Figure 1.
Scheme of the extraction methodology. After harvesting, separation of the leaves, seeds, skins and stems was carried out. Then, extraction with lyophilisation and homogenisation took place. Finally, the individual polyphenolic compounds were measured using LC-MS.
LC-MS, liquid chromatography-mass spectrometry.
Figure 2.
A schematic representation of the experiment with the addition of different plant materials before fermentation. After the processing of grapes, the must was taken away. Before fermentation, parts of the plant were added to each sample. After fermentation, the individual polyphenolic compounds were measured using LC-MS.
LC-MS, liquid chromatography–mass spectrometry.
Table 1.
Parameters of Souvignier gris.
| Parameters of Souvignier gris | |
|---|---|
| Sugar | 25°BX |
| Titratable acids | 7.6 g · L-1 |
| pH | 3.28 |
| YAN | 213 mg · L-1 |
| Village | Lednice |
Table 2.
Detector settings
| Q1 | Q3 | DT (ms) | Compound name | DP (V) | EP (V) | CEP (V) | CE (V) | CXP (V) |
|---|---|---|---|---|---|---|---|---|
| 169.0 | 125.0 | 10 | Gallic acid | -42.0 | -9.2 | -5.1 | -21.9 | -0.3 |
| 153.0 | 109.0 | 10 | Protocatechuic acid | -40.3 | -9.2 | -6.8 | -21.4 | -0.1 |
| 137.0 | 93.0 | 10 | 4-Hydroxybenzoic acid | -35.4 | -8.7 | -6.8 | -22.6 | -0.3 |
| 167.0 | 152.0 | 10 | Vanillic acid | -33.0 | -9.9 | -4.9 | -20.0 | -0.6 |
| 197.0 | 123.0 | 10 | Syringic acid | -32.1 | -10.2 | -6.1 | -33.6 | -0.3 |
| 311.0 | 179.0 | 10 | Caftaric acid | -34.3 | -4.2 | -9.1 | -22.2 | -0.3 |
| 616.1 | 149.0 | 10 | Grape reaction product | -63.0 | -6.0 | -18.0 | -48.0 | -0.9 |
| 179.0 | 135.0 | 10 | Caffeic acid | -38.9 | -10.7 | -9.8 | -24.2 | -0.5 |
| 207.0 | 135.0 | 10 | Ethyl caffeate | -55.0 | -4.8 | -11.0 | -32.0 | -0.6 |
| 295.0 | 163.0 | 10 | Coutaric acid | -41.0 | -4.1 | -16.5 | -22.0 | -0.9 |
| 163.0 | 119.0 | 10 | p-Coumaric acid | -37.5 | -8.7 | -8.8 | -22.5 | -0.3 |
| 191.0 | 117.0 | 10 | Ethyl coumarate | -50.0 | -4.2 | -9.0 | -42.0 | -1.0 |
| 325.0 | 193.0 | 10 | Fertaric acid | -44.0 | -3.3 | -9.0 | -23.0 | -0.3 |
| 193.0 | 134.0 | 10 | Ferulic acid | -40.0 | -10.2 | -5.9 | -24.4 | -0.5 |
| 221.0 | 206.0 | 10 | Ethyl ferulate | -55.0 | -7.9 | -15.2 | -25.0 | -0.5 |
| 227.0 | 143.0 | 10 | Resveratrol | -70.0 | -10.6 | -11.7 | -38.2 | -0.7 |
| 389.0 | 227.0 | 10 | Piceid | -70.0 | -10.0 | -17.3 | -28.1 | -0.3 |
| 243.0 | 159.0 | 10 | Piceatannol | -70.0 | -10.0 | -7.2 | -39.1 | -0.6 |
| 405.0 | 243.0 | 10 | Astringin | -63.0 | -5.6 | -20.0 | -31.0 | -0.3 |
| 289.0 | 123.0 | 10 | Catechin | -60.0 | -9.9 | -9.5 | -44.0 | -0.3 |
| 317.0 | 151.0 | 10 | Myricetin | -74.0 | -4.0 | -9.5 | -36.3 | -0.6 |
| 301.0 | 179.0 | 10 | Quercetin | -68.0 | -7.0 | -8.2 | -29.0 | -0.3 |
| 463.0 | 300.0 | 10 | Quercetin-3-glycosides | -81.0 | -4.0 | -13.0 | -41.0 | -0.9 |
| 609.0 | 300.0 | 10 | Rutin | -93.0 | -4.3 | -17.0 | -54.0 | -0.9 |
| 285.0 | 285.0 | 10 | Kaempferol | -120.0 | -8.8 | -16.0 | -17.0 | -0.5 |
| 315.0 | 300.0 | 10 | Isorhamnetin | -74.0 | -5.0 | -19.0 | -36.0 | -0.3 |
| 447.0 | 284.0 | 10 | Kaempferol-3-D-glucoside | -78.0 | -4.2 | -12.0 | -40.0 | -0.6 |
| 441.1 | 169.0 | 10 | Epicatechin-3 -g allate | -64.0 | -3.8 | -12.0 | -31.0 | -0.3 |
| 577.1 | 289.0 | 10 | Procyanidin B | -60.0 | -4.0 | -15.0 | -37.0 | -0.7 |
| 865.2 | 289.0 | 10 | Procyanidin C | -80.0 | -7.5 | -25.0 | -55.0 | -0.6 |
| 188.0 | 93.0 | 10 | α-Cyano-4-hydroxycinnamic acid | -36.0 | -9.4 | -10.7 | -32.6 | -0.1 |
| 249.0 | 205.0 | 10 | Trolox | -58.0 | -10.0 | -7.2 | -26.6 | -0.3 |
Table 3.
The measured results for each polyphenolic substance from different parts of the grapevine.
| Leaves (mg · L’1) | Skins (mg · L-1) | Seeds (mg · L-1) | Stems (mg · L-1) | p-value | |
|---|---|---|---|---|---|
| Myricetin | LOD | LOD | 0.24 ± 0.033 b | LOD | 0.001832 |
| Quercetin | 12.02 ± 3.50 d | 9.18 ± 0.48 c | 0.21 ± 0.053 a | 3.07 ± 0.178 b | 0.000000 |
| Quercetin-3-glucoside | 1586.05 ± 195.78 d | 1093.44 ± 46.97 c | 10.92 ± 0.920 a | 240.15 ± 10.32 b | 0.000000 |
| Quercetin-3-galactoside | 170.04 ± 33.81 c | 37.24 ± 1.91 b | 0.18 ± 0.022 a | 13.88 ± 0.33 a | 0.000039 |
| Rutin | 62.75 ± 13.36 d | 20.76 ± 1.01 b | 0.21 ± 0.032 a | 47.67 ± 2.31 c | 0.000000 |
| Kaempferol | 5.11 ± 1.04 c | 1.09 ± 0.16 b | 0.13 ± 0.088 a | 0.66 ± 0.098 ab | 0.000014 |
| Kaempferol-3-D-glucoside | 698.54 ± 95.79 b | 57.82 ± 4.32 a | 0.25 ± 0.018 a | 25.87 ± 1.02 a | 0.000000 |
| Isorhamnetin | 0.113 ± 0.08 a | 0.91 ± 0.08 c | 0.10 ± 0.087 a | 0.224 ± 0.082 b | 0.000001 |
| Gallic acid | 5.50 ± 0.77 a | 2.14 ± 0.17 a | 223.13 ± 21.73 c | 32.91 ± 2.27 b | 0.000160 |
| Protocatechuic acid | 7.03 ± 1.60 a | 8.46 ± 1.03 ab | 11.13 ± 0.745 c | 9.76 ± 1.23 bc | 0.000000 |
| 4-Hydroxybenzoic acid | 31.43 ± 7.35 b | 1.74 ± 0.48 a | 1.13 ± 0.216 a | 51.82 ± 4.92 c | 0.000001 |
| Vanillic acid | 0.176 ± 0.50 a | LOD | LOD | 0.000 ± 0.000 a | 0.324174 |
| Caftaric acid | 1975.65 ± 287.12 d | 403.73 ± 30.41 b | 9.35 ± 0.934 a | 684.42 ± 58.49 c | 0.000001 |
| Grape reaction product | 21.26 ± 5.43 c | 10.25 ± 1.050 a | 0.016 ± 0.044 b | 13.98 ± 0.93 a | 0.000000 |
| Caffeic acid | 12.20 ± 2.57 c | 1.28 ± 0.08 a | 0.85 ± 0.103 a | 3.75 ± 0.29 b | 0.000002 |
| Ethyl caffeate | 0.102 ± 0.08 b | 0.002 ± 0.007 a | 0.002 ± 0.005 a | 0.059 ± 0.039 ab | 0.000251 |
| Coutaric acid | 479.01 ± 93.55 d | 223.19 ± 15.22 b | 25.01 ± 2.070 a | 341.44 ± 29.60 c | 0.000000 |
| p-Coumaric acid | 4.27 ± 0.74 d | 0.50 ± 0.07 a | 3.48 ± 0.172 c | 2.44 ± 0.24 b | 0.000000 |
| Ethyl coumarate | LOD | LOD | 0.004 ± 0.010 a | LOD | 0.324174 |
| Fertaric acid | 76.06 ± 20.50 d | 19.64 ± 1.21 b | 1.00 ± 0.114 a | 53.42 ± 3.48 c | 0.000000 |
| Ferulic acid | 1.85 ± 0.65 b | 1.03 ± 0.14 a | 0.87 ± 0.157 a | 2.65 ± 0.27 c | 0.000000 |
| Ethyl ferulate | 9.42 ± 5.23 b | 3.60 ± 2.33 a | 3.61 ± 1.114 a | 4.59 ± 2.19 a | 0.000000 |
| Trans-resveratrol | 19.83 ± 2.57 a | 4.28 ± 0.47 a | 0.77 ± 0.158 a | 446.94 ± 59.15 b | 0.000903 |
| Cis-resveratrol | 12.33 ± 4.56 c | 1.16 ± 0.23 a | LOD | 6.07 ± 0.830 b | 0.000005 |
| Trans-piceid | 59.49 ± 12.95 b | 4.17 ± 0.44 a | 3.26 ± 0.245 a | 65.31 ± 6.58 b | 0.000000 |
| Cis-piceid | 186.94 ± 39.45 c | 6.28 ± 0.55 a | 0.09 ± 0.022 a | 33.07 ± 2.24 b | 0.000151 |
| Piceatannol | 1.35 ± 0.55 a | 0.04 ± 0.08 a | LOD | 69.08 ± 6.76 b | 0.001344 |
| Astringin | 7.38 ± 2.36 b | 0.10 ± 0.12 a | 0.296 ± 0.088 a | 10.50 ± 1.08 c | 0.000001 |
| Catechin | 187.73 ± 34.56 b | 13.50 ± 0.67 a | 1423.37 ± 91.967 d | 1175.31 ± 109.86 c | 0.000000 |
| Epicatechin | 15.98 ± 5.20 a | 8.56 ± 0.56 a | 1407.11 ± 83.606 c | 135.47 ± 24.64 b | 0.000384 |
| Epicatechin-3-gallate | 2.77 ± 0.75 a | 0.16 ± 0.07 a | 57.41 ± 4.970 b | 118.90 ± 23.12 c | 0.000006 |
| Procyanidin B1 | 95.91 ± 22.18 a | 34.51 ± 3.64 a | 365.06 ± 42.64 b | 3091.25 ± 174.01 c | 0.000198 |
| Procyanidin B2 | 19.48 ± 5.14 a | 8.03 ± 1.05 a | 611.43 ± 39.36 c | 157.25 ± 21.89 b | 0.000032 |
| Procyanidin C | 9.28 ± 2.03 a | 4.76 ± 0.51 a | 383.95 ± 39.36 c | 287.76 ± 36.08 b | 0.000001 |
Table 4.
Results of the measurement of polyphenols in wines, fermented with the individual parts of the grapevine.
| Skins (mg · L-1) | Stems (mg · L-1) | Leaves (mg · L-1) | Seeds (mg · L-1) | Control (mg · L-1) | p-value | |
|---|---|---|---|---|---|---|
| Myricetin | 0.002 ± 0.004 a | LOD | LOD | LOD | LOD | 0.325582 |
| Quercetin | 0.035 ± 0.008 b | 0.037 ± 0.008 a | 0.021 ± 0.009 c | 0.168 ± 0.023 d | 0.002 ± 0.004 a | 0.000081 |
| Quercetin-3-glucoside | 0.346 ± 0.038 b | 0.099 ± 0.007 a | 0.094 ± 0.013 a | 0.059 ± 0.005 c | 0.016 ± 0.004 d | 0.000004 |
| Quercetin-3-galactoside | 0.037 ± 0.002 ab | 0.027 ± 0.001 ab | 0.036 ± 0.012 a | 0.020 ± 0.002 b | 0.034 ± 0.011 a | 0.000000 |
| Kaempferol-3-D-glucoside | 0.014 ± 0.001 a | 0.004 ± 0.001 a | 0.005 ± 0.001 a | 0.003 ± 0.001 a | 0.003 ± 0.003 b | 0.000000 |
| Isorhamnetin | LOD | LOD | LOD | 0.031 ± 0.006 b | 0.003 ± 0.007 a | 0.007477 |
| Rutin | 0.001 ± 0.002 b | 0.002 ± 0.002 ab | 0.003 ± 0.002 ab | 0.003 ± 0.002 ab | 0.004 ± 0.001 a | 0.000000 |
| Gallic acid | 10.656 ± 1.483 a | 5.259 ±0.623 b | 0.079 ± 0.025 a | 31.48 ± 0.65 d | 0.165 ± 0.065 c | 0.000139 |
| Protocatechuic acid | 3.358 ± 0.091 a | 2.090 ± 0.178 b | 2.926 ± 0.299 c | 1.679 ± 0.025 a | 1.638 ± 0.100 d | 0.000000 |
| 4-Hydroxybenzoic acid | 0.709 ± 0.015 a | 0.611 ± 0.022 b | 0.869 ± 0.036 d | 0.405 ± 0.008 a | 0.432 ± 0.012 c | 0.000000 |
| Vanillic acid | 0.424 ± 0.030 ab | 0.360 ± 0.021 b | 0.352 ± 0.027 ab | 0.310 ± 0.017 a | 0.335 ± 0.024 c | 0.000000 |
| Syringic acid | 0.101 ± 0.009 a | 0.140 ± 0.015 d | LOD | 0.088 ± 0.013 ab | 0.072 ± 0.014 b | 0.000000 |
| Caftaric acid | 35.14 ± 2.90 ab | 44.56 ± 3.38 c | 4.339 ± 0.474 d | 42.14 ± 0.77 bc | 38.43 ± 1.41 a | 0.000000 |
| Grape reaction product | 0.527 ± 0.090 b | 5.376 ± 0.987 d | 0.189 ± 0.021 a | 3.996 ± 0.249 c | 2.950 ± 0.206 a | 0.000000 |
| Caffeic acid | 2.498 ± 0.215 b | 1.172 ± 0.117 d | 0.120 ± 0.017 a | 0.904 ± 0.044 c | 0.410 ± 0.034 e | 0.000000 |
| Ethyl caffeate | 1.861 ± 0.048 c | 0.572 ± 0.063 a | 0.065 ± 0.007 b | 0.509 ± 0.029 a | 0.162 ± 0.006 d | 0.000010 |
| Coutaric acid | 12.82 ± 1.23 a | 9.37 ± 0.18 b | 5.501 ± 0.038 a | 9.89 ± 0.13 b | 5.05 ± 0.06 c | 0.000000 |
| p-Coumaric acid | 1.188 ± 0.052 a | 0.954 ± 0.013 d | 0.524 ± 0.067 c | 0.325 ± 0.010 b | 0.211 ± 0.018 e | 0.000000 |
| Ethyl coumarate | 1.135 ± 0.035 a | 0.416 ± 0.022 d | 0.247 ± 0.020 c | 0.167 ± 0.010 b | 0.089 ± 0.007 e | 0.000002 |
| Fertaric acid | 3.046 ± 0.223 a | 3.88 ± 0.33 c | 0.865 ± 0.054 b | 5.67 ± 0.10 d | 2.86 ± 0.05 a | 0.000000 |
| Ferulic acid | 0.635 ± 0.027 b | 0.492 ± 0.029 d | 0.040 ± 0.002 a | 0.330 ± 0.013 c | 0.260 ± 0.036 e | 0.000000 |
| Ethyl ferulate | 0.022 ± 0.022 ab | 0.003 ± 0.007 ab | LOD | 0.002 ± 0.006 a | 0.003 ± 0.006 b | 0.034311 |
| Trans-resveratrol | 0.271 ± 0.055 a | 1.999 ± 0.110 c | 0.014 ± 0.015 a | 0.248 ± 0.037 b | 0.088 ± 0.019 b | 0.000707 |
| Cis-resveratrol | 2.621 ± 0.336 c | 1.038 ± 0.049 a | 0.049 ± 0.021 b | 0.833 ± 0.027 a | 0.403 ± 0.102 d | 0.000002 |
| Trans-piceid | 0.171 ± 0.011 a | 0.491 ± 0.019 a | 0.088 ± 0.012 b | 0.402 ± 0.008 d | 0.541 ± 0.095 c | 0.000000 |
| Cis-piceid | 0.772 ± 0.063 b | 0.634 ± 0.026 a | 0.046 ± 0.021 c | 1.114 ± 0.031 b | 0.965 ± 0.205 a | 0.000000 |
| Astringin | 0.004 ± 0.005 a | 0.001 ± 0.001 a | 0.001 ± 0.002 a | 0.002 ± 0.003 a | LOD | 0.009305 |
| Catechin | 29.30 ± 3.31 a | 12.37 ± 1.54 b | 0.119 ± 0.092 a | 107.08 ± 1.70 d | 0.342 ± 0.029 c | 0.000379 |
| Epicatechin | 19.24 ± 2.30 a | 1.778 ± 0.689 a | 0.053 ± 0.036 a | 64.64 ± 0.77 c | 0.111 ± 0.017 b | 0.000836 |
| Epicatechin-3-gallate | 0.041 ± 0.007 a | 0.024 ± 0.005 a | LOD | 1.084 ± 0.057 b | LOD | 0.007249 |
| Procyanidin B1 | 16.56 ± 2.690 a | 17.13 ± 1.13 b | 0.031 ± 0.026 a | 95.37 ± 2.12 c | 0.012 ± 0.017 b | 0.000526 |
| Procyanidin B2 | 11.80 ± 2.00 a | 2.174 ± 0.454 b | 0.020 ± 0.020 a | 79.40 ± 0.94 d | 0.025 ± 0.019 c | 0.002721 |
| Procyanidin C | 6.46 ± 0.989 a | 1.336 ± 0.302 a | LOD | 66.23 ± 2.84 c | LOD | 0.004521 |
1 Results (measured using LC-MS) are expressed as the mean value of nine measurements ± standard deviation, values are the result of ANOVA. The division into homogeneous groups (a, b, c, d) was based on Tukey’s test, the significance level is a = 0.05.
ANOVA, analysis of variance; LC-MS, liquid chromatography–mass spectrometry; LOD, limit of detection.