Separation and purification of anthocyanins by high-speed countercurrent chromatography and screening for antioxidant activity

The all-liquid chromatographic technique of high-speed countercurrent chromatography (HSCCC) has been applied for separations of anthocyanins. The biphasic mixture of tert-butyl methyl ether/n-butanol/acetonitrile/water (2:2:1:5) acidified with trifluoroacetic acid was found to be a suitable solvent system for anthocyanin separation. In some cases, enrichment of the pigments on Amberlite XAD-7 resin prior to HSCCC has been carried out. The anthocyanin mixtures from red cabbage, black currant, black chokeberry, and roselle were successfully fractionated using HSCCC. Peak purity control was done by nuclear magnetic resonance spectroscopy as well as electrospray ionization ion trap multiple mass spectrometry. Finally, antioxidant activity of the purified pigments was determined using the Trolox equivalent antioxidant capacity test.     

Fractionation of grape anthocyanin classes using multilayer coil countercurrent chromatography with step gradient elution

Grape anthocyanins from rosé wine pomace and grape skins were fractionated by multilayer coil countercurrent chromatography (MLCCC). Tert-butyl methyl ether/n-butanol/acetonitrile/water acidified with trifluoroacetic acid (2/2/x/5) was chosen as the solvent system because of its demonstrated efficiency in separating anthocyanins. A method combining MLCCC and step gradient elution has been developed. This method enabled the fractionation of anthocyanins as series of glucosides, and the corresponding acetylated, coumaroylated, and caffeoylated derivatives. The different anthocyanins were identified on the basis of their UV spectra, their elution time on reversed-phase HPLC as well as by mass spectrometry. A polar-colored fraction free of anthocyanin monomers and corresponding to the material remaining in the stationary phase after the completion of the gradient was recovered.     

New phenolic grape skin products from Vitis vinifera cv. Pinot Noir

Anthocyanins and their related compounds were extracted from grape skins of Pinot noir, using 50% aqueous methanol, and purified by solid phase extraction chromatography using XAD-7 resin to obtain a pigment-rich fraction. This fraction was subjected to multilayer coil countercurrent chromatography (MLCCC) using a quaternary solvent system consisting of tert-butyl methyl ether/n-butanol/acetonitrile/water acidified with 0.01% trifluoroacetic acid (2:2:0.1-1.8:5) (v/v/v/v) in a step gradient elution to separate anthocyanin oligomers from grape anthocyanins. In the process of the characterization of the MLCCC fractions by electrospray mass spectrometry, two noncolored anthocyanin derivatives were found and characterized on the basis of their mass spectral data. As a result, these compounds have been tentatively identified as coupling products between both hydrated malvidin-3-glucoside and peonidin-3-glucoside, with 2-S-glutathionyl caffeoyl tartaric acid (GRP). It is therefore proposed that grape skins contain this new class of coupling product, and a possible chemical pathway for their formation is suggested.     

Isolation and determination of anthocyanins in seed coats of black soybean (Glycine max (L.) Merr.).

Anthocyanin pigments in seed coats of black soybean (Glycine max (L.) Merr.) were extracted with 1% HCl-CH(3)OH, and the crude anthocyanin extract was purified by Shepadex LH-20 and Lichroprep RP-18 open-column chromatography. Three major anthocyanins were isolated, and their chemical structures were identified by spectroscopic methods (UV-visible, FABMS, (1)H and (13)C NMR, and by TLC). The complete structures of these anthocyanins were elucidated as delphinidin-3-glucoside, cyanidin-3-glucoside, and petunidin-3-glucoside. Among them, petunidin-3-glucoside was identified as a new anthocyanin in black soybeans. On the basis of RP-HPLC with a UV-vis detector, the contents of delphinidin-3-glucoside, cyanidin-3-glucoside, petunidin-3-glucoside, and total anthocyanins in seed coats of 10 black soybeans were found in the ranges of 0-3.71, 0.94-15.98, 0-1.41, and 1.58-20.18 mg/g, respectively. The results obtained in this study imply that the seed coats of black soybean can be used as a good source for cyanidin-3-glucoside and delphinidin-3-glucoside.     

Anthocyanins from black currants (Ribes nigrum L.)

Fifteen anthocyanin structures are reported from an extract of black currant berries (Ribes nigrum L.). These are the 3-O-glucosides and the 3-O-rutinosides of pelargonidin, cyanidin, peonidin, delphinidin, petunidin, and malvidin, cyanidin 3-O-arabinoside, and the 3-O-(6' '-p-coumaroylglucoside)s of cyanidin and delphinidin. The anthocyanins were characterized by means of size exclusion chromatography, high-performance liquid chromatography, UV-visible spectroscopy, and electrospray mass spectrometry. The four main pigments (the 3-O-glucosides and the 3-O-rutinosides of delphinidin and cyanidin) made up >97% of the total anthocyanin content. The minor pigments were enriched from the extract by successive partition against ethyl acetate and by gel fractionation. These chromatographic steps were successfully used to isolate the acylated anthocyanins from the ethyl acetate layer and to separate cyanidin 3-O-arabinoside from the mixture of anthocyanins. The amounts of anthocyanin rutinosides were found to be higher than the amount of the corresponding glucosides for all detected pigments having the same aglycon moiety.     

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.     

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.     

Bioavailability and tissue distribution of anthocyanins in bilberry (Vaccinium myrtillus L.) extract in rats

To clarify how structural diversity of anthocyanins relates to their in vivo function, bioavailability was precisely studied in rats using bilberry (Vaccinium myrtillus L.) extract (Bilberon 25) as an anthocyanin source that contains 15 different anthocyanins. The bilberry extract was orally or intravenously administered to rats, and the plasma levels of each anthocyanin were determined by high-performance liquid chromatography. As the result, all anthocyanins except peonidin 3-O-alpha-L-arabinoside were detectable in the blood plasma. The plasma concentration of anthocyanins as a whole reached the maximum level of 1.2 microM at 15 min after oral administration of 400 mg/kg bilberry extract (153.2 mg/kg as anthocyanins) and then decreased with time. Uptake and decay profiles of each anthocyanin in the plasma were almost the same for all anthocyanins except a few with their maximum after 30 min. Among the anthocyanins carrying the same aglycone, the plasma level after 15 min of oral administration was as follows: galactoside > glucoside > arabinoside. Plasma clearance of anthocyanins after intravenous administration clearly showed that arabinoside disappeared more rapidly than glucoside and galactoside. On the other hand, when anthocyanins carrying the same sugar moiety were compared, the half disappearance time of plasma anthocyanins was in the following order: delphinidin > cyanidin > petunidin = peonidin > malvidin. The bioavailability of anthocyanins was in the range of 0.61-1.82% and was 0.93% as the anthocyanin mixture. The bioavailability of anthocyanins carrying the same aglycone was in the following order: Galactoside showed the highest followed by glucoside and arabinoside for cyanidin and delphinidin, but arabinoside and galactoside showed a higher bioavailability than glucoside for petunidin and malvidin. Anthocyanins recovered in urine and bile during the first 4 h after intravenous administration were only 30.8 and 13.4%, respectively. Anthocyanin profiles in tissues were quite different from those in blood plasma. The major anthocyanins distributed in liver and kidney were the O-methyl anthocyanins such as peonidin, malvidin, and other O-methyl anthocyanins derived from delphinidin, cyanidin, and petunidin-glycosides.     

Development of analyses by high-performance liquid chromatography and liquid chromatography/tandem mass spectrometry of bilberry (Vaccinium myrtilus) anthocyanins in human plasma and urine

Anthocyanins are potent antioxidants that may possess chronic disease preventive properties. Here, rapid, reliable, and reproducible solid-phase extraction, high-performance liquid chromatography (HPLC), and mass spectrometry techniques are described for the isolation, separation, and identification of anthocyanins in human plasma and urine. Recoveries of cyanidin-3-glucoside (C3G) were 91% from water, 71% from plasma, and 81% from urine. Intra- and interday variations for C3G extraction were 9 and 9.1% in plasma and 7.1 and 9.1% in urine and were less than 15% for all anthocyanins from a standardized bilberry extract (mirtoselect). Analysis of mirtoselect by HPLC with UV detection produced spectra with 15 peaks compatible with anthocyanin components found in mirtoselect within a total run time of 15 min. Chromatographic analysis of human urine obtained after an oral dose of mirtoselect yielded 19 anthocyanin peaks. Mass spectrometric analysis employing multiple reaction monitoring suggests the presence of unchanged anthocyanins and anthocyanidin glucuronide metabolites.     

Comparison of anthocyanin pigment and other phenolic compounds of Vaccinium membranaceum and Vaccinium ovatum native to the Pacific Northwest of North America

Two huckleberry species, Vaccinium membranaceum and Vaccinium ovatum, native to Pacific Northwestern North America, were evaluated for their total, and individual, anthocyanin and polyphenolic compositions. Vaccinium ovatum had greater total anthocyanin (ACY), total phenolics (TP), oxygen radical absorbing capacity (ORAC), and ferric reducing antioxidant potential (FRAP) than did V. membranaceum. The pH and degrees Brix were also higher in V. ovatum. Berry extracts from each species were separated into three different fractions--anthocyanin, polyphenolic, and sugar/acid-by solid-phase extraction. The anthocyanin fractions of each species had the highest amount of ACY, TP, and antioxidant activity. Each species contained 15 anthocyanins (galactoside, glucoside, and arabinoside of delphinidin, cyanidin, petunidin, peonidin, and malvidin) but in different proportions. Their anthocyanin profiles were similar by high-performance liquid chromatography with photodiode array detection (LC-DAD) and high-performance liquid chromatography with photodiode array and mass spectrometry detections (LC-DAD-MS). Each species had a different polyphenolic profile. The polyphenolics of both species were mainly composed of cinnamic acid derivatives and flavonol glycosides. The major polyphenolic compound in V. membranaceum was neochlorogenic acid, and in V. ovatum, chlorogenic acid.     

Effect of anthocyanin fractions from selected cultivars of Georgia-grown blueberries on apoptosis and phase II enzymes

In recent years, considerable attention has been paid to anthocyanins due to their abilities to inhibit oxidative stress and cell proliferation. The regulations of apoptosis and the phase II enzymes glutathione-S-transferase (GST) and quinone reductase (QR) are other potential mechanisms through which flavonoids such as anthocyanins may prevent cancer. Our study confirmed that anthocyanin fractions from high bush blueberry cultivars increased apoptosis using two different methods: DNA fragmentation and caspase-3 activity. The effect of anthocyanins on the activity of the detoxifying enzymes GST and QR was also determined. Major anthocyanins identified were delphinidin, cyanidin, peonidin, petunidin, and malvidin. In Tifblue and Powderblue cultivars, DNA fragmentation increased at anthocyanin concentrations from 50 to 150 microg/mL, but cells treated with the anthocyanin fraction of Brightblue and Brightwell showed a prominent ladder at 50-100 microg/mL when compared to cells treated with 150 microg/mL. There was a significant difference in the caspase-3 activity (P < 0.05) between the control cells and the cells treated with anthocyanins from all of the cultivars. The response correlated positively with dose. The QR activity was lower in all cells treated with an anthocyanin fraction from Tifblue, Powderblue, Brightblue, and Brightwell cultivars than in control cells (P < 0.05). The activity decreased gradually when treated with increased concentrations of anthocyanin fractions (50-150 microg/mL) in the Tifblue and Powderblue cultivars. The GST activity was lower (P < 0.05) in cells treated with anthocyanin fractions from all of the cultivars and at all concentrations. These results indicated that apoptosis was confirmed in HT-29 cells when treated with anthocyanins from blueberry cultivars at 50-150 microg/mL concentrations, but these same concentrations decrease QR and GST activities rather than induce them.     

Analysis of anthocyanins in rat intestinal contents--impact of anthocyanin chemical structure on fecal excretion

Absorption of dietary anthocyanins is limited; however, fecal anthocyanin excretion has been rarely studied. We developed a method for extraction and analysis of fecal anthocyanins. Aqueous methanol (60%) maximized extraction efficiency (approximately 88%). Severe anthocyanin degradation (monitored by high-performance liquid chromatography) was observed in feces stored at -18 degrees C; therefore, storage time should be minimized and lower temperatures used. Fecal and cecal content samples were collected from 32 rats receiving either chokeberry, bilberry, grape-enriched (3.85 g monomeric anthocyanin per kg diet), or control diet for 14 weeks. Fecal anthocyanin concentrations were significantly different among groups (0.7/1.8/2.0 g/kg wet feces, chokeberry/bilberry/grape). Anthocyanin profiles of cecal contents and feces were similar. Losses in the intestinal contents were high for anthocyanin glucosides, moderate for galactosides, and negligible for arabinosides or xylosides. Acylation or diglucosylation enhanced anthocyanin stability. High anthocyanin concentration in the fecal content may favor anthocyanin absorption into the colon epithelial cells, resulting in potential health benefits.     

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 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.     

Composition and stability of anthocyanins in blue-grained wheat

Wheat grain is recognized as a good source of potentially health-enhancing components such as dietary fiber, phenolics, tocopherols, and carotenoids. Anthocyanins, another group of bioactive compounds, are found in blue and purple wheat grains. In the present study, a blue aleurone spring wheat line "Purendo 38" with relatively high content of total anthocyanins was used to investigate the composition and stability of anthocyanins over three crop years. Commercial cultivars of purple (Konini) and red (Katepwa) wheats were included in the study. Separation of anthocyanins by high-performance liquid chromatography (HPLC) showed that each wheat had a distinct anthocyanin profile. Four major anthocyanins were separated from blue wheat extracts as compared to five anthocyanins in purple wheat. Cyanidin 3-glucoside was the predominant anthocyanin in purple wheat, whereas it was the second major anthocyanin in blue wheat. The predominant anthocyanin in blue wheat, making up approximately 41% of the total anthocyanin content, remains to be structurally unidentified. Blue wheat anthocyanins were thermally most stable at pH 1. Their degradation was slightly lower at pH 3 as compared to pH 5. Increasing the temperature from 65 to 95 degrees C increased degradation of blue wheat anthocyanins. Addition of SO(2) during heating of blue wheat had a stabilizing effect on anthocyanin pigments. The optimal SO(2) concentrations were 500-1000 ppm for whole meals and 1000-3000 ppm for isolated anthocyanins. Further studies are underway to identify and verify individual anthocyanins in blue wheat and their potential end uses.     

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。     

Antioxidant activities and anthocyanin content of fresh fruits of common fig (Ficus carica L.)

Fig fruit has been a typical component in the health-promoting Mediterranean diet for millennia. To study the potential health-promoting constituents of fig fruits, six commercial fig varieties differing in color (black, red, yellow, and green) were analyzed for total polyphenols, total flavonoids, antioxidant capacity, and amount and profile of anthocyanins. Using reversed-phase liquid chromatography (RP-LC), various concentrations of anthocyanins but a similar profile was found in all varieties studied. Hydrolysis revealed cyanidin as the major aglycon. Proton and carbon NMR confirmed cyanidin-3-O-rhamnoglucoside (cyanidin-3-O-rutinoside; C3R) as the main anthocyanin in all fruits. Color appearance of fig extract correlated well with total polyphenols, flavonoids, anthocyanins, and antioxidant capacity. Extracts of darker varieties showed higher contents of phytochemicals compared to lighter colored varieties. Fruit skins contributed most of the above phytochemicals and antioxidant activity compared to the fruit pulp. Antioxidant capacity correlated well with the amounts of polyphenols and anthocyanins (R2 = 0.985 and 0.992, respectively). In the dark-colored Mission and the red Brown-Turkey varieties, the anthocyanin fraction contributed 36 and 28% of the total antioxidant capacity, respectively. C3R contributed 92% of the total antioxidant capacity of the anthocyanin fraction. Fruits of the Mission variety contained the highest levels of polyphenols, flavonoids, and anthocyanins and exhibited the highest antioxidant capacity.     

Mass spectrometric evidence for the existence of oligomeric anthocyanins in grape skins

The fractionation of a grape skin extract by multilayer countercurrent chromatography coupled with step gradient elution allowed the preparation of a fraction almost devoid of free anthocyanins. This fraction appeared to be almost exclusively polymeric, as judged by liquid chromatographic-mass spectrometric (LC-MS) analysis, color-bleaching tests with sulfur dioxide, and thiolysis. Electrospray mass spectrometric analysis indicated that the pigmented material in this fraction was chiefly composed of direct condensation products of anthocyanin extending up to trimers. With regard to their linkages, the anthocyanin units in the oligomers were possibly linked by either an A-type (by both carbon-carbon and ether bonds) or B-type (by carbon-carbon bond) linkage, like proanthocyanidins. The terminal anthocyanin unit of the oligomers is consistently in the flavylium form but the extension units are in the flavan form for the A-type oligomers and in the flavene form for the B-type oligomers. Although their linkages still need to be defined rigorously, this is the first mass spectrometric evidence confirming the existence of anthocyanin oligomers in the grape skin extract.     

effect of dose size on bioavailability of acylated and nonacylated anthocyanins from red cabbage (Brassica oleracea L. Var. capitata)

Recent studies indicate that anthocyanin intake conveys a variety of health benefits, which depend on absorption and metabolic mechanisms that deliver anthocyanins and their bioactive metabolites to responsive tissues. The anthocyanin bioavailability of red cabbage (Brassica oleracea L. var. capitata) was evaluated as reflected by urinary excretion of anthocyanins and anthocyanin metabolites. Twelve volunteers consumed 100, 200, and 300 g of steamed red cabbage (containing 1.38 micromol of anthocyanins/g of cabbage) in a crossover design. Anthocyanin concentration in cabbage extract and urine was measured by HPLC-MS/MS. Six nonacylated and 30 acylated anthocyanins were detected in red cabbage, and 3 nonacylated anthocyanins, 8 acylated anthocyanins, and 4 metabolites were present in urine. Mean 24 h excretion of intact anthocyanins increased linearly from 45 (100 g dose) to 65 nmol (300 g dose) for acylated anthocyanins and from 52 (100 g dose) to 79 nmol (300 g dose) for nonacylated anthocyanins. Urinary recovery of intact anthocyanins (percent of anthocyanin intake) decreased linearly from 0.041% (100 g dose) to 0.020% (300 g dose) for acylated anthocyanins and from 0.18% (100 g dose) to 0.09% (300 g dose) for nonacylated anthocyanins. Anthocyanin metabolites consisted of glucuronidated and methylated anthocyanins. The results show that red cabbage anthocyanins were excreted in both intact and metabolized forms and that recovery of nonacylated anthocyanins in urine was >4-fold that of acylated anthocyanins.     

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.