Industrial Crops and Products 43 (2013) 360364

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Industrial Crops and Products
j our nal home page: www. el sevi er . com/ l ocat e/ i ndcr op
Characterization of extracts from winery by-products with antifungal activity
against Botrytis cinerea
Leonora Mendoza

, Karen Ya nez, Marcela Vivanco, Ricardo Melo, Milena Cotoras

Facultad de Qumica y Biologa, Universidad de Santiago de Chile, Av. Libertador OHiggins 3363, Casilla-40, Correo-33, Santiago, Chile
a r t i c l e i n f o
Article history:
Received 21 March 2012
Received in revised form24 July 2012
Accepted 25 July 2012
Keywords:
Grape pomace
Antifungal activity
Botrytis cinerea
Phenolic compounds
a b s t r a c t
Phenolic compound proles of grape pomace extracts from a mixture of Chilean grape varieties (Cabernet
Sauvignon, Carmnere andSyrah) were analyzed. Two extraction methods were used: solvent andsoxhlet
extraction, followed by fractionation with hexane, chloroform and ethyl acetate. Phenolic compounds in
different fractions were identied by HPLC. Ethyl acetate phase of solvent extraction contained a wide
variety of phenolic compounds. Hexane phase of the extracts presented lowest diversity. Quercetin was
found in almost all fractions.
Also, the in vitro antifungal activity of these extracts against the phytopathogenic fungus Botrytis cinerea
was evaluated. Hexane or chloroform fractions from extracts obtained by solvent extraction showed the
highest inhibitory effect on mycelia growth of this fungus, with IC
50
value of 40 ppm. In general, ethyl
acetate fractions were less active against B. cinerea. Therefore, it can be concluded that grape pomace are
a good low cost source to obtain antifungal extracts.
2012 Elsevier B.V. All rights reserved.
1. Introduction
In the wine industry, a large amount of grape (Vitis vinifera)
residues (grape pomace) are obtained. Grape pomace consists
mainly of skin residues, broken cells with pulp remains, stalks,
and seeds (Ruggieri et al., 2009). This winery residue is produced
after pressing the crushing grapes in the white wine processing
or after fermentation in red wine production. In Chile, the wine
production was of 733,404,919L in 2010; 70.6% corresponded to
red wine reaching a production of 517,878,816L (Servicio Agrcola
y Ganadero, SAG, Chile). If it is considered that 18kg pomace are
produced by 100L of wine (Rockenbach et al., 2011), then, in Chile,
during 2010, the grape pomace production from red wine was
approximately of 93,000tons.
Grape pomace composition varies depending on grape variety,
climate and technology of vinication (Schieber et al., 2001) and it
has been used as a rich source to obtain high-value products such
as ethanol, tartrates and malates, citric acid, grape seed oil, and
phenolic compounds (Arvanitoyannis et al., 2006).
In grape pomace, the phenolic content is high because these
compounds are poorly extracted during vinication. Anthocyanins,
catechins, avonol glycosides, phenolic acids, alcohols and stil-
benes are the principal phenolic constituents of grape pomace

Corresponding authors. Tel.: +56 2 718 1094; fax: +56 2 6812108.

E-mail addresses: leonora.mendoza@usach.cl (L. Mendoza),
milena.cotoras@usach.cl (M. Cotoras).
(Corrales et al., 2008; Katalinic et al., 2010; Xia et al., 2010). Of these
biological properties described for phenolic compounds, the best
known and most widely studied is the antioxidant effect, which
has been the focus of a large amount of analysis, mostly of clini-
cal and nutritional nature (Meyer et al., 1998; Chidambara Murthy
et al., 2002; Gonzlez-Params et al., 2003; Rockenbachet al., 2011;
Sagdic et al., 2011). There are few reports on antifungal activity of
grape pomaces. Pomace of Gamay and Kalecik varieties inhibited
growth in vivo and in vitro of Zygosaccahromyces rouxii and Z. bailii
(Sagdic et al., 2011). Ontheother hand, redandwhitewinephenolic
extracts showed strong fungicidal activity against Candida albicans
(Papadopoulouet al., 2005; Junget al., 2007; Tehranifar et al., 2011).
Grey rot produced by Botrytis cinerea is an important disease of
grapevines intemperate climates as inChile andit causes extensive
economic losses in grape production. Grey rot produces, in grapes,
biochemical changes that reduce wine quality (Jacometti et al.,
2010), B. cinerea is managed through the use of synthetic fungicide.
These fungicides are becoming less accepted by the wine industry
becausetheincreaseof resistant strains, public disapproval for their
effect on human health and environmental pollution (Jacometti
et al., 2010). Therefore, the wine industry is beginning to require
alternative fungicides. Plant extracts are among the alternative
fungicides that have been successfully applied to control B. cinerea
in vineyards (Jacometti et al., 2010). Then, grape pomace extracts
could become an important alternative control method of this fun-
gus.
The objective of this work was to characterize the phenolic com-
pound prole of grape pomace extracts froma mixture of Chilean
0926-6690/$ see front matter 2012 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.indcrop.2012.07.048
L. Mendoza et al. / Industrial Crops and Products 43 (2013) 360364 361
varieties (Cabernet Sauvignon, Carmnere and Syrah) obtained by
organic solvent or soxhlet extraction. Secondly, the other objective
was to evaluate the effect of these extracts on mycelia growth and
conidia germination of B. cinerea.
2. Materials and methods
2.1. Grapes pomaces
Pomaces were obtained after fermentation from a mixture of
grape (V. vinifera) varieties (Cabernet Sauvignon, Carmnere and
Syrah) fromthe 2009 harvest season fromMiguel Torres vineyard
(Curic, Chile). Pomaces were maintained at 20

C until they were

used.
2.2. Extraction process
Grape pomaces were extracted by four different systems:
a. Methanol extraction: 450g or 300g of whole and ground grape
pomace, respectively were extracted with 1L of methanol/HCl
1% (v/v) for 4h with constant agitation at 4

C. Extracts were
concentrated in a rotary evaporator and 20mL of distilled
water were added and volume was decreased in a rotary
evaporator. Water addition was repeated three times. Finally,
aqueous suspension or crude extract was subjected to sequen-
tial liquidliquid extraction with hexane, chloroformand nally
ethyl acetate.
b. Ethanol extraction, method 1: 450g or 300g of whole and
ground grape pomace, respectively were extracted with 1L of
ethanol 70% (v/v) for 4h with constant agitation at 4

C. Extracts
were concentrated in a rotary evaporator to get crude extracts
whichweresubjectedtosequential liquidliquidextractionwith
hexane, chloroformand nally ethyl acetate.
c. Ethanol extraction, method 2: 450g or 300g of whole and
ground grape pomace, respectively were extracted with 1L of
ethanol 70% (v/v) for 4h with constant agitation at 4

C. Extracts
were concentrated in a rotary evaporator and 20mL of distilled
water were addedandthenthe volume was decreasedina rotary
evaporator. Water addition was repeated three times. Finally,
aqueous suspension or crude extract was subjected to sequen-
tial liquidliquid extraction with hexane, chloroformand nally
ethyl acetate.
d. Soxhlet extraction: 10g of whole grape pomace was submitted
to a soxhlet extraction using methanol in a solidliquid rate 1:20
at 40

C for 12h. Methanol was decreased at reduced pressure.

This methanol extract was subjected to liquidliquid extrac-
tion with hexane. Therefore, methanolic and hexane phase were
obtained.
In all extractions, residues were weighed to obtain the extrac-
tion yields.
2.3. Analysis of phenolic compounds in different extractions
Phenolic compound analysis was done by high performance
liquid chromatography (HPLC) by using a Waters 600 HPLC chro-
matograph (Waters, Mildford, MA, USA) equipped with a Waters
2990 diode array detector, and a Symmetry C-18 (5m) (Waters,
Milford, MA, USA) column (3.9mm150mm). Chromatography
was conducted at 25

C. Mobile phase was composed of 1.0% (v/v)

acetic acid in distilled water (solvent A) and acetonitrile (solvent
B). The system was run by 60min with a gradient program as
follows: 1020% B in 20min. This proportion was maintained dur-
ing 20min and then a linear gradient of 2050% B in 5min was
applied. Finally, this proportion was maintained during 15min.
The ow rate was 0.8mL/min and it was recorded at 280 and
360nm. For analysis, 5mg of extracts or standards were dis-
solved in 1mL of methanol and 20L of the sample solution
were injected. Identication of phenolic compound was done
by comparing their retention times and UVvis spectra with
standards. Gallic acid, protocatechuic acid, vanillic acid, sinapic
acid, syringic acid, ellagic acid, (+)-catechin, ()-epicatechin,
epigallocatechin, caffeic acid, vanillin, p-coumaric acid, 4-hydroxy-
3,5-dimethoxybenzaldehyde, 4-hydroxyphenylacetic acid, trans-
resveratrol, quercetin, kaempferol and myricetin were used as
standard.
2.4. Fungal isolate and culture conditions
In this study B. cinerea strain G29 was used. This strain was orig-
inally isolated from naturally infected grapes (V. vinifera) (Mu noz
et al., 2002) and was maintained on malt-yeast extract agar slants
(2%(w/v) malt extract, 0.2%(w/v) yeast extract and1.5%(w/v) agar)
at 4

C. The fungus was growninthe dark onmalt-yeast extract agar

medium or soft agar medium (2% (w/v) malt extract, 0.2% (w/v)
yeast extract and 0.6% (w/v) agar).
2.5. Antifungal activity
The fungitoxicity on mycelial growth of B. cinerea of differ-
ent extracts and phenolic compounds was assessed in vitro using
0
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
Methanol
extracon
Ethanol
extracon,
method 1
Ethanol
extracon,
method 2
Soxhlet
extracon
Y
i
e
l
d

p
e
r
c
e
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a
g
e

(
w
/
w
)
Ground
Whole
a
a
b
c
c
d
d
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1.5
2
2.5
H
e
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C
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Methanol
extracon
Ethanol extracon, Ethanol extracon,
method 1 method 2
Soxhlet
extracon
Y
i
e
l
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p
e
r
c
e
n
t
a
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e

(
w
/
w
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Ground
Whole
a a a a
a
a a
a
b
b
c
d
e e
e
f
b
c
c
g
A
B
Fig. 1. Yield (%) of grape pomace extraction. Yields were calculated based on dry
grape pomace. Grape pomaces were subjected to solidliquid or soxhlet extrac-
tion of grape pomace to obtain crude extracts (A). Then crude extracts were
extracted sequentially with different solvents (B). Extractions were realized using
whole (gray bar) and ground (black bar) grape pomace. All values are expressed as
meanstandard deviation fromat least three independent experiments. Different
letters indicate that the means are signicantly different at P0.05.
362 L. Mendoza et al. / Industrial Crops and Products 43 (2013) 360364
Table 1
Phenolic compounds in extracts obtained fromgrape pomace.
Extraction method Liquidliquid extraction solvent Phenolic compounds
Whole Ground
Methanol Hexane Quercetin Quercetin
Kaempferol Kaempferol
Chloroform Vanillic acid Vanillic acid
Syringic acid Syringic acid
Quercetin Quercetin
Kaempferol Kaempferol
Ethyl
acetate
Gallic acid Gallic acid
Protocatechuic acid Protocatechuic acid
Vanillic acid Vanillic acid
Syringic acid Syringic acid
Ellagic acid Ellagic acid
Quercetin Myricetin
Kaempferol Quercetin
Kaempferol
Ethanol,
method 1
Hexane n.d.
a
n.d.
a
Chloroform Vanillic acid Vanillic acid
Syringic acid Syringic acid
Quercetin Quercetin
Kaempferol
Ethyl
acetate
Gallic acid Gallic acid
Protocatechuic acid Vanillic acid
Vanillic acid Syringic acid
Syringic acid Quercetin
p-Coumaric acid Kaempferol
Ellagic acid
Ethanol,
method 2
Hexane n.d.
a
n.d.
a
Chloroform Vanillic acid Vanillic acid
Syringic acid Syringic acid
Kaempferol Kaempferol
Ethyl
acetate
Gallic acid Gallic acid
Quercetin ()Epicatechin
Syringic acid
Quercetin
Soxhlet Hexane n.d.
a
Methanol Gallic acid
Vanillic acid
Syringic acid
Quercetin
Kaempferol
a
Phenolic compounds not detected.
the radial growth test on malt-yeast extract agar (Mendoza et al.,
2009). Extracts, phenolic compounds or the fungicide iprodi-
one were dissolved in methanol at different nal concentrations.
Aliquots of these solutions (100L) were added to Petri dishes
containing malt-yeast extract agar medium (5mL). Final solvent
concentration was identical in the control and treatment assays.
Methanol was allowed to evaporate prior to inoculation. Petri
dishes wereinoculatedwithfreshmyceliumandincubatedat 22

C.
Results of antifungal effect were expressed as IC
50
(concentration
that reduced mycelial growth by 50%) determined by regressing
Table 2
Effect of extracts on mycelia growth of B. cinerea.
Extraction method Liquidliquid extraction solvent Antifungal activity IC
50
(g/mL)
Whole Ground
Methanol Hexane 486.6 21.3 i 40.09.8 a
Chloroform 460.8 25.5 i 40.02.0 a
Ethyl acetate 2832.0 105.3 k 108.317.9 c
Ethanol, method 1 Hexane 143.0 15.4 d 35.75.2 a
Chloroform 127.5 13.0 c 40.04.9 a
Ethyl acetate 209.7 25.0 f 85.310.0 b
Ethanol, method 2 Hexane 179.9 12.8 e 289.920.1 g
Chloroform 362.6 25.9h 236.78.5 f
Ethyl acetate 178.1 18.3 e 531.945.3 i
Soxhlet Hexane 1228.5 124.3 j ND
a
Methanol 126.8 25.6 c,d ND
a
a
Not determined.
All values are expressed as meanstandard deviation from at least three independent experiments. Different letters indicate that the means are signicantly different at
P 0.05.
L. Mendoza et al. / Industrial Crops and Products 43 (2013) 360364 363
the inhibition of radial growth values (percent control) at different
extract or compound concentrations after 48h of incubation. All
experiments were done at least in triplicate.
Also, the effect of extracts on conidial germination was deter-
mined. Conidial germinationassays werecarriedout onmicroscope
slides coatedwithsoft agar medium(2mmthickness). Extracts was
added dissolved in methanol at 160g/mL. Methanol was allowed
to evaporate prior to inoculation. The slides were inoculated with
dry conidia obtained fromsporulated mycelia (1 week old), placed
in a humid chamber (90% relative humidity), and incubated in the
dark at 22

C for 7h. Conidial germination was determined directly

on the slides at one-hour intervals. The percentage of germina-
tion was estimated by counting the number of germinated conidia
in ve microscope elds each containing approximately 40 coni-
dia. Conidia were judged to have germinated when the germtube
length was equal to or greater than conidial diameter. Each exper-
iment was done at least in triplicate.
2.6. Statistical analyses
The results were analyzed using analysis of variance (one-way
ANOVA). Means wereseparatedwiththeleast signicant difference
test (P0.05). All analyses were made using the statistical program
SPSS version 13.0.
3. Results and discussion
3.1. Extraction yields
Whole or ground grape pomaces were subjected to solvent
(solidliquidextraction) or soxhlet extractionandtheneachextract
was submitted to liquidliquid extraction using sequentially dif-
ferent solvents; extraction yields are presented in Fig. 1. Results
obtained show yields between 0.5 and 4.5%, based on dry grape
pomace (Fig. 1A). The yield depended on the extraction method
used, solidliquid or soxhlet extraction, and the substrate condi-
tions (whole or ground). In solidliquid extraction, highest yields
were obtained with ground substrate. When whole substrate was
used, similar yields were obtainedwithmethanol or soxhlet extrac-
tion (approx. 2%). However, these yields signicantly decreased at
the second extraction (liquidliquid), which showed yields from
0.1%to2%basedonthedrygrapepomace(Fig. 1B). Amongtheselast
values, it can be observed that extracts obtained by liquid extrac-
tion fromwhole substrates showed the lowest yields; whilst using
ethyl acetate as solvent and ground substrate highest yields were
obtained. Additionally, solvent extraction allowed obtaining more
polar compounds than the soxhlet extraction. As shown in liter-
ature, by using solidliquid extraction, values between 1 and 5%
are the most frequent yields (de Campos et al., 2008; Sagdic et al.,
2011).
3.2. Phenolic prole fromgrape pomaces
It has been largely reported that extraction method can affect
the phenolic prole of the extracts (Xia et al., 2010). Therefore, in
this work, two extraction methods and three solvent systems were
used. Phenolic compounds in different fractions were identied by
HPLC and are shown in Table 1. The phenolic composition in the
different fraction varied. Ethyl acetate phase of methanol extracts
contained a wider variety of phenolic compounds. Hexane phase of
all extracts presented lowest diversity. Quercetin was found in all
fractions except on the ethyl acetate fraction of extracts obtained
from whole grape pomace with ethanol method 1. Phenolic com-
pounds were not detected in hexane fractions of extracts obtained
fromwhole or ground grape pomace with ethanol methods 1 and
2 and soxhlet.
Table 3
Effect of phenolic compound on mycelia growth of B. cinerea.
Compound Antifungal activity IC
50
(g/mL)
Gallic acid 316.9 23.8 e
Protocatechuic acid 383.4 25.1 f
Vanillic acid 207.4 13.2 d
Ellagic acid 390.0 19.8 f
Quercetin 120.8 7.1 b
Kaempferol 100.9 10.8 a
p-Coumaric acid 95.8 3.6 a
Syringic acid 163.1 7.9 c
Epicatechin 208 15.7 d
Catechin 178.9 9.0 c
All values are expressed as meanstandard deviation fromat least three indepen-
dent experiments. Different letters indicatethat themeans aresignicantlydifferent
at P0.05.
Among simple phenolic compounds reported in skin or seed
grapes (Xia et al., 2010), resveratrol and (+)-catechin were not
detected in these extracts, although catequin has been reported
as a very abundant polyphenol in the seed extracts (Guendez et al.,
2005) andresveratrol very abundant inCabernet Sauvignonvariety
(Iacopini et al., 2008).
3.3. Antifungal activity
Effect of the extracts against mycelia growth of B. cinerea is
shown in Table 2. Hexane or chloroform fractions from methanol
or ethanol (method 1) extractions fromground grape pomace were
the more active extracts with IC
50
value of 40ppm. These fractions
didnot inhibit conidia germinationof B. cinerea (data not shown). In
general, ethyl acetatefractions wereless activeagainst B. cinerea. As
control, theeffect thecommercial fungicide, iprodione, onB. cinerea
mycelial growth was determined presenting an IC
50
of 4.9ppm.
Notably, although the extract concentration required to inhibit 50%
of B. cinerea mycelia growth is higher than the fungicide iprodione,
it is lower than those reported for other antifungal plant extracts
(Tegegne et al., 2008; Andrade Pinto et al., 2010; Mamoci et al.,
2011; Chen and Dai, 2012).
On the other hand, the antifungal activity of some of the com-
pounds identied in the fractions was evaluated (Table 3). More
active fractions (hexane or chloroformfractions frommethanol or
ethanol, method 1) contained vanillic and syringic acid, quercetin
and kaempferol. These compounds showed a lowantifungal effect,
with IC
50
value among 100 and 207ppm. These values are higher
than the extract values. Instead, the more active compound was
the p-coumaric acid that was present in fraction with lowantifun-
gal activity. Therefore, in the antifungal effect of the more active
fractions some other compounds couldbe implied. Compoundsuch
as fatty acid could be present in active fractions, since this kind of
compounds have been identied in grape pomace extracts from
Cabernet Sauvignon (de Campos et al., 2008). Some of these com-
pounds have showed antifungal activity against different fungi
(Avis and Blanger, 2001).
4. Conclusions
Comparing the techniques used in this study to obtain the
grape pomace extracts with antifungal activity against B. cinerea,
it was observed that more apolar fractions fromextracts obtained
with methanol or ethanol method 1 from ground grape pomace
presented the highest antifungal activities. These fractions, with
exception of hexane fraction from ethanol method 1, contained
phenolic compounds as kaempferol, quercetin, vanillic andsyringic
acid, however, these compounds presented lowantifungal activity.
Grape pomace canbe consideredas a goodlowcost source toobtain
extract with antifungal activity against B. cinerea.
364 L. Mendoza et al. / Industrial Crops and Products 43 (2013) 360364
Acknowledgements
This research was supported by the FONDECYT Grant No.
1090723 and the Departamento de Investigaciones Cientcas y
Tecnolgicas (DICYT) of the Universidad de Santiago de Chile.
References
Andrade Pinto, J.M., Souza, E.A., Oliveira, D.F., 2010. Use of plant extracts in the
control of common bean anthracnose. Crop Prot. 29, 838842.
Arvanitoyannis, I.S., Ladas, D., Mavromatis, A., 2006. Potential uses and applications
of treated wine waste: a review. Int. J. Food Sci. Technol. 41, 475487.
Avis, T.J., Blanger, R.R., 2001. Specicity and mode of action of the antifungal fatty
acidcis-9-heptadecenoic acidproducedbyPseudozyma occulosa. Appl. Environ.
Microb. 67, 956960.
Corrales, M., Toep, S., Butz, P., Knorr, D., Tauscher, B., 2008. Extraction of antho-
cyanins from grape by-products assisted by ultrasonics, high hydrostatic
pressure or pulsed electric elds: a comparison. Innov. Food Sci. Emerg. Technol.
9, 8591.
Chen, Y., Dai, G., 2012. Antifungal activity of plant extracts against Colletotrichum
lagenarium, the causal agent of anthracnose in cucumber. J. Sci. Food Agric. 92,
19371943.
ChidambaraMurthy, K.N., Singh, R.P., Jayaprakasha, G.K., 2002. Antioxidant activities
of grape (Vitis vinifera) pomace extracts. J. Agric. Food Chem. 50, 59095914.
de Campos, L.M.A.S., Leimann, F.V., Pedrosa, R.C., Ferreira, S.R.S., 2008. Free radical
scavenging of grape pomace extracts from Cabernet sauvingnon (Vitis vinifera).
Bioresour. Technol. 99, 84138420.
Gonzlez-Params, A.M., Esteban-Ruano, S., Santos-Buelga, C., de Pascual-Teresa, S.,
Rivas-Gonzalo, J.C., 2003. Flavanol content and antioxidant activity in winery
byproducts. J. Agric. Food Chem. 52, 234238.
Guendez, R., Kallithraka, S., Makris, D.P., Kefalas, P., 2005. Determination of low
molecular weight polyphenolic constituents in grape (Vitis vinifera sp.) seed
extracts: correlation with antiradical activity. Food Chem. 89, 19.
Iacopini, P., Baldi, M., Storchi, P., Sebastiani, L., 2008. Catechin, epicatechin, quercetin,
rutin and resveratrol in red grape: content, in vitro antioxidant activity and
interactions. J. Food Compos. Anal. 21, 589598.
Jacometti, M.A., Wratten, S.D., Walter, M., 2010. Review: alternatives to synthetic
fungicides for Botrytis cinerea management in vineyards. Aust. J. Grape Wine
Res. 16, 154172.
Jung, H.J., Seu, Y.B., Lee, D.G., 2007. Candicidal action of resveratrol isolated from
grapes on human pathogenic yeast C. albicans. J. Microbiol. Biotechnol. 17,
13241329.
Katalinic, V., Mozina, S.S., Skroza, D., Generalic, I., Abramovic, H., Milos, M.,
Ljubenkov, I., Piskernik, S., Pezo, I., Terpinc, P., Boban, M., 2010. Polyphenolic
prole, antioxidant properties and antimicrobial activity of grape skin extracts
of 14 Vitis vinifera varieties grown in Dalmatia (Croatia). Food Chem. 119,
715723.
Mamoci, E., Cavoski, I., Simeone, V., Mondelli, D., Al-Bitar, L., Caboni, P., 2011. Chem-
ical composition and in vitro activity of plant extracts fromFerula communis and
Dittrichia viscosa against postharvest fungi. Molecules 16, 26092625.
Mendoza, L., Espinoza, P., Urzua, A., Vivanco, M., Cotoras, M., 2009. In vitro anti-
fungal activity of the diterpenoid 7-Hydroxy-8(17)-labden-15-oic acid and its
derivatives against Botrytis cinerea. Molecules 14, 19661979.
Meyer, A.S., Jepsen, S.M., Srensen, N.S., 1998. Enzymatic release of antioxidants
for human low-density lipoprotein fromgrape pomace. J. Agric. Food Chem. 46,
24392446.
Mu noz, G., Hinrichsen, P., Brygoo, Y., Giraud, T., 2002. Genetic characterisation of
Botrytis cinerea populations in Chile. Mycol. Res. 106, 594601.
Papadopoulou, C., Soulti, K., Rousis, I., 2005. Potential antimicrobial activity of
red and white wine phenolic extarcts against stranis of Staphylococcus aureus,
Escherichia coli and Candida albicans. Food Technol. Biotech. 43, 4146.
Rockenbach, I.I., Rodrigues, E., Gonzaga, L.V., Caliari, V., Genovese, M.I., Gonc alves,
A.Ed.S.S., Fett, R., 2011. Phenolic compounds content and antioxidant activity in
pomace from selected red grapes (Vitis vinifera L. and Vitis labrusca L.) widely
produced in Brazil. Food Chem. 127, 174179.
Ruggieri, L., Cadena, E., Martnez-Blanco, J., Gasol, C.M., Rieradevall, J., Gabarrell, X.,
Gea, T., Sort, X., Snchez, A., 2009. Recoveryof organic wastes intheSpanishwine
industry. Technical, economic and environmental analyses of the composting
process. J. Clean. Prod. 17, 830838.
Sagdic, O., Ozturk, I., Ozkan, G., Yetim, H., Ekici, L., Yilmaz, M.T., 2011. RP-HPLC-DAD
analysis of phenolic compounds in pomace extracts from ve grape cultivars,
Evaluation of their antioxidant, antiradical and antifungal activities in orange
and apple juices. Food Chem. 126, 17491758.
Schieber, A., Stintzing, F.C., Carle, R., 2001. By-products of plant food processing
as a source of functional compoundsrecent developments. Trends Food Sci.
Technol. 12, 401413.
Servicio Agrcola Ganadero. http://www.sag.cl.
Tegegne, G., Pretorius, J.C., Swart, W.J., 2008. Antifungal properties of Agapan-
thus africanus L., extracts against plant pathogens. Crop Prot. 27, 1052
1060.
Tehranifar, A., Selahvarzi, Y., Kharrazi, M., Bakhsh, V.J., 2011. High potential
of agro-industrial by-products of pomegranate (Punica granatum L.) as the
powerful antifungal and antioxidant substances. Ind. Crops Prod. 34, 1523
1527.
Xia, E.-Q., Deng, G.-F., Guo, Y.-J., Li, H.-B., 2010. Biological activities of polyphenols
fromgrapes. Int. J. Mol. Sci. 11, 622646.