Comparison between HPLC and MALDI-TOF MS analysis of anthocyanins in highbush blueberries

High-performance liquid chromatography (HPLC) has been widely used as a reliable technique to quantify anthocyanins in food samples. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a new technique that is having a great impact on food analysis. This study is the first to compare HPLC and MALDI-TOF MS quantifications of anthocyanins. The analyses were carried out for highbush blueberries at different stages of anthocyanin formation. In general, both techniques provided comparable quantitative anthocyanin profiles for the samples. HPLC could distinguish anthocyanin isomers, whereas MALDI-TOF MS proved to be more rapid in the accurate identification and quantification of anthocyanins with different masses. A single MALDI-TOF MS run took just 4 min. MALDI-TOF MS analysis can serve as a rapid alternative to HPLC for the analysis of anthocyanins in fruits.     

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.     

Anthocyanins in wild blueberries of Quebec: extraction and identification

Anthocyanins were extracted from a mixture of berries of Vaccinium angustifolium and Vaccinium myrtillo?des at 7.7 degrees C, 26 degrees C, and 79 degrees C using ethanol alone or ethanol acidified with hydrochloric, citric, tartaric, lactic, or phosphoric acids at a solvent to solid ratio of 10. The effect of these parameters on extracted anthocyanins stability was investigated. The pH-differential and HPLC-DAD methods were used to determine anthocyanin contents. Extracted anthocyanins were purified on a C-18 solid-phase extraction cartridge and characterized by HPLC/electrospray ionization/mass spectrometry (HPLC-ESI-MS/MS). Anthocyanins were identified according to their HPLC retention times, elution order, and MS fragmentation pattern and by comparison with standards and published data. Anthocyanin extractions gave different yields depending on the type of added acid and the extraction temperature. High yields of monomeric and total anthocyanins (26.3 and 28.9 mg/g of dry matter) were obtained at 79 degrees C using phosphoric acid. Extraction using tartaric acid at 79 degrees C provided the lowest degradation index (1.05). Anthocyanins were stable and browning by polyphenol oxidase was inhibited under these conditions. Of the six common anthocyanindins, five were identified in the extracts, namely, delpinidin, cyanidin, peonidin, petunidin, and malvidin; pelargonidin was not found. In addition to well-known major anthocyanins, new anthocyanins were identified for the first time in extracts of wild blueberries from Quebec.     

Determination of anthocyanins from camu-camu (Myrciaria dubia) by HPLC-PDA, HPLC-MS, and NMR

Camu-camu [Myrciaria dubia (HBK) McVaugh] is a small fruit native to the Amazonian rain forest. Its anthocyanin profile has now been investigated for the first time. Fruits from two different regions of the S?o Paulo state, Brazil, were analyzed. The major anthocyanins were isolated by high-speed countercurrent chromatography. HPLC-PDA, HPLC-MS/MS, and 1H NMR were used to confirm the identity of the main anthocyanins of camu-camu. Cyanidin-3-glucoside was identified as the major pigment in the fruits from both regions, representing 89.5% in the fruits produced in Iguape and 88.0% in those from Mirandópolis, followed by the delphinidin-3-glucoside, ranging between 4.2 and 5.1%, respectively. Higher total anthocyanin contents were detected in the fruits from Iguape (54.0 +/- 25.9 mg/100 g) compared to those from Mirandópolis (30.3 +/- 6.8 mg/100 g), most likely because of the lower temperatures in the Iguape region.     

Anthocyanins present in selected tropical fruits: acerola, jambolão, jussara, and guajiru

Many tropical fruits are rich in anthocyanins, though limited information is available about the characterization and quantification of these anthocyanins. The identification and quantification of anthocyanin pigments in four tropical fruits were determined by HPLC-MS/MS. Fruits studied included acerola (Malphigia emarginata), jussara (Euterpe edulis), jambol?o (Syzygium cumini), and guajiru (Chrysobalanus icaco). All four fruits were found to contain anthocyanin pigments. Anthocyanidin backbones included cyanidin, delphinidin, peonidin, pelargonidin, petunidin, and malvidin. Guajiru contained several acylated forms, while acerola, jussara, and jambol?o contained only nonacylated glycosides. These results demonstrate that these tropical fruits are rich in anthocyanins and that the anthocyanins are widely ranging in anthocyanidin backbone, glycosylation, and acylation.     

小叶女贞果实花青素组分鉴定及色谱纯化技术

为提高小叶女贞果实的食用、药用价值,该文系统研究了果实中花青素种类构成及提取物的制备技术。试验采用紫外可见光谱法、高效液相色谱-质谱串联法、酸水解制备苷元等技术对小叶女贞果实花青素含量、单体种类进行了测定,并借助提取、萃取、柱层析等技术研究了花青素提取物的分离纯化过程。研究结果如下:测得每100 g小叶女贞果实中含花青素总量为(499±18.42)mg,从中鉴定出2种花青素单体,分别为矢车菊素-3-O-葡萄糖苷和牵牛花色素-3-O-葡萄糖苷,并以后者为主要存在形式;获得了纯天然、简单易行的花青素提取物制备技术,主要包括酸化乙醇提取、乙酸乙脂萃取、Amberlite XAD-7HP大孔树脂层析分离步骤,最终制得的花青素提取物纯度为35%、得率为0.6%。该研究为后期制备高纯度牵牛花素-3-O-葡萄糖苷单体提供了良好原料基础,为深入研究小叶女贞果实花青素功能活性及其在食品、药品领域潜在应用提供了参考。     

Probing anthocyanin profiles in purple sweet potato cell line (Ipomoea batatas L. Cv. Ayamurasaki) by high-performance liquid chromatography and electrospray ionization tandem mass spectrometry

A purple line cell line (PL) generated from the storage root of purple-fleshed sweet potato (Ipomoea batatas L.) cv. Ayamurasaki produces a complex mixture of anthocyanins, and seven major anthocyanins have been isolated and identified to date. All these anthocyanins are exclusively cyanidin or peonidin 3-sophoroside-5-glucosides and their acylated derivatives. High-performance liquid chromatography (HPLC) coupled to photodiode array (PDA) detection and electrospray ionization tandem mass spectrometry (ESI-MS/MS) on a triple quadrupole instrument was employed to further investigate the anthocyanin composition of the PL extract. Precursor-ion analysis, product-ion analysis, and selected reaction monitoring (SRM) MS/MS experiments were conducted sequentially to screen and characterize anthocyanins in the aqueous extract of the PL cell line. Precursor-ion analysis specifically detected the molecular cations of each category of anthocyanins by scanning the precursors of anthocyanidins (cyanidin, peonidin, and pelargonidin). The detected molecular cation of each anthocyanin was fragmented using product-ion analysis by collisionally activated dissociation (CAD). MS/MS using SRM detection was conducted to further confirm the fragmentation observed during product-ion analysis. In comparison to the commonly used product-ion analysis technique, the combined use of precursor-ion analysis, product-ion analysis, and SRM is particularly useful for positive identification of anthocyanins in complex matrixes and provides important information to confirm the proposed structures. Twenty-six anthocyanins were detected and characterized in the aqueous extract of the PL cell line. Several anthocyanins, including two pelargonidin derivatives, were tentatively identified for the first time in these cells.     

Characterization of anthocyanins and proanthocyanidins in some cultivars of Ribes, Aronia, and Sambucus and their antioxidant capacity

Anthocyanins and proanthocyanidins were characterized by HPLC-ESI-MS/MS coupled with a diode array and/or fluorescent detector in seven cultivars of Ribes nigrum (black currant) and Ribes rubrum (red currant, Red Lake), six cultivars of Ribes grossularia (gooseberries), Aronia melanocarpa(chokeberry), and Sambucus nigra (elderberry). Thirty-one different anthocyanins were detected in these berries, but not every anthocyanin was observed in each berry. A number of minor anthocyanins were identified from these berries for the first time. The concentrations of individual anthocyanins in all of the berries were quantified using relevant anthocyanidin 3-glucoside standards. Among the berries studied in this paper and in berries in general, chokeberry has the highest total anthocyanin concentrations [1480 mg/100 g of fresh weight (FW)], whereas the lowest total anthocyanin concentration in the berries studied was found in the gooseberry cv. Careless, which contained only 0.07 mg/100 g of FW. Two cultivars of gooseberries (Marigold and Leveller) did not contain any anthocyanins. Total proanthocyanidin concentrations in the berries studied ranged from 23 to 664 mg/100 g of FW in elderberry and chokeberry, respectively. Procyanidin or prodelphinidin polymers were the predominant components (>65% w/w) in most of the berries. The lipophilic and hydrophilic antioxidant capacities were measured by the oxygen radical absorbance capacity (ORAC(FL)) procedure. The total antioxidant capacity varied from 21 micromol of TE/g of FW in Careless gooseberry to 161 micromol of TE/g of FW in chokeberry. Total phenolics in the berries in general paralleled hydrophilic antioxidant capacity.     

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.     

红叶葡萄品种叶片花色苷种类的鉴定与分析

为解决葡萄园中由葡萄修剪产生的大量叶片残留问题,提高修剪产生的叶片的利用价值,本试验以8612、玫瑰香、红鸡心3个红叶葡萄品种和葡萄园皇后、玫瑰露、秋红3个绿叶葡萄品种不同发育时期的叶片为研究对象,利用MATLAB软件对整张叶片的图像数据进行提取,计算色差值;利用超高效液相-质谱法(LC-MS)检测花色苷的成分和含量,探讨不同发育时期叶片中花色苷不同组分的变化规律。结果表明,不同发育时期的红色和绿色叶片色差指数L^*、a^*、b^*变化趋势明显不同,共检测到18种花色苷组分,包括花青素类(4种)、甲基花青素类(4种)、花翠素类(4种)、甲基花翠素类(2种)和二甲基花翠素类(4种)。以上5类花色苷在红色叶片中均被检测到,绿叶品种中未发现花翠素类和甲基花翠素类花色苷。花色苷定量结果显示,红色叶片不同发育时期花色苷含量为123.468~855.001 mg·100g^-1,绿色叶片花色苷含量为4.407~44.517 mg·100g^-1。甲基化类花色苷占比随叶片发育均逐渐增大。花色苷酰化修饰类型分析结果发现,香豆酰化类型花色苷含量高于其他酰化类型花色苷,在花色苷总含量中所占比例较高,而阿魏酰化和糖酰化类花色苷含量非常少。色差和不同类型花色苷成分的相关性分析结果表明,红色叶片的色差指数与更多类型的花色苷含量存在相关关系。红色葡萄叶片中花色苷种类丰富、含量较高,是花色苷类化合物的潜在来源,具有很大的利用价值。本研究通过对叶片中花色苷成分和含量进行了详细调查,为今后葡萄园中叶片的加工再利用提供了依据。     

HPLC-PDA-MS/MS of anthocyanins and carotenoids from dovyalis and tamarillo fruits

Anthocyanins and carotenoids are natural pigments responsible for the color of vegetables and fruits, and they are also bioactive compounds, both demonstrating important biological, therapeutic, and preventative properties. Considering the biodiversity of edible fruits, high performance liquid chromatography coupled to photodiode array and mass spectrometry detectors (HPLC-PDA-MS) was used to establish the composition of carotenoids and anthocyanins from dovyalis and tamarillo fruits. Ten anthocyanins and 26 carotenoids were found in dovyalis, whereas tamarillo showed 3 anthocyanins and 17 carotenoids. Higher contents of anthocyanins and carotenoids were found in dovyalis, 42.0 and 6.6 mg/100 g, respectively, as compared to tamarillo fruits with 8.5 and 4.4 mg/100 g, respectively. Although these fruits belong to different families, delphinidin 3-rutinoside and beta-cryptoxanthin were found to be, respectively, the major anthocyanin and carotenoid in both fruits.     

Anthocyanins from new red leaf tea 'Sunrouge'

New red leaf tea cultivar 'Sunrouge' (Camellia taliensis × Camellia sinensis), for which an application for registration was made in 2009, is an anthocyanin-rich tea. The anthocyanin content of 'Sunrouge' was the highest among 4 tea cultivars, and was 8.4 times higher than that of 'Yabukita'. We purified and isolated 6 anthocyanins from 'Sunrouge' by chromatography, and identified them by LC/MS/MS and NMR analysis. As a result, the four anthocyanins were identified as delphinidin-3-O-β-D-(6-(E)-p-coumaroyl)galactopyranoside (2), delphinidin-3-O-β-D-(6-(E)-p-coumaroyl)glucopyranoside (3), cyanidin-3-O-β-D-(6-(E)-p-coumaroyl)galactopyranoside (4), and cyanidin-3-O-β-D-(6-(E)-p-coumaroyl)glucopyranoside (5), and the other two were estimated as delphinidin-(Z)-p-coumaroylgalactopyranoside (1), petunidin-(E)-p-coumaroylgalactopyranoside (6). Compound 3 was found in tea for the first time. In general, anthocyanins have various bioactivities, including relieving eyestrain and antioxidative effects, so it is expected that drinking 'Sunrouge' tea brings in similar bioactivities.     

Pigments in the fruit of red-fleshed kiwifruit (Actinidia chinensis and Actinidia deliciosa)

Kiwifruit cultivars (Actinidia chinensis and A. deliciosa) generally have fruit with yellow or green flesh when ripe. A small number of genotypes also have red pigments, usually restricted to the inner pericarp but varying in intensity and in distribution within the fruit. Carotenoids, chorophylls, and anthocyanins were extracted from the fruit pericarp of such red-fleshed kiwifruit selections. Pigments were analyzed by HPLC and identified by comparison with authentic standards and by liquid chromatography-mass spectroscopy to obtain a tentative identification of the major anthocyanins in red-fleshed kiwifruit. The yellow and green colors of the outer fruit pericarp are due to different concentrations and proportions of carotenoids and chlorophylls. The red color found mainly in the inner pericarp is due to anthocyanins. In the A. chinensis genotypes tested the major anthocyanin was cyanidin 3-O-xylo(1-2)-galactoside, with smaller amounts of cyanidin 3-O-galactoside. In the A. deliciosa genotypes analyzed, cyanidin 3-O-xylo(1-2)-galactoside was not detected; instead, the major anthocyanins identified were cyanidin 3-O-galactoside and cyanidin 3-O-glucoside. However, the two species did not differ consistently in anthocyanin composition.     

Anthocyanin metabolism in rats and their distribution to digestive area, kidney, and brain

Anthocyanins are present in human diet due to their wide occurrence in fruits and beverages. They possess antioxidant activities and could be involved in several health effects. The aim of this study was to investigate anthocyanin metabolism and distribution in the digestive area organs (stomach, jejunum and liver) and kidney, as well as a target tissue (brain) in rats fed with a blackberry (Rubus fruticosus L.) anthocyanin-enriched diet for 15 days. Identification and quantification of anthocyanin metabolites was carried out by HPLC-ESI-MS-MS and HPLC-DAD, respectively. The stomach exhibited only native blackberry anthocyanins (cyanidin 3-O-glucoside and cyanidin 3-O-pentose), while in other organs (jejunum, liver, and kidney) native and methylated anthocyanins as well as conjugated anthocyanidins (cyanidin and peonidin monoglucuronides) were identified. Proportions of anthocyanin derivatives differed according to the organ considered, with the liver presenting the highest proportion of methylated forms. Jejunum and plasma also contained aglycone forms. In the brain, total anthocyanin content (blackberry anthocyanins and peonidin 3-O-glucoside) reached 0.25 +/- 0.05 nmol/g of tissue (n = 6). The urinary excretion of total anthocyanins was low (0.19 +/- 0.02% of the ingested amount). Thus, organs of the digestive area indicated a metabolic pathway of anthocyanins with enzymatic conversions (methylation and/or glucurono-conjugation). Moreover, following consumption of an anthocyanin-rich diet, anthocyanins enter the brain.     

Identification of anthocyanins in berries by narrow-bore high-performance liquid chromatography with electrospray ionization detection

Qualitative determination of anthocyanins in extracts of red fruits by narrow-bore HPLC/ESI-MS was carried out. This method was used to investigate anthocyanin contents of black bilberry (Vaccinium myrtillus L.), blackberry (Rubus sp.), and mulberry (Morus nigra). An ultraviolet diode array and a mass spectrometer with ESI source were used for detection. Anthocyanin identifications were made by using retention time data and UV-vis and mass spectra and comparing them with those of commercially available standard compounds. The method allowed the identification of fourteen anthocyanins in black bilberry extract, six anthocyanins in blackberry extract, and five anthocyanins in mulberry extract.     

刺葡萄皮中花色苷的分离纯化与结构鉴定

为研究刺葡萄花色苷的结构及其纯化分离的柱层析法,将刺葡萄色素粗提液依次经大孔树脂HP-20、聚酰胺树脂、葡聚糖凝胶Sephadex LH-20吸附纯化,利用超高效液相色谱三重四级杆飞行时间质谱联用技术对分离所得花色苷进行结构鉴定,并运用荧光光度法探索荧光图谱与花色苷结构的关系。研究发现,聚酰胺树脂对部分花色苷产生了吸附作用,而Sephadex LH-20凝胶能起到较好的分离作用,最终得到3种色素,经鉴定,确定色素I为锦葵素-3,5-O-双葡萄糖苷,通过质谱信息初步确定色素III可能为锦葵素-3,5-O-双葡萄糖苷-香豆酰,色素IV可能为飞燕草素-3-O-芸香糖苷和锦葵素-3-O-芸香糖苷的混合物,经高效液相色谱以归一法计算峰面积,色素I和色素III的纯度分别达到了98.64%、98.33%,得率分别为0.114%和0.076%。研究结果为花色苷的分离及鉴定提供参考。     

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

Anthocyanin pigments in the berries of bagua?u (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 bagua?u berries. Therefore, the concomitant presence of six anthocyanins in a single plant species makes this product promising as a new pigment source.     

Peroxyl radical scavenging capacity, polyphenolics, and lipophilic antioxidant profiles of mulberry fruits cultivated in southern China

Twenty-seven cultivars of mulberry fruits ( Morus atropurpurea Roxb) were analyzed for their total phenolic content, total anthocyanin content, and peroxyl radical scavenging capacities. The proanthocyanidin contents of the fruit were also quantified using 4-dimethylamino-cinnamaldehyde assay, and characterization was attempted using electrospray ionization mass spectra. The phenolic compounds of mulberry fruits were characterized using HPLC with ESI-MS and diode array detection. Results showed that the content of mulberry fruits varied with different cultivars with total phenolic content, total anthocyanin content, total proanthocyanidin content, and peroxyl radical scavenging capacities ranging from 0.060-0.244, 0.001-0.056, 0.001-0.015, and 0.301-1.728, respectively. Good correlations were observed among the phenolic, anthocyanin, and proanthocyanidin contents and the radical scavenging capacities of mulberry fruits. Mulberry fruits were found to contain low amount of proanthocyanidins. The high total phenolic content of mulberry fruits were mainly contributed by anthocyanins, rutin, and chlorogenic acids. The lipid soluble antioxidants are profiled by an HPLC method developed in-house, and the results of selected mulberry fruits revealed significant amounts of lutein and delta- and gamma-tocopherols but low alpha-tocopherol. Our results provide useful antioxidant nutritional information of a mulberry cultivar that has potential for large scale plantations.     

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.     

Anthocyanin transformation in Cabernet Sauvignon wine during aging

Anthocyanins in Cabernet Sauvignon grapes and wines were elucidated by HPLC-MS/MS. Major anthocyanins in Cabernet Sauvignon grape extract are malvidin 3-O-glucoside and malvidin 3-O-acetylglucoside. In matured wine, anthocyanins are transformed to anthocyanin-vinyl derivatives, ethyl bridged anthocyanin-flavanol adducts, and anthocyanin-flavanol adducts. The major anthocyanin pigments are malvidin 3-O-glucoside-pyruvate, malvidin 3-O-acetylglucoside-pyruvate, malvidin 3-O-coumaroylglucoside-pyruvate, malvidin 3-O-glucoside-4-vinylphenol, malvidin 3-O-acetylglucoside-4-vinylphenol, and malvidin 3-O-coumaroylglucoside-4-vinylphenol. The presence of syringetin 3-O-glucoside and syringetin 3-O-acetylglucoside has been established for the first time in grape and wine.