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ARTICLE IN PRESS

JOURNAL OF
FOOD COMPOSITION
AND ANALYSIS
Journal of Food Composition and Analysis 20 (2007) 125132
www.elsevier.com/locate/jfca

Original Article

Polyphenolic content and in vitro antioxidant characteristics of wine


industry and other agri-food solid waste extracts
Dimitris P. Makrisa,b,, George Boskoub, Nikolaos K. Andrikopoulosb
a

Department of Food Quality Management & Chemistry of Natural Products, Mediterranean Agronomic Institute of Chania (M.A.I.Ch.),
P.O. Box 85, 73100, Chania, Greece
b
Laboratory of Food Chemistry, Biochemistry, Physical Chemistry, Department of Science of Dietetics-Nutrition, Harokopio University, 70,
El. Venizelou Str., 17671 Kallithea, Athens, Greece
Received 30 November 2005; received in revised form 7 February 2006; accepted 10 April 2006

Abstract
Solid by-products from white and red wine industry were subjected to evaluation as potential sources of antioxidant phytochemicals
on the basis of their content in phenolics and in vitro antioxidant activity. Furthermore, several other common plant solid wastes,
including apple, potato and onion peels, as well as carob pods and olive tree leaves were also considered, in order to carry out a
comparative assessment. The results showed that extracts from grape seeds (either white or red) contain exceptionally high amounts of
total polyphenols (10.311.1% on a dry weight basis), a great part of which is composed of avanols. Red grape pomace and stems
contained appreciable amounts of polyphenols, whereas potato and white grape peels were the tissues with the lowest polyphenol
content. The in vitro antiradical activity and reducing power were shown to be highly dependent on the total avonoid and total avanol
content Po0:001, but the hydroxyl free radical scavenging activity did not exhibit the same trend, suggesting dependence on particular
structural features. The results indicate that wine industry by-products, including grape seeds but also red grape pomace and stems, are
very rich sources of antioxidant polyphenols compared with other agri-food solid wastes, and therefore their exploitation as a source of
added-value products may be more cost-effective and merits a profounder investigation.
r 2006 Elsevier Inc. All rights reserved.
Keywords: Added-value products; Antiradical activity; Apples; Carobs; Flavonoids; Grape pomace; Olive tree leaves; Onions; Polyphenols; Potatoes;
Reducing power; Vinication by-products

1. Introduction
Large quantities of both liquid and solid wastes are
produced annually by the food processing industry. These
waste materials contain principally biodegradable organic
matter and their disposal creates serious environmental
problems. The waste loads at the processing plant can be
signicantly reduced through the use of new or modied
processing methods or through in-plant treatment and
Abbreviations: AAE, ascorbic acid equivalents; AAR , antiradical activity;
CTE, catechin equivalents; GAE, gallic acid equivalents; PR , reducing
power; QTE, quercetin equivalents; SAHFR , hydroxyl free radical
scavenging activity; TFl, total avanols; TFd, total avonoids; TP, total
polyphenols; TRE, Troloxs equivalents; S.D., standard deviation.
Corresponding author. Tel.: +3 28210 35056; fax: +3 28210 35001.
E-mail address: dimitris@maich.gr (D.P. Makris).
0889-1575/$ - see front matter r 2006 Elsevier Inc. All rights reserved.
doi:10.1016/j.jfca.2006.04.010

reuse, and a variety of processes are being developed


towards this direction, aiming at converting the waste
materials into bio-fuels, food ingredients and other addedvalue bio-products.
Wine industry wastes, which consist mainly of solid byproducts, include marcs, pomace, and stems, and may
account on average for almost 30% (w/w) of the grapes
used for wine production. All these by-products may bear a
considerable burden of phenolic components (GonzalezParamas et al., 2004), depending on the type of grape
(white or red), the part of the tissue (skins, seeds, etc.), as
well as the processing conditions (e.g., pomace contact).
Over the past few years, not only vinication by-products,
but also a number of other agricultural wastes of plant
origin have attracted considerable attention as potential
sources of bioactive phenolics, which can be used for

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D.P. Makris et al. / Journal of Food Composition and Analysis 20 (2007) 125132

various purposes in the pharmaceutical, cosmetic and food


industry. However, in many instances there is a rather
signicant lack of appropriate feasibility studies on the
exploitation of such wastes, and as a result their utilization
is still in its infancy.
Studies regarding vinication by-products are mainly
focused on the polyphenolic composition of seeds, which
are very rich in avanols (Yilmaz and Toledo, 2004;
Guendez et al., 2005), but pomace, which is composed of
seeds and skins, has also been evaluated as a potential
source of antioxidant polyphenols (Alonso et al., 2002;
Louli et al., 2004; Kammerer et al., 2005; Pinelo et al.,
2005). On the other hand, some other parts of grape
clusters that are discarded during the vinication process,
such as stems, have been given much less attention,
although they contain an important amount of polyphenols (Alonso et al., 2002; Souquet et al., 2000).
The examinations underlying value assessment of food
plant wastes are mainly based on the content and prole of
phenolics, as well as their in vitro antioxidant potency, but
the wide spectrum of analytical techniques employed for
both polyphenol analysis and antioxidant activity measurement make critical comparisons impractical or even
problematic. The objective of this study was an assessment
of several winery wastes as sources of polyphenolic
antioxidants on a comparative basis with several other
common food wastes. Comparisons were based on indices
pertaining to the polyphenolic composition, including the
determination of total polyphenols (TP), total avonoids
(TFd) and total avanols (TFl), and their association with
antioxidant activity, as this was revealed by three
representative in vitro tests.
2. Materials and methods
2.1. Chemicals
FolinCiocalteu phenol reagent and ascorbic acid were
from Fluka (Steinheim, Germany). Troloxs , gallic acid,
luminol, 2,4,6-tripyridyl-s-triazine (TPTZ), 2,2-diphenyl-

picrylhydrazyl DPPH stable radical, p-(dimethylamino)cinnamaldehyde (DMACA), quercetin and catechin were
from Sigma Chemical Co (St. Louis, MO, U.S.A.). Citric
acid, sodium nitrite, cobalt chloride CoCl2 .6H2 O, hydrogen peroxide, Na2 -EDTA and aluminium chloride hexahydrate AlCl3 .6H2 O were from Merck (Darmstadt,
Germany).
2.2. Plant solid wastes
Analytical details about the plant food by-products used
in this study are given in Table 1. White and red
vinication by-products were from Roditis and Agiorgitiko
cultivars (Vitis vinifera sp.) respectively, obtained from
wineries in the regions of Koropi and Nemea (prefectures
of Attica and Korinthia, Greece). Olive leaves were
harvested from an olive tree plantation (Attica). Deseeded
and chopped carob pods (kibbles), of approximately
1.52 mm diameter, were obtained from a carob-processing
factory (Chania, Crete). Potatoes, red onions and apples
were purchased from a local food store and peeled
immediately after receipt. All plant material was stored at
40  C.
2.3. Extraction procedure
A suitable quantity of tissue ranging from approximately
27 g was chopped into small pieces with a sharp, stainless
steel cutter to facilitate extraction. The chopped tissue was
ground with sea sand and a small portion of the extraction
solvent, which consisted of 0.1% HCl in methanol/acetone/
water (60/30/10, v/v/v), with a pestle and a mortar, and
then left to macerate for 30 min in the dark, covered with a
nylon membrane to minimize contact with air. The paste
that formed was placed in a 100 mL conical ask with
25 mL of solvent, and extraction was performed under
stirring at 700 rpm on a magnetic stirrer for 10 min. The
extract was ltered through a paper lter; this procedure
was repeated twice more. The extracts were then combined
in a 100 mL volumetric ask, made to the volume, and

Table 1
Agri-food wastes used in this study
Plant materiala

Tissue analysed

Moisture content (%)

White grape
Red grape
White grape
White grape
Red grape
White grape
Red grape
Olive tree leaves (Olea europaea)
Apples (red skinned) (Malus domestica)
Onion (red skinned) (Allium cepa)
Potato (brown skinned) (Solanum tuberosum)
Carob (Ceratonia siliqua)

Pomace (peels and seeds)


Pomace (peels and seeds)
Stems
Seeds
Seeds
Peels
Peels
Whole tissue
Peels
Outer dry and semi-dry layers and apical trims
Peels
Kibbles

71.56
55.62
60.38
45.04
41.56
75.28
55.50
48.79
81.68
88.73
81.83
11.51

All grape by-products used in this study came from Vitis vinifera cultivars.

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D.P. Makris et al. / Journal of Food Composition and Analysis 20 (2007) 125132

stored at 40  C until analysis. All extracts were centrifuged at 4500 rpm prior to analyses.
2.4. Determinations
Moisture content: Moisture was determined after drying
plant tissues in an air current-heated oven at 95 1C for 48 h.
Total polyphenols: Analysis was carried out employing
the FolinCiocalteu methodology, as described previously
(Arnous et al., 2002). Results were expressed as mg gallic
acid equivalents (GAE) per 100 g of fresh or dry weight.
Total flavonoids: A previously described protocol (Kim
et al., 2003), slightly modied, was used. A 0.1 mL aliquot
of extract appropriately diluted with MeOH was mixed
with 0.4 mL distilled water in a 2 mL microcentrifuge tube,
added 0.03 mL 5% NaNO2 , and allowed to react for
5 min. Following this, 0.03 mL 10% AlCl3 was added
and the mixture allowed to stand for a further 5 min.
Finally, 0.2 mL 1 M Na2 CO3 and 0.24 mL distilled water
were added to the reaction mixture and the absorbance at
510 nm was obtained against a blank that had been
prepared in a similar manner, by replacing the extract
with distilled water. Total avonoid content was calculated
from a calibration curve using catechin as standard, and
expressed as mg catechin equivalents (CTE) per 100 g fresh
tissue.
Total flavanols: Flavanols were determined after derivatization with p-DMACA, using an optimized methodology
(Nigel and Glories, 1991). Extract (0.2 mL) suitably diluted
with methanol was introduced into a 2 mL microcentrifuge
tube and 0.5 mL HCl (0.24 N in methanol) and 0.5 mL
DMACA solution (0.2% in MeOH) were added. The
mixture was allowed to react for 5 min at room temperature, and the absorbance was obtained at 640 nm. A
control sample was prepared by replacing sample with
methanol. Results were expressed as mg CTE per 100 g
fresh tissue.
Antiradical activity AAR : A procedure previously
reported (Arnous et al., 2002) was employed. Each extract
was diluted 1:20 with methanol immediately before the
.
analysis. Sample (0.025 mL) was added to 0.975 mL DPPH
solution (73 mM in methanol), and the absorbance at
t30
515 nm was read at t 0 At0
515 and t 30 min A515 .
s
Results were expressed as Trolox equivalents (mM TRE)
per g of fresh tissue using the following equation:


0:018  %DA 0:017
(1)
AAR
 FD
tw
as determined from linear regression, after plotting
%DA515 of known solutions of Troloxs against concentration, where
%DA515

At0
515

 At30
515
At0
515

 100,

tw the weight of fresh tissue (g), and F D the dilution


factor (20).

127

Reducing power PR : A modied protocol of that


described elsewhere (Arnous et al., 2002) was used. Sample
(0.05 mL) appropriately diluted with methanol was mixed
thoroughly with 0.05 mL FeCl3 solution (4.25 mM in 0.06 N
HCl), and incubated for 30 min in a water bath at 37 1C.
Following this, 0.9 mL TPTZ solution (1.07 mM in 0.06 N
HCl) were added, and the absorbance was recorded at
620 nm after exactly 5 min. PR was determined as mM
ascorbic acid equivalents (mM AAE) per g of fresh tissue
using the equation:


0:697  A620  0:012
(2)
PR
 F D,
tw
where tw is the weight of the tissue used and F D the
dilution factor.
Hydroxyl free radical scavenging activity SAHFR : A
highly sensitive, luminol-chemiluminescence assay was
employed (Parejo et al., 2000; Arnous et al., 2001). Samples
were diluted with borate buffer immediately before the
analysis to give a nal total polyphenol concentration of
approximately 1 mg L1 GAE. In a plastic tube, 0.9 mL
borate buffer (pH 9), 0.1 mL luminol solution 100 mg L1 ,
0.1 mL CoCl2 .6H2 O=Na2 -EDTA solution (2.26 and
9:92 g L1 , respectively), 0.025 mL sample and 0.05 mL
H2 O2 (50 mM) were added, and the tube was vortexed for
5 s to initiate in situ the chemiluminescence reaction. The
tube was then placed in a Junior LB 9509 luminometer
(Berthold Technologies, Bad Wildbad, Germany), which
was set to obtain the chemiluminescence intensity (ICL)
after 300 s, where it reaches a plateau (Parejo et al., 2000).
SAHFR was determined from the following equation:


0:0036  %DCL  0:1474
(3)
 FD
SAHFR
sw
as calculated from linear regression, after plotting known
amounts of quercetin against %DCL. Where
CLt0  CLt300
 100,
CLt0
sw is the weight of the sample used and F D the dilution
factor. Results were expressed as quercetin equivalents (mM
QTE) per g of tissue.
Statistical analyses: In all cases analyses were performed
in triplicate, unless elsewhere specied, and values averaged. The standard deviation (S.D.) was also calculated.
Correlations were established using regression analysis at a
95, 99, and 99.9% signicance level. Differences in content
of various polyphenol classes were assessed using Students
t-test at 95, 99, and 99.9%. All statistics were performed
with Microsoft ExcelTM 2000.
%DCL

3. Results
The comparative evaluation of the polyphenolic composition of the solid wastes was based on three representative
indices; the TP, TFd and TFl contents. Since some materials (e.g., onion and apple peels) contained signicantly

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D.P. Makris et al. / Journal of Food Composition and Analysis 20 (2007) 125132

128

avonoids. Red and white grape pomace were found to


have higher TP contents from all non-grape by-products,
but the analysis of peels indicated that a large part of
pomace polyphenols originated from seeds. It is worth
mentioning that white grape pomace had a TP level of
4826 mg per 100 g dry weight, whereas white grape peels
(seed-free pomace) contained only 970 mg of TP per 100 g
dry weight.
Regarding non-grape by-products, olive tree leaves had
the highest TP levels, followed by onion peels and apple
peels. Carobs and potato peels had TP content below
1500 mg per 100 g dry weight. In all these tissues avanol

higher moisture levels (Table 1), results were reported on


both fresh and dry weight basis for a more descriptive
expression.
Grape seeds were proven to bear the greatest TP burden,
which was 11.1% and 10.3% for white and red grape
samples, respectively, expressed on a dry weight basis
(Table 2). The TP content for these two by-products was
composed almost entirely of avonoids, as the TFd content
was over 99% of TP. In comparison with all other byproducts examined, grape seeds also contained exceptionally high amounts of TFl Po0:05. Grape stems had a TP
content of 5.8%, some 93% of which was attributed to

Table 2
Polyphenolic composition of the wine industry and other agri-food solid wastes tested
TPa

Plant material

White grape pomace


Red grape pomace
Stems
White grape seeds
Red grape seeds
White grape peel
Red grape peel
Olive tree leaves
Apple peels
Onionpeels
Potato peels
Carobs

TFdb

TFlc

fwb

dwb

fwb

dwb

fwb

dwb

1371731
23997118
2296770
60907499
64657487
23974
15027100
2058792
647712
42278
177717
1224762

48267111
54027266
57987178
111087909
103307837
970715
36257290
40277181
3522762
3727771
977796
1383770

100171
2349773
21387198
60807280
60737189
227723
14867175
858751
566755
25172
12773
817718

352275
52897334
53997399
110907511
102587323
922762
35877323
1678773
30827282
2212718
702718
923720

357.272.2
665.2715.4
772.8735.8
2486.97127.4
3372.07124.2
48.470.8
255.6712.6
1.170.3
20.672.4
0.0
0.0
18.470.5

1258.375.4
1509.6731.6
1977.0777.6
4605.17203.1*
5835.57209.6*
197.273.0
626.0726.6
2.170.5
110.679.6
0.0
0.0
20.470.6

TFd/TP
(%)d

TFl/TP
(%)d

73.0
97.9
93.1
99.8
99.3
95.1
99.0
41.7
87.5
59.4
71.9
66.7

26.1
28.0
34.1
41.5
56.5
20.3
17.3
0.1
3.1
0
0
1.5

Values represent means of triplicate determination ( S.D.), and are expressed on both fresh weight (fwb) and dry weight basis (dwb).
a
Total polyphenols (mg GAE per 100 g).
b
Total avonoids (mg CTE per 100 g).
c
Total avanols (mg CTE per 100 g).
d
Calculated on dry matter basis.
*
Values statistically signicant (Po0:05, students t-test).

Table 3
Antioxidant characteristics of the wine industry and other agri-food solid wastes tested
AAR a

Plant material

White grape pomace


Red grape pomace
Stems
White grape seeds
Red grape seeds
White grape peel
Red grape peel
Olive tree leaves
Apple peels
Onion peels
Potato peels
Carobs

PR b

SAHFR c

fwb

dwb

fwb

dwb

fwb

dwb

0.6370.05
1.0270.09
1.2670.01
3.2670.02
3.4870.06
0.4370.00
1.2770.00
0.6370.01
0.2870.01
0.1470.00
0.1570.00
0.8170.01

2.2270.17
2.3070.21
3.1770.03
5.9470.04
5.9470.11
1.7470.02
3.0670.01
1.2370.02
1.5170.07
1.2270.00
0.8170.02
0.9270.02

0.7770.04
1.5770.06
1.5370.01
4.0870.16
4.4470.04
0.1270.01
0.7270.05
0.7970.02
0.2470.02
0.0770.00
0.0170.00
0.7270.06

2.7170.13
3.5370.14
3.8670.02
7.4470.30
7.5870.07
0.5070.05
1.7470.11
1.5670.05
1.3270.08
0.6070.01
0.0770.00
0.8270.07

0.7370.01
0.6870.04
0.5670.02
0.7970.00
1.7470.15
0.1270.01
0.6170.14
0.9570.10
0.3970.01
0.5570.06
0.0570.01
0.3470.01

2.5770.20
1.5370.08
1.4270.06
1.4470.00
2.9770.25
0.4870.05
1.4870.34
1.8670.19
2.1370.07
4.8570.50
0.2970.05
0.3870.01

Values represent means of triplicate determination ( S.D.), and are expressed on both fresh weight (fwb) and dry weight basis (dwb).
a
Antiradical activity (mM TRE/g).
b
Reducing power (mM AAE/g).
c
Hydroxyl free radical scavenging activity (mM QTE/g).

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D.P. Makris et al. / Journal of Food Composition and Analysis 20 (2007) 125132

levels did not exceed 3.1% of TP content (Table 2), while


the percentage of TFd in relation to TP ranged from 41.7%
(olive leaves) to 87.5% (apple peels).
The estimation of the in vitro antioxidant activity
showed that grape seed extracts exhibit the highest
reducing power and antiradical activity (Table 3), but the
hydroxyl free radical scavenging activity of onion peels,
expressed on a dry weight basis, surpassed the activity of
all by-products. The uniformity of the expression of the
antioxidant activity based on the rst two tests was
demonstrated by simple linear regression analysis
(Table 4), which showed that PR and AAR are highly
dependent on the TFd and TFl contents Po0:001, but
SAHFR did not exhibit the same trend. This afnity was
further illustrated by the high correlation of values deriving
from PR and AAR tests (Fig. 1A). On the contrary, correlation of SAHFR values with either PR or AAR was poor
and statistically insignicant P40:05 (Fig. 1B and 1C).

PR (mM AAE/g dw)

8.0
y = 1.3759x - 0.8024
R2 = 0.9113

6.0

4.0

2.0

0.0
0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

AAR (mM TRE/g dw)

(A)

SAHFR (mM QTE/g dw)

0.6

4. Discussion

y = 0.1061x + 1.5175
R2 = 0.0219
0.4

0.2

0.0
0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

AAR (mM TRE/g dw)

(B)
0.6

SAHFR (mM QTE/g dw)

Fruit and vegetable residuals are rich in antioxidants and


their utilization for the production of natural food
additives has gained prominent attention, since polyphenols constitute one of the higher value options for byproduct exploitation. In this context several polyphenolcontaining materials have been examined for their efciency as food antioxidants (Laufenberg et al., 2003). In the
present study, wine industry solid wastes were compared
for their antioxidant ability on the basis of three
representative in vitro tests, which have different chemical
background, and therefore different aspects of antioxidant
expression may be revealed. To further stress the importance of vinication by-products as major sources of
antioxidant phytochemicals, several other food wastes were
examined, which contain phenolics with demonstrated
antioxidant capacity (Table 5).
From all grape by-products assessed, seeds were shown
to contain particularly high amounts of phenolics, an
important percentage of which was ascribed to avanols
(41.556.5%). Seeds were shown to contribute a signicant
part of polyphenols to pomace, since grape peels, especially
those originating from white grapes, were poorer polyphenol sources. Grape stems were also a prominent source

129

y = 0.0868x + 1.5538
R2 = 0.0304
0.4

0.2

0.0
0.0

0.4

0.2

0.6

0.8

PR (mM AAE/g dw)

(C)

Fig. 1. Correlation of values from different antioxidant tests, established


after linear regression analysis at 95% and 99.9% signicance level: (A)
reducing power versus antiradical activity; (B) hydroxyl free radical
scavenging activity versus antiradical activity; (C) hydroxyl free radical
scavenging activity versus reducing power. All values are expressed on a
dry weight basis.

Table 4
Statistical parameters calculated after linear regression analysis between polyphenol groups and antioxidant characteristics
PR

AAR

TPa
TFdb
TFlc
a

r2

slope

r2

slope

r2

slope

0.8084
0.9294
0.9222

1:61  105
4:46  107
7:25  107

5  104
5  104
9  104

0.9291
0.9668
0.9519

1:21  107
1:01  108
6:5  108

5  104
7  104
1:3  105

0.1218
0.0528
0.0308

0.2663
0.4726
0.5851

1  104
8  105
1  104

Total polyphenols (mg GAE per 100 g).


Total avonoids (mg CTE per 100 g).
c
Total avanols (mg CTE per 100 g).
b

SAHFR

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D.P. Makris et al. / Journal of Food Composition and Analysis 20 (2007) 125132

Table 5
Bibliographic data on the principal phenolics with demonstrated antioxidant activity in the by-products tested
Plant material

Phenolic(s)

Reference

White grape peels


Red grape peels

Flavanols, proanthocyanidins and hydroxycinnamates


Anthocyanins, avanols, proanthocyanidins, avonols
and hydroxycinnamates

Alonso et al. (2002)

Grape seeds

Gallic acid, proanthocyanidins and avanols

Grape stems

Flavanols, proanthocyanidins, avonols and hydroxycinnamates

Olive tree leaves

Oleuropein, avones, avonols, avonol derivatives

Apples (red skinned)

Flavanols, proanthocyanidins, avonols and hydroxycinnamates

Onion (red skinned)

Flavonols, anthocyanins

Potato (brown skinned)


Carob

Chlorogenic acid
Gallic acid, gallotannins, proanthocyanidins, avanols, avonols

of polyphenols, exhibiting levels comparable to those of


red grape pomace. In the same fashion, grape seed, pomace
and stem extracts had high PR and AAR , but their SAHFR
was 1.62.7 times lower than that of onion extracts. This
nding raised an issue concerning structure-activity relationships of the polyphenolic constituents in the byproducts tested.
Evaluation of the outcome from the antioxidant test
performed using linear regression analysis indicated that
both PR and AAR are linked with TFd and TFl in a very
signicant manner Po0:001. Such a correlation was
previously found for red wines (Makris and Kefalas, 2005)
and attributed to the relevance of the mechanisms underlying the antioxidant activity of certain polyphenols.
However, red wines exhibited the same trend for SAHFR
(Kefalas et al., 2003) as well, and it was concluded that in
matrices composed of similar phenolics, the antioxidant
activity may display uniformity in different test models. In
Table 5 it can be seen that the predominant antioxidants
in grape by-products, especially seeds, are avanols
(catechin and epicatechin) and proanthocyanidins (Gonzalez-Paramas et al., 2004; Yilmaz and Toledo, 2004;
Guendez et al., 2005), whereas onion skins contain mainly
avonols (Suh et al., 1999; Gennaro et al., 2002; Nuutila
et al., 2003; Furusawa et al., 2003). Increased potency was
also seen for apple peel and olive leaf extracts, which also
contain avonol glycosides (Lu and Foo, 2000; Schieber et
al., 2003; Chinnici et al., 2004; Bouaziz and Sayadi, 2005),
although other avonol-containing extracts, including
grape peels (Alonso et al., 2002; Bartolome et al., 2004;
Amico et al., 2004), stems (Alonso et al., 2002; Souquet

Kammerer et al. (2005)


Bartolome et al. (2004)
Amico et al. (2004)
Gonzalez-Paramas et al. (2004)
Yilmaz and Toledo (2004)
Guendez et al. (2005)
Alonso et al. (2002)
Souquet et al. (2000)
Bouaziz and Sayadi (2005)
Speroni et al. (1998)
Lu and Foo (2000)
Schieber et al. (2003)
Chinnici et al. (2004)
Suh et al. (1999)
Gennaro et al. (2002)
Nuutila et al. (2003)
Furusawa et al. (2003)
Kanatt et al. (2005)
Kumazawa et al. (2002)
Papagiannopoulos et al. (2004)

et al., 2000) and carobs (Kumazawa et al., 2002; Papagiannopoulos et al., 2004) were not as efcient in scavenging
hydroxyl radicals. In these cases, however, the presence of
other phenolics, i.e., anthocyanins, gallotannins, oleuropein, etc., must be pointed out, because interactions among
different classes of polyphenols may yield unpredictable
antioxidant efcacy, since phenomena of synergism as well
as antagonism can occur. It appears therefore that peculiar
constituents may dene to an important degree the
antioxidant behaviour, depending on the assay used, and
this evidence should be regarded as an additional criterion
for a more integrated assessment of plant by-products as
sources of antioxidant phenolics.

5. Conclusions
The comparison of several wine industry by-products
with other common agri-food solid wastes provided
evidence that grape seeds, red grape pomace and stems
are very rich sources of antioxidant phenolics, and thus a
deeper study of these products that aims at implementing
techniques for industrial exploitation merits further consideration. It was also shown that peels contain far fewer
important amounts of polyphenols than stems and seeds,
but do contain important moisture levels. This nding is of
prime importance, given that moisture plays a crucial role
regarding by-product preservation and drying costs. Thus
proper handling of grape pomace would minimize drying
time, increase preservation period and reduce the amount
of solvent required for optimal polyphenol recovery. Some

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D.P. Makris et al. / Journal of Food Composition and Analysis 20 (2007) 125132

more critical ndings of this investigation may be


summarized as follows:
1. Solid by-products from the wine industry contain a
larger burden of phenolics as compared with other
common food processing wastes.
2. Increased avonoid content is signicantly correlated
with increased antioxidant efciency, as this may be
expressed by radical scavenging and reducing capacity.
Hydroxyl free radical scavenging, however, appears to
be governed by components bearing specic structural
features. Therefore, the preparation of extracts with
improved antioxidant potency from vinication byproducts should be based on the estimation of other
polyphenolic classes, in addition to TP content, since the
overall antioxidant effect may be dened by the relative
amounts of the most active components.
3. Criteria for a credible assessment of plant by-products
as sources of polyphenolic antioxidants should be based
on the polyphenolic content, prole, and antioxidant
behaviour in more than one systems.

Acknowledgements
Dr. D.P. Makris wishes to thank the Greek Scholarships
Foundation for the nancial support in the form of postdoctoral scholarship. Mr. Dimitris Akrivos (Gaia Winery,
Nemea) and Mr. Vassilis Nikolou (Nikolou Winery,
Attica) are thanked for providing the red and white
vinication by-products, respectively. Mr. Christos Petrakis (Mediterranean Agronomic Institute of Chania, Crete)
is thanked for providing the carob pods.
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