e-SPEN, the European e-Journal of Clinical Nutrition and Metabolism 4 (2009) e41–e46

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e-SPEN, the European e-Journal of Clinical Nutrition and Metabolism
journal homepage: http://intl.elsevierhealth.com/journals/espen

Original Article

The key role of grape variety for antioxidant capacity of red wines
ˇ ˇ´ ˇ ´ ˇ ´ˇ ´ ´ ´ Alexey Kondrashov a, *, Rudolf Sevcık b, Hana Benakova c, Milada Kostırova c, Stanislav Stıpek a
a

Department of Medical Biochemistry, First Faculty of Medicine, Charles University in Prague, Katerinska 32, Prague 121 08, Czech Republic Department of Food Preservation, Faculty of Food and Biochemical Technology, Institute of Chemical Technology in Prague, Prague 166 28, Czech Republic c Department of Clinical Biochemistry and Laboratory Medicine, First Faculty of Medicine, Charles University in Prague, Prague 128 01, Czech Republic
b

a r t i c l e i n f o
Article history: Received 9 April 2008 Accepted 31 October 2008 Keywords: Polyphenols Red wine Antioxidant activity TEAC FRAP Grape variety Minerals

s u m m a r y
Background & aims: Recent epidemiological studies have supported the idea that food rich in plant bioactive compounds and polyphenols in particular may exert beneficial effects toward human health. One of the foodstuffs widely distributed in the human diet is red wine, a strong antioxidant that contains many bioactive compounds, such as: polyphenols, minerals, and vitamins. The objective of this study was therefore to assess the antioxidant capacity, total phenolics, selected vitamin and mineral content in red wine samples and also to elucidate the existence of a possible relationship between grape variety and all constituents of wines as mentioned above. Methods: The set of 10 red wines, six of Cabernet Sauvignon and four of Merlot was subjected to the study. In all samples total antioxidant capacity was measured by Trolox equivalent antioxidant capacity (TEAC) assay and the Ferric reducing ability of plasma (FRAP) assay simultaneously with total phenolic content, selected minerals and essential vitamins. Results: Both antioxidant capacity and phenolic content were higher in Cabernet Sauvignon wines compared to Merlot. The total antioxidant capacity correlated positively with total phenolic content (r ¼ 0.88, p < 0.001 for TEAC assay and r ¼ 0.89, p < 0.001 for FRAP assay respectively), while a significant relationship among antioxidant capacity, selected minerals and vitamins was not observed. Among the nine minerals analyzed, potassium, zinc and magnesium were the most abundant elements distributed throughout all wine samples. Our results suggest that antioxidant capacity is dependent mainly on total phenolics. Grape variety largely determines such components as phenolic content, antioxidant capacity and mineral content with the exception of vitamins. Ó 2008 European Society for Clinical Nutrition and Metabolism. Published by Elsevier Ltd. All rights reserved.

1. Introduction Of late, there has been a spate of reports considering dietary antioxidants as beneficial toward human health. Epidemiological studies in humans have found a positive correlation between incidence of chronic disease with the oxidative/nitrosative stress in underlying pathogenesis and the dietary patterns they have.1–5 Despite the difficulties in establishing the effects of diet from the other aspects of lifestyle most authorities agree that the benefits to human health of a diet rich in fruits and vegetables may have relations to bioactive compounds with strong antioxidant properties presented in it.6,7

Abbreviations: TEAC, Trolox equivalent antioxidant capacity; FRAP, Ferric reducing ability of plasma; TAC, Total antioxidant capacity; GAE, Gallic acid equivalents; TPC, Total phenolic content. * Corresponding author. Tel.: þ420 774190921; fax: þ420 224964280. E-mail address: alexey.kondrasov@lf1.cuni.cz (A. Kondrashov).

Among natural antioxidants red wine has attracted particular interest due to a high content of biologically active compounds.8These bioactive compounds can be divided into several groups, such as: plant polyphenols, carotenoids, vitamins.9,10 One of the relevant sources of polyphenols in diet throughout the world is wine. Dietary intake of plant polyphenols is inversely related to the development of cardiovascular diseases due to their direct free radical scavenging (antioxidant), anti-inflammatory, antiplatelet aggregation and hypolipemic activities.8,11–15 Polyphenols are the largest group among natural antioxidants, about 8000 compounds that includes mainly flavonoids, phenolic acids, lignans, coumarins, tannins, xantans and chromons.16 Plant polyphenols are non-nutritive, hydrophilic components found in small amounts (micrograms) in all kind of plant-derived food sources such as fruits and vegetables, drinks (wine, coffee, juices) and cereals. The daily intake of polyphenols could reach 1 g/d but broadly varies from one region to another and depends highly on dietary patterns of population.17 Polyphenols have attracted considerable interest from the scientific community due to the

1751-4991/$ - see front matter Ó 2008 European Society for Clinical Nutrition and Metabolism. Published by Elsevier Ltd. All rights reserved. doi:10.1016/j.eclnm.2008.10.004

Australia) for copper. Castel 9.31 Both assays expressed antioxidant power in Trolox (6-hydroxy-2. zinc. This program is appropriate to employ statistical tests for the small number of samples.1. Valencia France. Measurement of total phenolic content Total phenolic content was determined using spectrophotometric assay on a UV–vis spectrophotometer (PharmaSpec UV1700. Comparative analysis of 10 red wine samples of two different grape varieties Cabernet Sauvignon and Merlot was performed. Results 3. A scatter plot was used to graphically represent the data obtained. a key antioxidant enzyme. Measurement of total antioxidant capacity In the wine samples.5 monovarietal wines that are widespread in different parts of the world but not planted in the Czech Republic. Pyridoxine was measured directly with fluorescence detection using excitation and emission wavelengths at 370 nm and 470 nm respectively. Modular E 170.. selenium and lead. The similar program was used for finding correlations among minerals in wine samples. Western Cellary 4..6. selected vitamin and mineral content in red wine samples and also to elucidate the existence of a possible relationship between grape variety and all constituents of wines as mentioned above.18–22 Studies by Burns et al. The selecting criteria for the samples were to find Table 1 Samples of red wine subjected to the study. Santa Regina 6. Riboflavin was measured directly with fluorescence detection using excitation and emission wavelengths at 450 nm and 530 nm respectively.5 12. Castel 3.5 13.32 The absorbance was determined at 765 nm using gallic acid as the standard. Switzerland) for calcium..5% to 13. Chemicals Gallic acid. The mineral profile of wines is important because of their possible impact on enzymes involved in antioxidant defense system in humans. Czech Republic) was equipped with a pump (ECOM). ABTS radical and Trolox were purchased from Sigma–Aldrich. Determination of minerals The red wines elemental composition was determined by AAS method using acetylene/argon flame in Atomic Absorption Spectrometer (Varian Spectra AA 220 FS. Germany). TPC expressed in gallic acid equivalents (GAE).5 12.5 12. Pays d’Oc USA. / e-SPEN. Pays d’Oc Germany Italy Year 2005 2005 2004 2004 2003 2004 2005 2004 2005 2005 Alcohol content % 12. 2. Hardy’s 7.00-Photometric. . 2.5% and 0% in Carl Jung dealcoholised Merlot sample. Studies of DNA typing in the late 1990s have revealed the true origin of Cabernet Sauvignon as a progeny of Cabernet Franc and Sauvignon Blanc. Valencia France. Measurements of riboflavin (B2) and pyridoxine (B6) content Both vitamins determined by HPLC Fluorescent Detection using RECIPE complete set (Recipe Chemicals þ Instruments GmbH Munich. The statistical significance between the phenolic content and total antioxidant capacity was made by GraphPad Prism program version 4 for Windows using Pearson test (GraphPad Software Corp. 2. CA). San Diego. 2.5. Folin–Ciocalteau reagent. 3. they potentially may affect the metabolism in humans by their participation in the reactions of oxidation and reduction in the case of Riboflavin (B2) and transamination and decarboxylation of amino acids in the case of Pyridoxine (B6). All the other reagents were of analytical grade.2.4. total phenolics.23 have related the values of total antioxidant capacity (TAC) of red wines to their phenolic content. several minerals found in foodstuffs.5 13. Japan). The HPLC system (ECOM ltd. Pinewood Hill 5. Version 2.0 12. Samples were selected to be representative of the most consumed foreign wines in the Czech Republic. Cabernet Sauvignon and Merlot wines fit this criteria and comparative evaluation of their constituents was performed. 2. Positive effects of minerals in humans are related to their ability to enhance the activity of antioxidant defense system by catalyzing antioxidant enzymes.4.6–Tri(2-pyridyl)-s-triazine (TPTZ) was from FLUKA (Germany). The injection volume was 20 ml and the flow rate 1 ml/min. 2. California USA. Wine 1. such as copper and zinc are essential for activity of superoxide dismutase (SOD). TAC was determined using both Trolox equivalent antioxidant capacity (TEAC) assay30 and the ferric reducing ability of plasma (FRAP) assay.e42 A.5 13. Clarity software version 1. Despite the low concentrations of the B vitamins found in red wines.5 was used for quantification of the peak areas. The objective of this study was therefore to assess the antioxidant capacity. California Chile South-Eastern Australia Spain.27 Current investigation evaluates all possible sources of contribution to TAC among bioactive compounds presented in wine. TAC was measured using spectrophotometric assays on a UV–vis spectrophotometer (PharmaSpec UV1700. Total antioxidant capacity and phenolic content In present study.5. These activities of plant polyphenols have been extensively reviewed.24–26 Another constituent of wine is water-soluble vitamins. phosphorus and iron determination.28 Cabernet Sauvignon wine samples were chosen due to their fame as the premier red wine grape in the world and because of their wide-spread distribution in many countries that make it possible to compare wines independently. Japan) and was measured using Folin–Ciocalteau reaction. Statistics Data is presented as mean values Æ standard deviation (SD) (n ¼ 3).1. Finca del Mar 2. The wines examined (Table 1) were purchased in several local supermarkets and wine shops. Finca del Mar 8. Kondrashov et al. and Photometry (Roche equipment.29 Merlot wines despite their same ancestry as an offspring of Cabernet Franc differ from the Cabernet Sauvignon wines by taste and color. 10 red wines of two grape varieties were examined on total antioxidant capacity and total phenolic content. Shimadzu. fluorescence detector (ECOM) and HPLC column included in the kit. Materials and methods 2. the European e-Journal of Clinical Nutrition and Metabolism 4 (2009) e41–e46 multiple biological effects they exert. magnesium. The alcohol content ranged from 11. Shimadzu.8tetramethylchroman-2-carboxylic acid) equivalents (mmol/l). Carl Jung 10. Prague.3. Grape products and particularly red wine contain polyphenols in complexes with vitamins and minerals that exert higher biological effects than the sum of their individual effects.5 0 11.7. 2. The software parameters were as follows: Shimadzu UV-Probe. Cielo Grape variety Cabernet Sauvignon Cabernet Sauvignon Cabernet Sauvignon Ruby Cabernet Cabernet Sauvignon Cabernet Sauvignon Merlot Merlot Merlot Merlot Origin Spain. For instant. ISE (Ion Selective Electrode) for potassium determination.

We have found a strong positive correlation between the TAC values.2 Æ 0. / e-SPEN. Elemental content The concentrations of determined mineral elements are different among wines subjected to the study (Table 3).8 SD) than Merlot. the European e-Journal of Clinical Nutrition and Metabolism 4 (2009) e41–e46 e43 Total antioxidant capacity (TAC) and total phenolic content (TPC) of the Cabernet Sauvignon and Merlot red wine samples are presented in Table 2.2 – Total Phenolic Content (mg/l GAE) 2414 Æ 11 2912 Æ 26 1453 Æ 16 2118 Æ 19 2365 Æ 12 2168 Æ 27 1737 Æ 16 2100 Æ 25 2081 Æ 22 1447 Æ 21 1.0 Data are expressed as mean values Æ SD (n ¼ 3).9 Æ 0. 1.3 10.8 mmol/l in Merlot samples.9 Æ 0.2 15. the set of red wine samples was divided into two clusters of Cabernet Sauvignon and Merlot. Mean values were used in the comparative analysis of total polyphenol concentrations in Cabernet Sauvignon and Merlot red wine samples. Among other major determined elements zinc (Zn).8 Æ 0. The most predominant element detected in all wine samples was Potassium (K). evaluated by the TEAC and an FRAP assay. phenolics.2 mmol/l in Cabernet Sauvignon wines and 6.6 8.5 7. n ¼ 3) Limits of detection Total Antioxidant Capacity TEAC assay (Trolox mmol/l) 10. Merlot wines contained higher levels of Riboflavin (mean value of 68 mg/l Æ 16.1 Æ 0.2 Æ 0. The total antioxidant capacities based on TEAC and FRAP assays showed strong positive correlation with the Table 2 Total antioxidant capacity and total phenolic content of red wine samples. Iron (Fe) and copper (Cu) were in several cases below the detectable level.7 Æ 0.1 Æ 0.6 Æ 0. Discussion In this study of Cabernet Sauvignon and Merlot wine samples we discovered certain relationships between the antioxidant capacities.5 Æ 0.A.7 Æ 0.1 Æ 0.1 9.9 Æ 0. Cabernet Sauvignon had a higher concentration of Pyridoxine (mean value of 23 mg/l Æ 18.2 11.3 11. Relationship between antioxidant capacity and total polyphenols A significant positive relationship was observed between TAC and TPC values. California.34 3. This approach allows comparing TAC values between each of the wine samples.9 Æ 0.0 Æ 0. The set of examined wines includes six samples of Cabernet Sauvignon red wines of different origin from Spain. France.001 for FRAP assay respectively). Lowest concentrations were in Italian Merlot 1447 mg/l (wine #10) and Chilean Cabernet Sauvignon 1453 mg/l (wine #3).8–16. TAC was determined by two different assays TEAC and FRAP using Trolox equivalents to express the results.0 SD) in comparison to Cabernet Sauvignon. 3.9–9.2 16.1 3. The total phenolic content of the wine samples determined by using the Folin–Ciocalteau colorimetric method varied from 1447 mg/l in Merlot wines to 2912 mg/l of gallic acid equivalents (GAE) in Cabernet Sauvignon wine samples. In current research. For the results of this analysis see Table 4.23.1 9.5 Æ 0.88 and p < 0.2 9.7 Æ 0.6 mmol/l in Cabernet Sauvignon samples and 7. 3.3. At the same time. Vitamins Red wine is a source of water-soluble vitamins. Analysis of the mean values attributing to the each cluster of wines was used to discover the relevant relationships.5 9. According to FRAP assay TAC values ranged between 7.001 for TEAC assay) and (r ¼ 0. In order to justify and compare the TAC values evaluated by these two different assays a statistical analysis was performed. we measured higher zinc and pyridoxine content in Cabernet Sauvignon wines compared to Merlot. Kondrashov et al. For the purpose of finding an association between the grape variety and the content of bioactive compounds.4. This analysis showed that Cabernet Sauvignon wines contain higher levels of potassium (K). Wine 1 2 3 4 5 6 7 8 9 10 Grape variety Cabernet Sauvignon Cabernet Sauvignon Cabernet Sauvignon Ruby Cabernet Cabernet Sauvignon Cabernet Sauvignon Merlot Merlot Merlot Merlot Intra-assay repeatability (RSD in %.1 10. The analysis of the mean values was used in order to find differences in mineral content between red wines of two grape varieties investigated in the current study. We have found a strong positive relationship between the total antioxidant capacity (TAC) and the phenolic content in all wines independently of the grape sample. These results are consistent with the study by Vannucchi. These small differences in antioxidant values are caused by the TEAC and FRAP methods used for detection. A second method was used to assess the antioxidant capacity of red wine samples. Australia and four samples of Merlot from Spain.89 and p < 0. with the coefficient of correlation for the 10 pairs of samples (r ¼ 0.5 Æ 0. Folin–Ciocalteau’s total phenolic content (r ¼ 0. At the same time Potassium concentration was a little higher in Cabernet Sauvignon compared to Merlot.2. Germany and Italy.5 9. phosphorus (P) and magnesium (Mg).2 7.5–11. Antioxidant activity of red wines measured by TEAC assay ranged between 7.4 7. manganese (Mg). Results from FRAP assay were slightly lower compared to those from TEAC.2 mmol/l in Merlot wines. These results are consistent with the results published for red wines by other investigators.0–15.1 8.9963 and p < 0.0001). This relationship is illustrated in Fig. . phosphorus (P).5 3.1 6.33 4. Present research has established that the highest concentrations of polyphenols were detected in French Cabernet Sauvignon 2912 mg/l (wine #2) and Spanish Cabernet Sauvignon 2414 mg/l (wine #1).9 Æ 0.1 5.6 Æ 0. Cabernet Sauvignon and Merlot wines contain 2238 Æ 477 mg/l GAE in average Æ SD and 1841 Æ 311 mg/l GAE in average Æ SD respectively. Total antioxidant capacity showed resembled values higher in Cabernet Sauvignon samples compared to Merlot wines. copper (Cu) and zinc (Zn) compared to Merlot wines. Chile. France. mineral and vitamin content and the grape variety.1 – Total Antioxidant Capacity FRAP assay (Trolox mmol/l) 9.1 9. Current study determined Riboflavin (B2) and Pyridoxine (B6) in all red wine samples.

3 0. p < 0. Cu and Zn (wine #6).0 35.6 ND 3.8 31.7 P (mg/l) 121.7 56.7 4. ND.7 251. The strength of this study is that for the first time an approach of simultaneous analysis of the following: total phenolic content.7 8. Correlation between antioxidant capacity and total polyphenols. These relations are visualised in Fig.0 33.5 143. .3 7. Additionally.1 0.6 74.2 0.6 42. One possible explanation for the observed augmentation of TAC in Cabernet Sauvignon is in the interactions of the polyphenolic compound with the minerals and pyridoxine.1 94.1 Mg (mg/l) 102.5 1. phosphorus.6 0.0 35.5 9.5 296. affecting taste. Despite the existence of some publications with the evaluations of single components such as.0009553.3 82.8 0.1 57. However. Analysis of the wines of two grape varieties has revealed the fact that despite the similar ancestry.5 79.2 0.0 1.0 Pyridoxine (B6) (mg/l) 11.3 1105.6 89.7 122.005 Cu (mg/l) 0.0 1157.6 10.8 0.9 148.2 0. Statistical correlations between Total Antioxidant Capacity and single minerals were not significant.8 20.4 0. magnesium.1 96. In this study. minerals and vitamins was employed with the relation to grape variety.4 28.1 ND 0. despite the large spread in both antioxidant capacities and the phenolic content.1 0. Wine 1 2 3 4 5 6 7 8 9 10 Grape variety Cabernet Sauvignon Cabernet Sauvignon Cabernet Sauvignon Ruby Cabernet Cabernet Sauvignon Cabernet Sauvignon Merlot Merlot Merlot Merlot Intra-assay repeatability (RSD in %.7 106. Phenolic content of the examined Cabernet Sauvignon and Merlot red wines appears to be associated to the antioxidant capacities. / e-SPEN.6 53.4 1486. A positive correlation was found between zinc and magnesium content.0 64. Wine 1 2 3 4 5 6 7 8 9 10 Grape variety Cabernet Sauvignon Cabernet Sauvignon Cabernet Sauvignon Ruby Cabernet Cabernet Sauvignon Cabernet Sauvignon Merlot Merlot Merlot Merlot Intra-assay repeatability (RSD in %.8 101. antioxidant.6 42. We measured higher zinc and pyridoxine content in Cabernet Sauvignon wines compared to Merlot. whether determined by the TEAC assay or by the FRAP assay.5 3.5 146. zinc and vitamin B6 content compared to Merlot red wines. R2 ¼ 0.8 40. we could assume that the grape variety may predetermine the content of bioactive compounds including polyphenols.6 6.8 0. Scatter plot derived from Pearson test illustrates the statistical correlation between total antioxidant capacity and total phenolic content.39 This may be related to the synergy between all the constituents presented in wine.8 23.8 92.5 1054.1 2.8 11.6 1566.5 57. which as a result influence their antioxidant capacity. a Se and Pb were expressed in mg/l.06 Data are expressed as mean values (n ¼ 3).7 6.1 52.9 1.0 Pba(mg/l) 14.8 1.8 0. color and the content of all essential bioactive Table 4 Riboflavin and pyridoxine content in red wines.1 77. Our results show that wine with lower phenolic content has higher TAC if it simultaneously has the highest values of Mg. minerals and vitamins in red wine.5 9.35–38 Regarding the values of the wine constituents estimated.4 0.0 Zn (mg/l) 0.6 Data are expressed as mean values (n ¼ 3).7821.0 Fe (mg/l) ND ND 1. potassium.4 1092. mineral and phenolic content of wines.9 5.9 122. not detected.9 0.4 3. Cabernet Sauvignon wines have higher mean values of TAC and TPC compared to Merlot wines.3 0.5 1177.6 117.1 72.2 7.9 68. Our findings support results of previous studies where total antioxidant capacity of beverages was higher than simple addition of the antioxidant capacities of their individual bioactive components.6 89.2 245. FRAP assay: y ¼ 0.0008290.0 30.4 1.9 2.2 39.004645 Æ 0. R2 ¼ 0.8 122. 2 in which values of antioxidant capacities are plotted together with the phenolic content.4 3.1 13. Fig.05) between the magnesium (Mg) and zinc content (Zn) was found.4 1175. the selection process could lead to the appearance of different properties.8 1.6 4.2 31. TEAC assay: y ¼ 0. the European e-Journal of Clinical Nutrition and Metabolism 4 (2009) e41–e46 Table 3 Elemental profile of Cabernet Sauvignon and Merlot wine samples. In order to eliminate possible discrepancies total antioxidant capacity was evaluated by two assays.4 20.005119 Æ 0. Statistical analysis provided strong positive correlation for both assays.4 0.1 0.7 0.7970. n ¼ 3) Limits of detection K (mg/l) 1231.4 47.02 Sea (mg/l) 20.5 30.5 19. we can also assume that some minerals are able to contribute to the Total Antioxidant Capacity.1 9. copper.1 0. Moreover.5 62. Kondrashov et al. This was observed during comparative analysis of the mean values of the two clusters of wine samples: Cabernet Sauvignon and Merlot.7 1139.4 2.4 266.28.1 0. n ¼ 3) Limits of detection Riboflavin (B2) (mg/l) 48.9 0.1 Ca (mg/l) 57.9 20.9 131. This study adds to the existing literature by considering several constituents of wine with their relation to the total antioxidant capacity and a grape variety. Cabernet Sauvignon wines possessed higher total antioxidant capacity (TAC) together with the higher TPC. TEAC and FRAP.9 55.e44 A. 1.2 0. a positive correlation (r ¼ 0.2 ND 0.5 74.5 41.63.0 46. a sample high in phenolics did not demonstrate the highest TAC when having low mineral content (wine #1).5 76. there is lack of information about the major determinants and possible effects of interactions among polyphenolic compounds and minerals on antioxidant activity of wines.2 69.

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