To understand the anthocyanin characteristics of wine grape varieties, the anthocyanin composition and content of 31 wine grape varieties were analyzed to explore the use of anthocyanins as chemical fingerprints to distinguish varieties. Results showed that a total of 21 anthocyanins were detected in the skins, including cyanidin, delphinidin, petunidin, peonidin and malvidin 3-monoglucosides (or 3,5-diglucosides) along with the corresponding acetyl and p-coumaroyl derivatives. The highest and lowest total amount of anthocyanins were detected in ‘Ruby Cabernet’ and ‘Muscat Rouge’, respectively. In the 21
As the most important water-soluble pigments in plants, anthocyanins are widely found in the flowers and fruits of higher plants, which gives them different colors [
The anthocyanins in grapes are the glucoside forms of cyanidin (Cy), delphinidin (Dp), petunidin (Pt), peonidin (Pn), pelargonidin (Pg) and malvidin (Mv), along with the corresponding acetyl, p-coumaroyl, and caffeoyl derivatives in cultivars [
In this paper, 31 red wine grape varieties in Shanxi province of China were used as materials to determine the anthocyanin composition and content by high-performance liquid chromatography-mass spectrometry (HPLC-MS). The anthocyanin types in different grape varieties were compared. Cluster analysis and principal component analysis were carried out based on the anthocyanin content of each monomer to differentiate species by using anthocyanins as chemical fingerprints.
The fruits of 31 wine grape varieties were collected from Pomology Institute, Shanxi Academy of Agricultural Science, Taigu (Taigu) and the National Grape Germplasm Resources (Taigu) during the fruit ripening period from August to October in 2017. Each variety chose the same growth plants, three plants were 1 plot and three replicates, a total of nine plants. The fruits were sampled from the middle of dragon, and 4 clusters were randomly collected from each plot (considering the shaded and sunny side of the ear, 2 clusters on both sides of the fence, a total 12 clusters). After sampling, the fruits were taken back to the laboratory, all berries were mixed, then the skins were quickly peeled off with tweezers under 4°C, frozen in liquid nitrogen and stored at −80°C. The anthocyanins were extracted and analyzed in the laboratory from November to December in 2017. The names of the test materials and the sampling times are shown in
The anthocyanins in the skins were extracted and detected according to the method of Liu et al. [
The HPLC-MS analyses were carried out using a WATERS ACQUITY UPLC-PDA detector. Chromatographic separation was carried out using a C18 column (100 mm × 2.1 mm i.d. 1.7 μm) and thermostated at 45°C. The mobile phase was a linear gradient of water/formic acid (98:2) (solvent B) in acetonitrile (solvent A), at a flow rate of 0.3 mL·min−1. The following elution gradient was used: 0–20 min, 6%–16% A; 20–28 min, 16%–23% A; 28–35 min, 23%–50% A; 35–37 min, 50% A; 37–40 min, 50%–60% A. Then the anthocyanins were identified using WATERS MALDI SYNAPT QTOF-MS. The operating parameters were: Capillary voltage, 3 000 V; cone voltage, 30.0 V; cone gas flow, 10 L·h−1; desolvation gas flow, 700 L·h−1; ion source temperature, 100°C; desolvation temperature, 400°C. The instrument was operated in a positive ion mode scanning from 50 to 2 000 m·z−1.
The anthocyanin contents were identified based on the HPLC-UV-MS fingerprint library of grape and wine anthocyanins [
The data were reported as the mean ± standard error (S.E.) of three replicates. Analysis of significant differences was performed using SPSS 17.0 data analysis software at
The total anthocyanin contents in the skins of 31 grape varieties were significantly different, ranging from 36.793 to 3995.079 mg·kg−1 FW (
Varieties | Delphinidins | Petunidins | Malvidins | Cyanidins | Peonidins | Total anthocyanins |
---|---|---|---|---|---|---|
Cabernet Franc327 | 102.381 ± 8.092ijk | 95.329 ± 4.243hij | 1119.966 ± 28.480hij | 20.297 ± 1.917ijkl | 167.030 ± 2.050fgh | 1505.003 ± 12.177klm |
Cabernet Sauvignon169 | 254.313 ± 6.143f | 162.842 ± 2.054f | 1503.547 ± 24.785ef | 34.040 ± 1.584efg | 153.927 ± 2.659ghi | 2108.669 ± 28.738gh |
Merlot181 | 324.339 ± 2.335e | 258.314 ± 2.328de | 1451.5719 ± 63.624fg | 97.212 ± 1.066c | 311.686 ± 16.003b | 2443.123 ± 53.350e |
Cabernet Gernischt | 136.457 ± 3.829h | 160.062 ± 0.361f | 1934.229 ± 0.580c | 11.577 ± 0.907lmn | 109.292 ± 3.146j | 2351.618 ± 1.808ef |
Marselan c980 | 90.154 ± 7.046klm | 104.119 ± 6.966ghi | 1791.266 ± 6.045cd | – | 48.514 ± 2.884klm | 2034.053 ± 19.149ghi |
Ruby Cabernet | 608.144 ± 21.896b | 430.195 ± 15.710bc | 2804.578 ± 56.687a | 41.116 ± 0.517e | 111.046 ± 1.659j | 3995.079 ± 96.469a |
Meiyu | 189.825 ± 30.291g | 216.846 ± 27.358e | 2252.252 ± 128.098b | 11.673 ± 1.668lmn | 126.689 ± 10.373ij | 2797.284 ± 197.788d |
Carinena | 82.726 ± 37.376klm | 101.257 ± 18.195ghij | 643.695 ± 262.684k | 4.706 ± 0.487n | 227.942 ± 77.484de | 1060.326 ± 266.380pq |
Petit Verdot | 275.275 ± 6.555f | 257.098 ± 0.448de | 2205.892 ± 118.928b | 11.272 ± 2.271lmn | 70.318 ± 8.265k | 2819.855 ± 101.390cd |
Malbec | 195.080 ± 2.800g | 259.387 ± 1.952de | 2380.979 ± 92.490b | – | 40.288 ± 1.844klmn | 2875.734 ± 89.798cd |
Blue French | 36.963 ± 4.525op | 52.104 ± 6.017jk | 1105.866 ± 108.568hij | 11.479 ± 1.920lmn | 274.763 ± 29.284c | 1481.174 ± 150.314lmn |
Pinot Noir115 | 40.264 ± 1.913no | 45.171 ± 0.824k | 603.996 ± 26.921k | 18.662 ± 0.563jklm | 254.515 ± 12.351cd | 962.607 ± 16.744qr |
Syrah100 | 119.584 ± 19.154hij | 154.332 ± 17.216f | 1540.084 ± 129.568ef | 28.796 ± 7.023fghi | 197.372 ± 40.965ef | 2040.168 ± 213.925ghi |
Medoc Noir | 92.018 ± 11.839kl | 133.257 ± 20.635fgh | 1054.045 ± 158.42j | 16.897 ± 0.293klm | 68.746 ± 1.923k | 1364.963 ± 192.524mno |
Muscat Hamburg | 31.785 ± 3.515op | 25.600 ± 3.165k | 282.656 ± 26.735l | 23.596 ± 1.851hijk | 138.514 ± 0.920hij | 502.151 ± 30.644s |
Muscat Rouge | 3.586 ± 0.261q | – | – | 25.138 ± 1.286ghijk | 8.069 ± 0.488n | 36.793 ± 0.537t |
Gamay Teinturier | 27.770 ± 7.347opq | 41.116 ± 10.230k | 1087.36 ± 229.302ij | 6.077 ± 0.891n | 197.137 ± 14.458ef | 1359.460 ± 262.227mno |
Tempranillo | 392.975 ± 25.500d | 290.343 ± 17.351d | 1271.470 ± 29.987ghi | 65.013 ± 10.202d | 140.700 ± 18.249hij | 2160.501 ± 102.289fg |
Canepabn | 19.685 ± 1.689opq | 19.933 ± 0.760k | 632.775 ± 26.978k | 10.183 ± 1.286mn | 599.757 ± 42.269a | 1282.333 ± 72.983no |
Armenia | 108.070 ± 3.270ijk | 120.856 ± 2.083fgh | 1419.053 ± 16.327fg | 28.172 ± 3.009fghij | 173.615 ± 11.457fgh | 1849.765 ± 3.492ij |
Areni | 143.710 ± 0.248h | 145.316 ± 1.867fg | 1507.007 ± 31.363ef | 36.339 ± 7.042ef | 239.229 ± 37.929cd | 2071.600 ± 11.493gh |
Zhesexiang | 10.795 ± 1.612pq | – | – | 110.783 ± 12.536b | 5.631 ± 0.018n | 127.209 ± 14.166t |
Seibel Noir | 476.611 ± 18.826c | 580.397 ± 1.253a | 2650.154 ± 121.934a | – | 181.793 ± 11.377fg | 3888.955 ± 113.272a |
Beibinghong | 591.268 ± 6.383b | 471.857 ± 21.681b | 765.479 ± 48.349k | 1239.924 ± 198.56a | 25.909 ± 1.720opq | 3094.438 ± 85.223b |
Huapu 1 | 64.450 ± 11.150mn | 57.865 ± 1.455ijk | 711.513 ± 27.516k | – | 10.235 ± 2.428n | 844.063 ± 42.549r |
Gongniang 2 | 67.956 ± 5.138lm | 117.611 ± 6.638ghij | 1509.068 ± 23.131ef | – | – | 1694.635 ± 34.907jk |
Beihong | 695.520 ± 14.964a | 394.714 ± 3.873c | 1844.269 ± 34.960cd | 32.804 ± 1.761efgh | 55.902 ± 5.434kl | 3023.210 ± 12.449bc |
Beimei | 69.683 ± 1.639lm | 65.213 ± 0.595ijk | 1081.135 ± 4.48ij | – | – | 1216.031 ± 3.444op |
2--1-3 | 98.751 ± 0.705jk | 130.884 ± 5.909fgh | 1696.674 ± 42.596de | – | – | 1926.309 ± 49.210hi |
44-6-7-1 | 199.199 ± 68.580g | 412.150 ± 208.975c | 2402.220 ± 533.103b | 3.921 ± 0.130n | 27.613 ± 2.092lmn | 3045.103 ± 390.746bc |
Moldova | 129.155 ± 29.784hi | 149.890 ± 31.790fg | 1283.437 ± 132.042gh | – | 16.336 ± 1.946mn | 1578.819 ± 195.563kl |
In this study, the anthocyanins were comprised of cyanidin (Cy), delphinidin (Dp), petunidin (Pt), peonidin (Pn) and malvidin (Mv) 3-monoglucosides (or 3,5-diglucosides) along with the corresponding acetyl and p-coumaroyl derivatives in the skins of grape. Pt and Mv were not detected in the skins of ‘Muscat Rouge’ and ‘Zhesexiang’, while in the other 29 varieties, the contents of Mv accounted for 24.7%~89.0% of total anthocyanins, the highest and lowest contents of Mv were observed in ‘Seibel Noir’ and ‘Beibinghong’, respectively (
Varieties | Diglucoside |
Monoglucoside |
Acylated |
Coumarylated |
Methylated |
---|---|---|---|---|---|
Cabernet Franc327 | – | 1505.003 ± 12.177g | 28.420 ± 3.211ijk | 251.156 ± 6.308lm | 1382.325 ± 22.187lm |
Cabernet Sauvignon169 | – | 2108.669 ± 28.738d | 12.243 ± 0.479mn | 167.349 ± 2.710no | 1820.316 ± 24.179ghi |
Merlot181 | – | 2443.123 ± 53.350c | 25.601 ± 2.608jk | 313.497 ± 37.893kl | 2021.572 ± 49.949ef |
Cabernet Gernischt | – | 2351.618 ± 1.808c | 58.925 ± 20.307cd | 602.204 ± 31.241efg | 2203.583 ± 2.928de |
Marselan c980 | – | 2034.053 ± 19.149de | 30.245 ± 0.906ijk | 590.346 ± 40.903fg | 1943.899 ± 26.195fg |
Ruby Cabernet | – | 3995.079 ± 96.469a | 63.557 ± 1.495bc | 701.660 ± 25.123cd | 3345.819 ± 74.056a |
Meiyu | – | 2797.284 ± 197.788b | 50.718 ± 0.387de | 754.627 ± 32.225bc | 2595.786 ± 165.829c |
Carinena | – | 1060.326 ± 266.380i | 46.636 ± 5.269ef | 442.407 ± 56.555ij | 972.894 ± 229.364op |
Petit Verdot | – | 2819.855 ± 101.390b | 25.996 ± 2.069jk | 411.976 ± 40.244j | 2533.308 ± 110.216c |
Malbec | – | 2875.734 ± 89.798b | 102.778 ± 7.115a | 1087.759 ± 77.118a | 2680.654 ± 92.598bc |
Blue French | – | 1481.174 ± 150.314g | 3.794 ± 0.843n | 134.733 ± 27.509o | 1432.732 ± 143.869lm |
Pinot Noir115 | – | 962.607 ± 16.744i | – | 4.382 ± 2.889p | 903.682 ± 15.394pq |
Syrah100 | – | 2040.168 ± 213.925de | 48.472 ± 3.617ef | 644.407 ± 36.215def | 1891.788 ± 187.749fgh |
Medoc Noir | – | 1364.963 ± 192.524gh | 62.779 ± 9.827bc | 553.812 ± 74.587gh | 1256.048 ± 180.979mn |
Muscat Hamburg | – | 502.151 ± 30.644k | – | 56.835 ± 3.418p | 446.770 ± 28.980r |
Muscat Rouge | – | 36.793 ± 0.537n | – | – | 8.069 ± 0.488s |
Gamay Teinturier | – | 1359.460 ± 262.227gh | 13.573 ± 3.155lm | 426.114 ± 91.775j | 1325.613 ± 253.989lmn |
Tempranillo | – | 2160.501 ± 102.289d | 40.254 ± 4.753fgh | 328.566 ± 38.281k | 1702.513 ± 65.587ij |
Canepabn | – | 1282.333 ± 72.983h | – | 162.478 ± 21.191no | 1252.465 ± 70.007mn |
Armenia | – | 1849.765 ± 3.492f | 37.086 ± 1.737ghi | 646.349 ± 26.388def | 1713.524 ± 2.788hij |
Areni | – | 2071.600 ± 11.493de | 44.666 ± 1.430efg | 660.873 ± 12.354de | 1891.551 ± 4.699fgh |
Zhesexiang | 17.735 ± 3.049g | 109.475 ± 11.117mn | – | 7.032 ± 0.963p | 5.631 ± 0.018s |
Seibel Noir | 925.157 ± 94.193d | 2963.798 ± 19.079b | 43.159 ± 1.297efg | 791.740 ± 19.695b | 3412.345 ± 132.098a |
Beibinghong | 239.441 ± 18.798f | 2854.997 ± 66.425b | 60.713 ± 2.748bc | 337.266 ± 16.604k | 1263.245 ± 71.750mn |
Huapu 1 | 588.535 ± 15.292e | 255.528 ± 27.257lm | 11.554 ± 2.090mn | 150.486 ± 27.222o | 779.613 ± 31.399q |
Gongniang 2 | 1422.021 ± 29.579b | 272.613 ± 5.328lm | 24.905 ± 0.478jk | 409.384 ± 2.028j | 1626.679 ± 29.769jk |
Beihong | 1571.513 ± 122.168ab | 1451.697 ± 109.720gh | 32.275 ± 1.716hij | 256.825 ± 27.653lm | 2294.886 ± 25.652d |
Beimei | 886.350 ± 11.452d | 329.681 ± 8.008kl | 21.933 ± 1.111kl | 323.960 ± 18.984k | 1146.348 ± 5.084no |
2--1-3 | 1661.225 ± 42.915a | 265.084 ± 6.294lm | 23.840 ± 2.068jk | 499.666 ± 48.464hi | 1827.558 ± 48.505ghi |
44-6-7-1 | 1146.129 ± 402.710c | 1898.974 ± 11.964ef | 69.926 ± 9.108b | 743.065 ± 100.028bc | 2841.983 ± 322.036b |
Moldova | 826.488 ± 57.197d | 752.331 ± 138.366j | 28.039 ± 1.295jk | 217.432 ± 10.934mn | 1449.664 ± 165.779kl |
Note: Different small letters within the same column indicate significant difference at
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The cluster analysis of anthocyanin monomer characteristics showed that ‘Armenia’ and ‘Areni’ were the most similar among the 21 varieties, followed by ‘Ruby Cabernet’ and ‘Petit Verdot’. The monomer compositions of ‘Cabernet Gernischt’ and ‘Mei Yu’, ‘Merlot 181’ and ‘Tempranillo’ were also relatively similar, but ‘Muscat Rouge’ was very different from the other varieties.
The ten hybrid grape varieties included two
In ‘Zhesexiang’ and ‘Beibinghong’, the most abundant anthocyanin was M2, which accounted for 67.6% and 38.5% of the total anthocyanin contents, respectively, the second abundant anthocyanin was D2 and M5, respectively, which accounted for 13.9% and 18.3%; while M1 ranked third, which accounted for 8.5% and 14.8%, respectively. The top four monomers in ‘Seibel Noir’ were M5, M3, M1, and D9, which accounted for 43.4%, 11.8%, 10.1% and 9.8% of the total anthocyanin contents, respectively. Except for ‘Zhesexiang’, ‘Beibinghong’ and ‘Seibel Noir’, malvidin 3,5-O-diglucoside (D5) was the most abundant anthocyanin in the other seven varieties, which accounted for 30.0%~63.6% of the total anthocyanin contents. The second abundant anthocyanin in ‘Gongniang 2’, ‘Beimei’ and ‘2-1-3’ was malvidin-O-(trans-6-O-coumaroyl)-glucoside-5-glucoside (D9), which accounted for 14.6%, 15.9%, 16.5% of total anthocyanin content, respectively; however, M5 ranked second in ‘Huapu 1’, ‘Moldova’ and ‘44-6-7-1’, which accounted for 11.7%, 21.7%, 27.3% of the total anthocyanin contents. M12 ranked third in ‘Gongniang 2’, ‘2-1-3’ and ‘44-6-7-1’, which accounted for 5.3%, 4.8% and 14.8% of the total anthocyanin contents, respectively; while in ‘Huapu 1’, ‘Beimei’ and ‘Moldova’, D9, M5 and M3 ranked third, respectively. In ‘Beihong’, the monomers ranked second and third were M1 and M5, which accounted for 17.8% and 14.1%, respectively. D5, D9, M5 and M12 were the main anthocyanins in ‘Huapu 1’, ‘Gongniang 2’, ‘Beimei’ and ‘2-1-3’, the total contents of them accounted for 84.3%~88.9% of the total anthocyanin contents; however, in ‘44-6-7-1’ and ‘Moldova’, the main anthocyanins included D5, M5, M12 and M3, which accounted for 82.3% and 83.7% of the total anthocyanin content, respectively.
According to the cluster analysis of anthocyanin monomer characteristics, ‘Gongniang 2’ and ‘2-1-3’ were the closest among the 10 varieties, followed by ‘Huapu 1’ and ‘Beimei’. ‘Zhesexiang’ and ‘Seibel Noir’ were very different from the other varieties.
The principal component analysis (PCA) was carried out based on the ratios of different anthocyanin monomer content to total anthocyanin and the total anthocyanin of 21
PCA was also carried out based on the ratios of different anthocyanin monomer content to total anthocyanin and the total anthocyanin of the 10 other grape varieties (
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The anthocyanins of grapes are mainly found in the skins of colored grape varieties. The composition and contents of anthocyanins vary greatly among different grape varieties [
Many factors can influence the anthocyanin content of grape, such as variety, climate, soil and cultivation methods, and the composition and proportion of anthocyanins in different varieties and even different lines of the same variety are very different [
Studies have found that anthocyanins found in
A total of 5 to 15 anthocyanin monomers were detected in the skins of 10 hybrid grape varieties, all varieties contained di-glucoside anthocyanins. The most abundant monomer in ‘Zhesexiang’ and ‘Beibinghong’ was M2, while the ones in ‘Seibel Noir’ and other 7 varieties was M11 and D5, respectively. The anthocyanin ‘Moldova’ were consistent with Liu et al. [
There were significant differences in the anthocyanin types and contents of the skins between
Variety No. | Variety | Origin | Parents | Sampling date (day/month) |
---|---|---|---|---|
1 | Cabernet Franc 327 | – | 25/9 | |
2 | Cabernet Sauvignon 169 | Sauvignon Blanc × Cabernet Franc | 26/9 | |
3 | Merlot 181 | Magdeleine Noire des Charentes × Cabernet Franc | 19/9 | |
4 | Cabernet Gernischt | Carménère (natürliche Kreuzung von Moural × Cabernet Franc) | 26/9 | |
5 | Marselan c980 | Cabernet Sauvignon × Grenache | 14/9 | |
6 | Ruby Cabernet | Carinena × Cabernet Sauvignon | 25/9 | |
7 | Meiyu | Merlot × Petit Verdot | 11/9 | |
8 | Carinena | – | 26/9 | |
9 | Petit Verdot | – | 26/9 | |
10 | Malbec | Magdeleine Noire des Charentes × Prunelard | 26/9 | |
11 | Blue French | Gouais blanc × Blaue Zimmettraube | 11/9 | |
12 | Pinot Noir 115 | – | 31/8 | |
13 | Syrah 100 | Mondeuse Blanche × Dureza | 14/9 | |
14 | Medoc Noir | – | 25/8 | |
15 | Muscat Hamburg | Schiava Grossa × Muscat of Alexandria | 26/9 | |
16 | Muscat Rouge | Muscat Blanc a Petits Grains bud mutation | 13/9 | |
17 | Gamay Teinturier | Gamay Noir × Teinturier du Cher | 25/8 | |
18 | Tempranillo | Albillo Mayor × Benedicto | 15/9 | |
19 | Canepabn | – | 11/9 | |
20 | Armenia | – | 25/8 | |
21 | Areni | – | 4/8 | |
22 | Zhesexiang | Delaware |
13/9 | |
23 | Seibel Noir | – | 11/9 | |
24 | Beibinghong | Zouyouhong × 84-26-53 | 18/9 | |
25 | Huapu 1 | Zuoshan 1 × White Malaga | 13/10 | |
26 | Gongniang 2 | 11/9 | ||
27 | Beihong | Muscat × |
19/9 | |
28 | Beimei | Muscat × |
19/9 | |
29 | 2--1-3 | Muscat Rouge × |
18/9 | |
30 | 44-6-7-1 | Interspecific hybrids | 2-1-3 × Ruby Cabernet | 20/9 |
31 | Moldova | Interspecific hybrids | Guzal ikala × SV.12-375 | 26/9 |
Note:–means that its parents are unknown.
Code | Compounds detected in skins | MS and MS2 (m·z−1) |
---|---|---|
D1 | delphinidin 3,5-O-diglucoside | 627(465,303) |
D2 | cyanidin 3,5-O-diglucoside | 611(449,287) |
D3 | petunidin 3,5-O-diglucoside | 641(479,317) |
M1 | delphinidin 3-O-monoglucoside | 465(303) |
D4 | peonidin 3,5-O-diglucoside | 625(463,301) |
D5 | malvidin 3,5-O-diglucoside | 655(493,331) |
M2 | cyanidin 3-O-monoglucoside | 449(287) |
M3 | petunidin 3-O-monoglucoside | 479(317) |
M4 | peonidin 3-O-monoglucoside | 463(301) |
M5 | malvidin-3-O-monoglucoside | 493(331) |
D6 | delphinidin 3-O-(6-O-coumaroyl)-glucoside-5-glucoside | 773(611,465,303) |
D7 | petunidin 3-O-(6-O-coumaroyl)-glucoside-5-glucoside | 787(625,479,317) |
D8 | malvidin 3-O-(cis-6-O-coumaroyl)-glucoside-5-glucoside | 801(639,493,331) |
M6 | delphinidin-3-O-(trans-6-O-coumaroyl)-glucoside | 611(303) |
M7 | malvidin 3-O-(6-O-acetyl)-glucoside | 535(331) |
D9 | malvidin-O-(trans-6-O-coumaroyl)-glucoside-5-glucoside | 801(639,493,331) |
M8 | cyanidin 3-O-(6-O-coumaroyl)-glucoside | 595(287) |
M9 | petunidin 3-O-(6-O-acetyl)-glucoside | 625(317) |
M10 | malvidin 3-O-(cis-6-O-coumaroyl)-glucoside | 639(331) |
M11 | petunidin 3-O-(trans-6-O-coumaroyl)-glucoside | 609(301) |
M12 | malvidin 3-O-(trans-6-O-coumaroyl)-glucoside | 639(331) |