Optimisation of pulsed electric fields extraction of anthocyanin from Beibinghong Vitis Amurensis Rupr

Beibinghong Vitis amurensis Rupr has wide plantation area, high productivity and rich anthocyanin. Common hot-extraction has poor deficiency and destroys anthocyanin severely. For Beibinghong V. amurensis Rupr as materials, response surface-optimised electric fields were used, the structure of Beibinghong was observed by SEM, antioxidant activity was measured by DPPH, ABTS and reducing force, the component of anthocyanin was analyzed by HPLC-MS. We found the content of total anthocyanin extracted by pulsed electric fields was 166.65 ± 3.88 mg/100 g.FW. Total anthocyanin from Beibinghong had high antioxidant activity, also contained multiple steady anthocyanin of delphinidin 3-O-glucoside, cyanidin 3-O-glucoside, petunidin 3-O-glucoside, peonidin 3-O-glucoside, malvidin 3-O-glucoside, delphinidin-3-O-(6-O-acetyl) glucoside and delphinidin-3-O-(6-O-p-coumaroyl) glucoside et al. In conclusion, the optimised pulsed electric fields method can quickly and efficiently extract several kinds of anthocyanins from V. amurensis Rupr. This study promoted the intensive processing of V. amurensis Rupr and widened the practical application of pulsed electric field technology.     

Mass spectrometry in the study of anthocyanins and their derivatives: differentiation of Vitis vinifera and hybrid grapes by liquid chromatography/electrospray ionization mass spectrometry and tandem mass spectrometry

A mass spectrometric-based procedure for anthocyanin profiling was set up to distinguish authentic Vitis vinifera from hybrid red grapevine cultivars. 3-O-Monoglucoside and the related acetyl-, p-coumaryl- and caffeoyl-monoglucoside anthocyanins occurred only in Vitis vinifera, whereas 3,5-O-diglucoside and the substituted acetyl-, p-coumaryl-, feruloyl- and caffeoyl-diglucoside anthocyanins were the additional pigments in hybrid grapevines. The procedure was applied expressly to identify red grape cultivars based on the anthocyanin chemo-type determination. In particular, a red grape cultivar, having 3,5-O-diglucoside anthocyanins and a novel class of anthocyanin monoglucosides, such as cyanidin-3-O-, cyanidin-3-O-(6-O-acetyl)- and cyanidin-3-O-(6-O-p-coumaryl)pentoside, was classified as hybrid. A second vine cultivar, characterized exclusively by 3-O-monoglucoside anthocyanins, was included among the Vitis vinifera species. Anthocyanin profiling by mass spectrometry could represent the core of a chemotaxonomic procedure for distinguishing American and European grapevines based on the identification of post-synthetic anthocyanidin modification.     

Quality Characteristics and Anthocyanin Profiles of Different Vitis amurensis Grape Cultivars and Hybrids from Chinese Germplasm

To evaluate the important Vitis amurensis germplasm, the quality characteristics and anthocyanin profiles of the ripe berries of 20 V. amurensis grapes and 11 interspecific hybrids in two consecutive years were analysed. Compared with the V. vinifera grapes, V. amurensis grapes had small berries with low total soluble solids and high titratable acids, and were richer in phenolic compounds except for flanan-3-ols in their skins but had lower phenolic contents in their seeds and showed lower antioxidant activities. An outstanding feature of the V. amurensis grapes was their abundant anthocyanin contents, which was 8.18-fold higher than the three wine grapes of V. vinifera. The anthocyanin composition of V. amurensis was characterized by an extremely high proportion of diglucoside anthocyanins (91.71%) and low acylated anthocyanins (0.04%). Interestingly, a new type of speculated 3,5,7-O-triglucoside anthocyanins was first identified and only detected in V. amurensis grapes and hybrids. Based on the total phenolic and anthocyanin characteristics, V. amurensis grapes were set apart from V. vinifera cultivars and the interspecific hybrids, for the same qualities, fell between them, as assessed by principal component analysis.     

Purification and antioxidant capacity analysis of anthocyanin glucoside cinnamic ester isomers from Solanum nigrum fruits

In a recent study, anthocyanins, which have a strong free radical‐scavenging activity, were examined for their potential to effectively prevent cancer. However, clinical trials are limited by the purity of the anthocyanin. Multiple methods are used to extract and purify anthocyanins. Based on previous work on Solanum nigrum, which is a widely distributed plant, in this study, DM130 macroporous resin, Sephadex LH20, and a C18 column were used to separate cis–trans anthocyanin isomers. These anthocyanins constitute the majority of total S. nigrum anthocyanins. The results showed that this “DM130‐LH20‐C18 system” can be used to obtain a cinnamic acid‐derived cis–trans anthocyanin, petunidin‐3‐(p‐coumaroyl)‐rutinoside‐5‐glucoside, with a purity of 98.5%, for effective quantitation. In order to determine the antioxidant ability of the petunidin‐3‐(p‐coumaroyl)‐rutinoside‐5‐glucoside cis–trans isomers, three ordinary methods were adopted. The maximum antioxidant ability of the cis–trans anthocyanin was dozens of times higher than that of vitamin C.     

Inga edulis fruits: a new source of bioactive anthocyanins

Abstract

The extraction conditions and chromatographic analysis from seeds of Inga edulis were optimized and provided one anthocyanin from aqueous fraction and a mixture of three anthocyanins from methanolic fraction. The pure anthocyanin obtained was subjected to structural modifications and the products obtained were subjected to chemical and pharmacological assays, as well as quantum chemical calculations based on DFT and TD-DFT methods. Hence, the anthocyanin fractions were evaluated for their chemical-pharmacological potential through chemical and biological assays: antioxidant activity by the DPPH, determination of the Solar Protection Factor (SPF) and cytotoxic activity (hepatocellular carcinoma infected with hepatitis C virus). The results indicated that even the anthocyanin and derivatized compounds having high antioxidant potential showed an SPF lower than six, which is lower than the minimum accepted by current Brazilian legislation. In addition, none of compounds presented significant cytotoxic activity against the tumour cell line studied.     

General method for extraction of blueberry anthocyanins and identification using high performance liquid chromatography–electrospray ionization-ion trap-time of flight-mass spectrometry

A systematic approach for optimizing the extraction and identification of anthocyanins from blueberries was explored using HPLC-UV and HPLC–ESI-IT-TOF-MS. Sample homogenization effects, extraction solvent selection, type of acid, and amount used in extraction solvent were investigated. A mixture of methanol:water:trifluoroacetic acid (70:30:1, v/v/v) was found to be the best solvent system for blueberry anthocyanin extraction. Differences in total anthocyanin content due to commercial blueberry processing were explored as an application using the optimized extraction technique and HPLC-UV analysis. A methodical system for anthocyanin identification by HPLC–ESI-IT-TOF-MS without the use of standards was also reviewed and applied. Consideration was given to elution order by chromatographic separation with selective detection at 520 nm, high mass accuracy m/z values, tandem MS fragmentation, and previously published literature. Overall, 25 anthocyanins from a wild type highbush blueberry were identified and reported.     

Specific Poly-phenolic Compounds in Cell Culture of <Emphasis Type="Italic">Vitis vinifera</Emphasis> 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-p-coumaryl 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.     

Characterization and quantification of anthocyanins in selected artichoke (Cynara scolymus L.) cultivars by HPLC–DAD–ESI–MS n

The anthocyanin pattern of artichoke heads (Cynara scolymus L.) has been investigated by high-performance liquid chromatography–electrospray ionization mass spectrometry. For this purpose a suitable extraction and liquid chromatographic method was developed. Besides the main anthocyanins—cyanidin 3,5-diglucoside, cyanidin 3-glucoside, cyanidin 3,5-malonyldiglucoside, cyanidin 3-(3′′-malonyl)glucoside, and cyanidin 3-(6′′-malonyl)glucoside—several minor compounds were identified. Among these, two peonidin derivatives and one delphinidin derivative were characterized on the basis of their fragmentation patterns. To the best of our knowledge this is the first report on anthocyanins in artichoke heads consisting of aglycones other than those of cyanidin. Quantification of individual compounds was performed by external calibration. Cyanidin 3-(6′′-malonyl)glucoside was found to be the major anthocyanin in all the samples analyzed. Total anthocyanin content ranged from 8.4 to 1,705.4 mg kg⁻¹ dry mass.      

Identification and quantification of anthocyanins,flavonoids, and phenolic acids in flowers of Rhododendron arboreum and evaluation of their antioxidant potential

The current study aimed to investigate the anthocyanins, non-anthocyanins (flavonoids and phenolic acids), and free radicals scavenging potential in the flowers of Rhododendron arboreum using ultra high performance liquid chromatography with ion mobility quadrupole time of flight tandem mass spectrometry. A total of 25 constituents including nine anthocyanins, six phenolic acids, and ten flavonoids were identified in the flower extract. The major anthocyanins identified were cyanidin-3-O-β-galactoside ( 1 ), cyanidin-3-O-α-arabinoside ( 4 ), and cyanidin-3-O-rhamnoside ( 8 ), while quercetin glycosides were the main identified flavonoids in R. arboreum flowers. Additionally, ultra high performance liquid chromatography methods were developed and validated for the quantification of nine compounds (anthocyanins, flavonoid glycosides, and phenolic acids); five of them were quantified using internal standards. The extracts were analyzed for total phenolics (123.6 mg GAE/g), anthocyanin content (1.76% w/w), and evaluated for antioxidant properties against 2,2-diphenyl-1-picrylhydrazyl radical (IC₅₀: 102.06 and 96.92 μg/mL) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radical cation (112.25 and 45.59 μM TE/g) assays. The profiling of R. arboreum for anthocyanins is reported for the first time. The findings suggest that the flowers are a promising source of bioactive constituents and could be used as functional food, antioxidants, and nutraceuticals.     

Rapid quantitative analysis of individual anthocyanin content based on high‐performance liquid chromatography with diode array detection with the pH differential method

A new quantitative technique for the simultaneous quantification of the individual anthocyanins based on the pH differential method and high‐performance liquid chromatography with diode array detection is proposed in this paper. The six individual anthocyanins (cyanidin 3‐glucoside, cyanidin 3‐rutinoside, petunidin 3‐glucoside, petunidin 3‐rutinoside, and malvidin 3‐rutinoside) from mulberry (Morus rubra) and Liriope platyphylla were used for demonstration and validation. The elution of anthocyanins was performed using a C₁₈ column with stepwise gradient elution and individual anthocyanins were identified by high‐performance liquid chromatography with tandem mass spectrometry. Based on the pH differential method, the high‐performance liquid chromatography peak areas of maximum and reference absorption wavelengths of anthocyanin extracts were conducted to quantify individual anthocyanins. The calibration curves for these anthocyanins were linear within the range of 10–5500 mg/L. The correlation coefficients (r²) all exceeded 0.9972, and the limits of detection were in the range of 1–4 mg/L at a signal‐to‐noise ratio ≥5 for these anthocyanins. The proposed quantitative analysis was reproducible with good accuracy of all individual anthocyanins ranging from 96.3 to 104.2% and relative recoveries were in the range 98.4–103.2%. The proposed technique is performed without anthocyanin standards and is a simple, rapid, accurate, and economical method to determine individual anthocyanin contents.     

Anthocyanins HPLC-DAD and MS Characterization,Total Phenolics,and Antioxidant Activity of Some Berries Extracts

Extracts of indigenous wild blackberries, mulberries, bilberries, and blackthorns were analyzed for anthocyanin composition, anthocyanin content, total phenolics, and antioxidant capacity. Anthocyanins extraction with acidified methanol in ultrasonic condition (59 kHz, 60 min., 25°C) was carried out. The extracts were analyzed by high-performance liquid chromatography (HPLC) using a Dionex Ultimate 3000 apparatus equipped with photodiode array detector for qualitative characterization of the anthocyanins. The chromatograms revealed the presence of a large number of anthocyanins in fruits extracts: blackberries, 4 compounds; mulberries, 3 compounds; bilberries, 18 compounds; and blackthorns, 5 compounds. The most abundant anthocyanins were cyanidin-3-glucoside in blackberry, mulberry, and bilberry, and cyanidin-3-rutinoside in blackthorn extract. Structural information about anthocyanins was obtained by using a mass spectrometric method based on fully automated chip-nanoelectrospray ionization (nanoESI) high capacity ion trap (HCT). Anthocyanin content was quantified by the pH differential method and total phenolics were determined by Folin-Ciocalteu method. A Jasco V 530 UV-VIS spectrophotometer was used for absorbance measurements. The free radical scavenging activity of the berries extracts was performed by using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay. The reduction of DPPH was followed by a spectrophotometric method. Also, a correlation of the antioxidant capacities of the extracts with their anthocyanin content and total phenolics was attempted.     

Extraction of Anthocyanins from Borage (Echium amoenum) Flowers Using Choline Chloride and a Glycerol-Based,Deep Eutectic Solvent: Optimization,Antioxidant Activity,and In Vitro Bioavailability

Borage flower (Echium amoenum), an annual herb native to the Mediterranean region, is an excellent source of anthocyanins and is widely used in various forms due to its biological activities. In the present study, a choline chloride and glycerol (CHGLY)-based natural deep eutectic solvent (NADES) was applied in order to extract the anthocyanins from borage flowers. The traditional solvents, including water, methanol, and ethanol, were used to evaluate the efficiency of CHGLY. The results showed that CHGLY was highly efficient compared to the traditional solvents, providing the highest amounts of the total anthocyanin content (TAC), total phenolic content (TPC), total flavonoid content (TFC), individual anthocyanins, and antioxidant activity (DPPH radical scavenging (DPPH) and ferric-reducing antioxidant power (FRAP) assays). The most dominant anthocyanin found in studied borage was cyanidin-3-glucoside, followed by cyanin chloride, cyanidin-3-rutinoside, and pelargonidin-3-glucoside. The bioavailability % was 71.86 ± 0.47%, 77.29 ± 0.57%, 80.22 ± 0.65%, and 90.95 ± 1.01% for cyanidin-3-glucoside, cyanidin-3-rutinoside, by pelargonidin-3-glucoside and cyanin chloride, respectively. However, cyanidin-3-glucoside was the anthocyanin compound showing the highest stability (99.11 ± 1.66%) in the gastrointestinal environment. These results suggested that choline chloride and glycerol-based NADES is not only an efficient, eco-friendly solvent for the extraction of anthocyanins but can also be used to increase the bioavailability of anthocyanins.     

Hydrophilic Interaction Chromatography on Silica: Group Analysis of Grape Anthocyanins

A group control procedure is developed, in which the anthocyanin complex of a grape extract is divided into six groups without differentiation by aglycons: 3-glucosides (Zone III); 3-glucosides acylated with p-coumaric acid (Zone I); 3-glucosides acylated with acetic acid (Zone II); 3,5-diglucosides (Zone IV); 3,5-diglucosides acylated with p-coumaric acid (Zone IV); and 3,5-diglucosides acylated with acetic acid (Zone V). The anthocyanins were separated by hydrophilic interaction chromatography in a column packed with a silica using the mobile phases of acetonitrile–water (20–8 vol %) containing 0.5 vol % of orthophosphoric acid. Due to the low viscosity of the mobile phase and the complete separation of anthocyanin groups, the procedure suitable for determining the inheritance of genetic traits in the selection of grapes is transferred to a format of microcolumn chromatography using MiLiChrome chromatographs.     

The RP-HPLC analysis of anthocyanins

Summary Conditions were determined for the separation of a complex set of anthocyanins (free aglycones, mono- and multiglycosides and esterified forms) by HPLC. The optimised gradient elution method was then used to carry out qualitative and quantitative analysis of anthocyanin compounds present in the callus tissue ofRudbeckia hirta L. and the tubular flowers of the soil-based plant. The summary content of anthocyanin pigments and the content of the main pigment was identified in the analysed biomass. The method developed is useful for the purposes of monitoring the process of biosynthesis of anthocyanins in tissues obtained through in vitro cultures. The advantages of the method for anthocyanins and its application to other anthocyanin-rich materials are also discussed.     

Large‐scale isolation of high‐purity anthocyanin monomers from mulberry fruits by combined chromatographic techniques

Anthocyanins have attracted attention over the past several decades because of their beneficial health effects. In this research, a strategy combining column chromatography and high‐speed countercurrent chromatography was developed for the separation of high‐purity anthocyanin monomers from mulberry fruits. After purification using Amberlite XAD‐7HP column with 80% ethanol (0.1% HCl), a fraction of anthocyanins mixtures with a purity of 68.6% was obtained. High‐speed countercurrent chromatography with a biphasic solvent system of n‐butanol/methyl tert‐butyl ether/acetonitrile/water/trifluoroacetic acid (30:10:10:50:0.05, v/v) was used to separate the anthocyanin monomers. Three monomers of delphinidin‐3‐O‐ rutinoside, cyanidin‐3‐O‐ rutinoside, and cyanidin‐3‐O‐ glucoside were obtained, and identified by ¹H and ¹³C NMR spectroscopy and high‐performance liquid chromatography with electrospray ionization‐mass spectrometry. The method developed in this work can be used to conduct large‐scale separations of anthocyanin monomers from mulberry fruits and other plants.     

Synthesis and application of a nanoporous ion‐imprinted polymer for the separation and preconcentration of trace amounts of vanadium from food samples before determination by electrothermal atomic absorption spectrometry

A vanadium ion‐imprinted polymer was synthesized in the presence of V(V) and N‐benzoyl‐N‐phenyl hydroxyl amine using 4‐vinyl pyridine as the monomer, ethylene glycol dimethacrylate as the cross linker and 2,2’‐azobis(isobutyronitrile) as the initiator. The imprinted V(V) ions were completely removed by leaching the polymer with 5 mol/L nitric acid, and the polymer structure was characterized by Fourier transform infrared spectroscopy and scanning electron microscopy. The ion‐imprinted polymer was used as the sorbent in the development of the solid‐phase extraction method for V(V) prior to its determination by electrothermal atomic absorption spectrometry. The maximum sorption capacity for V(V) ions was 26.7 mg/g at pH 4.0. Under the optimum conditions, for a sample volume of 150.0 mL, an enrichment factor of 289.0 and a detection limit of 6.4 ng/L were obtained. The developed method was successfully applied to the determination of vanadium in parsley, zucchini, black tea, rice, and water samples.     

Synthesis of 3-Methoxy- and 3-(β-D-Glucopyranosyloxy)flavylium Ions. Influence of the flavylium substitution pattern on the reactivity of anthocyanins in aqueous solution

The synthesis of 3-glycosyloxylated flavylium ions (anthocyanins), in particular of callistephin ( 4 ), a natural anthocyanin, is described. The structural transformations in aqueous solution and molecular complexation with chlorogenic acid ( 7 ) and caffeine ( 8 ) of the synthesized pigments 3 and 4 are investigated and compared to those of the corresponding 3-methoxyflavylium ions 1 and 2 and to those of oenin ( 5 ) and malvin ( 6 ), two very common natural anthocyanins. The results are discussed in terms of the role played by the glycosyloxy residues in the chemical properties of anthocyanins. Anthocyanin molecular complexation (copigmentaion) is quantitatively investigated by UV/VIS spectroscopy and ¹H-NMR. In particular, the UV/VIS spectroscopic data are interpreted using a general theoretical treatment, which, e.g., allows to demonstrate the formation of molecular complexes between the colourless forms of an anthocyanin and 8 .     

Analysis and tentative structure elucidation of new anthocyanins in fruit peel of Vitis coignetiae Pulliat (meoru) using LC‐MS/MS: Contribution to the overall antioxidant activity

The skin of Vitis coignetiae Pulliat (meoru) grown wild in the Republic of Korea was analyzed for anthocyanins via HPLC coupled to ESI‐MS/MS in positive ion mode. Chromatographic separation was conducted via RP HPLC using a C₁₈ column, with a 50‐min gradient from 0 to 80% methanol in water containing 0.5% formic acid. A total of 18 anthocyanins were identified. Among them, nine compounds were newly determined by comparing the retention time (t_R) and mass fragmentation patterns with those of the previously reported anthocyanins for other grape varieties: malvidin hexose, peonidin 3‐galactoside, malvidin 3‐galactoside, cyanidin, petunidin, petunidin 3‐(6″‐coumaroyl)‐5‐diglucoside, peonidin, malvidin, and malvidin 3‐(6″‐coumaroyl)‐5‐diglucoside. The antioxidant activity of the V. coignetiae Pulliat anthocyanins was determined via 2,2‐diphenyl‐2‐picrylhydrazyl radical scavenging and 2,2′‐azinobis‐(3‐ethylbenzothiazoline‐6‐sulfonic acid) radical cation assays in a range of concentration from 25 to 500 mg/L. The capacity increased with concentration. The IC₅₀ values, defined as the concentration of sample required to scavenge 50% of free radicals, were calculated as follows: 189.63±11.31 mg/L and 141.29±6.70 mg/L for 2,2‐diphenyl‐2‐picrylhydrazyl and 2,2′‐azinobis‐(3‐ethylbenzothiazoline‐6‐sulfonic acid) radical cation, respectively. The antioxidant activity of the V. coignetiae Pulliat anthocyanins is substantially higher than that of ascorbic acid and is similar to the effects of the extracts obtained from other grape varieties.     

Redox Behavior of Anthocyanins Present in Vitis vinifera L.

Voltammetric techniques were employed to study the electrochemical behavior of several anthocyanins. The redox behavior of anthocyanins with the same basic structure, the influence of glycosylation on the redox behavior of anthocyanins derived from different anthocyanidins, and the influence of methoxylation were investigated. The anthocyanins used in this study were malvidin‐3‐O‐glucoside chloride, malvidin‐3,5‐di‐O‐glucoside chloride, cyanidin‐3‐O‐glucoside chloride, cyanidin‐3,5‐di‐O‐glucoside chloride, peonidin‐3‐O‐glucoside chloride, delphinidin‐3‐O‐glucoside chloride and the anthocyanidin petunidin chloride, all of them present in Vitis vinifera L. All hydroxyl groups of the anthocyanins can be electrochemically oxidized and the anthocyanins studied revealed a complex and pH dependent oxidation process, with the occurrence of adsorption and of oxidation products blocking the electrode surface.     

Screening for anthocyanins using high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry with precursor-ion analysis, product-ion analysis, common-neutral-loss analysis, and selected reaction monitoring

A systematic method for anthocyanin identification using tandems mass spectrometry (MS/MS) coupled to high-performance liquid chromatography (HPLC) with photo-diode array detection (PDA) was developed. Scan for the precursor ions of commonly found anthocyanidins (cyanidin, delphinidin, malvidin, pelargonidin, petunidin, and peonidin) using LC/MS/MS on a triple quadrupole instrument allows for the specific determination of each category of anthocyanins. Further characterization of each anthocyanin was performed using MS/MS product-ion analysis, common-neutral-loss analysis, and selected reaction monitoring (SRM). The method was demonstrated for analysis of anthocyanins in black raspberries, red raspberries, highbush blueberries, and grapes (Vitis vinifera). Previous reported anthocyanins in black raspberries and red raspberries are confirmed and characterized. Common-neutral-loss analysis allows for the distinction of anthocyanin glucosides or galactoside and arabinosides in highbush blueberries. Separation and identification of anthocyanin glucosides and galactosides were achieved by LC/MS/MS using SRM. Anthocyanin isomers such as cyanidin sophoroside and 3,5-diglucoside were differentiated by their fragmentation pattern during product-ion analysis. Fifteen anthocyanins (all possible combinations of five anthocyanidins and three sugars) were characterized in highbush blueberries. Pelargonidin 3-glucoside and pelargonidin 3,5-diglucoside were detected and characterized for the first time in grapes. The present approach allows mass spectrometry to be used as a highly selective detector for rapid identification and characterization of anthocyanins and can be used as a sensitive procedure for screening anthocyanins in fruits and vegetables.