Characterization Of Anthocyanins Research Articles

Identification and Characterization of Anthocyanins by High-Performance Liquid Chromatography−Electrospray Ionization−Tandem Mass Spectrometry in Common Foods in the United States: Vegetables, Nuts, and Grains

Anthocyanins in common foods in the United States, other than fruits and berries, were identified and characterized by high-performance liquid chromatography (HPLC)-electrospray ionization-tandem mass spectrometry coupled with diode array detection. Of all of the 40+ vegetables, nuts, and grains screened, seven vegetables, one nut, and one grain were found to contain anthocyanins; the number of anthocyanins detected varied from two in pistachio nuts to 34 in red radishes. The individual anthocyanins were identified by comparing their mass spectrometric data and retention times with those of standards, published data, and reference food samples. In all of the samples analyzed, except for sorghum, only six common anthocyanidins (delphinidin, cyanidin, pelargonidin, petunidin, peonidin, and malvidin) were found as their glycosides. Anthocyanins in certain vegetables such as red cabbage and red radish were highly conjugated with sugars and acylated groups, and thus, their structures were very complicated. Eight different either aliphatic or aromatic acylated groups (acetoyl, coumaroyl, malonoyl, p-hydroxybenzoyl, feruoyl, caffeoyl, sinapoyl, and oxaloyl) were identified in the anthocyanins. In addition to glucose, six other sugar moieties (galactose, xylose, rhamnose, rutinose, sambubiose, and laminaribiose) were observed. Three varieties of sorghum were found to contain 3-deoxyanthocyanidins and their derivatives as major anthocyanins. A number of new anthocyanins were identified in the foods studied. This paper presents complete HPLC profiles and MS spectrometric data, obtained under the same experimental conditions, for common vegetables, pistachio nuts, and sorghum that contain anthocyanins.

Systematic Identification and Characterization of Anthocyanins by HPLC-ESI-MS/MS in Common Foods in the United States: Fruits and Berries

Anthocyanins were systematically identified and characterized by HPLC-ESI-MS/MS coupled with diode array detection in common fruits from U.S. food markets and other commercial sources. Of the 25 different fruits that were screened, 14 fruits were found to contain anthocyanins; the number of anthocyanins varied from 2 in peaches and nectarines to 31 in Concord grape. The individual anthocyanins were identified by comparing their mass spectral data and retention times with those of standards and published data. In all of the samples analyzed, only 6 common anthocyanidins, delphinidin, cyanidin, pelargonidin, petunidin, peonidin and malvidin, were found. In addition to the well-known major anthocyanins, a number of minor anthocyanins were identified for the first time. Some possible guidelines that help to identify anthocyanins in foods with complex anthocyanin composition were deduced and discussed. For the first time, this paper presents complete anthocyanin HPLC profiles and MS spectral data of common fruits using the same uniform experimental conditions.

Anthocyanins, Flavonols, and Free Radical Scavenging Activity of Chinese Bayberry (Myrica rubra) Extracts and Their Color Properties and Stability

Characterization of anthocyanins and flavonols and radical scavenging activity assays of extracts from four Chinese bayberry (Myrica rubra) varieties with different fruit colors were carried out. One dominant anthocyanin and three major flavonols were isolated by HPLC, and cyanidin-3-O-glucoside and two of three flavonols, myricetin and quercetin-3-O-rutinoside, were identified by cochromatography with authentic standards. Both DPPH* and ABTS*(+) cation assays indicated that the black varieties (Biji and Hunan) demonstrated much higher radical scavenging activities than the pink (Fenhong) and yellow (Shuijing) varieties, which may be attributed to much higher levels of anthocyanins, flavonoids, and total phenolics in the black varieties. Biji and Hunan had 6.49 and 6.52 mM Trolox equivalent antioxidant capacity (TEAC) per 100 g of fresh weight, whereas the pink (Fenhong) and yellow (Shuijing) bayberries had 1.32 and 1.31 mM TEAC/100 g. Different fruit color was reflected by the surface color and pigment extract color. Color stability of the pigment was dependent on pH, and the pigment was more stable at low pH (pH approximately 1.5). The lightness (L) increased while the chroma (C value) decreased with increase of pH until pH 5, but higher pH caused a small decrease for L and an increase for C.

Natural occurrence of free anthocyanin aglycones in beans ( Phaseolus vulgaris L.)

The application of an improved method of extraction and purification has allowed the characterization of anthocyanins and free anthocyanidins in beans, Phaseolus vulgaris L. using LC-MS determination. HPLC analysis of the final extract showed the existence of seven compounds when 520 nm was the selected wavelength. Major peaks were identified, according to their retention times and UV–Vis spectra, as cyanidin and pelargonidin monoglucosides when they were compared to those of the in-house library. Taking into account mass spectrometric data, together with elution time and spectral features, it was possible to identify peaks corresponding to cyanidin and pelargonidin diglucosides. As far as we know, these anthocyanins have not been detected in P. vulgaris L. before. The last three peaks in the chromatograms were thought to be acylated anthocyanins, due to their smaller polarity. However, MS spectra showed signals at m/ z 287, 301 and 271, which would be in accordance with molecular ions corresponding to anthocyanidins: cyanidin, peonidin and pelargonidin, respectively. Moreover, these compounds had the same retention times and UV–Vis spectral features as those of aglycones arising from acid hydrolysis of the final extract. These results demonstrate, for the first time, the presence, in natural form, of free aglycones in vegetable samples. Until now, the bibliographical data have indicated that, in nature, this type of compound is only found in heteroside forms.

Caractérisation et mesure des activités anti-radicalaires d'anthocyanes de plantes du Burkina Faso

Characterisation and antioxidative scavenging activities of anthocyanins in plants of Burkina Faso. Nine anthocyanins were isolated from three plants and their structures elucidated using chemical, GC, MS and NMR methods. Adenium obesum flowers anthocyanins were identified as cyanidin 3- O-(4- O-α- l-rhamnosyl)- β- d-galactopyranoside. The cyanidin 3- O-β- d-glucopyranoside, the cyanidin 3- O-(6- O-α- l-rhamnopyranosyl)- β- d-glucopyranoside, the pelargonidin 3- O-β- d-glucopyranoside and the pelargonidine 3- O-(6- O-α- l-rhamnopyranosyl)- β- d-glucopyranoside were isolated and identified from the flowers of Jatropha integuerrima. From flower chalices of H. sabdariffa, four compounds were established as cyanidin 3- O-β- d-glucopyranoside, cyanidin 3- O-(2- O-β- d-xylopyranosyl)- β- d-glucopyranoside, delphinidin 3- O-β- d-glucopyranoside and delphinidin 3- O-(2- O-β- d-xylopyranosyl)- β- d-glucopyranoside. The DPPH assay allowed us to determine that the antioxidative activities of the crude extract of H. sabdariffa and the Cyanidin 3- O-β- d-glucopyranoside of J. integuerrima were similar to those of ascorbic acid used as reference. To cite this article: É. Palé et al., C. R. Chimie 7 (2004) .

Characterization and quantification of anthocyanins and polyphenolics in bluehHoneysuckle (Lonicera caerulea L.).

Anthocyanins and phenolics of 10 blue honeysuckle (Lonicera caerulea L.) genotypes were characterized and quantified by HPLC-DAD. Peak assignments were confirmed by low-resolution electrospray mass spectrometry. Six anthocyanins were detected with the major peak identified as cyanidin 3-glucoside. Five additional anthocyanins were characterized as cyanidin 3,5-diglucoside, cyanidin 3-rutinoside, pelargonidin 3-glucoside, peonidin 3-glucoside, and peonidin 3-rutinoside. Four polyphenolics were identified as chlorogenic acid, neochlorogenic acid, quercetin 3-rutinoside, and quercetin 3-glucoside. Two additional unidentified phenolics were characterized as flavonol and hydroxycinnamic derivatives based on UV-vis spectra. Hydroxycinnamate levels ranged from 30.4 to 156.2 mg/100 g, whereas the flavonol content ranged from 12.6 to 32.8 mg/100 g. The L. caerulea subspecies boczkarnikovae contained the highest amounts of hydroxycinnamic derivatives and flavonols.

Characterization of Anthocyanins by NMR

Current Protocols in Food Analytical ChemistryVolume 9, Issue 1 p. F1.4.1-F1.4.23 UNIT Characterization of Anthocyanins by NMR Øyvind M. Andersen, Øyvind M. Andersen University of Bergen, Bergen, NorwaySearch for more papers by this authorTorgils Fossen, Torgils Fossen University of Bergen, Bergen, NorwaySearch for more papers by this author Øyvind M. Andersen, Øyvind M. Andersen University of Bergen, Bergen, NorwaySearch for more papers by this authorTorgils Fossen, Torgils Fossen University of Bergen, Bergen, NorwaySearch for more papers by this author First published: 01 February 2004 https://doi.org/10.1002/0471142913.faf0104s09Citations: 12Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature RelatedInformation

Characterization of anthocyanins from the fruits of baguaçu (Eugenia umbelliflora Berg).

Anthocyanin pigments in the berries of baguaçu (Eugenia umbelliflora Berg), a tropical fruit from Brazil, were extracted with 0.1% HCl in ethanol, and the crude anthocyanin extract was purified by Amberlite XAD-7 open-column chromatography. Six major anthocyanins were isolated by preparative HPLC, and their chemical structures were identified by spectroscopic methods (TLC, UV-vis, MS, and (1)H NMR). Delphinidin 3-O-beta-glucopyranoside, cyanidin 3-O-beta-glucopyranoside, petunidin 3-O-beta-glucopyranoside, pelargonidin 3-O-beta-glucopyranoside, peonidin 3-O-beta-glucopyranoside, and malvidin 3-O-beta-glucopyranoside were identified. On the basis of chromatographic data the total anthocyanin content was 342 mg/100 g of fresh baguaçu berries. Therefore, the concomitant presence of six anthocyanins in a single plant species makes this product promising as a new pigment source.

Characterization of anthocyanins in grape juices by ion trap liquid chromatography-mass spectrometry.

A reverse phase HPLC and electrospray interface with ion trap mass spectrometer method was developed for the characterization of anthocyanins in Concord, Rubired, and Salvador grape juices. Rubired and Salvador grapes are hybrids from Vitis vinifera and Vitis rupestris. Concord grape is a grape from the native American cultivar Vitis labrusca. Individual anthocyanins in these three varieties were identified on the basis of UV-vis and MS spectra and further elucidated by MS/MS spectra. Anthocyanins in Salvador and Concord grapes were 3-O-glucosides, 3-O-(6' '-O-p-coumaroyl)glucosides, 3-O-(6' '-O-p-acetyl)glucosides, 3,5-O-diglucosides, and 3-O-(6' '-O-p-coumaroyl)-5-O-diglucosides of delphinidin, cyanidin, petunidin, peonidin, and malvidin. Vitisin B was detected in Salvador grape juice. Anthocyanins in Rubired grape juice were primarily anthocyanin diglucosides: peonidin 3,5-O-diglucoside, malvidin 3,5-O-diglucoside, peonidin 3-O-(6' '-O-p-coumaroyl)-5-O-diglucoside, and malvidin 3-O-(6' '-O-p-coumaroyl)-5-O-diglucoside are the four major anthocyanins. The presence of pelargonidin 3-O-glucoside, not previously reported, has been established for the first time in all three juices.

Characterization of Anthocyanins in Flowers of Japanese Garden Iris, Iris ensata Thunb.

The flower colors of 15 cultivars and a line in Japanese garden iris, Iris ensata Thunb. were examined by CIELAB scatter diagram. The cultivars classified as anthocyanin types, malvidin 3RGac5G-petunidin 3RGac5G, malvidin 3RG5G-petunidin 3RG5G and peonidin 3RGac5G showed color distribution from purple to red-purple, while the cultivar and line in peonidin 3RG5G and peonidin 3RG5G-cyanidin 3RG5G types were purplish red to red. This indicates that cyanidin 3RG5G and peonidin 3RG5G are good anthocyanin source for the breeding of red flower color. The anthocyanins of 83 cultivars, 13 lines and 5 wild forms of I. ensata were analyzed by HPLC, and these plants were classified into 17 types of major anthocyanins. Among these, 7 new types, delphinidin 3RGac5G-delphinidin 3RG, delphinidin 3RG-petunidin 3RGac5G, petunidin 3G-delphinidin 3G, petunidin 3RGac, peonidin 3RG5G, cyanidin 3RG5G and malvidin 3RGac5G-peonidin 3RGac5G-petunidin 3RGac5G were detected, and a novel anthocyanin source of red flower, cyanidin 3RG5G was obtained for the first time.

76(P-63) チョウマメの青色花弁におけるポリアシル化アントシアニン生合成経路 : フラボノイド組成及びアントシアニン3'-グルコシルトランスフェラーゼの解析(ポスター発表の部)

76(P-63) チョウマメの青色花弁におけるポリアシル化アントシアニン生合成経路 : フラボノイド組成及びアントシアニン3'-グルコシルトランスフェラーゼの解析(ポスター発表の部)

Rapid characterization of anthocyanins in red raspberry fruit by high-performance liquid chromatography coupled to single quadrupole mass spectrometry

Anthocyanins from red raspberries were extracted from the fruit by homogenizing in acidified methanol. The methanolic extract was centrifuged and the supernatant analyzed by reversed-phase HPLC. The eluent was monitored at 371 and 520 nm before being introduced into a single quadrupole mass spectrometer through an atmospheric pressure chemical ionization probe operating in positive ion mode. This method allowed the identification of eight anthocyanins. In the absence of readily available reference compounds, approaches that can be taken to analyse anthocyanins by HPLC with absorbance and mass spectrometric detection are discussed.

Characterization of major anthocyanins and the color of red-fleshed Budd Blood orange (Citrus sinensis).

High-performance liquid chromatography (HPLC) with photodiode array detection was applied for the characterization of anthocyanins in red-fleshed Budd Blood (Citrus sinensis) orange. More than seven anthocyanin pigments were separated within 30 min by using a binary gradient (0.1% H(3)P0(4) in water and 0.1% H(3)PO(4) in acetonitrile) elution on a Prodigy ODS column. Separations by reversed-phase HPLC and semipreparative HPLC on a Prodigy 10-microm ODS Prep column, and acid and alkali hydrolyses were used for identification of anthocyanins. The primary anthocyanins in Budd Blood orange grown in Florida were cyanidin-3-(6"-malonylglucoside) (44.8%) followed by cyanidin-3-glucoside (33.6%). Two other minor pigments were also acylated with malonic acid. Malonated anthocyanins represented the major proportion (>51%) of anthocyanins in Budd Blood orange. Total anthocyanin contents and juice color parameters (CIE L,a,b) were compared with six other Florida-grown blood oranges.

Modification of flower color and fragrance by antisense suppression of the flavanone 3-hydroxylase gene

Anthocyanins are the major pigments contributing to carnation flowercoloration. Most carnation varieties are sterile and hence molecular breedingis an attractive approach to creating novel colors in this commercially importantcrop. Characterization of anthocyanins in the flowers of the modern carnationcv. Eilat revealed that only the orange pelargonidin accumulates, due to a lackof both flavonoid 3′,5′-hydroxylase and flavonoid3′-hydroxylase activities. To modify flower color in cv. Eilat, we usedantisense suppression to block the expression of a gene encoding flavanone3-hydroxylase, a key step in the anthocyanin pathway. The transgenic plantsexhibited flower color modifications ranging from attenuation to complete lossof their original orange/reddish color. In the latter, only traces ofpelargonidin were detected. Dramatic suppression of flavanone 3-hydroxylaselevel/activity in these transgenes was confirmed by northern blot, RT-PCR andenzymatic assays. The new phenotype has been stable for over 4 years ofvegetative propagation. Moreover, transgenic plants with severe colormodification were more fragrant than control plants. GC-MS headspace analysesrevealed that transgenic anti-f3h flowers emit higherlevels of methyl benzoate. The possible interrelation between pathways leadingto anthocyanin and fragrance production is discussed.

Use of modern nuclear magnetic resonance spectroscopy in wine analysis: determination of minor compounds

In our work, the capacity of 1D and 2D 1 H and 13 C homo and heteronuclear nuclear magnetic resonance (NMR) experiments were examined for the characterization of amino acids and anthocyanins, the “minor” but important compounds of wine. The compositions of amino acids and anthocyanins influence the flavour of wines and their composition could be used to differentiate wines according to the vine variety, geographical origin and year of production. Anthocyanins are also the major components responsible for the red wine color and they have a wide range of biochemical and pharmacological effects such as antimicrobial and antioxidant activities. We use the signal intensities in 1D 1 H NMR spectra in the chemometric analysis of white wines from Slovenia. The use of signals of seven amino acids gives a good separation of wines according to the vine variety, while adding also the signal intensities of glycerol, butylene glycol and succinic acid, the separation of wines between the Coastal and continental part of Slovenia was achieved. By using 1D and 2D NMR experiments the reliable assignment of 1 H and 13 C resonances of anthocyanins, peonidine 3-glucoside, petunidine 3-glucoside, cyanidine 3-glucoside, delphinidine 3-glucoside and malvidine 3-glucoside in CD 3OD was established, which will serve as a tool for the identification of a particular fraction in liquid chromatographic analysis.

Separation and Characterization of Anthocyanins by HPLC

Separation and Characterization of Anthocyanins by <scp>HPLC</scp>

Characterization and Measurement of Anthocyanins by UV‐Visible Spectroscopy

Current Protocols in Food Analytical ChemistryVolume 00, Issue 1 p. F1.2.1-F1.2.13 UNIT Characterization and Measurement of Anthocyanins by UV-Visible Spectroscopy M.Mónica Giusti, University of Maryland, College Park, MarylandSearch for more papers by this authorRonald E. Wrolstad, Oregon State University, Corvallis, OregonSearch for more papers by this author M.Mónica Giusti, University of Maryland, College Park, MarylandSearch for more papers by this authorRonald E. Wrolstad, Oregon State University, Corvallis, OregonSearch for more papers by this author First published: 01 August 2001 https://doi.org/10.1002/0471142913.faf0102s00Citations: 510Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinked InRedditWechat Citing Literature RelatedInformation

Isolation and characterization of anthocyanin 5- O-glucosyltransferase from flowers of Dahlia variabilis

In the cyanic flowers ofDahlia variabilis (Asteraceae), an enzyme was demonstrated which catalyzes a glucosyl group transfer from UDP-glucose to the 5 position of anthocyanidin 3-O-glucoside and 3-O-malonylglucoside. The anthocyanin 5-O-glucosyltransferase (5GT) was purified 88-fold at 8 percnt; yield by (NH4)2SO4 precipitation followed by successive chromatography on DEAE-cellulose, Sephacryl S-200 and Mono P. 5GT exhibited a pH optimum at 8.0 and a pI of 4. 2. Its apparent molecular weight calculated from Sephacryl S-200 was 53 kDa. Its activity was stimulated by 2-ME and DTE but strongly inhibited by PCMB and NEM. It was slightly activated by Mg2+ and Ca2+ but strongly inhibited by Hg2+, Zn2+, Cu2+, Mn2+, Fe3+ and Al3+. No effect of EDTA was observed. The apparent Km values for cyanidin 3-O-glucoside, cyanidin 3-O-(6′′-O-malonyl)glucoside and UDP-glucose were 120 μmol/L, 75 μmol/L and 250 μmol/L, respectively. Pelargonidin 3-O-glucoside and malonylglucoside were also considerable substrates, but low relative activity was observed for delphinidin 3-O-glucoside which has yet not been found inDahlia flowers. Dahlia 5GT showed substrate specificities different from those reported forSilene, Petunia, Matthiola andPerilla. Neither ADP-glucose nor UDP-galactose could serve as glycosyl donor.

Characterisation of anthocyanins derived from carrot (Daucus carota) cell culture

Abstract Two anthocyanin pigments were isolated from cell cultures of Carrot, Nentes scarlet-104 local variety. Chemical hydrolysis, column and paper chromatography, HPLC, proton and 13C NMR and mass spectroscopic studies indicated the presence of cyanidin-3-lathyroside [cyanidin-3-0{β- d -xylopyranosyl (1→2) β- d -galacto pyranoside}] (90%) and cyanidin-3-β- d -glucopyranoside (10%) in the callus cultures, whereas only cyanidin-3-lathyroside (0.05%) was found in the explant carrot. The significant difference was that there was no acylated anthocyanin present as reported in other varieties of carrot.

Electrospray and tandem mass spectroscopy as tools for anthocyanin characterization.

The utility of electrospray and tandem mass spectroscopy (ES-MS and MS-MS) in anthocyanin characterization was tested using different anthocyanin extracts. Anthocyanins were semipurified by using a C-18 resin, washed with acidified water followed by ethyl acetate, and recovered with acidified methanol. Samples were directly injected into a mass spectrometer in either aqueous or methanolic solutions. The positive charge of anthocyanins favored fast and effective ES-MS detection of intact molecular ions. Little interference from other compounds was observed when the ethyl acetate cleaning procedure was used. Tandem mass spectroscopy provided clear and characteristic fragmentation patterns. The voltage used affected only the proportions at which these fragments were present. ES-MS may be used as a fast procedure for identification of anthocyanins, requiring minimal sample preparation. In combination with HPLC, ES-MS and MS-MS could be very powerful tools for anthocyanin characterization and monitoring the authenticity of anthocyanin-containing fruit juices and vegetable extracts.