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449

Since the 1990s, the wines of Uruguay have been ac-quiring an increasing importance in the world wine market.In the 2004, the annual wine production (red, white, rosé,and sparkling wines) of Uruguay was ~113 million L(www.inavi.com.uy), and 36% of this production was redwine. Among these Uruguayan wines, those made fromVitis vinifera cv. Tannat grapes are the most well-known.The Tannat variety has its origins in southern France.However, today Uruguay is one of the few countries inwhich this variety is found. These grapes produce a color-ful wine, with high acidity and high tannins, which allowfor long-term aging. Although it is the second mostwidely cultivated red grape in Uruguay (www.inavi.com.uy), other grape varieties have been and are now be-ing introduced and are favored by climatic conditions,which are similar to those of the Mediterranean region.

Some of these varieties have their origins in Europe, whileothers result from crosses.

Caladoc is a recently introduced variety in Uruguayand originates from a cross between Vitis vinifera cvs.Grenache and Malbec. It produces a wine of intense color,nonbitter and soft tannins, and raspberry-spicy flavors.Marselan results from a cross between Cabernet Sau-vignon and Grenache varieties achieved in 1961 by re-searchers at the INRA and ENSA (France). Although Mar-selan grapes were initially considered of no value to thewine industry because of moderate yield, the wines can beof excellent quality, which has led to its cultivation incountries such as Uruguay. The grapes produce a complexwine that is colorful, with soft tannins and juicy black-currant flavors. Marzemino is a variety that originated inItaly and has also been introduced into Uruguay. Itswines are slightly tannic, with sweet violet overtones.Cheveñasco has been cultivated in Uruguay for manyyears, but its origin remains unknown.

The aim of this work was to determine by HPLC-DAD-MS the pigment profiles and different anthocyanin-derivedpigment formation in young red wines produced in Uru-guay from these four grape varieties and compare them tothat of a young Tannat red wine produced in the samevintage. Until now, only the major anthocyanins(monoglucosides and acyl derivatives originally present ingrapes) were determined to characterize grapes and thewines made from them (Cacho et al. 1992, Mazza et al.1999, Burns et al. 2002, González-Neves et al. 2004a), theanthocyanin-derived pigments not being determined asthey are present in lower concentrations. The use of mass

Pigment Profiles in Monovarietal WinesProduced in Uruguay

Cristina Alcalde-Eon,1 Eduardo Boido,2* Francisco Carrau,2

Eduardo Dellacassa,2 and Julián C. Rivas-Gonzalo1

1Unidad de Nutrición y Bromatología, Facultad de Farmacia, Universidad deSalamanca, Campus Miguel de Unamuno, 37007 Salamanca, Spain; 2Secciónde Enología, Facultad de Química, Universidad de la República, Gral. Flores2124, 11800 Montevideo, Uruguay.

*Corresponding author [email: eboido@fq.edu.uy]

Acknowledgments: This work was supported by grant AGL-2002-00167from the Ministerio de Ciencia y Tecnología, Spain, and is a part of thePrograma Iberoamericano de Ciencia y Tecnología para el Desarrollo(proyect IV.10). The authors thank the Ministerio de Educación, Cultura yDeporte, Spain, for a F.P.U. predoctoral scholarship to C. Alcalde-Eon.

The authors also thank Mr. G.H. Jenkins for revising the English version ofthe manuscript.

Manuscript submitted March 2006; revised July 2006

Copyright © 2006 by the American Society for Enology and Viticulture.All rights reserved.

Abstract: The aim of this study was to determine by HPLC-DAD-MS the pigment profiles and the differentanthocyanin-derived pigment formation in four monovarietal young red wines produced in Uruguay from dif-ferent grape varieties (Caladoc, Marselan, Marzemino, and Cheveñasco) and compare them to that of a youngVitis vinifera cv. Tannat wine produced in the same vintage. In quantitative terms, Tannat wine had the highesttotal pigment content whereas Cheveñasco had the lowest. Results showed differences not only in anthocyanincomposition but also in the chemical nature of the major anthocyanin-derived pigments present in each wine.Tannat had the lowest content in derived pigments, vitisin A being the most representative. Caladoc and Marselanwere characterized by the highest percentages of malvidin-3-glucoside and its acyl derivatives. The 4-vinylphenolderivative of malvidin-3-glucoside was the major pyranoanthocyanin in both wines, along with vitisin A in Caladocand vitisin B in Marselan. The major anthocyanin-derived pigments of Marzemino were B-type vitisins. Marzeminoalso contained anthocyanin-flavanol acetaldehyde-mediated condensation products, which might indicate higherlevels of acetaldehyde in this wine than in the others. Cheveñasco had the highest percentage of derived pig-ments, with the 4-vinylphenol derivative of malvidin-3-glucoside the most representative and the second mostabundant compound after malvidin-3-glucoside.

Key words: anthocyanins, pyranoanthocyanins, anthocyanin-flavanol condensation products, red wines

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spectrometry coupled to HPLC-DAD has allowed a moredetailed study of anthocyanin-derived pigment fractions(Vivar-Quintana et al. 2002, Wang et al. 2003, Monagas etal. 2003, Alcalde-Eon et al. 2004a), which could also beuseful in characterization of wines, as both the anthocya-nin fraction and anthocyanin-derived pigments contributeto final wine color. Thus, it is important to know whichtype of anthocyanin derivatives are primarily formed fromeach grape variety during the winemaking process. Sincethe grapes of four of the analyzed wines (Caladoc,Marselan, Marzemino, and Cheveñasco) were collectedfrom the same vineyard in the same vintage and have un-dergone the same process, differences in the derived pig-ment composition might be due, above all, to the variety.Moreover, knowledge of pigment composition can be in-formative for better management of vintage conditions inorder to preserve the characteristic properties.

The anthocyanin profiles of Caladoc, Marselan, Mar-zemino, and Cheveñasco grape varieties and the wineshave not previously been described. For Tannat grapesand wines, phenolic potential, main anthocyanins, and to-tal pigment content and their variations with vintage andvineyard treatments have been reported (González-Neveset al. 2001, 2004a,b), although a more complete study ofpigment profile has still to be performed. For Caladoc andMarselan grapes, some of the parent varieties have beenstudied. Knowledge of pigment composition can be use-ful in determining the profile of Marselan, as it should besimilar to its parents. Particularly, Cabernet Sauvignongrapes and wines have been well studied (González-Neves et al. 2001, Arozarena et al. 2002, Burns et al. 2002,Wang et al. 2003, Monagas et al. 2003, Núñez et al. 2004).While the phenolic and anthocyanin composition ofGrenache has been studied to a lesser extent than Caber-net Sauvignon, there are studies of the influence of vin-tage, climatic variables, and quality of the vineyard in thecomposition of these grapes (Cacho et al. 1992, Bergqvistet al. 2001), but only major anthocyanins or total antho-cyanin content were considered. Although the concentra-tions of each individual component in one grape varietycan be affected by different factors (such as vintage, soilconditions, climatic variables), the pigment profile (deter-mined by the proportions of the different anthocyanins orpigment families) is characteristic of the variety (Aroza-rena et al. 2002, González-Neves et al. 2004a).

Materials and Methods

Wine samples. Fresh grapes (Vitis vinifera L. cv. Tan-nat) were acquired from a vineyard located in northeasternUruguay (Cerro Chapeu) and delivered to the winemakingfacilities. Batches of 10 tons were processed in the 2003vintage. The grapes were destemmed, crushed, and a sub-sample analyzed for sugar content, total acidity, and pH.Sulfur dioxide (SO2) was added to the must (50 mg/L)which was then inoculated with reactivated dry yeast(Saccharomyces cerevisiae, strain D 254; Lallemand, Can-

ada). Fermentation was carried out at 20 to 22°C in openstainless steel tanks, and when the must density hadreached 1000 g/L, the wine was separated from the skins.The skins were then pressed and both wine fractionscombined and stored in 225-L French oak barrels (50%new barrel), where fermentation was completed. Uponcompletion of alcoholic fermentation, malolactic fermenta-tion (MLF) was completed by inoculation with Oenoccocusoeni strain DSM 7008 (Viniflora, Chr. Hansen, Horsholm,Denmark) at 16°C. MLF was followed by determining theconcentrations of malic and lactic acids by thin-layerchromatography (Boido et al. 1999). Upon completion ofMLF, the wines were treated with 50 mg/L SO2. Sampleswere stabilized at 4°C for 20 days and sterile-filtered (0.45μm cellulose acetate membrane). Free SO2 content was ad-justed to 35 mg/L, and finally, the samples were bottledand maintained at 10°C for three months, until high-per-formance liquid chromatography–diode array detection–mass spectrometry (HPLC-DAD-MS) analysis.

Fresh grapes of the other four varieties (Caladoc, Mar-selan, Marzemino, and Cheveñasco) were acquired from avineyard located in southern Uruguay. Samples of 30 kg ofeach variety were microvinified during the same vintage asTannat grapes. The winemaking process was similar tothat described above for Tannat grapes. However, therewere some differences in the strains employed in the alco-holic fermentation (Saccharomyces cerevisiae, strain CIVC8130; Gist Brocades, Chile) and MLF (Oenoccocus oeni,Lalvin 31, Lallemand, Canada), and glass containers wereused instead of barrels in the MLF. Stabilization and bot-tling processes were identical to those undergone byTannat wine. The samples were diluted (1/5) with acidifiedwater (HCl at pH 0.5) and filtered through a 0.45-μm Millexsyringe-driven filter unit (Millipore Corporation, Bedford,MA) before HPLC-DAD-MS analysis.

HPLC-DAD-MS analysis. The analysis was performedin a Hewlett-Packard 1100 series liquid chromatograph, anddetection was carried out using a photodiode detector andan AQUA C18 reversed-phase, 5-μm, 150 mm x 4.6 mm col-umn (Phenomenex, Torrance, CA) thermostated at 35°C.

The HPLC-DAD conditions were previously employedwith satisfactory results in our laboratory in the analysisof wine samples (Alcalde-Eon et al. 2004a). The solventsused were (A) an aqueous solution (0.1%) of trifluoro-acetic acid (TFA) and (B) 100% HPLC-grade acetonitrile,establishing the following gradient: isocratic 10% B for 5min, from 10 to 15% B for 15 min, isocratic 15% B for 5min, from 15 to 18% B for 5 min, and from 18 to 35% B for20 min, at a flow rate of 0.5 mL/min. Detection was carriedout at 520 nm as the preferred wavelength. Spectra wererecorded from 220 to 600 nm.

The mass analyses were performed using a FinniganLCQ ion-trap instrument (Thermoquest, San Jose, CA)equipped with an electrospray ionization (ESI) interface.The LC system was connected to the probe of the massspectrometer via the UV cell outlet. Nitrogen was used assheath and auxiliary gas. The sheath gas flow was 1.2 L/

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min and the auxiliary gas flow was 6 L/min. The capillaryvoltage was 4V and capillary temperature 195°C. Sourcevoltage was 4.5 kV. Spectra were recorded in positive ionmode between 120 and 1500 m/z. The mass spectrometerwas programmed for a series of three consecutive scans:full mass, MS2 scan of the most abundant ion in the fullmass, and MS3 of the most abundant ion in the MS2. Thenormalized energy of collision was 45%.

Results and Discussion

Total pigment. Total pigment was calculated from thetotal area of the peaks obtained in the chromatograms ofeach wine recorded at 520 nm (Figure 1) and expressed asmalvidin-3-glucoside equivalents (Table 1) using a calibra-tion curve generated from a standard isolated in our labo-ratory as previously described (Heredia et al. 1998).Tannat wine had the most total pigment (762.4 mg/L),whereas Cheveñasco had the least (214.5 mg/L). Caladoc,Marselan, and Marzemino wines had similar total pigment(Table 1). The high pigment content of the Tannat sampleagreed with results from other studies in which total an-thocyanin content in Tannat grapes and wines was com-pared with Cabernet Sauvignon and Merlot (González-Neves et al. 2001, 2004b). Tannat samples also showedthe most total pigment in all these studies.

Qualitative and quantitative analyses. Identification ofpigments was by HPLC-DAD-MS analyses. Althoughwine possesses a wide variety of pigments, the HPLCconditions used in this study allowed a pigment separa-tion that differentiated a high number of compounds bytheir chromatographic features and UV-visible (vis) andmass spectra (Table 2, see page 453). Compounds 1 to 78were all found in the Tannat sample, while compounds ato n were found in samples other than Tannat and notfound in Tannat (Table 2). Numbers and letters were as-signed in accordance with retention times. Figure 1shows the compounds that represented more than 0.1%of the total pigment as well as those compounds specificto one or more than one wine. Retention times and num-bers listed in Table 2 can be used to locate minor com-pounds.

Individual concentrations were expressed as a percent-age of each compound over the total area (Table 2). Tak-ing into account that in the conditions used some of thedetected compounds coeluted, making it difficult to as-sign the chromatographic area corresponding to each, theareas obtained in the MS analyses were employed in thiscalculation. Thus, for each detected compound (eitheridentified or not), the areas of their respective molecularions were obtained by representing the abundance ofeach m/z ratio versus time. Integration of the peaksformed in this plot determined their areas, overcoming theproblem of coelution. The total area was calculated bysumming all the individual areas. The area values wereused instead of concentration values because of the lackof available standards for each family of compounds.

Figure 1 Chromatograms recorded at 520 nm corresponding to (A)Tannat, (B) Caladoc, (C) Marselan, (D) Marzemino, and (E) Cheveñascowines. Numbers correspond to those reported in Table 2.

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Pigment profiles of all five wines, expressed as a percent-age of each major family of compounds, are shown inTable 1.

Anthocyanins. Delphinidin, cyanidin, petunidin, peoni-din, and malvidin 3-glucoside (peaks 7, 10, 12, 16, and19), 3-(6’’-acetylglucosides) (peaks 22, 31, 32, 40, and42), and 3-(6’’-p-coumaroylglucosides) (peaks 43, 55, 56,67, and 69) were identified in the samples by retentiontime, elution order, UV-vis spectrum, molecular and frag-ment ions, and data reported in previous works (Vivar-Quintana et al. 2002, Burns et al. 2002, Monagas et al.2003, Alcalde-Eon et al. 2004a). All these compounds werepresent in all varieties analyzed, except cyanidin-3-acetylglucoside and peonidin-3-(6’’-p-coumaroylglucoside)in Cheveñasco and cyanidin-3-(6’’-p-coumaroylglucoside)in Marselan.

The molecular ions of peaks 59 and 61 had the samem/z ratio and fragmentation pattern as peaks 67 and 69,peonidin and malvidin 3-(6’’-p-coumaroylglucosides), re-spectively, and were identified as the cis isomers of thelatter compounds, taking into account existing data in dif-ferent types of samples (Alcalde-Eon et al. 2004b), in-cluding wine (Monagas et al. 2003). Peak 61 was presentin all five wines, whereas the cis isomer of peonidin-3-(6’’-p-coumaroylglucoside) was only present in Tannatand Caladoc. This compound has also been reported inGraciano and Tempranillo grapes (Núñez et al. 2004).

The UV-vis spectrum of peak 49 had a shoulder at 328nm, which indicated acylation of the molecule with caf-feic acid. Its molecular ion and fragmentation pattern(Table 2) confirmed the presence of this acid in the mol-ecule and identified the compound as malvidin-3-(6’’-caffeoylglucoside). This compound was found in allsamples.

Peaks f and 28 possessed the same m/z ratio as peaks32 and 42 (acetyl derivatives of petunidin and malvidin,respectively); their fragmentation pattern was similar tothat of the former ones, but they eluted earlier. Given thataglycon was the same for peaks f and 32 (petunidin) andfor peaks 28 and 42 (malvidin) and that in all samplesthe presence of acetylglycoside was detected (loss of 204amu) (arbitrary mass units) in the m/z signal of the mo-

lecular ion), it was proposed that each pair of compoundsdiffered only in the glycosidic pattern. Grape and wineanthocyanins are always reported to be glucosides. How-ever, Wang et al. (2003) found malvidin-3-O-galactoside,malvidin-3-O-acetylgalactoside, and malvidin-3-O-couma-roylgalactoside in Cabernet Sauvignon grape skin extractsand confirmed the identity of malvidin-3-O-galactoside bycomparing its chromatographic and mass spectral fea-tures to a standard. To our knowledge, no other hexoseshave been reported in grape and wine anthocyanins. Inaddition, anthocyanin 3-O-galactosides elute earlier on re-versed-phase HPLC than their corresponding glucosides(Dugo et al. 2001, Wang et al. 2003). Given these data,compounds f and 28 were proposed to be petunidin andmalvidin 3-(6’’-acetylgalactoside), respectively. Compoundf was found only in Marzemino, but compound 28 waspresent in all analyzed wines except Cheveñasco.

As expected for young red wines, the highest percent-age of the total area corresponded to the monoglucosidesof the anthocyanins in all samples. Cheveñasco had themost at 46.9%, whereas Marzemino had the least (33.7%)(Table 1). Among monoglucosides, malvidin-3-glucosidewas most abundant, especially in Marselan and Caladocwines (88.3 and 83.9% of the monoglucosides, respec-tively). Cyanidin-3-glucoside was detected in these twosamples at less than 0.05%. Acetyl derivatives of themonoglucosides were present in all the samples, but indifferent amounts. Cheveñasco had the least in relationto the other samples (only 10.3%), while Marzemino hadthe most (33.6%). In the Marzemino sample, monogluco-sides of the anthocyanins and their corresponding acetylderivatives had very similar percentages (see Table 2). Inall the samples analyzed, the coumaroyl derivatives of theanthocyanins were less abundant than the acetyl. Thisfeature is characteristic of some red grape varieties, in-cluding Tannat (González-Neves et al. 2001, 2004a),Cabernet Sauvignon (Burns et al. 2002, Arozarena et al.2002, Núñez et al. 2004), which is a parent of Marselan,Cabernet franc (Mazza et al. 1999), and Merlot (González-Neves et al. 2001). Cheveñasco wine again had the lowestpercentage of coumaroyl derivatives. As reported above,the cis isomer of the malvidin-3-(6’’-p-coumaroyl) gluco-

Table 1 Total pigment content (expressed as mg of malvidin-3-glucoside per liter of wine) and percentages of the mainpigment families found in Tannat, Caladoc, Marselan, Marzemino, and Cheveñasco wines.

Tannat Caladoc Marselan Marzemino Cheveñasco

Total pigment content (mg/L) 762.4 469.1 445.7 497.1 214.5

Anthocyanin monoglucosides (%) 45.2 42.6 40.6 33.7 46.9

Acetyl derivatives of monoglucosides (%) 27.0 28.7 30.9 33.6 10.3

Coumaroyl derivatives of monoglucosides (%) 13.7 14.7 11.3 8.8 7.8

Caffeoyl derivatives of monoglucosides (%) 0.4 1.7 1.0 0.2 0.2

Pyranoanthocyanins (%) 5.5 5.5 9.5 18.5 26.2

Direct condensation products (%) 1.3 1.5 1.2 0.8 4.4

Acetaldehyde-mediated condensation products (%) 1.1 1.5 0.8 1.5 0.6

Other compounds (%) 5.8 3.8 4.7 2.9 3.6

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side was present in all samples, but Caladoc had the most(1.2% of the total area) and the highest cis isomer/transisomer ratio (~11%). As usually occurs in red wines, thecaffeoyl derivatives of the monoglucosides were presentin small amounts, with Caladoc having the most (1.7%).

Pyranoanthocyanins. Different types of pyranoantho-cyanins were found in all samples. There were severalcompounds with molecular ions that possessed the samem/z ratios and similar retention times, and only the differ-ence in their UV-vis spectra allowed the assignment of theidentity.

The UV-vis spectra of peaks 4, 11, 14, 15, 18, 23, and41 corresponded to pyranoanthocyanins originating inthe cycloaddition of pyruvic acid and different anthocya-nins (A-type vitisins or vinylformic adducts). This cy-cloaddition increases the m/z ratio of the anthocyaninmolecular ions in 68 amu (Bakker and Timberlake 1997,Fulcrand et al. 1998). The molecular and fragment ions ofcompounds 4, 11, 15, and 18 confirmed that they origi-nated from the 3-monoglucosides of delphinidin, petun-idin, peonidin, and malvidin, respectively. In peaks 14,23, and 41, the presence of acylation was detected bythe losses in the MS2 analyses of 204 amu from the mo-lecular ions of peaks 14 and 23 (petunidin and malvidinderivatives, respectively) and 308 amu in peak 41(malvidin derivative), indicating the acylation with aceticand p-coumaric acids, respectively.

Unlike A-type vitisins, which were present in differentvarieties, B-type vitisins or vinyl adducts (originating inthe cycloaddition between acetaldehyde and anthocya-nins) were found almost exclusively in Marzemino. Theirpresence in red wines has been reported (Bakker andTimberlake 1997, Hayasaka and Asenstorfer 2002), and inreversed-phase HPLC they are generally retained longerthan the corresponding A-type vitisins (Vivar-Quintana etal. 1999). The UV-vis spectrum of this class of pyrano-anthocyanins has a similar shape to that of the A-typevitisins, but with the absorption maximum hypsochrom-ically shifted in relation to the latter. Thus, under theconditions used in this study, the A-type vitisin of malvi-din-3-glucoside (vitisin A) possessed a visible maximum at510 nm and the B-type vitisin of the same anthocyanin(vitisin B) at 490 nm (Figure 2, see page 456). Further-more, the molecular ions of the B-type compounds have24 additional amu in the m/z ratio when compared withthose of the anthocyanins from which they originate.Peaks c, d, 17, g, 24, j, 35, k, and 62 had these features.Peaks c, 17, and 24 originated from the 3-glucosides ofdelphinidin, petunidin, and malvidin, respectively. Themolecular ions and fragmentation patterns of peaks d, g,j, and 35 showed the presence of an acetyl moiety intheir parent molecules and were identified as the B-typevitisins of delphinidin, petunidin, peonidin, and malvidin3-acetylglucosides. Similarly, peaks k and 62 were identi-fied as B-type vitisins of petunidin and malvidin 3-p-coumaroylglucosides. Peak i yielded a signal in the MSspectrum at the same m/z as the B-type vitisin of

malvidin-3-acetylglucoside, but its retention time waslower. This compound was present in a low concentration(<0.05%) only in the Marselan sample, and it probablyoriginated from the cycloaddition of the acetaldehydewith malvidin 3-acetylgalactoside (peak 28). Since Mar-selan wine possessed the highest percentage of com-pound 28, it appears reasonable that compound i wasonly present in this sample. Compounds d and i are de-scribed here for the first time.

The UV-vis spectra of compounds 48, 65, 72, 73, 77,and 78 were typical of compounds first described else-where (Fulcrand et al. 1996) which originate in a reactionbetween the different anthocyanins and 4-vinylphenol.Their presence in wine has been described (Hayasaka andAsenstorfer 2002, Vivar-Quintana et al. 2002, Wang et al.2003, Alcalde-Eon et al. 2004a). Two different mechanismsof formation have been proposed: (1) the anthocyanin re-acts with the vinylphenol formed by decarboxylation ofp-coumaric acid by yeasts (Fulcrand et al. 1996) and (2)the reaction occurs directly between anthocyanin and theintact hydroxycinnamic acid (Schwarz et al. 2003). Dataobtained in the MS, MS2, and MS3 analyses helped con-firm the nature of the compounds and allowed their iden-tification. Peaks 48, 65, 72, and 73 were the 4-vinyl-phenol derivatives of the 3-glucosides of delphinidin,petunidin, peonidin, and malvidin, respectively, whereaspeaks 77 and 78 corresponded to the 4-vinylphenol de-rivatives of malvidin 3-(6’’-acetyl) and 3-(6’’-p-coumaroyl)glucosides (Table 2). These compounds were present in allthe wine samples, but Cheveñasco had the highest per-centages.

Also related to the 4-vinylphenol derivatives of the an-thocyanins are 4-vinylcatechol and 4-vinylguaiacol, inwhich the additional aromatic ring in the pyranoantho-cyanin structure bears two substitutions in the ortho po-sition instead of the single hydroxyl group of the 4-vinylphenol derivatives. Their UV-vis spectra are slightlydifferent when compared to the 4-vinylphenol derivatives.Thus, the UV-vis spectra were helpful in the assignmentof identities since the retention times, m/z ratios, andfragmentation patterns of all these compounds in theconditions employed in this study were at times identical.Peaks l, 68, 74, and n were identified as the 4-vinyl-catechol derivatives of petunidin 3-glucoside, malvidin-3-glucoside, malvidin 3-acetylglucoside, and malvidin 3-p-coumaroylglucoside, respectively. They were less abun-dant than the vinylphenol derivatives, and Cheveñascoalso contained the highest percentages. Peaks 75 and mcorresponded to the 4-vinylguaiacol derivatives ofmalvidin-3-glucoside and 3-acetylglucoside, and no morepeaks corresponding to these types of compounds werefound in any sample. Their presence in red wine has beenreported (Hayasaka and Asenstorfer 2002, Wang et al.2003, Alcalde-Eon et al. 2004a).

Peak 66 possessed a UV-vis spectrum similar to allthese vinyl derivatives, and its molecular ion produced asignal in the MS spectrum at m/z 805. Taking into account

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Table 2 Chromatographic, UV-vis, mass spectral data, identities, and percentages of compoundsdetected in Tannat, Caladoc, Marselan, Marzemino, and Cheveñasco wines.

M+ MS2 MS3 λmax

Peak Rta (m/z) fragm fragm (nm) Compound TAN CAL MARS MARZ CHE

Anthocyanins

7 22.1 465 303 303 277, 342, 524 Delphinidin-3-glucoside 3.9 0.8 0.6 3.2 2.2

10 26.4 449 287 287 279, 516 Cyanidin-3-glucoside 0.6 *b * 0.1 0.2

12 28.6 479 317 317 277, 347, 525 Petunidin-3-glucoside 8.6 3.7 2.4 5.4 5.1

16 34.5 463 301 301 280, 517 Peonidin-3-glucoside 4.4 2.3 1.8 1.9 2.9

19 36.0 493 331 331 277, 348, 527 Malvidin-3-glucoside 27.7 35.8 35.9 23.2 36.4

f 37.1 521 317 317 Petunidin-3-(6”-acetylgalactoside) ndc nd nd 0.3 nd

22 38.7 507 303 303 276, 346, 527 Delphinidin-3-(6”-acetylglucoside) 2.0 0.3 0.3 3.0 0.4

28 41.1 535 331 331 278, 531 Malvidin-3-(6”-acetylgalactoside) 0.6 1.1 1.3 1.0 nd

31 41.5 491 287 287 280, 523 Cyanidin-3-(6”-acetylglucoside) 0.5 * * 0.4 nd

32 42.0 521 317 317 270, 529 Petunidin-3-(6”-acetylglucoside) 5.0 1.4 1.2 5.0 0.9

40 44.3 505 301 301 280, 522 Peonidin-3-(6”-acetylglucoside) 2.1 2.0 1.6 1.7 0.5

42 44.6 535 331 331 278, 350, 530 Malvidin-3-(6”-acetylglucoside) 17.3 23.8 27.6 23.4 8.5

43 45.0 611 303 303 280, 313, 531 Delphinidin-3-(pcoumglc) 0.9 0.3 0.1 0.5 0.3

49 46.2 655 331 331 282, 328, 436, 534 Malvidin-3-(6”-caffeoylglucoside) 0.4 1.7 1.0 0.2 0.2

55 46.9 595 287 287 Cyanidin-3-(pcoumglc) 0.3 0.1 nd 0.1 0.1

56 47.2 625 317 317 282, 313, 532 Petunidin-3-(pcoumglc) 2.1 1.2 0.3 0.8 0.6

59 47.9 609 301 301 283, 300, 535 Peonidin-3-(pcoumglc) (cis isomer) 0.1 0.1 nd nd nd

61 48.1 639 331 331 280, 303, 537 Malvidin-3-(pcoumglc) (cis isomer) 0.5 1.2 0.5 0.4 0.6

67 49.1 609 301 301 283, 313, 526 Peonidin-3-(pcoumglc) (trans isomer) 1.0 0.9 0.7 0.4 nd

69 49.4 639 331 331 282, 313, 532 Malvidin-3-(pcoumglc) (trans isomer) 8.1 11.0 8.3 5.3 6.2

Pyranoanthocyanins

4 20.3 533 371 371 297, 368, 507 A-type vitisin of Dp-3-glc 0.1 nd nd 0.1 0.1

c 26.0 489 327 327 B-type vitisin of Dp-3-glc nd nd nd 0.3 nd

11 27.4 547 385 385 299, 371, 508 A-type vitisin of Pt-3-glc 0.2 * * 0.1 0.2

14 32.1 589 385 385 A-type vitisin of Pt-3-acetylglc 0.1 nd nd 0.1 nd

d 32.4 531 327 327 495 B-type vitisin of Dp-3-acetylglc nd nd nd 0.3 nd

15 34.0 531 369 369 503 A-type vitisin of Pn-3-glc 0.1 nd nd * 0.2

17 35.0 503 341 341 492 B-type vitisin of Pt-3-glc 0.1 nd 0.1 0.8 nd

18 35.7 561 399 399 299, 372, 510 Vitisin A 1.4 0.8 0.6 0.8 2.4

23 38.8 603 399 399 A-type vitisin of Mv-3-acetylglc 0.6 0.4 0.4 0.7 0.3

g 39.3 545 341 341 B-type vitisin of Pt-3-acetylglc nd nd nd 0.8 nd

24 39.8 517 355 355 294, 358, 490 Vitisin B 0.3 0.4 1.6 4.5 0.8

i 42.2 559 355 355 B-type vitisin of Mv-3-acetylgalactoside nd nd * nd nd

33 42.3 531 369 369 480 Acetone derivative of Mv-3-glc 0.1 0.2 0.3 0.1 0.3

j 42.4 529 325 325 B-type vitisin of Pn-3-acetylglc nd nd * 0.5 nd

35 42.7 559 355 355 298, 361, 494 B-type vitisin of Mv-3-acetylglc 0.1 0.2 1.1 4.9 0.3

41 44.4 707 399 399 517 A-type vitisin of Mv-3-pcoumglc 0.5 0.4 0.2 0.3 0.8

44 45.1 573 369 369 Acetone derivative of Mv-3-acetylglc * 0.2 0.3 0.2 0.1

k 45.8 649 341 341 B-type vitisin of Pt-3-pcoumglc nd nd nd 0.1 nd

48 46.1 581 419 419 503 Vinylphenol derivative of Dp-3-glc 0.4 0.3 0.2 0.3 1.5

l 46.7 611 nd nd Vinylcatechol derivative of Pt-3-glc nd nd nd nd 0.5

62 48.2 663 355 355 279, 307, 497 B-type vitisin of Mv-3-pcoumglc 0.1 0.2 0.3 1.2 0.2

65 48.7 595 433 433 503 Vinylphenol derivative of Pt-3-glc 0.1 0.1 * 0.1 1.0

66 49.0 805 nd nd 510 Vinylepicatechin derivative of Mv-3-glc * * 0.1 * 0.2

68 49.2 625 463 463 511 Vinylcatechol derivative of Mv-3-glc 0.1 0.1 0.2 0.1 1.9

72 50.8 579 417 417 275, 403, 500 Vinylphenol derivative of Pn-3-glc 0.1 0.1 0.1 * 0.9

73 51.1 609 447 447 294, 336, 408, 504 Vinylphenol derivative of Mv-3-glc 0.5 1.1 1.7 0.9 9.2

74 51.3 667 463 463 513 Vinylcatechol derivative of Mv-3-acetylglc 0.1 nd 0.2 0.1 0.3

75 51.6 639 477 477 511 Vinylguaiacol derivative of Mv-3-glc 0.1 0.3 0.3 0.1 1.6

77 53.4 651 447 447 298, 416, 505 Vinylphenol derivative of Mv-3-acetylglc 0.2 0.6 1.1 0.7 1.5

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Table 2 continued

M+ MS2 MS3 λmax

Peak Rta (m/z) fragm fragm (nm) Compound TAN CAL MARS MARZ CHE

Pyranoanthocyanins, cont’d.

m 53.8 681 477 477 514 Vinylguaiacol derivative of Mv-3-acetylglc nd nd 0.2 0.1 0.2

n 53.8 771 nd nd 514 Vinylcatechol derivative of Mv-3-pcoumglc nd nd nd nd 0.3

78 55.9 755 447 447 289, 313, 416, 505 Vinylphenol derivative of Mv-3-pcoumglc 0.1 0.3 0.3 0.2 1.5

Direct condensation products

1 10.9 797 635 467 281, 440, 531 Mv-3-glc-GC 0.2 0.2 0.2 0.2 0.8

2 11.0 753 591 439 Dp-3-glc-C 0.1 nd nd nd 0.2

3 16.3 767 605 453 Pt-3-glc-C 0.1 0.1 0.1 0.1 0.3

5 20.4 751 589 437 279, 459, 530 Pn-3-glc-C 0.1 0.1 0.1 0.1 0.2

6 21.0 781 619 467 288, 455, 532 Mv-3-glc-C 0.5 0.8 0.6 0.3 2.7

21 37.3 823 619 467 Mv-3-acetylglc-C 0.2 0.2 0.3 0.2 0.3

Acetaldehyde-mediated condensation products

25 40.0 795 343 343 539 Pt-3-glc-ethyl-(E)C 0.1 * * * 0.1

27 41.0 809 357 357 278, 350, 446, 536 Mv-3-glc-ethyl-EC 0.1 * 0.1 0.1 *30 41.5 809 357 357 Mv-3-glc-ethyl-C 0.1 0.2 0.1 0.1 0.1

34 42.5 809 357 357 281, 352, 445, 541 Mv-3-glc-ethyl-C 0.3 0.4 0.2 0.3 0.1

38 43.4 809 357 357 278, 362, 446, 542 Mv-3-glc-ethyl-EC 0.1 0.1 0.1 0.4 0.1

50 46.3 851 357 357 281, 539 Mv-3-acetylglc-ethyl-C 0.1 0.1 0.1 0.3 nd

60 48.0 851 357 357 Mv-3-acetylglc-ethyl-EC * 0.1 * 0.1 nd

63 48.2 955 nd nd Mv-3-pcoumglc-ethyl-(E)C 0.2 0.3 0.1 0.1 0.2

64 48.6 955 nd nd Mv-3-pcoumglc-ethyl-(E)C 0.1 0.1 * 0.1 nd

Other compounds

8 23.4 655 331 331 Mv-3,5-diglc 0.1 0.1 0.1 0.7 0.1

9 24.8 641 nd nd Pt-3,7-diglc * nd nd nd nd

13 31.2 655 331 331 278, 526 Mv-3,7-diglc 0.2 0.4 0.6 0.3 0.4

29 41.4 463 331 331 276, 533 Mv-3-pentose? 0.4 0.3 0.5 0.2 nd

46 45.6 355 nd nd 508 Vitisin B aglycone? 0.8 nd nd nd nd

53 46.6 301 301 286 Peonidin aglycone 0.2 nd nd nd nd

54 46.9 331 331 316 Malvidin aglycone 0.1 0.2 0.3 0.2 0.1

70 49.9 369 369 369 506 Acetone derivative of Mv aglycone 2.2 0.2 0.1 0.1 nd

71 50.3 355 355 355 508 Vitisin B aglycone 0.1 0.3 0.8 0.2 0.5

a 17.9 753 nd nd Unknown nd nd * nd nd

b 17.9 783 nd nd Unknown nd nd * nd nd

e 36.7 697 nd nd Unknown nd nd nd * nd

20 36.9 971 nd nd Unknown * nd nd nd nd

h 40.0 653 635 447 Unknown nd 0.7 0.8 nd nd

26 40.8 819 657 531 Unknown 0.3 0.1 0.1 0.1 0.4

36 43.2 549 345 345 Unknown * nd 0.1 0.1 nd

37 43.3 853 nd nd Unknown * nd nd nd nd

39 44.0 1027 823 449 Unknown 0.4 0.2 0.5 0.5 0.2

45 45.2 1117 809 647 Unknown 0.1 0.1 0.1 0.1 0.2

47 46.1 1131 823 661 Unknown 0.5 0.7 0.4 0.3 0.8

51 46.3 847 685 531 450 Unknown 0.2 0.1 0.3 0.2 0.4

52 46.4 1131 823 661 Unknown * nd nd nd nd

57 47.4 803 495 331 450 Unknown 0.1 0.2 0.1 nd 0.1

58 47.7 685 523 331 440, 532 Unknown 0.2 0.2 0.1 0.1 0.3

76 51.6 587 425 343 284, 441, 539 Unknown 0.2 nd nd nd nd

aAbbreviations: Rt: retention time. TAN: Tannat. CAL: Caladoc. MARS: Marselan. MARZ: Marzemino. CHE: Cheveñasco. Dp: delphinidin.Cy: cyanidin. Pt: petunidin. Pn: peonidin. Mv: malvidin. Glc: glucose. acetylglc: (6’’-acetylglucoside). pcoumglc: (6’’-p-coumaroylglucoside).C: catechin. EC: epicatechin. (E)C: (epi)catechin. GC: gallocatechin.

b*: detected, but not quantified.cnd: not detected.

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Am. J. Enol. Vitic. 57:4 (2006)

its retention time, spectral data, and other data (Alcalde-Eon et al. 2004a, Rivas-Gonzalo et al. 1995, Hayasaka andAsenstorfer 2002, Monagas et al. 2003), this compoundwas identified as the product resulting from a cycloaddi-tion reaction between the monoglucoside of malvidin andvinylepicatechin. Two compounds of this type can befound (Monagas et al. 2003, Alcalde-Eon et al. 2004a): oneoriginated in the reaction between the anthocyanin andvinylcatechin and the other between the anthocyanin andvinylepicatechin. NMR studies have allowed differentia-tion of these two isomers (Mateus et al. 2002) and showthat the vinylepicatechin isomer is more retained than thevinylcatechin isomer in reversed-phase. In addition, re-tention time of peak 66 was similar to the former whendetected in other red wines under the same chromato-graphic conditions (Alcalde-Eon et al. 2004a). This com-pound was detected in all wines, but in low amounts.

The molecular ion corresponding to peak 33 pos-sessed the same m/z ratio as that of peak 15 (A-typevitisin of peonidin-3-glucoside). Nevertheless, their UV-vis spectra were different, and that of peak 33 had amaximum at a lower wavelength than that of peak 15(Table 2). In addition, the compound corresponding topeak 33 was more retained in reversed-phase than that ofpeak 15, indicating that the former was less polar. Weidentified it as a pyranoanthocyanin originating in the re-action between malvidin-3-glucoside and acetone (vinyl-methyl adduct). Although the possibility of formation ofthis type of compound has been demonstrated (Benab-deljalil et al. 2000, Lu and Foo 2001), there are few refer-ences to its presence in wine (Hayasaka and Asenstorfer2002, Wang et al. 2003). The molecular ion of peak 44 had42 additional amu compared to peak 33, and its retentiontime was higher. Thus, peak 44 was identified as thevinylmethyl adduct of malvidin-3-(6’’-acetylglucoside). Toour knowledge, this is the first time this compound hasbeen described in red wines.

The Cheveñasco and Marzemino samples showed thehighest percentages (26.2 and 18.5%, respectively) of to-tal pyranoanthocyanins (Table 1). Nevertheless, the chemi-cal profile was different in both samples. In Cheveñasco,

~78% of the pyranonanthocyanins corresponded to the 4-vinylphenol, 4-vinylcatechol, 4-vinylguaiacol, and vinyl-catechin derivatives of the anthocyanins. Among them,the 4-vinylphenol derivative of malvidin-3-glucoside ac-counted for 9.2% of the total area. The A-type vitisinswere also important (15% of the pyranoanthocyanins) andthat of malvidin-3-glucoside accounted for 2.4% of the to-tal area. In Marzemino, ~72% of the pyranoanthocyaninscorresponded to B-type vitisins, in whose formation ac-etaldehyde is involved. The most representative com-pounds were the B-type vitisins of malvidin-3-glucosideand malvidin-3-acetylglucoside, which accounted for 4.5%and 4.9% of the total peak area, respectively.

The Tannat and Caladoc samples had the same per-centage of total pyranoanthocyanins (5.5%), but again,the families of pyranoanthocyanins were different. InTannat, the pyruvic acid adducts of the anthocyanins (A-type vitisins) represented almost 60% of the total pyrano-anthocyanins. 4-Vinylphenol, 4-vinylcatechol, 4-vinylgua-iacol, and vinylcatechin derivatives were also important(~30% of the pyranoanthocyanins). In Caladoc, the per-centages were inverted: 27% for A-type vitisins and 52%for vinyl derivatives.

The Marselan sample had 9.5% total pyranoantho-cyanins, 47% of which were vinyl derivatives and 32% B-type vitisins. As could be predicted, acetylated pyrano-anthocyanins were more abundant than coumaroylated inall wines, since the acetyl derivatives of the monoglu-cosides were more abundant than the coumaroyl and thepyranoanthocyanins that originated from them.

Flavanol-anthocyanin direct and acetaldehyde-mediatedcondensation products. The chromatographic and spec-tral features of peak 6 corresponded to those of the com-pound originating in direct condensation between mal-vidin-3-glucoside and catechin when observed under thesame chromatographic conditions (Alcalde-Eon et al.2004a). This compound has been reported in wines (Vivar-Quintana et al. 1999, 2002, Remy et al. 2000, Monagas etal. 2003, Wang et al. 2003). Similarly, peaks 2, 3, and 5were identified as direct condensation products of cat-echin with delphinidin, petunidin, and peonidin 3-gluco-sides, respectively. To our knowledge, compound 3 is re-ported here in wines for the first time, peak 2 waspreviously identified in wine (Alcalde-Eon et al. 2004a),and peak 5 in red grapes and wines (González-Paramás etal. 2006; Alcalde-Eon et al. 2004a). The molecular andfragment ions of compound 21 (Table 2), which was moreretained than the former compounds, helped in its identi-fication as the direct condensation product of malvidin-3-(6’’-acetylglucoside) and catechin, here reported for thefirst time.

The UV-vis spectrum of peak 1 was similar to those ofthese peaks, but its earlier elution indicated substitutionin the molecule with more polar substituents. The molecu-lar ion of this compound gave a signal at m/z 797. Themajor fragment ion in the MS2 corresponded to the agly-cone (m/z 635) and was fragmented in the MS3 analysis

Figure 2 UV-vis specta and structures corresponding to vitisins A andB (peaks 18 and 24).

Pigment Profiles in Monovarietal Wines – 457

Am. J. Enol. Vitic. 57:4 (2006)

yielding five major signals (Figure 3). The presence of theion at m/z 331 indicated that the compound was a mal-vidin derivative. The losses of 168 and 304 amu were in-dicative of the presence of one moiety of gallocatechin inthe compound, since the first loss results in the retroDiels–Alder cleavage and the second corresponds tothe loss of the entire gallocatechin moiety when it islinked to another gallocatechin or anthocyanin by aninterflavanic linkage at C4. The fragment ion of 373 isalso indicative of the presence of malvidin in the com-pound, since this ion is also present in the fragmentationof compound 6 and originates in cleavage of bonds 2 and4 in the C ring of the flavanol (Figure 3). The fragmenta-tion patterns of compound 1 and compound 6 (Alcalde-Eon et al. 2004a) are the same in both cases. The fragmen-tation pattern of direct condensation products always hasthe same steps independent of the anthocyanin andflavanol involved (González-Paramás et al. 2006). Thus,compound 1 was identified as the product of direct con-densation between malvidin-3-glucoside and gallocatechin.As far as we know, this is the first time it has been de-scribed in wines.

Peaks 27, 30, 34, and 38 had molecular ions with thesame m/z ratios (809) and fragmentation patterns. Theirspectral and chromatographic features corresponded tothose of the dimers originating in the acetaldehyde-medi-ated condensation between (epi)catechin and malvidin-3-glucoside (Alcalde-Eon et al. 2004a). In this reaction twodiasteroisomers may be formed for each pair of com-pounds (catechin-malvidin-3-glucoside and epicatechin-malvidin-3-glucoside), as was demonstrated in model so-lutions (Rivas-Gonzalo et al. 1995). The presence of allfour isomers was detected in each analyzed wine. Thus,peaks 30 and 34 were identified as the two possible iso-mers of malvidin-3-glucoside-ethyl-catechin and peaks 27and 38 were identified as the isomers of malvidin-3-glu-coside-ethyl-epicatechin.

The molecular ions of compounds 50 and 60 had 42additional amu in comparison to those of peaks 27, 30,34, and 38, and the fragment ions originated in the MS2

and MS3 analyses were the same as in the latter ones.These compounds were identified as malvidin-3-acetyl-glucoside-ethyl-(epi)catechin dimers, and peak 50 likelycorresponds to the most abundant of the two catechin

Figure 3 MS2 and MS3 spectraand the fragmentation schemeof compound 1 (m/z 797).

458 – Alcalde-Eon et al.

Am. J. Enol. Vitic. 57:4 (2006)

derivatives and peak 60 to the most abundant of theepicatechin derivatives, but further studies are necessaryfor complete identification. Similarly, the molecular ionsof peaks 63 and 64 were identified as the products re-sulting from acetaldehyde-mediated condensation be-tween malvidin-3-p-coumaroylglucoside and (epi)catechin,but their complete identities have yet to be established.

The chromatographic and spectral data of compound25 were similar to those of flavanol-anthocyanin acetalde-hyde-mediated condensation products, and it was identi-fied as the product derived from petunidin-3-glucoside.The identity of the flavanol involved (either catechin orepicatechin) has yet to be established.

The abundance of these two families of compoundswas not very high. In Tannat and Caladoc, the ratio be-tween direct and acetaldehyde-mediated condensationproducts was close to 1, which means that the relativeabundance of both families was similar. In Marselan andCheveñasco, the direct condensation products were moreabundant than the acetaldehyde-mediated products. ForCheveñasco, the ratio was much higher (5.5). Moreover,the Cheveñasco sample showed the most direct conden-sation products. In contrast, in the Marzemino sample,the acetaldehyde-mediated condensation products weremore abundant than the direct ones (ratio: 0.5). The pref-erential formation of this type of compound and the pres-ence of B-type vitisins as major pyranoanthocyanins inthis sample might be related to greater acetaldehyde pro-duction in Marzemino with respect to the other wines(Vivar-Quintana et al. 1999).

Diglucosides, aglycones, and unknown compounds.Peak 8 was identified as malvidin-3,5-diglucoside. Thiscompound was observed in all five wines analyzed but inlow percentages, except in Marzemino, where it repre-sented 0.7% of the total area and its retention time wascoincident with that of a commercial standard of malvin(Sigma, St Louis, MO). Peak 13 showed the same m/z ra-tio as peak 8, but its UV-vis spectra was different. Thepresence of a shoulder in the 440 nm region was indica-tive of the absence of substitution in the hydroxyl groupin position 5 of the anthocyanin. The fragmentation pat-tern of the molecular ion (fragment ions at m/z 331 and493) indicated that the substitutions should be in differentpositions of the molecule (Giusti et al. 1999). Thus, com-pound 13 is proposed to be malvidin-3,7-diglucoside.Similarly, peak 9 is proposed to be petunidin-3,7-diglu-coside.

Peaks 53, 54, 70, and 71 corresponded to aglycones(peonidin, malvidin, pyranoanthocyanins resulting fromthe cycloaddition between malvidin and acetone and be-tween malvidin and acetaldehyde). Peak 46 showed thesame m/z ratio as peak 71 (aglycone of vitisin B) and itsidentity has yet to be established, as do the identities ofpeaks a, b, e, h, 20, 26, 29, 36, 37, 39, 45, 47, 51, 52,57, 58, and 76. Some of these compounds were exclusiveto one or two samples, while others were present in all ofthem.

ConclusionThe Tannat sample was characterized by the large pro-

portion of anthocyanins (86%), the monoglucosides beingthe most abundant compounds. Acetyl derivatives of an-thocyanins were more abundant than p-coumaroyl. Com-pared to the other wines, the formation of anthocyanin-derived pigments was not very important, vitisin A beingthe most representative.

The Caladoc sample had the same total anthocyaninsas Tannat. Unlike Tannat, in which malvidin-3-glucoside,3-acetylglucoside, and 3-(6’’-p-coumaroylglucoside) repre-sented 60% of the anthocyanins, these three compoundscomprised 80% of the anthocyanins in Caladoc. Conse-quently, the anthocyanin-derived pigments were almost allmalvidin derivatives. The percentages of these derivedpigments were similar to those of Tannat, but the majorcompounds were different. Among the pyranoantho-cyanins, the 4-vinylphenol derivative of malvidin-3-glu-coside was the most abundant. A and B-type vitisinswere also found.

The Marselan sample had less total anthocyanins(83%), but the three malvidin derivatives were dispropor-tionately abundant (86% of the anthocyanins, 70% of thetotal area). The 4-vinylphenol derivative of malvidin-3-glucoside and vitisin B, together with their correspondingacetyl derivatives, were the most abundant anthocyanin-derived pigments.

In the Marzemino sample, the anthocyanins only ac-counted for 76.1% of the total pigments, and monoglu-cosides of the anthocyanins and their acetylderivativeswere equally abundant. The percentage of anthocyanidinsother than malvidin was higher in Marzemino than inCaladoc and Marselan, but lower than in Tannat.Marzemino is well characterized by its pyranoanthocyanincomposition (18.5% of the total area), and B-type vitisinswere the major anthocyanin-derived pigments. Presence ofthese vitisins and anthocyanin-flavanol acetaldehyde-me-diated condensation products might indicate higher levelsof acetaldehyde in this wine than in the others.

Although Cheveñasco had the least anthocyanin(65%), the content of anthocyanin monoglucosides wassimilar to Tannat (46.9%). Nevertheless, Cheveñasco hadless acylated compounds when compared to the othersamples. The second most abundant group of compoundswas the pyranoanthocyanin family (26.2%). The 4-vinylphenol derivative of malvidin-3-glucoside accountedfor 9.2% of the total area, being the second most impor-tant compound after malvidin-3-glucoside. The anthocya-nin-flavanol direct condensation products were also im-portant in this sample, showing the highest contentamong all the wines.

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