Aoki, H.

Fujiwara, M.

Hirano, K.

Ichi, T.

Okamoto, G.

Ueki, K.

English

JKI Open Journal Systems (Julius Kühn-Institut)

Vitis 41 (3), 161162 (2002)Research NoteJuice constituents and skin pigmentsin Vitis coignetiae Pulliat grapevinesG. OKAMOTO1), K. UEKI2), T. ICHI3), H. AOKI1), M. FUJIWARA1)and K. HIRANO1)1) Faculty of Agriculture, Okayama University, Okayama, Japan2) The Graduate School of Natural Science and Technology,Okayama University, Okayama, Japan3) San-Ei Gen F. F. I., Inc., Toyonaka, JapanK e y   w o r d s :  Vitis coignetiae, juice constituents, anthocy-anin, malvidin-3,5-diglucoside.Introduction: Vitis coignetiae Pulliat is a wild grape spe-cies widely distributed in mountainous regions of JapaneseIslands. In the Hiruzen Highlands, located in the northernOkayama Prefecture, V. coignetiae grapevines have beendomesticated during the last two decades mainly for winemaking.Vines had been raised from cuttings of dormant canescollected from several wild vines during 1980s. Nowadays,the cultivated area is about 10 ha. The berries contain apeculiar flavor and abundant red pigments, but a very highacidity and low sugar level which are thought to be deterio-rating the wine's taste. In the Hiruzen vineyards, grapes areusually harvested in late September, i.e. 6 weeks afterveraison, primarily to avoid fungal damages of berries suchas ripe rot and downy mildew (UEKI et al. 2001). However, ithas not yet been established when V. coignetiae berriesreach full ripeness and what are the constituents of juiceand skins at this stage.The aim of this study was to elucidate the optimum har-vest time for wine making as well as the characteristics ofjuice and skin constituents in V. coignetiae grape berries ascompared to Cabernet Sauvignon (V. vinifera L.).Material and Methods: Six and five 15-20-year-oldV. coignetiae vines, exhibiting the typical color in youngleaves and cluster stems, were chosen from 6 commercialvineyards in the Hiruzen Highlands in 2000 and 2001. Theywere planted 4x2.5 m and trained to a multiple scaffold branchsystem on a horizontal trellis (year 2000) and to a bilateralcordon system on a vertical trellis (year 2001). 20-30 berrieswere sampled from each vine at 2 to 3-week intervals fromearly August (before veraison) until late October. Berries of8-year-old vines of Cabernet Sauvignon (V. vinifera), culti-vated in the same district, were also sampled in early Octo-ber 2001, i.e. at the usual harvest time of Hiruzen.J u i c e   c o n s t i t u e n t s :  Berry samples were pressedby hand through doubled gauze to obtain the juice. Totalsoluble solids (TSS) were measured by a refractometer(Atago-30) and titratable acidity (TA) by titrating the juicewith 0.1 N NaOH until neutralization. Sugar and acid con-tents were determined by GC (Shimadzu GC-14A) after puri-fying the sample juice by ion exchange resin and silylatingwith hexamethyldisilazane and trimethylchlorosilane. Foramino acid analysis, 0.5 ml of juice was filtered through af 0.45 mm filter and injected into a HPLC (JLC-300).S k i n   a n t h o c y a n i n :  10 mg of lyophilized skinsample was homogenized with 25 ml of 80 % methanol con-taining 1 % HCl. Anthocyanins were extracted for 1 h at25 °C twice. OD535 of the extracts was determined with aspectrophotometer (Beckmann DU530). For the identifica-tion of skin anthocyanins, 1 kg of fresh V. coignetiae berrieswere extracted with 4000 ml of 50 % ethanol containing 0.1 %trifluoroacetic acid (TFA) for 1 d. The extract was concen-trated under reduced pressure at 40 °C and applied to a1-liter HP-20 column. The adsorbed pigments were elutedwith 60 % EtOH containing 0.1 % TFA. The eluent was con-centrated under reduced pressure at 40 °C and 204.3 g ofberry colorant were obtained. The colorant was purified withYMC-ODS-L80 (30 i.d.x400 mm, YMC) column and SephadexLH-20 (40 i.d.x600 mm, Pharmacia) column. HPLCchromatograms of the colorant revealed that it consists ofabout 7 pigments (Figure). Three main pigments, Pig-1, Pig-2 and Pig-3, were isolated, and the structures of these pig-ments were identified using ESI-LC-MS and NMR (1D and2D NMR, HMBC, HMQC).Correspondence to: Dr. G. OKAMOTO, Okayama University, Fac-ulty of Agriculture, Tsushima-naka, Okayama 700-8530, Japan.Fax: +81-86-251-8388. E-mail: gokamoto@cc.okayama-u-ac.jpFigure: HPLC chromatogram of the colorant extracted fromV. coignetiae berry skins.Results and Discussion: In 2000, TSS of berry juice,sampled from 6 V. coignetiae vines, were about 15 °Brix inlate September and 18-20 °Brix in late October (Table). Thesedata suggest that in Hiruzen V. coignetiae grapes are gener-ally harvested before the stage of full ripeness. GC analysesof sugars revealed that the levels of fructose were signifi-cantly higher than of glucose. TA, expressed as tartaric acidequivalent, decreased rapidly after veraison. However, thefinal TA level was higher than 1.6 g 100 ml-1 for each vine.Malate decreased continuously until late October whiletartarate did not decrease after late September. Total aminoacid content increased significantly until late October, butthe final levels were about 5 mmol l-1 or lower, much lowerthan in berries of V. vinifera wine grapes (HUANG and OUGHAbsorbance (525nm)162 G. OKAMOTO, K. UEKI, T. ICHI, H. AOKI, M. FUJIWARA and K. HIRANO1991). Furthermore, the major constituents were proline andalanine indicating that the amino acid profiles of V. coignetiaeberries resemble those of V. labrusca varieties rather thanV. vinifera grapes (SPAYD and ANDERSEN-BAGGE 1996).V. coignetiae berries contain far higher TA and lowerpH levels than Cabernet Sauvignon berries. The extraordi-nary high level of tartaric acid, compared to that of mostwine grape cultivars (LAVEE and NIR 1986), might be specificfor this species. It is generally accepted that grape berriesfor wine making should contain 20-25 °Brix of TSS and 0.6-0.8 mg 100 ml-1 of TA (WINKLER 1974). The lower levels ofTSS in both V. coignetiae and Cabernet Sauvignon berriesharvested in the Hiruzen vineyards may have resulted frominsufficient sunshine and frequent rainfall during autumn.The significantly higher level of skin anthocyanins is acharacteristic of V. coignetiae berries. The molecular for-mula of three main constituents, Pig-1, Pig-2 and Pig-3, shownin the Figure, were determined as C29H35O17, C23H25O12,C38H41O19, respectively, by positive-ion ESI-LC-MS (mo-lecular ion m/z 655, 493, 801). ESI-LC-MS spectra of the threepigments indicated that malvidin (m/z 331) is a common agly-cone unit in those pigments and that there are two glucoseunits in Pig-1 (m/z 655), one glucose unit in Pig-2 (m/z 493),and two glucoses and one p-coumarate in Pig-3 (m/z 801).On the basis of these results and spectral data of NMR, thestructures of Pig-1, Pig-2 and Pig-3 were finally confirmed asmalvidin-3,5-diglucoside, malvidin-3-glucoside, andmalvidin-3,5-diglucoside-coumarate, respectively. The otherpigments, denoted as a, -b, -c, and -d in the Figure can beestimated to be delphinidin-3-glucoside, petunidin-3-gluco-side, malvidin-3-glucoside-acetate, and malvidin-3-glucside-coumarate, respectively, by referring to WULF and NAGEL(1978). The major anthocyanins in V. coignetiae berry skin,identified als malvidin-3,5-diglucoside and malvidin-3,5-diglucoside-p-coumarate are usually not found in berries ofV. vinifera cultivars but are common in V. labrusca and hy-brids between V. vinifera and V. labrusca (GALVI and FRANCIS1978; YOKOTSUKA and SINGLETON 1997). IGARASHI et al. (1986)have already shown the presence of malvidin-3,5-glucosidein V. coignetiae berries and noted a high antioxidative activ-ity of this pigment. As shown in our previous report (UEKIet al. 2002), the total anthocyanin content in V. coignetiaeberry skins is 2-4 times higher than that in V. vinifera cultivarssuch as Cabernet Sauvignon, Cabernet Franc, and Pinot noir.The anti-oxidative activity of V. coignetiae juice should befurther investigated.There may be some genetic diversity in the V. coignetiaevines cultivated in Hiruzen, because they have been propa-gated by cuttings collected from several wild vines. Juiceand skin constituents among the vines should be examinedand they should be genetically identified.CALVI, J. P.; FRANCIS, F. J.; 1978: Stability of Concord grape (Vitis lab-rusca) anthocyanins in model systems. J. Food Sci. 43,1448-1456.HUANG, Z.; OUGH, C. S.; 1991: Amino acid profiles of commercialgrape juices and wines. Am. J. Enol. Vitic. 42, 261-267.IGARASHI, K.; TAKANASHI, K.; MAKINO, M.; YASUI, T.; 1989: Antioxidativeactivity of major anthocyanin isolated from wild grapes (Vitiscoignetiae). Nippon Shokuhin Kogyo gakkaishi 36, 852-856.LAVEE, S.; NIR, G.; 1986: Grape - maturation and accumulation ofspecific components. In: S. P. MONSELISE (Ed.): Handbook ofFruit Set and Development, 171-175. CRC Press, Florida, USA.SPAYD, S. E.; ANDERSEN-BAGGE, J.; 1996: Free amino acid composition ofgrape juice from 12 Vitis vinifera cultivars in Washington. Am. J.Enol. Vitic. 47, 389-402.UEKI, K.; AOKI, H.; OKAMOTO, G.; HIRANO, K.; 2001: Effect of leaf numberon berry ripening and characterization of juice composition inVitis coignetiae Pulliat grapevines. (in Japanese with Englishsummary). J. ASEV Jpn. 12, 58-65.YOKOTSUKA, K.; SINGLETON, V.; 1993: Disappearance of anthocyanins asgrape juice is prepared and oxidized with PPO and PPO substrates.Am. J. Enol. Vitic. 48, 13-25.WINKLER, A. J.; COOK, J. A.; KLIEWER, W. M.; LIDER, L. A.; 1974: GeneralViticulture. Univ. California Press, Berkeley, USA.WULF, L. W.; NAGEL, C. W.; 1978: High-pressure liquid chromato-graphic separation of anthocyanins of Vitis vinifera. Am. J. Enol.Vitic. 29, 42-50.T a b l eSeasonal changes in the juice constituents and skin anthocyanins of Vitis coignetiae berries in comparison to V. viniferaSpecies Date of Berry Juice Skincultivar harvest weight TSS TA pH Total Anthocyaninz(g) (°Brix) (g 100 ml-1) amino acid (O.D. 535)(mmol 1-1)V. coignetiaey              2000 Sep. 12 1.10 b 13.8 c 2.38 a 2.63 b 1.8 c 0.36 cSep. 26 1.13 ab 15.1 b 2.19 ab 2.68 ab 2.4 b 0.65 bcOct. 10 1.16 a 17.8 ab 1.86 b 2.72 ab 3.4 ab 0.90 bOct. 10 1.18 a 19.2 a 1.76 b 2.84 a 4.5 a 1.16 aV. coignetiaex              2001 Oct. 2 1.10 b 18.8 1.89 a 2.64 b 5.1 b 0.86 aCabernet Sauvignon (V. vinifera)x                    2001 Oct. 10 1.40 a 18.5 0.82 b 3.08 a 12.0 a 0.44 bz Anthocyanins extracted from 1 g of lyophilized samples were suspended into 100 ml of 80 % ETOH.y Berries were harvested from 6 V. coignetiae vines. Means were separated by DMRT at p<0.05.x Berries were harvested from 5 V. coignetiae and 3 Cabernet Sauvignon vines. Means were separated byDMRT at p<0.05.

Juice constituents and skin pigments in Vitis coignetiae Pulliat grapevines

https://ojs.openagrar.de/index.php/VITIS/article/download/4473/4407

Juice constituents and skin pigments in Vitis coignetiae Pulliat grapevines

Abstract

Similar works

Full text

Available Versions

JKI Open Journal Systems (Julius Kühn-Institut)