Minor Anthocyanin Research Articles

Flavonoids from the flowers of Adenium obesum (Forssk.) Roem. & Schult., Mandevilla sanderi (Hemsl.) Woodson, and Nerium oleander L. (Apocynaceae)

Twenty-two ornamental flowers from different Adenium obesum, Mandevilla sanderi, and Nerium oleander cultivars/seedlings were analyzed for the presence of anthocyanins, flavonols, and chlorogenic acid using nuclear magnetic resonance (NMR) and mass spectrometry (MS). Cyanidin 3-O-[6-O-(rhamnosyl)-galactoside] and cyanidin 3-O-(galactoside) were identified as the major and minor anthocyanins, respectively, in three A. obesum seedlings that had red and red-purple flowers.Cyanidin 3-O-[2-O-(xylosyl)-galactoside] was identified as the major anthocyanin, whereas cyanidin 3-O-[6-O-(rhamnosyl)-galactoside] and cyanidin 3-O-(galactoside) were identified as the minor anthocyanins in 8 M. sanderi cultivars that had red and red-purple flowers. Cyanidin 3-O-[6-O-(rhamnosyl)-galactoside] and cyanidin 3-O-(galactoside) were identified as the major anthocyanins, whereas cyanidin 3-O-[2-O-(xylosyl)-galactoside] was identified as the minor anthocyanin in 8 N. oleander cultivars with red and red-purple flowers. Low levels of anthocyanins were detected in the N. oleander and M. sanderi cultivars that had white flowers, and there were no anthocyanins detected in the N. oleander cultivars with yellow flowers. Chlorogenic acid and four flavonols, quercetin 3-O-[6-O-(rhamnosyl)-galactoside], quercetin 3-O-[6-O-(rhamnosyl)-glucoside], kaempferol 3-O-(galactoside), and kaempferol 3-O-[6-O-(rhamnosyl)-galactoside], were identified in the flowers from all 22 cultivars/seedlings investigated.

Detection of Quantitative Trait Loci Controlling the Content of Phenolic Compounds in an Asian Plum (Prunus salicina L.) F1 Population.

Consumption of fresh fruit is known to protect against non-communicable diseases due to the fruit's content in compounds with an antioxidant capacity, among them is polyphenols. Asian plums (Prunus salicina L.) accumulate more than 40 phenolic compounds, with a remarkable diversity in their profiles, depending on the variety and environmental conditions. Although candidate genes have been indicated to control this trait, the loci controlling its phenotypic variation have not yet been defined in this species. The aim of this work was to identify the quantitative trait Loci (QTL) controlling the phenolic compounds content in the Asian plum skin and flesh. Using UHPLC-DAD-Orbitrap-MS, we determined that cyanidin-3-glucoside and cyanidin-3-rutinoside are the main anthocyanins in Asian plums. Other anthocyanins found to a lesser extent were tentatively identified as cyanidin bound to different sugar and procyanidin moieties. Then we phenotyped fruits of 92 and 80 F1 seedlings from the cross < “98.99” × “Angeleno”> (98 Ang) for two harvest seasons. We used HPLC-DAD to quantify single anthocyanin and spectrophotometric techniques to determine the total content of phenols, flavonoids, procyanidins, and antioxidant activity (DPPH and FRAP). To determine the phenotype-genotype association of phenolic compounds content, phenotypic values (adjusted by linear mixed-effects models), genotypic data and linkage maps were analyzed with the multiple QTL model (MQM) approach. We found a total of 21 significant trait-marker associations: 13 QTLs segregating from “98.99” and 8 QTLs from “Angeleno.” From these associations, 8 corresponded to phenolic compound content in the flesh and 13 in the skin. Phenotype variance was explained by the detected loci, ranging from 12.4 to 27.1%. The identified loci are related to the content of cyanidin-3-glucoside (LG4), cyanidin-3-rutinoside (LG4), total flavonoids and procyanidins (LG5 and LG8), and minor anthocyanin compounds (LG3 and LG4). These results will help improve the efficiency of breeding programs for the generation of Asian plum varieties with high phenolic compound content.

Discovery of a natural cyan blue: A unique food-sourced anthocyanin could replace synthetic brilliant blue.

The color of food is critical to the food and beverage industries, as it influences many properties beyond eye-pleasing visuals including flavor, safety, and nutritional value. Blue is one of the rarest colors in nature's food palette-especially a cyan blue-giving scientists few sources for natural blue food colorants. Finding a natural cyan blue dye equivalent to FD&C Blue No. 1 remains an industry-wide challenge and the subject of several research programs worldwide. Computational simulations and large-array spectroscopic techniques were used to determine the 3D chemical structure, color expression, and stability of this previously uncharacterized cyan blue anthocyanin-based colorant. Synthetic biology and computational protein design tools were leveraged to develop an enzymatic transformation of red cabbage anthocyanins into the desired anthocyanin. More broadly, this research demonstrates the power of a multidisciplinary strategy to solve a long-standing challenge in the food industry.

Flavonoids of the rose-pink, blue, and white flowers of Nigella damascena L. (Ranunculaceae)

Nigella damascena L. is an annual plant belonging to the buttercup family Ranunculaceae, commonly known as love-in-a-mist. It is used as a garden plant. The flowers are blue, pink, or white. In order to determine the pigments responsible for the flower colors of N. damascena, the anthocyanins and flavonols in the rose-pink, blue, and white flowers of N. damascena were investigated using cultivars of the ‘Miss Jekyll’ series. The identities and structures of compounds were determined by chemical and spectroscopic analyses. Four cyanidin 3-glycosides were detected and identified from the rose-pink flowers. In the blue flowers, one major anthocyanin and several minor anthocyanins were detected. The identities of the major and one minor anthocyanin were determined. The main anthocyanins in the rose-pink and the blue flowers were cyanidin 3-[2-(xylosyl)-6-(rhamnosyl)-glucoside] and 7-methyldelphinidin 3-[2-(xylosyl)-6-(rhamnosyl)-glucoside], respectively. Petunidin 3-[2-(xylosyl)-6-(rhamnosyl)-glucoside] had been previously reported as the major anthocyanin from the blue flowers, however, any petunidin-type anthocyanins were not found from the blue flowers. Cyanidin 3-[2-(glucuronosyl)-6-(rhamnosyl)-glucoside], one of minor anthocyanins in the rose-pink flowers, and 7-methyldelphinidin 3-[2-(xylosyl)-6-(rhamnosyl)-glucoside] are reported for the first time. This is the second report of an anthocyanin of the 7-methyldelphinidin type, following that from Catharanthus roseus flowers. The white-flower cultivar contained anthocyanins analogous to those of the blue flowers but at very low levels. The same three flavonols were detected in the rose-pink, blue, and white flowers.

Characteristic phenolic composition of the Greek variety Mavrokountoura grape and wine

Mavrokountoura, a clone of the Mandilaria grape cultivar, was studied for the first time. The morphological characteristics and composition of berries were determined, as well as total phenols and tannins in skins, flesh, and seeds. The skin anthocyanins were identified using HPLC-MS/MS and quantified using HPLC-DAD. Two main anthocyanins were identified as malvidin-3-O-glucoside and malvidin-3-O-(6″-p-coumaroyl)-glucoside. One minor anthocyanin was identified as malvidin-3-O-(6″-acetyl)-glucoside. Glycosides of delphinidin, petunidin, and peonidin were detected in traces. Young Mavrokountoura wine was also studied. Extraction of total phenols from the grape to the young wine amounted to 32%, composing mainly non-tannic phenols. The individual anthocyanin components exhibited different extractability values from the skin to wine, ranging between 10–94%.

The blue anthocyanin pigments from the blue flowers of Heliophila coronopifolia L. (Brassicaceae)

Six acylated delphinidin glycosides (pigments 1– 6) and one acylated kaempferol glycoside (pigment 9) were isolated from the blue flowers of cape stock ( Heliophila coronopifolia) in Brassicaceae along with two known acylated cyanidin glycosides (pigments 7 and 8). Pigments 1– 8, based on 3-sambubioside-5-glucosides of delphinidin and cyanidin, were acylated with hydroxycinnamic acids at 3-glycosyl residues of anthocyanidins. Using spectroscopic and chemical methods, the structures of pigments 1, 2, 5, and 6 were determined to be: delphinidin 3- O-[2- O-(β-xylopyranosyl)-6- O-(acyl)-β-glucopyranoside]-5- O-[6- O-(malonyl)-β-glucopyranoside], in which acyl moieties were, respectively, cis- p-coumaric acid for pigment 1, trans-caffeic acid for pigment 2, trans- p-coumaric acid for pigment 5 (a main pigment) and trans-ferulic acid for pigment 6, respectively. Moreover, the structure of pigments 3 and 4 were elucidated, respectively, as a demalonyl pigment 5 and a demalonyl pigment 6. Two known anthocyanins (pigments 7 and 8) were identified to be cyanidin 3-(6- p-coumaroyl-sambubioside)-5-(6-malonyl-glucoside) for pigment 7 and cyanidin 3-(6-feruloyl-sambubioside)-5-(6-malonyl-glucoside) for pigment 8 as minor anthocyanin pigments. A flavonol pigment (pigment 9) was isolated from its flowers and determined to be kaempferol 3- O-[6- O-( trans-feruloyl)-β-glucopyranoside]-7- O-cellobioside-4′- O-glucopyranoside as the main flavonol pigment. On the visible absorption spectral curve of the fresh blue petals of this plant and its petal pressed juice in the pH 5.0 buffer solution, three characteristic absorption maxima were observed at 546, 583 and 635 nm. However, the absorption curve of pigment 5 (a main anthocyanin in its flower) exhibited only one maximum at 569 nm in the pH 5.0 buffer solution, and violet color. The color of pigment 5 was observed to be very unstable in the pH 5.0 solution and soon decayed. In the pH 5.0 solution, the violet color of pigment 5 was restored as pure blue color by addition of pigment 9 (a main flavonol in this flower) like its fresh flower, and its blue solution exhibited the same three maxima at 546, 583 and 635 nm. On the other hand, the violet color of pigment 5 in the pH 5.0 buffer solution was not restored as pure blue color by addition of deacyl pigment 9 or rutin (a typical flower copigment). It is particularly interesting that, a blue anthocyanin–flavonol complex was extracted from the blue flowers of this plant with H 2O or 5% HOAc solution as a dark blue powder. This complex exhibited the same absorption maxima at 546, 583 and 635 nm in the pH 5.0 buffer solution. Analysis of FAB mass measurement established that this blue anthocyanin–flavonol complex was composed of one molecule each of pigment 5 and pigment 9, exhibiting a molecular ion [M+1] + at 2102 m/z (C 93H 105O 55 calc. 2101.542). However, this blue complex is extremely unstable in acid solution. It really dissociates into pigment 5 and pigment 9.

Specific Poly-phenolic Compounds in Cell Culture of Vitis vinifera L. cv. Gamay Fréaux

Cell cultures established from plants represent an attractive alternative to whole plants for effective production of bioactive secondary metabolites. Cell culture from Vitis vinifera L. cv. Gamay Fréaux accumulated high amounts of hydroxycinnamic acid derivatives and anthocyanins. Two new compounds were identified: 3-O-glucosylresveratrol, a stilbene derivative, abundant in cell suspension culture, and a hydroxyphenol, 4-(3,5-dihydroxyphenyl)-phenol, abundant in callus culture. The major anthocyanin monoglucosides present in cell suspension culture were cyanidin 3-O-glucoside and peonidin 3-O-glucoside, and the major cinnamoyl derivatives were cyanidin 3-O-p-coumaryl glucoside and peonidin 3-O-p-coumaryl glucoside. Three minor anthocyanin compounds were found in V. vinifera cell culture: delphinidin 3-O-glucoside, petunidin 3-O-glucoside, and delphinidin 3-O-pcoumaryl glucoside. Anthocyanin levels of cell suspension cultures increased significantly--about eight fold--after 4-day cultivation in new medium. Salicylic acid at a concentration of 50 μM did not enhance anthocyanin accumulation in cell suspension culture, and similar levels of jasmonic acid significantly reduced the anthocyanin content.

Anthocyanins from red cabbage – stability to simulated gastrointestinal digestion

Anthocyanins were the main polyphenol components in extracts of fresh and pickled red cabbage. The composition of anthocyanins in red cabbage was studied using liquid chromatography mass–spectrometry. Eleven major peaks absorbing at 520 nm were discerned, which represented 18 different anthocyanin structures. Another five minor anthocyanin components could be identified by searching at their respective m/ z values but only in anthocyanin-enriched concentrates produced by sorption to solid phase extraction matrices. The predominant anthocyanins were constructed of cyanidin-3-diglucoside-5-glucoside “cores” which were non-acylated, mono-acylated or di-acylated with p-coumaric, caffeic, ferulic and sinapic acids. Pelargonidin-3-glucoside and novel forms of cyanidin-3- O-triglucoside-5- O-glucoside di-acylated with hydroxycinnamic acids were also detected in extracts of raw red cabbage, commercially pickled red cabbage and anthocyanin-enriched concentrates. The stability of the anthocyanins to simulated gastrointestinal digestion was assessed. The anthocyanins were effectively stable in the acidic gastric digestion conditions but the total recovery after simulated pancreatic digestion was around 25% compared to around 100% recovery of phenol content. As anthocyanins make up the majority of red cabbage polyphenols, this suggested that anthocyanins broke down to form new phenolic components. The recovery of the individual anthocyanins was monitored by LC–MS n . All of the anthocyanins were reduced in content after pancreatic digestion but acylated forms were notably more stable than non-acylated forms. There was also a relationship between the type of acylated hydroxycinnamic acid and stability to pancreatic digestion.

Black Flower Coloration in Wild Lisianthius nigrescens: Its Chemistry and Ecological Consequences

The major pigments responsible for the flower color within the black flowered Gentianaceae, Lisianthius nigrescens, were characterized by HPLC and chemical analyses HPLC analysis showed one major and one minor anthocyanin and 3 major and 3 minor flavone glycosides. The anthocyanins [delphinidin-3-O-rhamnol(1-6)galactoside and its 5-O-glucoside] comprised an extraordinary 24% of the dry weight of wild collected L. nigrescens corallas, and were accompanied in a 1:1 ratio by a range of apigenin and luteolin 8-C-glucosides and their 7-O-methyl ethers. The high levels of anthocyanins and flavones (and their co-pigmentation) is thought to account for the almost complete absorption of both UV and visible wavebands observed by reflectance photography.

Relationship of light quantity and anthocyanin production in Pennisetum setaceum Cvs. rubrum and red riding hood.

Pennisetum setaceum cvs. Rubrum and Red Riding Hood are purple-pigmented ornamental grasses when grown in high-light environments. In low-light environments, foliage appears light purple or green, and as a result, aesthetic appeal is reduced. The impact of light on anthocyanin pigmentation was compared for P. setaceum Rubrum foliage and flowers and Red Riding Hood foliage grown under different light intensities and light sources. Light environments included UV supplemental light in the greenhouse, high-pressure sodium supplemental light in the greenhouse, cool-white fluorescent light in a growth chamber, and full sun outside. Anthocyanins in two cultivars of P. setaceum were analyzed by HPLC and characterized by (1)H and (13)C NMR spectral experiments. Two anthocyanins, cyanidin 3-glucoside and cyanidin 3-rutinoside, were identified in the leaves and flowers of both cultivars and quantified by HPLC analysis. The major anthocyanin in both cultivars was cyanidin 3-glucoside and had highest concentration (0.199% fresh weight) in Rubrum leaves grown under fluorescent lights in the growth chamber with a photoperiod of 24 h and a daily light integral (DLI) of 13.3 mol m(-)(2) day(-)(1) and in Rubrum and Red Riding Hood leaves and flowers (0.097 and 0.12% fresh weight) from plants grown outside in full sun with a photoperiod ranging from 15 to 13.5 h and DLI of 42 mol m(-)(2) day(-)(1). The minor anthocyanin, cyanidin 3-rutinoside, had the highest quantity in plants grown in low-light-intensity greenhouse environments with a photoperiod ranging from 15 to 13.5 h and DLI of 2.3-7.0 mol m(-)(2) day(-)(1). The functional significance of anthocyanins in P. setaceum Rubrum is discussed.

(Delphinidin 3-Gentiobiosyl) (Luteorin 7-Glucosyl) Malonate from the Flowers of Eichhornia crassipes

An additional new pigment exhibiting anthocyanin and flavone disubstituted malonate structure, was isolated from the blue-purple flowers of Eichhornia crassipes as a minor anthocyanin. This pigment was identified as (6'''-O-(delphinidin 3-O-(6''-O-(β-D-glucopyranosyl)-β-D-glucopyranosyl))) (6''-O-(luteolin 7-O-(β-D-glucopyranosyl))) malonate by chemical and spectroscopic methods. The λmax of this pigment is 547 nm in 0.1% HCl-MeOH, where the negative cotton effect is also observed as similar to that of Eichhornia anthocyanin A, (delphinidin 3-gentiobiosyl) (apigenin 7-glucosyl) malonate previously isolated from the same flowers. This result supports that the new pigment also takes a folding conformation between delphinidin (chromophore) and luteorin (co-pigmnet).

Separation and characterization of simple and malonylated anthocyanins in red onions, Allium cepa L.

The colour of red onions is due primarily to anthocyanins present in the epidermal cells of the scale leaves of the bulb. The occurrence of the four major anthocyanins, cyanidin 3-glucoside, cyanidin 3-laminaribioside, cyanidin 3-malonylglucoside and cyanidin 3-malonyllaminaribioside, was confirmed in four red onion cultivars, namely ‘Mambo’, ‘Red Jumbo’, ‘Red Bone’, and ‘Red Granex’. In addition, four new minor anthocyanin pigments were isolated and characterized from red onions. Two structures were determined by chemical and spectroscopic methods to be peonidin 3-glucoside, and peonidin 3-malonylglucoside. This is the first report of the occurrence of simple and malonylated peonidin glucosides in red onions.

Pelargonidin 3-glucoside-5-acetylglucoside in Verbena flowers

A new acylated anthocyanin was isolated from deep pink flowers of Verbena hybrida cv ‘Tropic’ as a minor anthocyanin and identified as pelargonidin 3-O-β- d-glucoside -5-O-(6-O- (acetyl)-β- d-glucoside ) with chromatographic and spectral methods.

Characterization, Quantitation, and Distribution of Anthocyanins and Colorless Phenolics in Sweet Cherries

The anthocyanins and colorless phenolics in 11 cultivars and hybrids of sweet cherries were characterized and quantified by high-performance liquid chromatography and gas chromatography. All of the dark-colored cherry genotypes were found to contain 3-rutinoside and 3-glucoside of cyanidin as the major anthocyanins and the same glycosides of peonidin as minor anthocyanins. Another minor anthocyanin, pelargonidin 3-rutinoside, was characterized in sweet cherries for the first time. The major colorless phenolics were characterized as neochlorogenic acid and p-coumaroylquinic acid. The total anthocyanin content ranged from 82 to 297 mg/100 g of pitted cherry for the dark cherries and from 2 to 41 mg/100 g of pitted fruit for the light-colored cherries. The neochlorogenic acid ranged from 24 to 128 mg/100 g of pitted cherry and p-coumaroylquinic acid from 23 to 131 mg/100 g, respectively. The relative amounts of the two phenolic acids varied widely across the cherry cultivars examined in this study.

A minor anthocyanin from cultured cells of Aralia cordata

A minor anthocyanin accumulated in the cultured cells of Aralia cordata has been identified as peonidin 3- O-β- d-xylopyranosyl (1→2)-β- d-galactopyranoside, which is the 3′- O-methyl ether of the major anthocyanin, on the basis of the FABMS and 1H, 13CNMR spectra. Effects of culture conditions on the anthocyanin composition accumulated in the cells were investigated. Light irradiation, high sucrose or low nitrate appears to stimulate the methylation of the major anthocyanin.

An acylated cyanidin 3-sophoroside-5-glucoside in the violet-blue flowers of Pharbitis nil

A new acylated anthocyanin was isolated from the violet-blue flowers of Pharbitis nil as a minor anthocyanin. The structure of this pigment was determined to be cyanidin 3- O-(2- O-(β- d- glucopyranosyl)-6- O-( trans-4- O-(β- d-glucopyranosyl)caffeyl)-β- d-glucopyranosyl)- 5- O-(β- d-glucopyranoside) based on spectroscopic methods.

Unusual Koshu Grape Skin Anthocyanins

Two unusual fumaric acid acylated peonidin-3-monoglucosides were fractionated and purified from the skins of the Koshu grape, native to Japan, as minor anthocyanin pigments. The differentiation of Koshu from <i>Vitis vinifera</i> and American species is discussed. Diglucoside pigments are absent; therefore, Koshu cannot be directly derived from <i>V. labrusca</i> or other American species or <i>V. amurensis</i>. The pigments are, nevertheless, different from those reported for <i>V. vinifera</i>.

Pigments of lily flowers I

1. This experiment was practiced to identify anthocyanin pigments in flowers of 19 lily varieties by paper chromatographic and spectrophotometric methods.2. The anthocyanins were identified in 15 varieties out of 19, and they were keracyanin (Cyanidin-3-rhamnoglucoside) and minor anthocyanin.3. Four varieties were devoid of anthocyanin pigments and in 2 varieties with white flowers no leuco-anthocyanins were detected.