The aim of the study was to investigate changes in the content of biologically active compounds during the fermentation and aging of natural meads with the addition of three Cornelian cherry juices from three cultivars: 'Koralovyi', 'Podolski' and 'Yantarnyi', in the amount of 10% v/v. After the fermentation process the content of gallic and ellagic acids significantly increased, in relation to wort. Whereas the greatest losses were observed among unstable anthocyanins. The three-month aging process also reduced the content of the analyzed compounds except for ellagic acid, the content of which increased by up to 90%. The content of biologically active compounds, including iridoids and antioxidant phenolics, are constantly changing in the process of fermentation and aging of fruit meads. The studies proved that the addition of Cornelian cherry juice allows significantly enriched classic meads with new biologically active compounds, such as: exceptional iridoids (loganic acid, cornuside, loganine, sweroside), flavonols, phenolic acids and anthocyanins.

Anthocyanin Accumulation in the Leaves of the Purple Sweet Potato (Ipomoea batatas L.) Cultivars.

Sweet potato anthocyanins are water-soluble pigments with many physiological functions. Previous research on anthocyanin accumulation in sweet potato has focused on the roots, but the accumulation progress in the leaves is still unclear. Two purple sweet potato cultivars (Fushu No. 23 and Fushu No. 317) with large quantities of anthocyanin in the leaves were investigated. Anthocyanin composition and content were assessed with ultra-performance liquid chromatography diode-array detection (UPLC-DAD) and ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS), and the expressions of genes were detected by qRT-PCR. The two cultivars contained nine cyanidin anthocyanins and nine peonidin anthocyanins with an acylation modification. The acylation modification of anthocyanins in sweet potato leaves primarily included caffeoyl, p-coumaryl, feruloyl, and p-hydroxy benzoyl. We identified three anthocyanin compounds in sweet potato leaves for the first time: cyanidin 3-p-coumarylsophoroside-5-glucoside, peonidin 3-p-coumarylsophoroside-5-glucoside, and cyanidin 3-caffeoyl-p-coumarylsophoroside-5-glucoside. The anthocyanidin biosynthesis downstream structural genes DFR4, F3H1, anthocyanin synthase (ANS), and UDP-glucose flavonoid 3-O-glucosyltransferase (UFGT3), as well as the transcription factor MYB1, were found to be vital regulatory genes during the accumulation of anthocyanins in sweet potato leaves. The composition of anthocyanins (nine cyanidin-based anthocyanins and nine peonidin-based anthocyanins) in all sweet potato leaves were the same, but the quantity of anthocyanins in leaves of sweet potato varied by cultivar and differed from anthocyanin levels in the roots of sweet potatoes. The anthocyanidin biosynthesis structural genes and transcription factor together regulated and controlled the anthocyandin biosynthesis in sweet potato leaves.

Caracterização quí­mica de tomate roxo e estudo da metabolização de flavonoides in vitro e in vivo / Chemical characterization of purple tomato and in vitro and in vivo study of metabolism of flavonoids

Tendo em vista a importância dos compostos bioativos (CBAs) para a promoção da saúde, foram desenvolvidos, a partir de cruzamentos do tomate cereja com espécies selvagens, os tomates laranja (rico em ß-caroteno) e roxo (rico em antocianinas), por meio da técnica de introgressão de alelos. Diante disso, o objetivo deste trabalho foi caracterizar o perfil de compostos bioativos e voláteis dos tomates enriquecidos e avaliar a estabilidade e metabolização de flavonoides do tomate roxo durante digestão in vitro e em modelo animal. Os tomates foram caracterizados quanto ao conteúdo de compostos fenólicos totais; capacidade antioxidante por DPPH e ORAC; ácidos orgânicos; açúcares solúveis; perfil de carotenoides por CLAE/DAD; flavonoides por CLAE/DAD e LC/ESI/MS/MS e compostos voláteis por CG/MS. Avaliou-se ainda a estabilidade dos flavonoides da casca do tomate roxo por simulação da digestão in vitro, utilizando o Simulator of the Human Intestinal Microbial Ecosystem (SHIME), bem como a formação de AGCC por GC-MS, e excreção em ratos Wistar, com posterior identificação dos compostos fenólicos por LC/Q-TOF/MS. O tomate roxo apresentou aumento no conteúdo de fenólicos totais, capacidade antioxidante e vitamina C, com destaque para casca. A rutina foi o principal flavonol identificado em todos os frutos, e na casca do tomate roxo foi encontrado alto teor de petunidina (p-coumaroil)-rutinosídeo-hexosídeo, além da superexpressão de outros flavonoides como a quercetina-3-O-rutinosídeo e kaempferol. Não houve alteração no perfil de flavonoides do fruto laranja. Este, por sua vez, apresentou acúmulo de ß-caroteno, importante pró-vitamina A, ao passo que o tomate roxo também teve seus conteúdos de ß-caroteno e licopeno aumentados. Os frutos apresentaram perfil de compostos voláteis diferentes entre si, o que foi relacionado à degradação dos diferentes CBAs característicos de cada um. O extrato fenólico da casca de tomate roxo, submetido à digestão in vitro, se manteve estável na primeira porção, relativo às condições estomacais. Contudo, o conteúdo de flavonoides apresentou redução significativa (p<0,05) nas porções que simulam as condições do duodeno e do colón, com a formação de catabólitos pela ação da microbiota intestinal e/ou pela degradação química espontânea. Foi observado o aparecimento de novos ácidos fenólicos não presentes inicialmente na matriz, dentre eles o ácido 3-O-metilgálico e o ácido homovanílico, supostamente derivados da degradação da petunidina e da quercetina, respectivamente. Houve aumento na produção total de AGCC, com excessão do butirato. Na urina dos animais foram detectados diversos outros compostos fenólicos derivados do metabolismo de fase II, dentre eles o ácido hipúrico e o 3-O-metilcatecol. Nas fezes foram identificados cerca de metade dos compostos presentes na fermentação in vitro. Dessa forma, o melhoramento convencional pode ser uma alternativa para o enriquecimento, com CBAs, de alimentos amplamente consumidos pela população, como o tomate. Além disso, durante a passagem pelo trato gastrointestinal, os flavonoides presentes na casca do tomate roxo sofrem intensa degradação pela microbiota intestinal, com formação de catabólitos com reconhecido potencial benefício à saúde

Considering the importance of bioactive compounds (BACs) for health promotion, the orange (ß-carotene-rich) and purple (anthocyanin-rich) tomatoes were developed from cherry tomato interspecific crossing with wild species, using the technique of allele introgression. The objective of this work was to characterize the profile of bioactive and volatile compounds of enriched tomatoes, and to evaluate the stability and metabolism of purple tomato's flavonoids during in vitro and in vivo digestion. The tomatoes were characterized by its content of total phenolic compounds; antioxidant capacity (DPPH and ORAC); organic acids; soluble sugars; carotenoids (HPLC/DAD); flavonoids (HPLC/DAD and LC/ESI/MS/MS) and volatile compounds (GC/MS). The stability of the flavonoids from purple tomato peel was assessed by using the Simulator of the Human Intestinal Microbial Ecosystem (SHIME), as well as the formation of AGCC by GC-MS, and metabolism and excretion in Wistar rats, with subsequent identification of phenolic compounds by LC/Q-TOF/MS. The purple tomato showed an increase in the total phenolic content, antioxidant capacity and vitamin C, with highlight for the peel. The rutin was the main flavonol identified in all fruits, and it was found high content of petunidin (p-coumaryl)-rutinoside-hexoside in the purple tomato peel, in addition to overexpression of other flavonoids such as quercetin-3-O-rutinoside and kaempferol. There was no change in the flavonoid profile of the orange fruit. This one, in turn, presented accumulation of ß-carotene, important pro-vitamin A, while the purple tomato also showed an increase in its ß-carotene and lycopene contents. The fruits presented different volatile compounds profile among them, which was related to the degradation of the different BACs composition of each one. In the in vitro digestion, the phenolic extract of the purple tomato peel remained stable in the first portion, relative to the stomach conditions. However, the content of flavonoids presented a significant reduction (p<0.05) in the portions simulating the duodenum and colon, with the formation of catabolites by the action of intestinal microbiota and/or spontaneous chemical degradation. It was observed the appearance of new phenolic acids that was not initially present in the matrix, among them 3-O-methylgalic acid and homovanilic acid, supposedly derived from the degradation of petunidin and quercetin, respectively. There was an increase in the total production of SCFAs, with the exception of butyrate. In the urine of the animals several other phenolic compounds derived from phase II metabolism were detected, among them hippuric acid and 3-O-methylcatechol. About half of the compounds present in the in vitro fermentation were identified in the feces. Conventional breeding may be an alternative for the enrichment, with BACs, of foods that are widely consumed by the population, such as tomatoes. In addition, during the passage through the gastrointestinal tract, the flavonoids present in the purple tomato peel are severely degraded by the intestinal microbiota, with formation of catabolites with recognized potential health benefit

Classification of fruits based on anthocyanin types and relevance to their health effects.

Anthocyanins are a group of water-soluble pigments that confer the blue, purple, and red color to many fruits. Anthocyanin-rich fruits can be divided into three groups based on the types of aglycones of their anthocyanins: pelargonidin group, cyanidin/peonidin group, and multiple anthocyanidins group. Some fruits contain a major anthocyanin type and can serve as useful research tools. Cyanidin glycosides and peonidin glycosides can be metabolically converted to each other by methylation and demethylation. Both cyanidin and peonidin glycosides can be metabolized to protocatechuic acid and vanillic acid. Pelargonidin-3-glucoside is metabolized to 4-hydroxybenoic acid. On the other hand, phenolic acid metabolites of delphinidin, malvidin, and petunidin glycosides are unstable and can be further fragmented into smaller molecules. A literature review indicates berries with higher cyanidin content, such as black raspberries, chokeberries, and bilberries are more likely to produce an antiinflammatory effect. This observation seems to be consistent with the hypothesis that one or more stable phenolic acid metabolites contribute to the antiinflammatory effects of anthocyanin-rich fruits. More studies are needed before we can conclude that fruits rich in cyanidin, peonidin, or pelargonidin glycosides have better antiinflammatory effects. Additionally, fruit polyphenols other than anthocyanins could contribute to their antiinflammatory effects. Furthermore, blueberries could exert their health effects with other mechanisms such as improving intestinal microbiota composition. In summary, this classification system can facilitate our understanding of the absorption and metabolic processes of anthocyanins and the health effects of different fruits.

Anthocyanins from the flowers of Nagai line of Japanese garden Iris (Iris ensata).

Six anthocyanins were isolated from the flowers of the Nagai line of Iris ensata Thunb. They were identified as petunidin and malvidin 3-O-beta-[(4"'-Z-p-coumaroyl-alpha-rhamnopyranosyl)-(1-->6)-beta-glucopyranoside]-5-O-beta-glucopyranosides (1 and 3) and their E-forms (2 and 4), and petunidin and malvidin 3-O-rutinoside-5-O-glucosides (5 and 6). Though the E-form of petunidin 3-O-[(4"'-p-coumaroylrhamnosyl)-(1-->6)-glucoside]-5-O-glucoside has been reported, its Z-form was found for the first time. The presence of Z- and E-forms of malvidin 3-O-[(4'''-p-coumaroylrhamnosyl)-(1-->6)-glucoside]-5-O-glucoside is also reported for the first time. Fifty-one cultivars of Nagai line and their wild form (I. ensata var. spontanea) were divided into four anthocyanin patterns, i.e. 1) the presence of 1-4, 2) the presence of 2 and 4, 3) the presence of 5 and 6, and 4) no anthocyanin.

Systematic qualitative and quantitative assessment of anthocyanins, flavones and flavonols in the petals of 108 lotus (Nelumbo nucifera) cultivars.

Petal colour is one of the major characteristics that determine the ornamental value of lotus. To assess the contribution of different flavonoids to this character, composition and content of anthocyanins, flavones and flavonols were analysed through high performance liquid chromatography coupled with photodiode array detection tandem electrospray ionisation triple quad mass spectrometry in 108 lotus cultivars with red, pink, yellow, white and red/white pied petal colours. Totally, five anthocyanins and fourteen flavones and flavonols were detected and quantified. In general, the yellow, white and pied species hardly contained any anthocyanins; red cultivars contain more than pink cultivars. Among the five anthocyanins, malvidin 3-O-glucoside was the most abundant one in all the cultivars that contain anthocyanin. The fourteen flavones and flavonols belonged to four groups based on their aglycones. Except for the yellow cultivars, kaempferol-derivatives were the most abundant one. These data might be helpful in lotus breeding for different colours.

Molecular genetic study on the anthocyanin chemotypes of Perilla frutescens var. crispa.

The chemotypes found in various plant species are the good subjects for the studies to understand the regulatory mechanism in secondary metabolism. The biochemistry and molecular biology of flavonoid biosynthesis was studied using chemotypes of Perilla frutescens var. crispa differing anthocyanin accumulation. The expression of the most of structural genes for anthocyanin biosynthesis was coordinately regulated in chemotype-specific manner and by light. However, the genes for shared enzymes between anthocyanin and flavone pathway were expressed both chemotypes. Biochemical characteristics of enzymes involved in anthocyanin biosynthesis were investigated in this plant. Furthermore, the candidates of regulatory factors, members of MYB-bHLH-WD complex, of anthocyanin production were characterized in this plant.

Metabolomics-oriented isolation and structure elucidation of 37 compounds including two anthocyanins from Arabidopsis thaliana.

In order to conduct metabolomic studies in a model plant for genome research, such as Arabidopsis thaliana (Arabidopsis), it is a prerequisite to obtain structural information for the isolated metabolites from the plant of interest. In this study, we isolated metabolites of Arabidopsis in a relatively non-targeted way, aiming at the construction of metabolite standards and chemotaxonomic comparison. Anthocyanins (5 and 7) called A8 and A10 were isolated and their structures were elucidated as cyanidin 3-O-[2-O-(beta-D-xylopyranosyl)-6-O-(4-O-(beta-D-glucopyranosyl)-E-p-coumaroyl)-beta-D-glucopyranoside]-5-O-[6-O-(malonyl)-beta-D-glucopyranoside] and cyanidin 3-O-[2-O-(2-O-(E-sinapoyl)-beta-D-xylopyranosyl)-6-O-(4-O-(beta-D-glucopyranosyl)-E-p-coumaroyl)-beta-D-glucopyranoside]-5-O-[beta-D-glucopyranoside] from analyses of 1D NMR, 2D NMR ((1)H NMR, NOE, (13)C NMR, HMBC and HMQC), HRFABMS, FT-ESI-MS and GC-TOF-MS data. In addition, 35 known compounds, including six anthocyanins, eight flavonols, one nucleoside, one indole glucosinolate, four phenylpropanoids and a derivative, together with three indoles, one carotenoid, one apocarotenoid, three galactolipids, two chlorophyll derivatives, one steroid, one hydrocarbon, and two dicarboxylic acids, were also isolated and identified from their spectroscopic data.

Anthocyanins protect against A2E photooxidation and membrane permeabilization in retinal pigment epithelial cells.

The pyridinium bisretinoid A2E, an autofluorescent pigment that accumulates in retinal pigment epithelial cells with age and in some retinal disorders, can mediate a detergent-like perturbation of cell membranes and light-induced damage to the cell. The photodynamic events initiated by the sensitization of A2E include the generation of singlet oxygen and the oxidation of A2E at carbon-carbon double bonds. To assess the ability of plant-derived anthocyanins to modulate adverse effects of A2E accumulation on retinal pigment epithelium (RPE) cells, these flavylium salts were isolated from extracts of bilberry. Nine anthocyanin fractions reflecting monoglycosides of delphinidin, cyanidin, petunidin and malvidin were obtained and all were shown to suppress the photooxidation of A2E at least in part by quenching singlet oxygen. The anthocyanins tested exhibited antioxidant activity of variable efficiency. The structural characteristics relevant to this variability likely included the ability to form a stable quinonoidal anhydro base at neutral pH, a conjugated diene structure in the C (pyrane) ring, the presence of hydroxyl groups on the B (benzene) ring and the relative hydrophobicity conferred by the arrangement of substituents on the B ring. Cells that had taken up anthocyanins also exhibited a resistance to the membrane permeabilization that occurs as a result of the detergent-like action of A2E.

Anthocyanic vacuolar inclusions (AVIs) selectively bind acylated anthocyanins in Vitis vinifera L. (grapevine) suspension culture.

Anthocyanic vacuolar inclusions (AVIs) appear as dark red-to-purple spheres of various sizes in vacuoles of grapevine (Vitis vinifera L.) cell suspension culture due to their interaction with anthocyanins. AVIs were purified and the bound anthocyanins extracted and analysed by HPLC from two lines of V. vinifera isolated from the same callus accumulating anthocyanin in the dark, yet varying in their anthocyanin profiles and accumulation. An intermediate-pigmented line (FU-1) with a 1.3:1 ratio of acylated:non-acylated anthocyanins, a colour value of 0.84 units and cyanidin and peonidin as the dominant species was compared with a high-pigmented line (FU-2) with a 1.2:1 ratio of acylated:non-acylated anthocyanins, a colour value of 3.72 units and malvidin predominating. The profile of AVI-bound anthocyanins showed an increase in acylated anthocyanins in both lines of approx. 28-29%, with no apparent preference for anthocyanin species. This resulted in a ratio of acylated:non-acylated anthocyanins of 6.2:1 for FU-1 and 4.9:1 for FU-2. The reasons for the selectivity of the AVIs for acylated (specifically p-coumaroylated) species compared with the whole cell profile are discussed.

Unusual carotenogenesis in fruits with pronounced anthocyanic ripening (Olea europaea Var. Arbequina).

Fruits of the Olea europaea Arbequina variety showed a carotenogenic stage overlapping the biosynthesis of anthocyanins. At the start of ripening, the carotenoid concentration increases (20%) or is maintained (besides the esterification of xanthophylls found in this variety), compared with a decrease (30%) found in the same period in the Picual variety (used as a control that was representative of the rest of the varieties). Both the beta,beta and beta,epsilon carotenoid series were found to be implicated in this carotenogenic process. Differences in both varieties for the chlorophyll a/b ratio, chlorophylls/carotenoids ratio, and relative carotenoid composition may indicate the existence of a different metabolism of chloroplast pigments, but also indicates a different structure and function of the photosynthetic apparatus. This fact shows that the photosynthetic behavior in the Picual variety is similar to that of shaded leaves, and in the Arbequina variety is similar to that of sun leaves.

Natural pigments: carotenoids, anthocyanins, and betalains--characteristics, biosynthesis, processing, and stability.

Pigments are present in all living matter and provide attractive colors and play basic roles in the development of organisms. Human beings, like most animals, come in contact with their surroundings through color, and things can or cannot be acceptable based on their color characteristics. This review presents the basic information about pigments focusing attention on the natural ones; it emphasizes the principal plant pigments: carotenoids, anthocyanins, and betalains. Special considerations are given to their salient characteristics; to their biosynthesis, taking into account the biochemical and molecular biology information generated in their elucidation; and to the processing and stability properties of these compounds as food colorants.