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
Alexey Kondrashov a, *, Rudolf Ševčı́k b, Hana Benáková c, Milada Koštı́řová c, Stanislav Štı́pek a
a
Department of Medical Biochemistry, First Faculty of Medicine, Charles University in Prague, Katerinska 32, Prague 121 08, Czech Republic
b
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

a r t i c l e i n f o s u m m a r y

Article history: Background & aims: Recent epidemiological studies have supported the idea that food rich in plant
Received 9 April 2008
bioactive compounds and polyphenols in particular may exert beneficial effects toward human health.
Accepted 31 October 2008
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
Keywords:
therefore to assess the antioxidant capacity, total phenolics, selected vitamin and mineral content in red
Polyphenols
wine samples and also to elucidate the existence of a possible relationship between grape variety and all
Red wine
Antioxidant activity constituents of wines as mentioned above.
TEAC Methods: The set of 10 red wines, six of Cabernet Sauvignon and four of Merlot was subjected to the
FRAP study. In all samples total antioxidant capacity was measured by Trolox equivalent antioxidant capacity
Grape variety (TEAC) assay and the Ferric reducing ability of plasma (FRAP) assay simultaneously with total phenolic
Minerals 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 Among natural antioxidants red wine has attracted particular

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 proper-
ties 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).
interest due to a high content of biologically active com-
pounds.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
e42 A. Kondrashov et al. / e-SPEN, the European e-Journal of Clinical Nutrition and Metabolism 4 (2009) e41–e46
multiple biological effects they exert. These activities of plant
polyphenols have been extensively reviewed.18–22
Studies by Burns et al.,23 have related the values of total anti-
oxidant capacity (TAC) of red wines to their phenolic content.
The mineral profile of wines is important because of their
possible impact on enzymes involved in antioxidant defense
system in humans. Positive effects of minerals in humans are
related to their ability to enhance the activity of antioxidant
defense system by catalyzing antioxidant enzymes. For instant,
several minerals found in foodstuffs, such as copper and zinc are
essential for activity of superoxide dismutase (SOD), a key antiox-
idant enzyme.24–26
Another constituent of wine is water-soluble vitamins. Despite
the low concentrations of the B vitamins found in red wines, they
potentially may affect the metabolism in humans by their partici-
pation 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).27
Current investigation evaluates all possible sources of contri-
bution to TAC among bioactive compounds presented in wine.
Comparative analysis of 10 red wine samples of two different grape
varieties Cabernet Sauvignon and Merlot was performed. 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.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. 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.29
Merlot wines despite their same ancestry as an offspring of
Cabernet Franc differ from the Cabernet Sauvignon wines by taste
and color.
The objective of this study was therefore to assess the antioxi-
dant 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.
2. Materials and methods
2.1. Chemicals
Gallic acid, Folin–Ciocalteau reagent, ABTS radical and Trolox
were purchased from Sigma–Aldrich. 2,4,6–Tri(2-pyridyl)-s-triazine
(TPTZ) was from FLUKA (Germany). All the other reagents were of
analytical grade.
The wines examined (Table 1) were purchased in several local
supermarkets and wine shops. Samples were selected to be
representative of the most consumed foreign wines in the Czech
Republic. The selecting criteria for the samples were to find
Table 1
Samples of red wine subjected to the study.
Wine Grape variety Origin
1. Finca del Mar Cabernet Sauvignon Spain, Valencia
2. Castel Cabernet Sauvignon France, Pays d’Oc
3. Western Cellary Cabernet Sauvignon USA, California
4. Pinewood Hill Ruby Cabernet USA, California
5. Santa Regina Cabernet Sauvignon Chile
6. Hardy’s Cabernet Sauvignon South-Eastern Australia
7. Finca del Mar Merlot Spain, Valencia
8. Castel Merlot France, Pays d’Oc
9. Carl Jung Merlot Germany
10. Cielo Merlot Italy
monovarietal wines that are widespread in different parts of the
world but not planted in the Czech Republic. Cabernet Sauvignon
and Merlot wines fit this criteria and comparative evaluation of
their constituents was performed. The alcohol content ranged from
11.5% to 13.5% and 0% in Carl Jung dealcoholised Merlot sample.
2.2. Measurement of total antioxidant capacity
In the wine samples, TAC was measured using spectrophoto-
metric assays on a UV–vis spectrophotometer (PharmaSpec UV-
1700, Shimadzu, Japan). The software parameters were as follows:
Shimadzu UV-Probe, Version 2.00-Photometric. TAC was deter-
mined using both Trolox equivalent antioxidant capacity (TEAC)
assay30 and the ferric reducing ability of plasma (FRAP) assay.31 Both
assays expressed antioxidant power in Trolox (6-hydroxy-2,5,7,8-
tetramethylchroman-2-carboxylic acid) equivalents (mmol/l).
2.3. Measurement of total phenolic content
Total phenolic content was determined using spectrophoto-
metric assay on a UV–vis spectrophotometer (PharmaSpec UV-
1700, Shimadzu, Japan) and was measured using Folin–Ciocalteau
reaction.32 The absorbance was determined at 765 nm using gallic
acid as the standard. TPC expressed in gallic acid equivalents (GAE).
2.4. Determination of minerals
The red wines elemental composition was determined by AAS
method using acetylene/argon flame in Atomic Absorption Spec-
trometer (Varian Spectra AA 220 FS, Australia) for copper, zinc,
selenium and lead, ISE (Ion Selective Electrode) for potassium
determination, and Photometry (Roche equipment, Modular E 170,
Switzerland) for calcium, magnesium, phosphorus and iron
determination.
2.5. 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, Germany). The HPLC system (ECOM ltd., Prague, Czech
Republic) was equipped with a pump (ECOM), fluorescence detector
(ECOM) and HPLC column included in the kit. The injection volume
was 20 ml and the flow rate 1 ml/min. Riboflavin was measured
directly with fluorescence detection using excitation and emission
wavelengths at 450 nm and 530 nm respectively. Pyridoxine was
measured directly with fluorescence detection using excitation and
emission wavelengths at 370 nm and 470 nm respectively. Clarity
software version 1.5 was used for quantification of the peak areas.
2.6. Statistics
Data is presented as mean values  standard deviation (SD)
(n ¼ 3). 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.,
Year Alcohol San Diego, CA). This program is appropriate to employ statistical tests
content % for the small number of samples. A scatter plot was used to graphi-
2005 12.5 cally represent the data obtained. The similar program was used for
2005 12.5 finding correlations among minerals in wine samples.
2004 12.5
2004 13.5
2003 13.5
3. Results
2004 13.0
2005 12.5 3.1. Total antioxidant capacity and phenolic content
2004 12.5
2005 0
In present study, 10 red wines of two grape varieties were
2005 11.5
examined on total antioxidant capacity and total phenolic content.
 A. Kondrashov et al. / e-SPEN, the European e-Journal of Clinical Nutrition and Metabolism 4 (2009) e41–e46
Total antioxidant capacity (TAC) and total phenolic content (TPC) of
the Cabernet Sauvignon and Merlot red wine samples are pre-
sented in Table 2. The set of examined wines includes six samples of
Cabernet Sauvignon red wines of different origin from Spain,
France, Chile, California, Australia and four samples of Merlot from
Spain, France, Germany and Italy.
TAC was determined by two different assays TEAC and FRAP
using Trolox equivalents to express the results. This approach
allows comparing TAC values between each of the wine
samples.
Antioxidant activity of red wines measured by TEAC assay
ranged between 7.8–16.6 mmol/l in Cabernet Sauvignon samples
and 7.5–11.2 mmol/l in Merlot wines. A second method was used
to assess the antioxidant capacity of red wine samples. According
to FRAP assay TAC values ranged between 7.0–15.2 mmol/l in
Cabernet Sauvignon wines and 6.9–9.8 mmol/l in Merlot samples.
Results from FRAP assay were slightly lower compared to those
from TEAC. These small differences in antioxidant values are
caused by the TEAC and FRAP methods used for detection. In
order to justify and compare the TAC values evaluated by these
two different assays a statistical analysis was performed. We have
found a strong positive correlation between the TAC values,
evaluated by the TEAC and an FRAP assay, with the coefficient of
correlation for the 10 pairs of samples (r ¼ 0.9963 and
p < 0.0001).
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.
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). Lowest concentrations were in Italian Merlot 1447 mg/l
(wine #10) and Chilean Cabernet Sauvignon 1453 mg/l (wine #3).
Mean values were used in the comparative analysis of total
polyphenol concentrations in Cabernet Sauvignon and Merlot red
wine samples. Cabernet Sauvignon and Merlot wines contain
2238  477 mg/l GAE in average  SD and 1841  311 mg/l GAE in
average  SD respectively. Total antioxidant capacity showed
resembled values higher in Cabernet Sauvignon samples compared
to Merlot wines.
3.2. Relationship between antioxidant capacity and total
polyphenols
A significant positive relationship was observed between TAC
and TPC values. 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.
Wine Grape variety Total Antioxidant Capacity
TEAC assay (Trolox mmol/l)
1 Cabernet Sauvignon 10.5  0.2
2 Cabernet Sauvignon 16.6  0.4
3 Cabernet Sauvignon 7.7  0.3
4 Ruby Cabernet 10.9  0.5
5 Cabernet Sauvignon 9.9  0.3
6 Cabernet Sauvignon 11.7  0.6
7 Merlot 8.9  0.2
8 Merlot 11.2  0.5
9 Merlot 9.9  0.2
10 Merlot 7.5  0.1
Intra-assay repeatability (RSD in %, n ¼ 3) 5.1
Limits of detection – –
Data are expressed as mean values  SD (n ¼ 3).
e43
Folin–Ciocalteau’s total phenolic content (r ¼ 0.88 and p < 0.001 for
TEAC assay) and (r ¼ 0.89 and p < 0.001 for FRAP assay respec-
tively). This relationship is illustrated in Fig. 1.
3.3. Elemental content
The concentrations of determined mineral elements are
different among wines subjected to the study (Table 3).
The most predominant element detected in all wine samples
was Potassium (K). At the same time Potassium concentration was
a little higher in Cabernet Sauvignon compared to Merlot. Among
other major determined elements zinc (Zn), phosphorus (P) and
magnesium (Mg). Iron (Fe) and copper (Cu) were in several cases
below the detectable level. 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. This
analysis showed that Cabernet Sauvignon wines contain higher
levels of potassium (K), manganese (Mg), phosphorus (P), copper
(Cu) and zinc (Zn) compared to Merlot wines.
3.4. Vitamins
Red wine is a source of water-soluble vitamins. Current study
determined Riboflavin (B2) and Pyridoxine (B6) in all red wine
samples. For the results of this analysis see Table 4. Cabernet Sau-
vignon had a higher concentration of Pyridoxine (mean value of
23 mg/l  18.8 SD) than Merlot. At the same time, Merlot wines
contained higher levels of Riboflavin (mean value of 68 mg/l  16.0
SD) in comparison to Cabernet Sauvignon.
In current research, we measured higher zinc and pyridoxine
content in Cabernet Sauvignon wines compared to Merlot. These
results are consistent with the study by Vannucchi.33
4. Discussion
In this study of Cabernet Sauvignon and Merlot wine samples
we discovered certain relationships between the antioxidant
capacities, phenolics, mineral and vitamin content and the grape
variety. For the purpose of finding an association between the grape
variety and the content of bioactive compounds, the set of red wine
samples was divided into two clusters of Cabernet Sauvignon and
Merlot. Analysis of the mean values attributing to the each cluster
of wines was used to discover the relevant relationships.
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 results published for red wines by other investigators.23,34
Total Antioxidant Capacity Total Phenolic
FRAP assay (Trolox mmol/l) Content (mg/l GAE)
9.5  0.2 2414  11
15.2  0.5 2912  26
7.0  0.1 1453  16
9.6  0.1 2118  19
9.1  0.1 2365  12
10.8  0.1 2168  27
8.1  0.1 1737  16
9.7  0.2 2100  25
9.1  0.1 2081  22
6.9  0.1 1447  21
3.2 1.5
3.0
e44 A. Kondrashov et al. / e-SPEN, the European e-Journal of Clinical Nutrition and Metabolism 4 (2009) e41–e46

Table 3
Elemental profile of Cabernet Sauvignon and Merlot wine samples.

Wine Grape variety K (mg/l) Mg (mg/l) P (mg/l) Ca (mg/l) Fe (mg/l) Cu (mg/l) Sea (mg/l) Pba(mg/l) Zn (mg/l)
1 Cabernet Sauvignon 1231.7 102.6 121.5 57.7 ND 0.2 20.2 14.0 0.4
2 Cabernet Sauvignon 1139.4 89.7 143.2 56.5 ND 0.1 31.5 46.4 0.8
3 Cabernet Sauvignon 1175.4 122.7 245.5 74.5 1.9 0.1 30.0 28.5 0.8
4 Ruby Cabernet 1486.6 106.9 296.7 62.1 1.1 ND 33.6 9.6 0.8
5 Cabernet Sauvignon 1566,0 122.5 251.4 72.1 0.9 0.2 42.0 4.9 0.8
6 Cabernet Sauvignon 1157.4 146.8 266.9 77.0 2.6 0.2 35.0 5.4 1.4
7 Merlot 1092.5 101.8 148.8 64.9 ND ND 35.8 20.5 0.7
8 Merlot 1054.5 92.6 122.5 68.1 3.1 0.1 23.8 41.1 0.6
9 Merlot 1177.3 117.1 76.9 94.6 0.4 0.1 40.0 9.6 0.3
10 Merlot 1105.0 96.5 131.8 89.4 2.7 0.2 30.7 42.4 0.6
Intra-assay repeatability (RSD in %, n ¼ 3) 1.2 1.4 1.4 1.1 6.3 7.2 4.5 3.5 6.8
Limits of detection 39.1 0.7 3.1 2.0 0.005 0.02 9.0 3.0 0.06

Data are expressed as mean values (n ¼ 3). ND, not detected.

a
Se and Pb were expressed in mg/l.

Phenolic content of the examined Cabernet Sauvignon and
Merlot red wines appears to be associated to the antioxidant
capacities, whether determined by the TEAC assay or by the FRAP
assay. In order to eliminate possible discrepancies total antioxidant
capacity was evaluated by two assays, TEAC and FRAP. Statistical
analysis provided strong positive correlation for both assays.
Despite the existence of some publications with the evaluations
of single components such as, antioxidant, mineral and phenolic
content of wines, there is lack of information about the major
determinants and possible effects of interactions among poly-
phenolic compounds and minerals on antioxidant activity of
wines.35–38
Regarding the values of the wine constituents estimated, we
could assume that the grape variety may predetermine the content
of bioactive compounds including polyphenols, minerals and vita-
mins in red wine, which as a result influence their antioxidant
capacity. This was observed during comparative analysis of the
mean values of the two clusters of wine samples: Cabernet Sau-
vignon and Merlot.
These relations are visualised in Fig. 2 in which values of anti-
oxidant capacities are plotted together with the phenolic content.
In this study, despite the large spread in both antioxidant
capacities and the phenolic content, Cabernet Sauvignon wines
have higher mean values of TAC and TPC compared to Merlot wines.
Moreover, we can also assume that some minerals are able to
contribute to the Total Antioxidant Capacity. Our results show that
wine with lower phenolic content has higher TAC if it simulta-
neously has the highest values of Mg, Cu and Zn (wine #6). Addi-
tionally, a positive correlation (r ¼ 0.63, p < 0.05) between the
magnesium (Mg) and zinc content (Zn) was found. However,
a sample high in phenolics did not demonstrate the highest TAC
when having low mineral content (wine #1). Statistical correlations
between Total Antioxidant Capacity and single minerals were not
significant.
Cabernet Sauvignon wines possessed higher total antioxidant
capacity (TAC) together with the higher TPC, potassium, magne-
sium, phosphorus, copper, zinc and vitamin B6 content compared to
Merlot red wines.
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.
A positive correlation was found between zinc and magnesium
content. We measured higher zinc and pyridoxine content in
Cabernet Sauvignon wines compared to Merlot.
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 compo-
nents.28,39 This may be related to the synergy between all the
constituents presented in wine.
The strength of this study is that for the first time an approach of
simultaneous analysis of the following: total phenolic content,
minerals and vitamins was employed with the relation to grape
variety.
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. Analysis of the wines of two grape
varieties has revealed the fact that despite the similar ancestry, the
selection process could lead to the appearance of different prop-
erties, affecting taste, color and the content of all essential bioactive

Table 4
Riboflavin and pyridoxine content in red wines.

Wine Grape variety Riboflavin Pyridoxine

(B2) (mg/l) (B6) (mg/l)
1 Cabernet Sauvignon 48.2 11.8
2 Cabernet Sauvignon 69.6 31.6
3 Cabernet Sauvignon 74.5 10.1
4 Ruby Cabernet 57.6 57.5
5 Cabernet Sauvignon 53.4 19.7
6 Cabernet Sauvignon 47.3 8.8
7 Merlot 82.9 20.3
8 Merlot 55.1 7.8
9 Merlot 52.5 11.1
10 Merlot 79.9 13.2
Fig. 1. Correlation between antioxidant capacity and total polyphenols. Scatter plot
Intra-assay repeatability (RSD in %, n ¼ 3) 0.9 0.9
derived from Pearson test illustrates the statistical correlation between total antioxi-
Limits of detection 20.0 0.6
dant capacity and total phenolic content. TEAC assay: y ¼ 0.005119  0.0009553,
Data are expressed as mean values (n ¼ 3). R2 ¼ 0,7821; FRAP assay: y ¼ 0.004645  0.0008290, R2 ¼ 0,7970.
 A. Kondrashov et al. / e-SPEN, the European e-Journal of Clinical Nutrition and Metabolism 4 (2009) e41–e46
Fig. 2. Relationship among total antioxidant capacity and total phenolic content of examined red wines. Antioxidant capacity determined by two methods, the TEAC assay and FRAP
assay. Both assays expressed antioxidant power in Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) equivalents (mmol/l). Total phenolic content was determined
by Folin–Ciocalteau colorimetric assay [results expressed as mg/l gallic acid equivalents (GAE)]. All data are expressed as mean values  SD.
compounds. Inclusion of the alcohol free Merlot wine sample into
this study was made to confirm the statement that antioxidant
capacity and TPC are independent of alcohol content. Similarly,
Stein et al.40 have found that purple grape juice exerts their anti-
oxidant activities independent of alcohol content.
It may be concluded that Cabernet Sauvignon wines might be of
a particular nutritional interest due to their high antioxidant
capacities.
Conflict of interest
There is no conflict of interests for all of listed authors of this
article.
Acknowledgments
This work was supported by the Ministry of Education, Youth
and Sports of the Czech Republic (research plan MSM 0021620807)
and by the Grant Agency of the Czech Republic (grant 525/06/
0268). We would like to thank prof. Tomaš Zima (Charles University
in Prague, Czech Republic) for his support of this research and prof.
Zdenka Ďuračková (Comenius University, Bratislava, Slovak
Republic) for her guidance during the several experimental
measurements.
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