Int. J. Mol. Sci. 2010, 11, 622-646; doi:10.

3390/ijms11020622
OPEN ACCESS

International Journal of
Molecular Sciences
ISSN 1422-0067
www.mdpi.com/journal/ijms
Review

Biological Activities of Polyphenols from Grapes

En-Qin Xia, Gui-Fang Deng, Ya-Jun Guo and Hua-Bin Li *

Department of Nutrition, School of Public Health, Sun Yat-Sen University, Guangzhou 510080,
China; E-Mails: enqinxia@163.com (E.X.); misyfly@163.com (G.D.); guoyajunleo@163.com (Y.G.)

* Author to whom correspondence should be addressed; E-Mail: lihuabin@mail.sysu.edu.cn;

Tel.: +86-20-8733-2391; Fax: +86-20-8733-0446.

Received: 1 December 2009; in revised form: 26 January 2010 / Accepted: 29 January 2010 /
Published: 4 February 2010

Abstract: The dietary consumption of grape and its products is associated with a lower
incidence of degenerative diseases such as cardiovascular disease and certain types of
cancers. Most recent interest has focused on the bioactive phenolic compounds in grape.
Anthocyanins, flavanols, flavonols and resveratrol are the most important grape polyphenols
because they possess many biological activities, such as antioxidant, cardioprotective,
anticancer, anti-inflammation, antiaging and antimicrobial properties. This review
summarizes current knowledge on the bioactivities of grape phenolics. The extraction,
isolation and identification methods of polyphenols from grape as well as their
bioavailability and potential toxicity also are included.

Keywords: grape; polyphenol; bioactivity; antioxidant activity; cardioprotective action;

anticancer activity; anti-inflammation activity; antimicrobial effect

1. Introduction

Grapes have a long and abundant history. During the ancient Greek and Roman civilizations, grapes
were revered for their use in winemaking. Nowadays, there are three main species of grapes: European
grapes (Vitis vinifera), North American grapes (Vitis labrusca and Vitis rotundifolia) and French
hybrids. Grapes are classified as table grapes, wine grapes (used in viniculture), raisin grapes, and so
on, with edible seeds or seedless. People often enjoy the various grape products, such as fruit, raisins,
juice and wine. Grape fruit contains various nutrient elements, such as vitamins, minerals,
carbohydrates, edible fibers and phytochemicals. Polyphenols are the most important phytochemicals
Int. J. Mol. Sci. 2010, 11 623

in grape because they possess many biological activities and health-promoting benefits [1–3]. The
phenolic compounds mainly include anthocyanins, flavanols, flavonols, stilbenes (resveratrol) and
phenolic acids [4–6]. Anthocyanins are pigments, and mainly exist in grape skins. Flavonoids are
widely distributed in grapes, especially in seeds and stems, and principally contain (+)-catechins,
(−)-epicatechin and procyanidin polymers. Anthocyanins are the main polyphenolics in red grapes,
while flavan-3-ols are more abundant in white varieties [7–9].
From the clue of "French paradox", polyphenolics from grapes and red wines attracted the attention
of scientists to define their chemical composition and their properties for human health [10]. The
reported evidences of beneficial health effects of phenolic compounds include inhibiting some
degenerative diseases, such as cardiovascular diseases [11–14], and certain types of cancers [15–17],
reducing plasma oxidation stress and slowing aging [18,19]. Phenolic compounds are also regarded as
preservatives against microbes and oxidation for food [20,21]. What’s more, in vivo assays showed
that phenolic compounds are bioavailable [10,22]. Therefore, besides wine and juice, grape diet
supplements would be promising functional foods worthy of popularization. However, some reports
have also shown that at higher concentrations the effect of phenolic compounds on health was negative
and some structures in particular promoted the negative effects [23]. In addition, some high molecular
weight phenolics could not be absorbed [24,25]. Apparently, research on direct ingestion of different
doses and compositions of grape products are the urgent task in the field.
This review summarizes current knowledge on extraction, isolation and identification methods,
bioactivities, bioavailability and potential toxicity of grape phenolics. Special attention is paid to the
bioactivities, including antioxidant, cardioprotective, anticancer, anti-inflammation, antiaging and
antimicrobial properties. Finally, this paper tries to show some directions for further research and
applications of grapes.

2. The Distribution of Phenolic Compounds in Grape

Grape is a phenol-rich plant, and these phenolics are mainly distributed in the skin, stem, leaf and
seed of grape, rather than their juicy middle sections (Table 1) [26,27]. Total concentration of phenolic
compounds were about 2178.8, 374.6, 23.8, and 351.6 mg/g GAE (gallic acid equivalent) in seed, skin,
flesh, and leaf, respectively [26]. The total phenolic content of grape skins varied with cultivar, soil
composition, climate, geographic origin, and cultivation practices or exposure to diseases, such as
fungal infections [28]. The compounds mainly included proanthocyanidins, anthocyanins, flavonols,
flavanols, resveratrols and phenolic acids [4,5,29,30]. Proanthocyanidins are the major phenolic
compounds in grape seed and skin of grape [30]. Anthocyanins are pigments and responsible for the
color of grape fruits, and flesh did not contain anthocyanins [4,13]. In red wine, anthocyanins and
flavonoids are the major two groups of phenolic compounds, and (+)-catechin is an abundant
flavonoid [31].
Int. J. Mol. Sci. 2010, 11 624

Table 1. The phenolic compounds in different parts of grape and its products.

Resource Phenolic compounds References

seed gallic acid, (+)-catechin, epicatechin, dimeric procyanidin, [26,30–32]
proanthocyanidins
skin Proanthocyanidins, ellagic acid, myricetin, quercetin, [26,30]
kaempferol, trans-resveratrol
leaf myricetin, ellagic acid, kaempferol, quercetin, gallic acid [26]
stem rutin, quercetin 3-O-glucuronide, trans-resveratrol, astilbin [27]
raisin hydroxycinnamic acid, hydroxymethylfurfural [34]
red wine malvidin-3-glucoside, peonidin-3-glucoside, cyanidin-3- [35–37]
glucoside, petunidin-3-glucoside, catechin, quercetin,
resveratrol, hydroxycinnamic acid

3. Extraction, Purification and Identification of Phenolic Compounds from Grape

Liquid-liquid extraction is usually used for extraction of phenolic compounds from grapes. The
extraction solvent is often ethanol, methanol, acetone or formic acid and water in different ratios. For
grape skin, the crude extract mainly contained anthocyanins and flavonols. Grape seeds could be
extracted by pressurizing and heating, and flavanols and hydroxycinnamic derivatives were
obtained [38]. Although the solvent extraction offers high recovery of phenolic compounds from
grapes, the use of large amounts of organic solvents poses health and safety risks to researchers, and is
environmentally unfriendly. Thus, several improved methods have been developed to extract phenolics
from grapes, such as microwave-assisted extraction [39], ultrasound-assisted extraction [5,40,41],
supercritical fluid extraction [42,43], subcritical water extraction [44]. These extraction methods could
significantly eliminate or reduce the use of organic solvents. In addition, a Lichroprep RP-18 column
was employed to isolate catechin, oligomeric and polymeric procyanidin fractions from the crude
extract of grape seeds using the distilled water adjusted to pH 7.0 to eliminate phenolic acids, followed
by ethyl acetate to elute catechins and oligomeric fraction. The polymeric procyanidins absorbed at the
top of the bed were eluted with methanol [45–47].
Total phenolic content was analyzed by a colorimetric assay using Folin–Ciocalteu’s phenol
reagent [32]. Ferulic acid or gallic acid was used as standard, and the total phenolic content was
expressed as mg/L of ferulic acid equivalent, or GAE against the fresh weight of the sample (mg/g)
[48,49]. In the literature, much attention has been paid to the determination of anthocynins and
flavonoids in grapes. The methods were mainly high-performance liquid chromatography (HPLC) with
different detectors, in which HPLC-UV detection was a common tool [41,46,50], followed by
HPLC-mass spectrometry (MS) detection [51]. Some complex devices have been employed by more
than one MS. Before injection into the HPLC, the crude extract could be purified by solid-phase
extraction (SPE) or improved liquid chromatography employed in order to obtain a more perfect
profile of phenolic compounds in grape than ever possible before[4,6,53,54].
In the literature, chemical structures of many phenolic compounds from grapes have been reported.
The chemical structures of some important phenolic compounds are shown in Figure 1.
Int. J. Mol. Sci. 2010, 11 625

Figure 1. The chemical structures of some phenolic compounds from grapes.

OH HO
OH
OH
HO O

OH
OH
OH
Catechin resveratrol

OH
R3

HO O
R4

R2
R1 O
flavonols

R1 R2 R3 R4
quercetin OH OH OH -
rutin OH O-Rutinose OH -
morin OH OH - OH
myricetin OH OH OH OH
fisetin OH OH OH -

R1
R2
+
HO o
R3

O-Gluc
OH O
anthocyanins

R1 R2 R3
peonidin-3-O-glucoside OCH3 OH -
petunidin-3-O-glucoside OH OH OCH3
malvidin-3-O-glucoside OCH3 OH OCH3
cyaniding-3-O-glucoside OH OH -
delphinidin-3-O-glucoside OH OH OH
 Int. J. Mol. Sci. 2010, 11 626

4. Bioactivity of Phenolic Compounds from Grape

Recently, growing interests on phenolic compounds from grapes have focused on their biological
activities linking to human health benefits, such as antioxidant, cardioprotective, anticancer, anti-
inflammation, antiaging and antimicrobial properties.

4.1. Antioxidant Activities

Being most the notable bioactivity of phenolic compounds from grapes, the antioxidative
characteristics have been widely studied, including scavenging of free radicals, inhibition of lipid
oxidation, reduction of hydroperoxide formation, and so on [18,19]. Several methods were employed
to evaluate the antioxidant capacities of phenolic compounds extracted from various grapes or
different parts of grapes, such as the 1,1-diphenyl-2-picryhidrazyl (DPPH) method [55], oxygen
radical absorbance capacity (ORAC) assay [56], crocin bleaching assay (CBA) [57], 2,2’-azino-bis-(3-
ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay [58], the thiobarbituric acid reactant substances
(TBARS) [59], Trolox equivalent antioxidant capacity (TEAC) assay [60], and the ferric reducing
antioxidant power (FRAP) assay [61].

Table 2. The antioxidant capacities of the extracts from different parts of grape and
its products.
Resource TEACa FRAP DPPH ORAC Ref.
grape pomace 0.91 g/L (EC50 ) - 0.20 - [51]
g/L (EC50 )
grape seed - - >663 μmol TE/g - [62]
defatted grape seed 36.36 mol TE/100 g 21.6 mol TE/100 g - -
[47]
whole seed 76.3 mol TE/100 g 58.04 mol TE/100 g - -
grape seed - - 16.8 to 92 mmol TE/g 42.18 mmol TE/g
[63]
grape skin - - 15.7 to 113.3 mmol TE/g 36.40 mmol TE/g

grape seed 281.3 μmol TE/g - - -

grape leaf 236.1 μmol TE/g - - -
[26]
grape skin 12.8 μmol TE/g - - -
grape flesh 2.4 μmol TE/g - - -
2+
grape juice 25 mmol TE/L 32 mmol Fe /L 15 mmol TE/L - [48]
grape wine - 8.8 μmol TE/g 22.9 to 26.7 μmol TE/g - [64]
grape wine - 3.098 mg TE/L 70.7% inhibition 10.724 μmol/L [65]
a
TE is Trolox® antioxidant equivalent.

Using these methods above, notable antioxidant activities were found for grape wine and juice and
the extracts from different parts of grapes. The values of antioxidant capacities were very different
(Table 2). Seen from the Table 2, juice and wine, and even pomace from grapes had high antioxidant
capacities. The extracts of defatted grape seeds expressed half less antioxidant capacity than that of
whole grape seeds, which indicates that the process of oil extraction removed or damaged some
Int. J. Mol. Sci. 2010, 11 627

antioxidant compounds. In different parts of grape, the highest antioxidant capacity was found in grape
seeds, followed by skin, and the flesh displayed the lowest antioxidant capacity [26]. Therefore, the
extracts from grape seeds are a promising antioxidant for dietary supplement.
The antioxidant activities of the extracts from grape and its products have been widely studied in
different biological or food system. Seen from Table 3, the extracts from grape and its products could
reduce oxidative stress of biological system and prevent food spoil.

Table 3. Antioxidant activities of the extracts from grapes and its products.

Resource Antioxidant activity References

grape seed decreasing the oxidated LDL in plasma [69]
juice reducing oxidative stress in serum [48]
red wine protection against membrane oxidation of [70]
Saccharomyces cerevisiae induced by H2O2
fruit beverage protecting mitochondrial and the antioxidant [71]
(grape+orange+apricot) system against oxidative stress induced by H2O2
grape wine protecting hypercholesterolemic hamsters against [37]
aortic fatty streak accumulation
defatted milled grape seed dealing with the oxidant stress induced by [72]
chemical anticancer adriamycin; reducing TBAS
and elevating the levels of GSH and ATP
grape seed extract food preservatives for fish flesh and oil [62]
white grape dietary fiber antioxidation for polyunsaturated fatty acid in oil [73]
concentrate

Many researchers have tried to discover which phenolic compounds and chemical structure(s) are
mainly responsible for the antioxidant activities of grape extracts. For same phenolic compounds, 50%
and 25% (v/v) concentrations showed the same antioxidant activities, both being better than that of the
10% (v/v) concentration. The result suggested that perhaps the antioxidant capacity of phenolics has a
concentration saturation limit, and above this limit, the activity could not increase further with the
concentration [66]. However, the relationship between phenolic compounds and antioxidant capacity
was inconsistent among the results from different studies, which indicated that, besides the
concentration, the antioxidant capacities of phenolic compounds were affected by other factors [49,67].
In a study, malvidin-3-glucoside showed the highest antioxidant capacity in wine anthocyanins [35].
Although total phenolic index was lower in grape flesh than in grape skin because anthocyanins were
absent in the flesh, they possessed equal amounts of reactivity to hydroxyl radicals [13]. In another
study, the results also showed that the anti-radical activity was due to the flavanols, rather than
anthocyanins [68].
The results showed that procyanidin polymers with higher degrees of polymerization had higher
antioxidant activities [46]. However, Faria et al. [74] showed that in five fractions of different degrees
of procyanidins polymers, the second degree fraction displayed the highest antioxidant capacity
(scavenging peroxyl radicals). A similar result was obtained by Soobratteea et al. [75], who showed
Int. J. Mol. Sci. 2010, 11 628

that the most antioxidative compound in various phenolics was procyanidin dimer, and the decrease in
antioxidant capacity was in order of procyanidin dimer, flavanol, flavonol, hydroxycinnamic acids and
simple phenolic acids. Diphenols are more effectively antioxidant than simpler phenols due to
stabilization of the phenoxy-radical through hydrogen bonding [50]. The high molecule weight
compounds might be as important as the monomer flavanols such as catechin, which have been
demonstrated high antioxidant potential in phenolic compounds [76]. Furthermore, the antioxidant
activity of a sample could be synergic effect among several compositions, rather than a single
compound [47,77].
Pinelo et al. studied the impact of solvent on the antioxidant activity of catechin, resveratrol and
grape extracts dissolved in ethanol, methanol and water. The maximum antiradical activity was in
ethanol, then in methanol, and the minimum was in water [45]. By in vitro physiological procedure
such as digestive enzymatic extraction, phenolic compounds from grape seed displayed a higher
phenolic content and antioxidant capacity than by chemical procedure [78], which could be employed
for the aim of getting dietary supplements from grapes.
The antioxidative characteristics of phenolic compounds are mainly ascribed to their free radical
scavenging and metal chelating properties, as well as their effects on cell signaling pathways and on
gene expression [75,79]. Arora et al. [80] found that flavonoids displayed higher antioxidant capacity
against metal-ion-induced peroxidation than peroxyl-radical-induced peroxidation. The mechanism
was mainly speculated to react directly to generate phenoxyl radicals [81], which was stable and cuts
off the reaction chains. The chemical functional group and structure is OH for antioxidant capacity of
phenolic compounds. The number of OH group and its position on the ring of molecule determined the
antioxidant capacity of flavonols [80]. When the OH added onto the flavonoid nucleus, the activity
enhanced, while substituted by the OCH3 groups, the activity diminished. The results were proved by
Majo et al. [67,82]. The o-diphenoxyl groups in resveratol were determined to exhibit higher
antioxidant activity than other compositions [83].

4.2. Cardioprotection Action

Postprandial hyperlipemia and oxidative stress, a well-defined risk factor for atherosclerosis, could
be reduced by grape seed extracts or phenolic-rich grape juice. These oxidative stress factors refer to
plasma lipid hydroperoxides, serum lipid peroxidation products, malondialdehyde-modified-LDL
(MDA-LDL). The lipid-bound polyphenols increasing in serum were found even two hours after
intake of phenolics, and MDA-LDL was detected after six weeks [48,69,84]. Grape seed extracts
protected the rat liver against oxidative damage induced by irradiation in vivo, and remained the
activities of superoxide dismutase and catalase at normal level [85].
Grape seed extracts (5–50 μg/mL) rich in polyphenols displayed reduction of platelet adhesion and
aggregation and generation of superoxide anion, and were more effective than pure resveratrol [12].
Shanmuganayagam et al. [11] employed rabbits to investigate the potential of phenolic compounds to
defend the hypercholesterolemic-induced platelet aggregation. After intake of the grape juice
(225 mL/day), which was rich in polyphenolics, with hypercholesterolemic diet for 96 days, platelet
aggregation in rabbits was significantly ameliorated and the development of atheroma was near 30%
lower than that of the control group. Aortic fatty streak areas of hamster also showed significant
Int. J. Mol. Sci. 2010, 11 629

reduction in the groups receiving catechin (84%) or quercetin (80%) or resveratrol (76%) in
comparison to the controls [37,86]. Dell Agli et al. [79] showed anthocyanins from wine and grape
skin inhibited phosphodiesterase-5 activity, which reduced the risk of cardiovascular diseases by
vasorelaxation. Falchi et al. [13] made ischemic to isolated heart of rats for 30 min followed by two
hours of reperfusion, and found that the ischemic reperfusion injury were significantly inhibited in the
rats after 30 days consumption of the extracts of flesh and skin of grapes, and flesh and skin of grapes
exhibited equal effect of cardioprotection.
Castilla et al. [87] found that phenolic compounds significantly ameliorated plasma lipid levels.
After drinking 100 mL red grape juice/day for 14 days, the concentration of cholesterol-standardized-
tocopherol and antioxidant capacity of plasma were significantly increased, and oxidized LDL and
LDL were significantly reduced. The plasma level of HDL and apolipoprotein A-I were also elevated.
In addition, consumption of red wine resulted to high concentrations of HDL cholesterol [14], which
linked to control of the risk of coronary heart diseases. Ardevol et al. [88] reported that treatment of
differentiated 3T3-L1 cells with procyanidin extracts reduced HSL in the mRNA levels, and inhibited
triacylglycerol synthesis and boost its hydrolysis. After feeding to hamsters at a moderate dose of
grape extracts, the plasma cholesterol was reduced 11% on average [86]. Moreover, plasma
apolipoprotein A1 concentration was increased 26%, 22%, and 19%, induced by catechin, quercetin,
and resveratrol, respectively [37].
For hemodialysis patients, phenolics of grapes are offered to prevent from inflammation. Red grape
juice significantly reduced plasma monocyte chemoattractant protein 1, an inflammatory factor
involved with cardiovascular disease risk, after three weeks’ consumption [87]. Tsang et al. [14]
showed that after two weeks of daily red wine consumption (375 mL), the maximum concentrations of
cunjugated dienes and TBAES in Cu-oxidised LDL were reduced. It was reported that red wine
consumption reduced oxidative stress induced by Cu-oxidised LDL and increased HDL cholesterol
concentrations. Grape juices showed complete inhibition of copper-induced oxidation of human LDL
at the concentration of 0.01% [89]. Phenolic compounds in grapes have showed effective power to
regulate the plasma lipid and oxidative stress.

4.3. Anticancer Activities

Many evidences have shown that the extracts from grapes and its products had anticancer activity.
Hudson et al. [90] reported that the grape skin extract induced prostate tumor cell lines apoptosis with
high rates. The extract from pomace remaining after wine production inhibited activities of matrix
metalloproteinases-2 and -9, and expressed a significant antiproliferative effect on human colon
adenocarcinoma cells (Caco-2), which implied by-product of wine would help to fight against
carcinogenesis [15,91]. Phenolics of grape juice also significantly inhibited carcinogen-induced DNA
adduct formation in rat model [17], and inhibited DNA synthesis in breast cancer cells [16].
Anticancer activities of phenolic compounds from grapes have been studied widely, and the results
are summarized in Table 4. Phenolic compounds had dual effects on cells, and modulated cell
proliferation was notablely dose-dependent [92]. At high concentration, they were attributed to direct
toxic effect and induced cells to death [93].
Int. J. Mol. Sci. 2010, 11 630

Table 4. Anticancer activities of phenolic compounds from grapes.

Phenols Subject Effects References

proanthocyanidins mouse mammary inhibited breast cancer metastasis
[94]
carcinoma cell line
anthocyanin rat liver clone 9 cells activated antioxidant response
[95]
element upstream of genes
colon cancer cell lines induced 2–4 times increase in
[96]
(HT-29 and Caco-2) DNA fragmentation
vascular tumor biology repaired and protected genomic
DNA integrity and retard blood [97]
vessel growth in some tumors
procyanidin, mice spleen cells inhibited DNA damage induced by
catechin or gallic hydrogen peroxide [98]
acid
catechin human breast cancer cell decreased cell viability and
line proliferation at 30 and 60 μg/mL
procyanidins decreased cell viability and [74]
proliferation at 30, but not
60 μg/mL
flavone human colon carcinoma reduced cell proliferation with an
HT-29 cells EC50 value of 54.8 ± 1.3 µmol/L,
induced differentiation and
[99]
apoptosis
flavonoid HT-29 cells more effectively induced apoptosis
than antitumor agent camptothecin
resveratrol prostate cancer cell lines induced apoptotic and
antiproliferative effects at [100]
≥ 15 µmol/L and above 24 hours
human mammary inhibited cyclooxygenase-2
[101]
epithelial cells transcription

The relationship between anticancer activity and structure of phenolic compounds was also
investigated. The regulation target of grape skin extracts to cell apoptosis was the phosphatidylinositol
3-kinase–Akt and mitogen-activated protein kinase survival pathways. The extracts reduced Akt
transcription, and enhanced proteosome degradation [90]. Resveratrol was determined mainly bearing
o-diphenoxyl groups, which displayed inhibiting DNA damage induced by ROS, and accelerating
DNA damage induced by cupric ions, as well as inducing apoptosis of HL-60 cells, while the
composition without such groups did not display the capacity [83].
Int. J. Mol. Sci. 2010, 11 631

4.4. Anti-inflammation Activities

Phenolic compounds in grapes, especially in grape seeds, have showed significant anti-
inflammation effects on rats, mice and human [7,36,102,102], and the contributive molecules may be
flavonols, flavanols and procyanidins (oligomeric flavonoids) [7,36,102]. Bralley et al. [103] found
that extracts from grape skins and seeds inhibited mouse ear inflammation, edema, and
polymorphonuclear leukocyte infiltration induced by 12-O-tetradecanoylphorbol 13-acetate, after
treated with the extracts for 30 minutes. Moreover, the effect of the combination of grape seeds and
skins almost paralleled to that of indomethacin, a common drug against degenerative diseases of joint.
These findings indicated that phenolic compounds in grapes possessed obviously anti-inflammatory
activity.
The mechanism of anti-inflammation of procyanidins was investigated, and the results showed that
it might inhibit releasing proinflammation factors. Immunomodulation was the main pathway, and
antioxidative action was another pathway for the anti-inflammation effect of grape phenolics
[7,36,104]. Panico et al. [36] employed human chondrocytes assays to prove this. After treatment with
a combination of extract of grape wine and IL-1b, a notable decrease was detected in the concentration
of nitric oxide, prostaglandins E2 and reactive oxygen species in human chondrocytes culture,
compared to control groups, and the effects were equal or super to that of indomethacin. Li et al. [104]
demonstrated that proanthocyanidins could prevent the increase of MDA in rat paws with arthritis
induced by carrageenan at the concentration of 10 mg/kg by injection. Nitric oxide synthase activity
and N-acetyl-β-D- glucosaminidase were also successfully inhibited by proanthocyanidins.
Inhibition or reduction of the cytokine gene expression may be a basic pathway to anti-
inflammation for grape phenolics [7,102,104]. After pre-treated with extracts of grape seed
procyanidins, human adipocytes and macrophage-like cell lines produced less IL-6 and MCP-1
induced by inflammatory stimulus, and increase in anti-inflammatory adipokine and adiponectin
appears. The results demonstrated that grape seeds procyanidins might modulate adipokine and
cytokine gene expression related to anti-inflammation [7]. Terra et al. [102] reported that grape seed
procyanidins inhibited the increase of C-reaction protein in rat plasma induced by high fat feed, and
the same trend in IL-6 and TNF-α was detected in the mesenteric white adipose tissue (WAT). Further
research demonstrated that CRP mRNA expression was decreased in the liver and mesenteric WAT,
while adiponectin mRNA expression was increased in the mesenteric WAT. Then, lipid metabolic
disorder and inflammation were availably inhibited. The results indicated that procyanidins in grapes
inhibited inflammation at mRNA levels, and major health benefits brought by them involved in
decreasing the risk of diseases link to high fat diets and obesity, such as cardiovascular and
metabolic disorders.

4.5. Antiaging Effects

It was found that polyphenolics presented in foods might be beneficial in reversing the course of
neuronal and behavioral aging. Due to their notable antioxidant activity, such as scavenging free
radical, they could prevent organs and tissues from oxidative damage, and modify the body negative
mechanism of redox status. The evidences were obtained by observing the behaviors of rats, from age
19 to 21 months. After drinking the 10% grape juice, improvements were detected on release of
Int. J. Mol. Sci. 2010, 11 632

dopamine from striatal slices, as well as cognitive performance in the Morris water maze, while the
50% grape juice improved action capacity [105]. Further research discovered that supplement with
grape seed extracts (100 mg/kg b.wt.) for 30 days, phenolic compounds from the extracts inhibited the
accumulation of age-related oxidative DNA damages in neural tissue [106]. Balu et al. [107] reported
the decreased incidence of free radical-induced lipid peroxidation in the central nervous system of
aged rats.

4.6. Antimicrobial Effects

Plant polyphenols have been demonstrated potential antibacterial [48,107,108], antifungal [28,110]
and antiviral [111,112] activities. Rodriguez-Vaquero et al. [113] have showed that grape wine
inhibited microbial, especially Escherichia coli growth, and the inhibition increased as the polyphenol
concentration increased, and clarified wines were inactive against all bacteria tested. The extracts of
alcohol-free red and white wine exhibited antimicrobial activity to some pathogens such as
Staphylococcus aureus, Escherichia coli and Candida albicans [114]. The results suggested that
polyphenolic compounds contained in red wines were responsible for the antimicrobial effects. Some
studies reported phenolic compounds inhibited other food-borne species such as Salmonella
typhimurium [115] and Listerial monocytogenes [62].
Various bacterial species exhibit different sensitivities towards phenolic compounds. Papadopoulou
et al. [114] demonstrated Staphylococcus aureus were most sensitive to wine extracts, followed by
Escherichia coli and the least effect of inhibition was detected in Candida albicans. The same results
were obtained by Radovanovic et al. [49], the diameter of the inhibition growth zone for
Staphylococcus aureus and the zone for Escherichia coli were 16–22 mm and 12–20 mm, respectively,
and the later exhibited less sensitive to phenolic compounds. Rotava et al. [116] showed that phenolic
compounds from defatted grape (Vitis vinifera) seed extract inhibited the growth of Staphylococcus
aureus and Escherichia coli, while they showed no effects on Salmonella sp. Rodriguez-Vaquero et al.
[21] showed that Flavobacterium sp. was not inhibited by all any phenolic compounds tested. The time
of reaction were also different, for example, Karapinar et al. [117] demonstrated that koruk (unripe
grape from Vitis vinifera) juice immediately decreased the initial populations of Salmonella
typhimurium at 1–3.5 log cfu/g. But for some microbial species, the antibacterial activity acted too
slowly. Baydar et al. [109] showed that grape seed extract acted against Staphylococcus aureus after
48 hours and Aeromonas hydrophila after one hour.
The phenolic compounds from different parts of grapes displayed different antimicrobial effects.
The antimicrobial activity of fermented pomace was either as effective as or significantly better than
whole fruit grape extracts [118]. Some researches showed that seed extracts were more effectively
antimicrobial than other parts of grapes. The experimental study showed the minimum inhibition
concentration (MICs) of seed and stem extracts for antilisterial were 0.26 and 0.34 mg GAE/L,
respectively [119]. The extracts from whole grape fruit inhibited bacterial growth at concentrations of
680 mg GAE/L and 1360 mg GAE/L for Gram(+) and Gram(−) bacteria, respectively. Jayaprakasha et
al. [109] showed grape seed extracts inhibited bacterial growth at 340−390 mg GAE/L and
475−575 mg GAE/L for Gram(+) and Gram(−) bacteria, respectively. The extract of grape leaves also
exhibited less antimicrobial activity than seed extracts. The extract from grape flesh did not exhibit
Int. J. Mol. Sci. 2010, 11 633

any antimicrobial effect at all [120]. Brown et al. [121] showed that the grape skin possessed the
strongest activity in anti-Helicobacter pylori, followed by grape synergy (skin and seed) and seed. The
increase order of the antimicrobial activity was flesh, whole fruit grape extracts, fermented pomace,
skin, leave and seed.
Phenolic compounds in grape such as resveratrol displayed potent antifungal activity against the
human pathogenic fungi Candida albicans at concentrations of 10–20 μL. The notable benefit of
phenolics was no induction of hemolytic activity against human erythrocytes, compared to chemical
medicines [110]. Anastasiadi et al. [119] suggested that high concentration of flavonoids and their
derivatives in grape seeds and flavonoids, stilbenes, and phenolic acids in grape stem were responsible
for the antimicrobial activity. Rodriguez–Vaquero et al. [20] concluded that the non-flavonoid caffeic
acid and the flavonoids rutin and quercetin were the compounds with higher inhibitory activities on
Listerial monocytogenes growth. Rhodes et al. [21] showed that polymeric phenolic fractions acted the
highest inhibition activity for all Listerial species, but not for other bacteria, such as Bacillus cereus,
Salmonella Menston, Escherichia coli, Staphylococcus aureus or Yersinia enterocolitica. The red-
pigmented polymeric phenolics from juice and skin showed pH-dependent antilisterial activity, while
the unpigmented polymeric phenolics from the seed showed antilisterial activity which was
independent of pH, as some phenolic acids acted.
The relationship between compound structure and antimicrobial activity has been investigated. The
core structures with 3,4,5-trihydroxyphenyl groups found in epigallocatechin, epigallocatechin-3-O-
gallate, castalagin and prodelphinidin might be important for antibacterial activity. This indicated that
the number of hydroxyls and the degree of polymerizzation might be pivotal for antimicrobial activity
of phenolic compounds [122]. According to anti-rabies activity of 24 phenolic compounds, Chavez et
al. [112] considered that free hydroxyl and ether groups mainly influenced the anti-rabies activity.
Employing herpes simplex virus (HSV) and human immunodeficiency virus (HIV), De Bruyne et al.
[111] found epicatechin-containing dimer and the presence of ortho-trihydroxyl groups in the B-ring
were important for anti-HSV, radical-scavenging and immunological activities. Thtmothe et al. [118]
demonstrated that the different concentration of anthocyanins and flavonols notablely decreased the
activity of glucosyltransferases B and C (70%-85%) in Streptococcus mutans cells at total
concentrations 62.5 μg/mL. At the same time, F-ATPase activity was reduced 30–65% at 125 μg/mL.
The result suggested that conjugation of phenolic and protein in microorganism, especially key
enzyme might be major pathway to inhibit the growth of microorganism.
The application of phenolic compounds could be better in food preservation than in medical
field [54,62], and the potent function of phenolics as perfect nature preservative and antimicrobial
agents for food is very promising. In Turkish diet, koruk juice is used as flavoring and acidifying agent.
It has acted as a practicable antimicrobial agent for salad vegetables unconsciously due to its
immediate inhibition against Salmonella typhimurium [117]. In order to check the effect of protection
food from microbial infecting, Sivarooban et al. [115] exposed several species of microorganism to the
soy protein isolate film with GSE 1%, nisin 10,000 IU/g, and EDTA 0.16%. This film reduced
Listerial monocytogenes populations by 2.9 log CFU/mL, and Escherichia coli O157:H7 and
Salmonella typhimurium were reduced by 1.8 and 0.6 log CFU/mL, respectively. This finding
suggested the potential applications of phenolic compounds to maintain shelf life, and improve safety
of ready-to-eat food products.
Int. J. Mol. Sci. 2010, 11 634

The antioxidant, cardioprotective, anticancer, anti-inflammation, antiaging and antimicrobial

activities of grapes and its products have been discussed above. Finally, the bioactivities of phenolic
compounds from grapes are summarized in Table 5. As shown in Table 5, the phenolic compounds
have a variety of bioactivities.

Table 5. Bioactivities of some phenolic compounds from grapes.

Phenolic compound Bioactivity References

resveratrol free radical scavenging [76,81]
antiproliferation [83,100]
enhancing plasma NO level [123]
regulating lipid metabolism [37]
protection against membrane oxidation [124]
quercetin antibacterial [20]
enhancing plasma NO level [123]
catechin anticancer [74]
free radical scavenging [13,68,83]
antibacterial [119]
anti-inflammation [36]
protection against membrane oxidation [124]
flavone antiproliferation [99]
flavonol free radical scavenging [75,81]
procyanidin anticancer [74,94]
free radical scavenging [75]
anti-inflammation [8,102,125]
antioxidant [89]
anthocyanin vasorelaxation [79]
free radical scavenger [97]
antibacterial [118,119]
antioxidant [89]
inducing apoptosis [126]
gallic acid free radical scavenger [76]
epicatechin antibacterial [76,118]

5. Bioavailability

Several studies showed rapid absorption of the polyphenolics, such as procyanidins, quercetin and
flavanols from grapes into plasma, with plasma concentrations peaking at two or three hours after
ingestion [31,48,87,127–129]. The increase of lipid-bound polyphenolics in serum could be detected,
and as a result of the bioactivity of polyphenolics, significant decrease was detected on lipid
peroxidation in serum [48]. Moreover, after two weeks of daily red wine consumption (375 mL),
plasma levels of total phenolic concentrations increased significantly, and trace levels of metabolites,
mainly glucuronides and methyl glucuronides of (+)-catechin and (−)-epicatechin, were detected in
Int. J. Mol. Sci. 2010, 11 635

plasma, which could not be found in a control group [14]. These results indicated that phenolic
compounds could be absorbed by human digestion system, and entered the blood successfully. The
phenolic compounds in the extracts from defatted mill grape seed acted bioactive function by
protecting the isolated rat hepatocytes from oxidative stress induced by anticancer drugs. In order to
research the mechanisms involved in pathways of phenolic compounds entering into cell, Laurent
et al. [33] employed an in vitro digestion/Caco-2 cell culture model. However, no phenolic compounds
were detected in the basal compartment of transwells or in cell monolayers. They also showed that the
availability of phenolic compounds was not affected by salivary and gastric incubations but decreased
during intestinal digestion.
The mechanisms involved in the process of digestion and absorption of phenolic compounds in
gastrointestinal lumen are complex and not very clear. Some results showed that phenolic compounds
were able to chelate to iron. Presence of iron and phenolic compounds has been found in the lumen
during digestion, during which iron–polyphenol interacted and formed iron-chelating complexes [22],
which provoked a more marked decrease in the concentration of hydroxycinnamic derivatives,
flavones and flavan-3-ols compared to the control assays during in vitro gastointestinal digestion. In
vitro digestion, Argyri et al. [130] demonstrated that red wine decreased the concentration of digest
phenolics attributable to the formation of iron-polyphenolic chelates. By molecule analyzing, the
interaction of iron and polyphenolic involved the chemical structures of hydroxyl groups, as reported
in flavonoids: ortho-dihydroxyl groups, the presence of 5- OH and/or 3-OH in conjunction with a C4
keto group, and a large number of OH groups [131].
It was found that phenolic compounds have affinities with some proteins after absorbed
[24,25,132,133]. Employing in vitro digestion/ Caco-2 cell culture model, Laurent et al. [33] found
about 43.9% of catechin, 85.3% of epicatechin and all dimers disappeared at the end of 2 h of
intestinal incubation, associating with a decrease of some cells enzyme activities, such as alkaline
phosphatase and sucrase-isomaltase aminopeptidase N. The results showed that phenolics had
interacted with pancreatic proteins, which were detected by unmasked by acetonitrile extraction.
Polyphenols also seemed to have affinities with enterocyte brush border enzymes [132]. Some
researchers showed that phenolic compounds had strong affinities with proteins and particularly with
human salivary prolinerich proteins and histatins [24,25,133] to form both non-covalent and covalent
associations according to the phenolic compound size. Flavonoids were strongly affected by the
presence of milk, especially after the digestion process [129]. Procyanidins from grape seed extracts
strongly combined to milk protein attributing to the higher degree of polymerization. The insoluble
complexes, such as protein-tannins, were stable throughout the digestive tract [131,133]. However, the
fate of the complexes of low molecule weight phenolics and protein is still unclear.
Decomposition of phenolic compounds caused by pH changes has been shown in digestion lumen.
After two hours of in vitro incubation, monomers and dimers were quite stable at pH 7 in intestine
medium, but 20% dimers were degraded at pH 7.4, and all dimers disappeared in pH 8.5. 15%–34% of
epicatechin were degraded at pH 7.5 with incubation for two hours, while catechin was stable [134].
At pH 2, decomposition of high polymerized oligomers (>trimers) of procyanidins might occur and the
slight increase in dimers procyanidins was observed through gastric step [135]. Flavanols and
flavonols monomers and dimers were stable at acidic condition [136,137]. Anthocyanins could be
digested completely in the contents of large intestine of freshly slaughtered pigs after six-hour
Int. J. Mol. Sci. 2010, 11 636

incubation, and the metabolites were mainly 3–O-methylgallic acid, syringic acid and 2,4,6-
trihydroxybenzaldehyde [138].

6. Potential Toxicity

The potential toxicity of some polyphenols from grape, such as epicatechin to the fibroblast, and
keratinocyte cell lines, has been investigated. After exposing the two cell lines to epicatechin for
24 hours or more time, the notablely negative effects were observed when the concentration was 3−7
fold higher than that of expressing positively antioxidant activity. Moreover, the compounds with a
gallate group exhibited more potential toxicity than those without the gallate group [23]. In addition,
noticeable DNA damage was induced in mice spleen cells by incubating with higher concentration
(150 μmol/L) of catechin [98]. Grape extracts was also found to promote mitomycin C inducing sister
chromatid exchange at concentration from 75 to 300 μg/mL in human peripheral blood
lymphocytes [139]. The compounds with polyphenols, caffeic acid, gallic acid, and rutin hydrate
enhanced MMC-induced clastogenicity at accordant concentrations. The results suggested that
negative effects of phenolic compounds were related to the synergistic effect of some molecules, and
the concentration was not always a crucial factor. Therefore, the dose and composition of grape
extracts should be investigated further for secure and healthy application of grape products.

7. Conclusions and Future Prospects

Grape and products from grape have been consumed for a long time. The studies have demonstrated
an inverse association between intake of grape and products from grape and mortality from age-related
diseases such as coronary heart diseases. The health benefits of grapes are thought to arise mainly from
bioactivities of their polyphenols. Anthocyanins, flavonoids and resveratrol are the major functional
components that are responsible for most of biological activities of grape. Tremendous progress has
been obtained for the extraction, analysis and biological activities of polyphenols in grape. The
bioactive compounds were usually extracted from grape using the liquid-liquid extraction, and high-
performance liquid chromatography with UV or MS detection could be applied to analysis of active
components in grape. The grape and its main components anthocyanins, flavonoids and resveratrol
have a variety of bioactivities, such as antioxidant, cardioprotective, anticancer, anti-inflammation,
antiaging and antimicrobial activities, which are closely related to the prevention against disease and
promotion of health, making greater potential for grape in the field of food and pharmaceutical
application. The structure-activity relationships of some polyphenols have been studied, and the results
obtained could be used to modify structure of polyphenol as well as to design and synthesize novel
polyphenols with special function. Most of phenolic compounds were bioavailable, but some high
molecular weight phenolics could not be absorbed. In addition, the effect of some phenolic compounds
was negative on health at higher concentration, and some structures promoted the negative effect.
In the future, the extraction methods of polyphenols from grape should be improved, and the
by-products of wine industry should be utilized effectively. The crude extracts from grape could be
used as diet supplements for health-protection after defining the levels or limits to make sure the dose
is safe for health, but bioactive components at high purity should be used instead of crude extracts in
medicinal preparations from grape. In order to explore more effective functional food or
Int. J. Mol. Sci. 2010, 11 637

pharmaceutical products based on grape, more wide pharmacological studies should be carried out to
determine new pharmacodynamic effects, such as anti-influenza, anti-obesity and antidiabetic
activities. The relationship of structure-activity should be studied further, and the key mechanisms of
bioactivities should be understood clearly. In addition, more attention should be paid to minor
components in grape because special pharmacodynamic effects could be found from minor
components. The structural diversities and pronounced biological activities of compounds in grape
indicate that grape are worthy of further studies that may lead to the identification of new functional
constituents. The polyphenols from grape will widely be employed to prevent and treat these diseases
in association with reactive oxygen species, such as atherosclerosis, coronary heart diseases and cancer.

Acknowledgements

This research was supported by the Hundred-Talents Scheme of Sun Yat-Sen University.

References and Notes

1. Shrikhande, A.J. Wine by-products with health benefits. Food Res. Internat. 2000, 33, 469–474.
2. Silva, R.C.; Rigaud, J.; Cheynier, V.; Chemina, A. Procyanidin dimers and trimers from grape
seeds. Phytochemistry 1991, 30, 1259–1264.
3. Wada, M.; Kido, H.; Ohyama, K; Ichibangas, T.; Kishikaw, N.; Ohba, Y.; Nakashima, M.N.;
Kurod, N; Nakashima, K. Chemiluminescent screening of quenching effects of natural colorants
against reactive oxygen species: evaluation of grape seed, monascus, gardenia and red radish
extracts as multi-functional food additives. Food Chem. 2007, 101, 980–986.
4. Dopico-Garcia, M.S.; Fique, A.; Guerra, L.; Afonso, J.M.; Pereira, O.; Valentao, P.; Andrade,
P.B.; Seabra, R.M. Principal components of phenolics to characterize red Vinho Verde grapes:
anthocyanins or non-coloured compounds? Talanta 2008, 75, 1190–1202.
5. Novaka, I.; Janeiroa, P.; Serugab, M.; Oliveira-Brett, A.M. Ultrasound extracted flavonoids from
four varieties of Portuguese red grape skins determined by reverse-phase high-performance
liquid chromatography with electrochemical detection. Anal. Chim. Acta 2008, 630, 107–115.
6. Spacil, Z.; Novakova, L.; Solich, P. Analysis of phenolic compounds by high performance
liquidchromatography and ultra performance liquid chromatography. Talanta 2008, 76, 189–199.
7. Chacona, M.R.; Ceperuelo-Mallafrea, V.; Maymo-Masipa, E.; Mateo-Sanzb, J.M.; Arolac, L.;
Guitierreza, C.; Fernandez-Reald, J.M.; Ardevolc, A.; Simona, I.; Vendrella, J. Grape-seed
procyanidins modulate inflammation on human differentiated adipocytes in vitro. Cytokine 2009,
47, 137–142.
8. Bagchi, D.; Bagchi, M.; Stohs, S.J.; Das, D.K.; Ray, C.A.; Kuszynski, S.S.; Joshi, H.G. Free
radicals and grape seed proanthocyanidin extract: importance in human health and disease
prevention. Toxicology 2000, 148, 187–197.
9. Cantos E.; Espin J.C.; Tomas-Barberan F.A. Varietal differences among the polyphenol profiles
of seven table grape cultivars studied by LC-DAD-MS-MS. J. Agric. Food Chem. 2002, 50,
5691–5696.
10. Urpi-Sarda, M.; Monagas, M.; Khan. N.; Lamuela-Raventos, R.M.; Santos-Buelga, C.;
Sacanella, E.; Castell, M.; Permanyer, J.; Andres-Lacueva, C. Epicatechin, procyanidins, and
Int. J. Mol. Sci. 2010, 11 638

phenolic microbial metabolites after cocoa intake in humans and rats. Anal. Bioanal. Chem.
2009, 394, 1545–1556.
11. Shanmuganayagam, D.; Warner, T.F.; Krueger, C.G.; Reed, J.D.; Folts, J.D. Concord grape juice
attenuates platelet aggregation, serum cholesterol and development of atheroma in
hypercholesterolemic rabbits. Atherosclerosis 2007, 190, 135–142.
12. Olas, B.; Wachowicz, B.; Tomczak, A.; Erler, J.; Stochmal, A.; Oleszek, W. Comparative anti-
platelet and antioxidant properties of polyphenol-rich extracts from: berries of Aronia
melanocarpa, seeds of grape and bark of Yucca schidigera in vitro. Platelets 2008, 19, 70–77.
13. Falchi, M.; Bertelli, A.; Scalzo, R.L.; Morassut, M.; Morelli, R.; Das, S.; Cui, J.H.; Das, D.K.
Comparison of cardioprotective abilities between the flesh and skin of grapes. J. Agric. Food
Chem. 2006, 54, 6613–6622.
14. Tsanga, C.; Higginsa, S.; Duthiea, G.G.; Duthiea, S.J.; Howiea, M.; Mullena, W.; Leana, M.E.J.;
Crozier, A. The influence of moderate red wine consumption on antioxidant status and indices of
oxidative stress associated with CHD in healthy volunteers. Br. J. Nutr. 2005, 93, 233–240.
15. God, J.M.; Tate, P.; Larcom, L.L. Anticancer effects of four varieties of muscadine grape. J.
Med. Food. 2007, 10, 54–59.
16. Singletary, K.W.; Stansbury, M.J.; Giusti, M.; Breemen, R.B.V.; Wallig, M.; Rimando, A.
Inhibition of rat mammary tumorigenesis by concord grape juice constituents. J. Agric. Food
Chem. 2003, 51, 7280–7286.
17. Jung, K.; Wallig, M.; Singletary, K. Purple grape juice inhibits 7,12-dimethylbenz- [a]anthracene
(DMBA)-induced rat mammary tumorigenesis and in vivo DMBA-DNA adduct formation.
Cancer Lett. 2006, 233, 279–288.
18. Meyer, A.S.; Yi, O.S.; Pearson, D.A.; Waterhouse, A.L.; Frankel, E.N. Inhibition of human low-
density lipoprotein oxidation in relation to composition of phenolic antioxidants in grapes (Vitis
vinifera). J. Agric. Food Chem. 1997, 45, 1638–1643.
19. Sato, M.; Ramarathnam, N.; Suzuki, Y.; Ohkubo, T.; Takeuchi, M.; Ochi, H. Varietal differences
in the phenolic content and superoxide radical scavenging potential of wines from different
sources. J. Agric. Food Chem. 1996, 44, 37–41.
20. Rodriguez-Vaquero, M.J.; Alberto, M.R.; Manca-de-Nadra, M.C. Antibacterial effect of phenolic
compounds from different wines. Food Control. 2007, 18, 93–101.
21. Rhodes, P.L.; Mitchell, J.W.; Wilson, M.W.; Melton, L.D. Antilisterial activity of grape juice
and grape extracts derived from Vitis vinifera variety Ribier. Int. J. Food Microbiol. 2006, 107,
281–286.
22. Hurrell, R.F.; Reddy, M.; Cook, J.D. Inhibition of non-haem iron absorption in man by
polyphenolic-containing beverages. Br. J. Nutr. 1999, 81, 289–295.
23. Ugartondo, V.; Mitjans, M.; Lozano, C.; Torres, J.L.; Vinardell, M.P. Comparative study of the
cytotoxicity induced by antioxidant epicatechin conjugates obtained from grape. J. Agric. Food
Chem. 2006, 54, 6945–6950.
24. Lu, Y.;Bennick, A. Interaction of tannin with human salivary proline-rich proteins. Arch. Oral.
Biochem. 1998, 43, 717–728.
Int. J. Mol. Sci. 2010, 11 639

25. Wroblewski, K.; Muhandiram, R.; Chakrabartty, A.; Bennick, A. The molecular interaction of
human salivary histatins with polyphenolic compounds. Eur. J. Biochem. 2001, 268,
4384–4397.
26. Pastrana-Bonilla, E.; Akoh, C.C.; Sellappan, S.; Krewer, G. Phenolic content and antioxidant
capacity of muscadine grapes. J. Agric. Food Chem. 2003, 51, 5497–4503.
27. Makris, D.P.; Boskou, G.; Andrikopoulos, N.K.; Kefalas, P. Characterisation of certain major
polyphenolic antioxidants in grape (Vitis vinifera) stems by liquid chromatography-mass
spectrometry. Eur. Food Res. Technol. 2008, 226, 1075–1079.
28. Bruno, G.; Sparapano, L. Effects of three esca-associated fungi on Vitis vinifera L : V. Changes
in the chemical and biological profile of xylem sap from diseased cv. Sangiovese vines. Physiol.
Mol. Plant Pathol. 2007, 71, 210–229.
29. Mazza, G.J. Anthocyanins and heart health. Ann. Ist Super Sanita. 2007, 43, 369–374.
30. Hernandez-Jimenez, A.; Gomez-Plaza, E.; Martinez-Cutillas, A.; Kennedy, J.A. Grape skin and
seed proanthocyanidins from Monastrell x Syrah grapes. J. Agric. Food Chem. 2009, 57,
10798–10803.
31. Bell, J.R.C.; Donovan, J.L.; Wong, R.; Waterhouse, A.L.; German, J.B.; Walzem, R.L.; Kasim-
Karakas, S.E. (+)-Catechin in human plasma after ingestion of a single serving of reconstituted
red wine. Am. J. Clin. Nutr. 2000, 71, 103–108.
32. Huang, D.; Ou, B.; Prior, R.L. The chemistry behind antioxidant capacity assays. J. Agric. Food
Chem. 2005, 53, 1841–1856.
33. Laurent, C.; Besancon, P.; Caporiccio, B. Flavonoids from a grape seed extract interact with
digestive secretions and intestinal cells as assessed in an in vitro digestion/Caco-2 cell culture
model. Food Chem. 2007, 100, 1704–1712.
34. Karadeniz F.; Durst R.W.; Wrolstad R.E. Polyphenolic composition of raisins. J. Agric. Food
Chem. 2000, 48, 5343–5350.
35. Rivero-Perez, M.D.; Muniz, P.; Gonzalez-Sanjose, M.L. Contribution of anthocyanin fraction to
the antioxidant properties of wine. Food Chem. Toxicol. 2008, 46, 2815–2822.
36. Panico, A.M.; Cardile, V.; Avondo, S.; Garufi, F.; Gentile, B.; Puglia, C.; Bonina, F.; Santagati,
N.A.; Ronsisvalle, G. The in vitro effect of a lyophilized extract of wine obtained from Jacquez
grapes on human chondrocytes. Phytomedicine 2006, 13, 522–526.
37. Auger, C.; Teissedre, P.L.; Gerain, P.; Lequeux, N.; Bornet, A.; Serisier, S.; Besançon, P.;
Caporiccio, B.; Cristol, J.P.; Rouanet, J.M. Dietary wine phenolics catechin, quercetin, and
resveratrol efficiently protect hypercholesterolemic hamsters against aortic fatty streak
accumulation. J. Agric. Food Chem. 2005, 53, 2015–2021.
38. Guerrero, R.F.; Liazid, A.; Palma, M.; Puertas, B.; Gonzalez-Barrio, R.; Gil-Izquierdo, A.;
Garcia-Barroso, C.; Cantos-Villar, E. Phenolic characterisation of red grapes autochthonous to
Andalusia. Food Chem. 2009, 112, 949–955.
39. Hong N.; Yaylayan V.A.; Raghavan G.S.V.; Pare J.R.J.; Belanger J.M.R. Microwave-assisted
extraction of phenolic compounds from grape seed. Nat. Prod. Lett. 2001, 15, 197–204.
40. Ghafoor K.; Choi Y.H.; Jeon J.Y.; Jo I.H. Optimization of ultrasound-assisted Extraction of
phenolic compounds, antioxidants, and anthocyanins from grape (Vitis vinifera) seeds. J. Agric.
Food Chem. 2009, 57, 4988–4994.
Int. J. Mol. Sci. 2010, 11 640

41. Chafer, A.; Pascual-Marti, M.C.; Salvador, A.; Berna, A. Supercritical fluid extraction and
HPLC determination of relevant polyphenolic compounds in grape skin. J. Sep. Sci. 2005, 28,
2050–2056.
42. Fiori, L.; de Faveri, D.; Casazza, A.A.; Perego, P. Grape by-products: extraction of polyphenolic
compounds using supercritical CO2 and liquid organic solvent—a preliminary investigation.
CYTA—J. Food 2009, 7, 163–171.
43. Vatai, T.; Skerget, M.; Knez, Z. Extraction of phenolic compounds from elder berry and different
grape marc varieties using organic solvents and/or supercritical carbon dioxide. J. Food Eng.
2009, 90, 246–254.
44. Ju, Z.Y.; Howard, L.R. Subcritical water and sulfured water extraction of anthocyanins and other
phenolics from dried red grape skin. J. Food Sci. 2005, 70, 270–276.
45. Pinelo, M.; Rubilar, M.; Sineiro, J.; Nunez, M.J. A thermal treatment to increase the antioxidant
capacity of natural phenols: catechin, resveratrol and grape extract cases. Eur. Food Res.
Technol. 2005, 221, 284–290.
46. Spranger, I.; Sun, B.; Mateus, A.M.; de Freitas, V.; Ricardo-da-Silva, J.M. Chemical
characterization and antioxidant activities of oligomeric and polymeric procyanidin fractions
from grape seeds. Food Chem. 2008, 108, 519–532.
47. Maier, T.; Schieber, A.; Kammerer, D.R.; Carle, R. Residues of grape (Vitis vinifera) seed oil
production as a valuable source of phenolic antioxidants. Food Chem. 2009, 112, 551–559.
48. Garcia-Alonso, J.; Ros, G.; Vidal-Guevara, M.L.; Periago, M.J. Acute intake of phenolic-rich
juice improves antioxidant status in healthy subjects. Nutr. Res. 2006, 26, 330–339.
49. Radovanovic, A.; Radovanovic, B.; Jovancicevic, B. Free radical scavenging and antibacterial
activities of southern Serbian red wines. Food Chem. 2009, 117, 326–331.
50. Amico, V.; Chillemi, R.; Mangiafico, S.; Spatafora, C.; Tringali, C. Polyphenol-enriched
fractions from Sicilian grape pomace: HPLC–DAD analysis and antioxidant activity. Bioresour.
Technol. 2008, 99, 5960–5966.
51. Rubilar, M.; Pinelo, M.; Shene, C.; Sineiro, J.; Nunez, M.J. Separation and HPLC-MS
identification of phenolic antioxidants from agricultural residues: almond hulls and grape
pomace. J. Agric. Food Chem. 2007, 55, 10101–10109.
52. Serra, A.; Macia, A.; Romero, M.P.; Salvado, M.J.; Bustos, M.; Fernandez-Larrea, J.; Motilva,
M.J. Determination of procyanidins and their metabolites in plasma samples by improved liquid
chromatography-tandem mass spectrometry. J. Chromatogr. B 2009, 877, 1169–1176.
53. Jeffery, D.W.; Mercurio, M.D.; Herderich, M.J.; Hayasaka, Y.; Smith, P.A. Rapid isolation of
red wine polymeric polyphenols by solid-phase extraction. J. Agric. Food Chem. 2008, 56,
2571–2580.
54. Serra, A.T.; Matias, A.A.; Nunes, A.V.M.; Leitao, M.C.; Brito, D.; Bronze, R.; Silva, S.; Pires,
A.; Crespo, M.T.; Romao, M.V.S.; Duarte, C.M. In vitro evaluation of olive- and grape-based
natural extracts aspotential preservatives for food. Inno. Food Sci. Emerg. Technol. 2008, 9,
311–319.
55. Brand-williams, W.; Cuvelier, M.E.; Berset, C. Use of a free radical method to evaluate
antioxidant activity. LWT - Food Sci. Technol. 1995, 28, 25–30.
Int. J. Mol. Sci. 2010, 11 641

56. Prior, R.L.; Hoang, H.; Gu, L.W.; Wu, X.L.; Bacchiocca, M.; Huang, D.J.; Ou, B.X.; Jacob, R.
Assays for hydrophilic and lipophilic antioxidant capacity (oxygen radical absorbance capacity)
of plasma and other biological and food samples. J. Agric. Food Chem. 2003, 51, 3273–3279.
57. Chatterjee, S.; Poduval, T.B.; Tilak, J.C.; Devasagayam, T.P. A modified, economic, sensitive
method for measuring total antioxidant capacities of human plasma and natural compounds using
Indian saffron (Crocus sativus). Clin. Chim. Acta 2005, 352, 155–165.
58. Cano, A.; Hernandez-Ruiz, J.; Garcia-Canovas, F.; Acosta, M.; Arnao, M.B. An end-point
method for estimation of the total antioxidant activity in plant material. Phytochem. Anal. 1998,
9, 196–202.
59. de Ruiter, N.; Ottenwalder, O.; Muliawan, H.; Kappus, H. Lipid peroxidation in isolated rat
hepatocytes measured by ethane and n-pentane formation. Arch. Toxicol. 1982, 49, 265–273.
60. Wang, C.C.; Chu, C.Y.; Chu, K.O.; Choy, K.W.; Khaw, K.S.; Rogers, M.S.; Pang, C.P. Trolox-
equivalent antioxidant capacity assay versus oxygen radical absorbance capacity assay in plasma.
Clinic. Chem. 2004, 50, 952–954.
61. Benzie, I.F.; Strain, J.J. The ferric reducing ability of plasma (FRAP) as a measure of
"antioxidant power": the FRAP assay. Anal. Biochem. 1996, 239, 70–76.
62. Luther, M.; Parry, J.; Moore, J.; Meng, J.H.; Zhang, Y.F.; Cheng, Z.H.; Yu, L. Inhibitory effect
of chardonnay and black raspberry seed extracts on lipid oxidation in fish oil and their radical
scavenging and antimicrobial properties. Food Chem. 2007, 104, 1065–1073.
63. Poudel, P.R.; Tamura, H.; Kataoka, I.; Mochioka, R. Phenolic compounds and antioxidant
activities of skins and seeds of five wild grapes and two hybrids native to Japan. J. Food Comp.
Anal. 2008, 21, 622–625.
64. Hogan, S.; Zhang, L.; Li, J.; Zoecklein, B.; Zhou, K. Antioxidant properties and bioactive
components of Norton (Vitis aestivalis) and Cabernet Franc (Vitis vinifera) wine grapes. LWT—
Food Sci. Technol. 2009, 42, 1755.
65. Feliciano, R.P.; Bravo, M.N.; Pires, M.M.; Serra, A.T.; Duarte, C.M.; Boas, L.V.; Bronze, M.R.
Phenolic content and antioxidant activity of moscatel dessert wines from the setubal region in
Portugal. Food Anal. Meth. 2009, 2, 149–161.
66. Dani, C.; Oliboni, L.S.; Vanderlinde, R.; Pra, D.; Dias, J.F.; Yoneama, M.L.; Bonatto, D.;
Salvador, M.; Henriques, J.A.P. Antioxidant activity and phenolic and mineral content of rose
grape juice. J. Med. Food. 2009, 12, 188–192.
67. Majo, D.D.; Guardia, M.L.; Giammanco, S.; Neve, L.L.; Giammanco, M. The antioxidant
capacity of red wine in relationship with its polyphenolic constituents. Food Chem. 2008, 111,
45–49.
68. Arnous, A.; Makris, D.P.; Kefalas, P. Correlation of pigment and flavanol content with
antioxidant properties in selected aged regional wines from Greece. J. Food Compos. Anal. 2002,
15, 655–665.
69. Sano, A.; Uchida, R.; Saito, M.; Shioya, N.; Komori, Y.; Tho, Y.; Hashizume, N. Beneficial
effects of grape seed extract on malondialdehyde-Modified LDL. J. Nutr. Sci. Vitaminol. 2007,
53, 174–182.
Int. J. Mol. Sci. 2010, 11 642

70. Dani, C.; Bonatto, D.; Salvador, M.; Pereira, M.D.; Henriques, J.P.; Eleutherio, E. Antioxidant
protection of resveratrol and catechin in Saccharomyces cerevisiae. J. Agric. Food Chem. 2008,
56, 4268–4272.
71. Cilla, A.; Laparra, J.M.; Alegria, A.; Barbera, R.; Farre, R. Antioxidant effect derived from
bioaccessible fractions of fruit beverages against H2O2-induced oxidative stress in Caco-2 cells.
Food Chem. 2008, 106, 1180–1187.
72. Valls-Belles, V.; Torres, M.C.; Muniz, P.; Beltran, S.; Martinez-Alvarez, J.R.; Codoner-Franch,
P. Defatted milled grape seed protects adriamycin-treated hepatocytes against oxidative damage.
Eur. J. Clin. Nutr. 2006, 45, 251–258.
73. Sanchez-Alonso, I.; Borderias, J.; Larsson, K.; Undeland, I. Inhibition of hemoglobin-mediated
oxidation of regular and lipid-fortified washed cod mince by a white grape dietary fiber. J. Agric.
Food Chem. 2007, 55, 5299–5305.
74. Faria, A.; Calhau, C.; De Freitas, V.; Mateus, N. Procyanidins as antioxidants and tumor cell
growth modulators. J. Agric. Food Chem. 2006, 54, 2392–2397.
75. Soobrattee, M.A.; Neergheena, V.S.; Luximon-Rammaa, A.; Aruomab, O.I.; Bahoruna, T.
Phenolics as potential antioxidant therapeutic agents: Mechanism and actions. Mut. Res. Fund.
Mol. Mech. Mutagen. 2005, 579, 200–213.
76. Yilmaz, Y.; Toledo, R.T. Major flavonoids in grape seeds and skins: Antioxidant capacity of
catechin, epicatechin, and gallic acid. J. Agric. Food Chem. 2004, 52, 255–260.
77. Monagas, M.; Hernandez-Ledesma, B.; Garrido, P.; Martin-alvarez, P.J.; Gomez-Cordoves, C.;
Bartolome B. Quality assessment of commercial dietary antioxidant products from Vitis vinifera
L. grape seeds. Nutr Cancer. 2005, 53, 244–254.
78. Li, H.; Wang, X.Y.; Li, P.H.; Li, Y.; Wang, H. Comparative study of antioxidant activity of
grape (Vitis vinifera) seed powder assessed by different methods. J. Food. Drug Anal. 2008, 16,
67–73.
79. Dell Agli, M.; Galli, G.V.; Vrhovsek, U.; Mattivi, F; Bosisio, E. In vitro inhibition of human
cGMP-specific phosphodiesterase-5 by polyphenols from red grapes. J. Agric. Food Chem. 2005,
53, 1960–1965.
80. Arora, A.; Nair, M.G.; Strasburg, G.M. Structure-activity relationships for antioxidant activities
of a series of flavonoids in a liposomal system. Free Radic. Biol. Med. 1998, 24, 1355–1363.
81. Yoshimura, Y.; Nakazawa, H.; Yamaguchi, F. Valuation of the NO scavenging activity of
procyanidin in grape seed by use of the TMA-PTIO/NOC 7 ESR system. J. Agric. Food Chem.
2003, 51, 6409–6412.
82. Majo, D.D.; Giammanco, M.; Guardia, M.L.; Tripoli, E.; Giammanco, S.; Finotti, E. Flavanones
in Citrus fruit: Structure–antioxidant activity relationships. Food Res. Int. 2005, 38, 1161–1166.
83. Qian, Y.P.; Cai, Y.J.; Fan, G.J.; Wei, Q.Y.; Yang, J.; Zheng, L.F.; Li, X.Z.; Fang, J.G.; Zhou, B.
Antioxidant-based lead discovery for cancer chemoprevention: the case of resveratrol. J. Med.
Chem. 2009, 52, 1963–1974.
84. Natella, F.; Belelli, F.; Gentili, V.; Ursini, F.; Scaccini, C. Grape seed proanthocyanidins prevent
plasma postprandial oxidative stress in humans. J. Agric. Food Chem. 2002, 50, 7720–7725.
Int. J. Mol. Sci. 2010, 11 643

85. Cetin, A.; Kaynar, L.; Kocyigit, I.; Hacioglu, S.K.; Saraymen, R.; Ozturk, A.; Sari, I.; Sagdic, O.
Role of grape seed extract on methotrexate induced oxidative stress in rat liver. Am. J. Chin.
Med. 2008, 36, 861–872.
86. Auger, C.; Gerain, P.; Laurent-Bichon, F.; Portet, K.; Bornet, A.; Caporiccio, B.; Cros, G.;
Teissedre, P.; Rouanet, J.M. Phenolics from commercialized grape extracts prevent early
atherosclerotic lesions in hamsters by mechanisms other than antioxidant effect. J. Agric. Food
Chem. 2004, 52, 5297–5302.
87. Castilla, P.; Echarri, R.; Davalos, A.; Cerrato, F.; Ortega, H.; Teruel, J.L.; Lucas, M.F.; Gomez-
Coronado, D.; Ortuno, J.; Lasuncion, M.A. Concentrated red grape juice exerts antioxidant,
hypolipidemic, and antiinflammatory effects in both hemodialysis patients and healthy subjects.
Am. J. Clin. Nutr. 2006, 84, 252–262.
88. Ardevol, A.; Blade, C.; Salvado, M.J.; Arola, L. Changes in lipolysis and hormone-sensitive
lipase expression caused by procyanidins in 3T3-L1 adipocytes. Int. J. Obes. 2000, 24, 319–324.
89. Kulisic-Bilusic, T.; Schnabele, K.; Schmoller, I.; Dragovic-Uzelac, V.; Krisko, A.; Dejanovic,
B.; Milos, M.; Pifat, G. Antioxidant activity versus cytotoxic and nuclear factor kappa B
regulatory activities on HT-29 cells by natural fruit juices. Eur. Food Res. Technol. 2009, 228,
417–424.
90. Hudson, T.S.; Hartle, D.K.; Hursting, S.D.; Nunez, N.P.; Wang, T.T.Y.; Young, H.A.; Arany, P.;
Green, J.E. Inhibition of prostate cancer growth by muscadine grape skin extract and resveratrol
through distinct mechanisms. Cancer Res. 2007, 67, 8396–8405.
91. Lazze, M.C.; Pizzala, R.; Pecharroman, F.J.G.; Garnica, P.G.; Rodriguez, J.M.A.; Fabris, N.;
Bianchi, L. Grape waste extract obtained by supercritical fluid extraction contains bioactive
antioxidant molecules and induces antiproliferative effects in human colon adenocarcinoma cells.
J. Med. Food. 2009, 12, 561–568.
92. Ramos, S.; Alia, M.; Bravo, L.; Goya, L. Comparative effects of food-derived polyphenols on the
viability and apoptosis of a human hepatoma cell line (HepG2). J. Agric. Food Chem. 2005, 53,
1271–1280.
93. Saleem, A.; Husheem, M.; Harkonen, P; Pihlaja, K. Inhibition of cancer cell growth by crude
extract and the phenolics of Terminalia chebula retz. fruit. J. Ethnopharmacol. 2002, 81,
327–336.
94. Mantena, S.K.; Baliga, M.S.; Katiyar, S.K. Grape seed proanthocyanidins induce apoptosis and
inhibit metastasis of highly metastatic breast carcinoma cells. Carcinogenesis 2006, 27,
1682–1691.
95. Shih, P.H.; Yeh, C.T.; Yen, G.C. Anthocyanins induce the activation of phase II enzymes
through antioxidant response element pathway against oxidative stress-induced apoptosis. J.
Agric. Food Chem. 2007, 55, 9427–9435.
96. Yi, W.G.; Fischer, J.; Akoh, C.C. Study of anticancer activities of muscadine grape phenolics in
vitro. J. Agric. Food Chem. 2005, 53, 8804–8812.
97. Bagchi, D.; Sen, C.K.; Bagchi, M.; Atalay, M. Anti-angiogenic, antioxidant, and anti-
carcinogenic properties of a novel anthocyanin-rich berry extract formula. Biochemistry 2004,
69, 75–80.
Int. J. Mol. Sci. 2010, 11 644

98. Fan, P.; Lou, H.X. Effects of polyphenols from grape seeds on oxidative damage to cellular
DNA. Mol. Cell Biochem. 2008, 267, 67–74.
99. Wenzel, U.; Kuntz, S.; Brendel, M.D.; Daniel, H. Dietary flavone is a potent apoptosis inducer in
human colon carcinoma cells. Cancer Res. 2000, 60, 3823–3831.
100. Kuwajerwala, N.; Cifuentes, E.; Gautam, S.; Menon, M.; Barrack, E.R.; Reddy, G.P.V.
Resveratrol induces prostate cancer cell entry into S phase and inhibits DNA synthesis. Cancer
Res. 2002, 62, 2488–2492.
101. Subbaramaiah, K.; Chung, W.J.; Michaluarti, P.; Telang, N.; Tanabe, T.; Hiroyasu, I.; Jang, M.;
Pezzuto, J.M.; Dannenberg, A,J. Resveratrol inhibits cyclooxygenase-2 transcription and activity
in phorbol ester-treated human mammary epithelial cells. J. Biol. Chem. 1998, 273,
21875–21882.
102. Terra, X.; Montagut, G; Bustos, M.; Llopiz, N.; Ardevol, A.; Blade, C.; Fernandez-Larrea, J.;
Pujadas, G.; Salvado, J.; Arola, L. Grape-seed procyanidins prevent low-grade inflammation by
modulating cytokine expression in rats fed a high-fat diet. J. Nutr. Biochem. 2009, 20, 210–218.
103. Bralley, E.E.; Hargrove, J.L.; Greenspan, P.; Hartle, D.K. Topical anti-inflammatory activities of
vitis rotundifolia (Muscadine Grape) extracts in the tetradecanoylphorbol acetate model of ear
inflammation. J. Med. Food. 2007, 10, 636–642.
104. Li, W.G.; Zhang, X.Y.; Wu, Y.J.; Tian, X. Anti-inflammatory effect and mechanism of
proanthocyanidins from grape seeds. Acta Pharmacol. Sin. 2001, 22, 1117–1120.
105. Shukitt-Hale, B.; Carey, A.; Simon, L.; Mark, D.A.; Joseph, J.A. Effects of Concord grape juice
on cognitive and motor deficits in aging. Nutrition 2006, 22, 295–302.
106. Balu, M.; Sangeetha, P.; Murali, G.; Panneerselvam, C. Modulatory role of grape seed extract on
age-related oxidative DNA damage in central nervous system of rats. Brain Res. Bull. 2006, 68,
469–473.
107. Balu, M.; Sangeetha, P.; Haripriya, D.; Panneerselvam, C. Rejuvenation of antioxidant system in
central nervous system of aged rats by grape seed extract. Neurosci. Lett. 2005, 383, 295–300.
108. Baydar, N.G.; Sagdic, O.; Ozkan, G.; Cetin, S. Determination of antibacterial effects and total
phenolic contents of grape (Vitis vinifera) seed extracts. Int. J. Food Sci. 2006, 41, 799–804.
109. Jayaprakasha, G.K.; Tamil, S.; Sakartah, K.K. Antibacterial and antioxidant activities of grape
(Vitis vinifera) seed extracts. Food Res. Int. 2003, 36, 117–122.
110. Jung, H.J.; Hwang, I.A.; Sung, W.S.; Kang, H.; Kang, B.S.; Seu, Y.B.; Lee, D.G. Fungicidal
effect of resveratrol on human infectious fungi. Arch. Pharm. Res. 2005, 2, 557–560.
111. de Bruyne, T.; Pieters, L.; Witvrouw, M.; de Clercq, E.; Berghe, D.V.; Vlietinck, A.J. Biological
evaluation of proanthocyanidin dimers and related polyphenols. J. Nat. Prod. 1999, 62, 954–958.
112. Chavez, J.H.; Leal, P.C.; Yunes, R.A.; Nunes, R.J.; Barardi, C.R.; Pinto, A.R.; Simoes, C.M.;
Zanetti, C.R. Evaluation of antiviral activity of phenolic compounds and derivatives against
rabies virus. Vet. Microbiol. 2006, 116, 53–59.
113. Rodriguez-Vaquero, M.J.; Alberto, M.R.; Manca-de-Nadra, M.C. Influence of phenolic
compounds from wines on the growth of Listeria monocytogenes. Food Control. 2007, 18,
587–593.
Int. J. Mol. Sci. 2010, 11 645

114. Papadopoulou, C.; Soulti, K.; Roussis, I.G. Potential antimicrobial activity of red and white wine
phenolic extracts against strains of Taphylococcus aureus, Escherichia coli and Candida
albicans. Food Technol. Biotechnol. 2005, 43, 41–46.
115. Sivarooban, T.; Hettiarachchy, N.S.; Johnson, M.G. Physical and antimicrobial properties of
grape seed extract, nisin, and EDTA incorporated soy protein edible films. Food Res. Int. 2008,
41, 781–785.
116. Rotava, R.; Zanella, I.; da Silva, L.P.; Manfron, M.P.; Ceron, C.S.; Alves, S.H.; Karkow, A.K.;
Santos, J.P.A. Antibacterial, antioxidant and tanning activity of grape by-product. Cienc. Rural
2009, 39, 941–944.
117. Karapinar, M.; Sengun, I.Y. Antimicrobial effect of koruk (unripe grape—Vitis vinifera) juice
against Salmonella typhimurium on salad vegetables. Food Control 2007, 18, 702–706.
118. Thtmothe, J.; Bonsi, I.A.; Padilla-Zakour, O.I. Chemical characterization of red wine grape(Vitis
vinifera and Vitis Interspecific Hybrids)and pomace phenolic extracts and their biological
activity against Streptococcus mutans. J. Agric. Food Chem. 2007, 55, 10200–10207.
119. Anastasiadi, M.; Chorianopoulos, N.G.; George-John, E.N.; Haroutounian, S.A. Antilisterial
activities of polyphenol-rich extracts of grapes and vinification byproducts. J. Agric. Food Chem.
2009, 57, 457–463.
120. Yigit, D.; Yigit, N.; Mavi, A.; Yildirim, A.; Guleryuz, M. Antioxidant and antimicrobial
activities of methanol and water extracts of fruits, leaves and seeds of Vitis vinifera L. cv.
Karaerik. Asian J. Chem. 2009, 21, 183–194.
121. Brown, J.C.; Huang, G.; Haley-Zitlin, V.; Jiang, X.P. Antibacterial effects of grape extracts on
Helicobacter pylori. Appl. Environ. Microbiol. 2009, 75, 848–852.
122. Tagurt, T.; Tanaka, T.; Kouno, I. Antimicrobial activity of 10 different plant polyphenols against
bacteria causing food-borne disease. Biol. Pharm. Bull. 2004, 27, 1965–1969.
123. Appeldoorn M.M.; Venema D.P.; Peters T.H.F.; Koenen M.E.; Arts I.C.W.; Vincken J.P.;
Gruppen H.; Keijer J.; Hollman P.C.H. Some phenolic compounds increase the nitric oxide level
in endothelial cells in vitro. J. Agric. Food Chem. 2009, 57, 7693–7699.
124. Halliwell, B. Oxidative stress, nutrition and health. Experimental strategies for optimization of
nutritional antioxidant intake in humans. Free Radic. Res. 1996, 25, 57–74.
125. Romay, C.; Ledon, N.; Gonzalez, R. Further studies on anti-inflammatory activity of
phycocyanin in some animal models of inflammation. Inflamm. Res. 1998, 47, 334–338.
126. Lazze, M.C.; Savio, M.; Pizzala, R.; Cazzalini, O.; Perucca, P.; Scovassi, A.I.; Stivala, L.A.;
Bianchi, L. Anthocyanins induce cell cycle perturbations and apoptosis in different human cell
lines. Carcinogenesis 2004, 25, 1427–1433.
127. Baba S, Osakabe N, Natsume M, Muto Y,Takizawa T,Terao J. Absorption and urinary excretion
of (−)-epicatechin after administration of different levels of cocoa powder or (−)-epicatechin in
rats. J. Agric. Food Chem. 2001, 49, 6050–6056.
128. Baba, S.; Osakabe, N.; Natsume, M.; Muto, Y.; Takizawa, T.; Terao, J. In vivo comparison of the
bioavailability of (+)-catechin, (-)-epicatechin and their mixture in orally administered rats. J.
Nutr. 2001, 131, 2885–2891.
129. Cilla, A.; Gonzalez-Sarrias, A.; Tomas-Barberan, F.A.; Espin, J.C.; Barbera, R. Availability of
polyphenols in fruit beverages subjected to in vitro gastrointestinal digestion and their effects on
Int. J. Mol. Sci. 2010, 11 646

proliferation, cell-cycle and apoptosis in human colon cancer Caco-2 cells. Food Chem. 2009,
114, 813–820.
130. Argyri, K.; Proestos, C.; Komaitis, M.; Kapsokefalou, M. Phenolic compounds in red wine
digested in vitro in the presence of iron and other dietary factors. Int. J. Food Sci. Nutr. 2005, 56,
213–222.
131. Khokhar, S.; Richard, K.; Apenten, O. Iron binding characteristics of phenolic compounds: some
tentative structure-activity relations. Food Chem. 2003, 81, 133–140.
132. Tebib, K.; Rouanet. J.M.; Besançon, P. Effect of grape seed tannins on the activity of some rat
intestinal enzyme activities. Enzyme Prot. 1994–1995, 48, 51–60.
133. de Freitas, V.; Mateus, N. Structural features of procyanidin interactions with salivary proteins. J.
Agric. Food Chem. 2001, 49, 940–945.
134. Zhu, Q.Y.; Holt, R.R.; Lazarus, S.A.; Ensunsa, J.L.; Hammerstone, J.F.; Schmitz, H.H. Stability
of the flavan-3-ols epicatechin and catechin and related dimeric procyanidins derived from cocoa.
J. Agric. Food Chem. 2002, 50, 1700–1705.
135. Beart, J.E.; Lilley, T.H.; Haslam, E. Polyphenol interactions. Part 2. Covalent binding of
procyanidins to proteins during acid-catalysed decomposition; observations on some polymeric
proanthocyanidins. J. Chem. Soc. Perkin. Trans. 1985, 2, 1439–1443.
136. Record, I.; Lane, J.M. Simulated intestinal digestion of green and black teas. Food Chem. 2001,
73, 481–486.
137. Spencer, J.P.E.; Pannala, A.S.; Srai, S.K.; Debnam, E.; Rice-Evans, C.; Chaudry, F.
Decomposition of cocoa procyanidins in the gastric milieu. Biochem. Biophys. Res. Commun.
2000, 272, 236–241.
138. Forester, S.C.; Waterhouse, A.L. Identification of Cabernet Sauvignon anthocyanin gut
microflora metabolites. J. Agric. Food Chem. 2008, 56, 9299–9304.
139. Stagos, D.; Spanou, C.; Margariti, M.; Stathopoulos, C.; Mamuris, Z.; Kazantzoglou, G.;
Magiatis, P.; Kouretas, D. Cytogenetic effects of grape extracts (Vitis vinifera) and polyphenols
on mitomycin C-induced sister chromatid exchanges (SCEs) in human blood lymphocytes. J.
Agric. Food Chem. 2007, 55, 5246–5252.

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