Vous êtes sur la page 1sur 9

ARTICLE IN PRESS

LWT 41 (2008) 42–50


www.elsevier.com/locate/lwt

Antioxidant protection of white grape pomace on


restructured fish products during frozen storage
I. Sánchez-Alonso, A. Jiménez-Escrig, F. Saura-Calixto, A.J. Borderı́as
Instituto del Frı´o (CSIC), José Antonio Novais 10, E-28040 Madrid, Spain
Received 1 September 2006; received in revised form 26 January 2007; accepted 1 February 2007

Abstract

White grape antioxidant dietary fibre (WGDF) was obtained from white grape (Vitis vinifera, var. Airén) pomace from wine
production. The antioxidant capacity of WGDF was determined in minced fish muscle (MFM) during frozen storage. Concentrations of
0, 2, and 4% WGDF ((0-WGDF), (2-WGDF), and (4-WGDF) respectively) were added to MFM samples. Analyses were carried out
immediately after preparation of samples and over 6 months of storage at 20 1C. WGDF was characterized in terms of dietary fibre
(DF) (insoluble and soluble), total polyphenols and antioxidant capacity, and multifunctional antioxidant assays were done on all the
MFM samples. The addition of white grape DF considerably delayed lipid oxidation in minced horse mackerel muscle during the frozen
storage. Vacuum-packing the sample with 2% WGDF significantly enhanced the antioxidant properties of WGDF.
r 2007 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.

Keywords: Antioxidant dietary fibre; Grape polyphenols; Lipid oxidation; Conjugated hydroperoxides; Total reducing power; Free radical scavenging
activity

1. Introduction faster particularly in fatty and semi-fatty species like horse


mackerel (Trachurus trachurus), in whose muscle large
Dietary supplements and food fortification are a amounts of haemoglobin (a well-known activator of lipid
potential alternative route to the consumption of minor oxidation) and lipids coexist (Richards & Hultin, 2002).
plant components and dietary fibre (DF) that may have Among fruits, grapes constitute one of the major sources
health benefits (Schieber, Stintzing, & Carle, 2001). of phenolic compounds, and grape pomace is rich in
Bioactive components of foods exert their health benefits phenols (Yildrim, Akcay, Guvenc, Altindisli, & Sozmen,
partly through antioxidant activities. Considerable interest 2005). Even after contact with the fermenting wine, grape
has recently been focused on the addition of natural pomace contains high concentrations of both DF and
antioxidants to foods to replace synthetic antioxidants, due phenols (Valiente, Arrigoni, Esteban, & Amado, 1995)
to their potential to prolong the shelf life of food products with potential antioxidant activity (Saura-Calixto, 1998).
by inhibiting and delaying lipid oxidation. Seafoods White grape pomace has also been characterized as a
contain high concentrations of polyunsaturated fatty acids promising source of polyphenolics (Larrauri, Ruperez, &
(PUFA), eicosapentaenoic acid and docosahexaenoic acid Saura-Calixto, 1996; Larrauri, Sánchez-Moreno, & Saura-
(Ackman, 1999); because of this high unsaturated lipid Calixto, 1998; Kammerer, Claus, Carle, & Schieber, 2004).
content, fish products are very susceptible to loss of quality Most of the phenolic compounds in grape by-products are
through lipid oxidation; the development of off-flavours flavonoids (Escribano-Bailón, Guerra, Rivas-Gonzalo, &
and rancidity in these products is the main stumbling-block Santos-Buelga, 1995; Mazza, 1995), which have been
in their production and commercialization (Frankel, Satué- reported to have beneficial effects on lipid metabolism
Gracia, Meyer, & German, 2002). The onset of rancidity is (Teissedre, Frankel, Waterhouse, Peteg, & German, 1996).
In food systems, flavonoids can act as free radical
Corresponding author. Tel.: +34915492300; fax: +34915493627. scavengers and terminate the radical chain reactions that
E-mail address: isblsa@hotmail.com (I. Sánchez-Alonso). occur during the oxidation of triglycerides. There are many

0023-6438/$30.00 r 2007 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.lwt.2007.02.002
ARTICLE IN PRESS
I. Sánchez-Alonso et al. / LWT 41 (2008) 42–50 43

references in the literature to the composition and (Vitis vinifera, var. Airén) obtained from a Spanish winery
antioxidant properties of grape polyphenols (González- (Vinı́cola de Castilla, S.A., Manzanares, Ciudad Real,
Paramás, Esteban-Ruano, Santos-Buelga, Pascual-Teresa, Spain) was processed following a patented procedure
& Rivas-Gonzalo, 2004; Yilmaz & Toledo, 2004), and (Saura-Calixto & Larrauri, 1997), freeze-dried, milled to
studies have recently been published on their effectiveness a particle size less than 0.25 mm and stored at 20 1C. The
in delaying lipid oxidation in minced fatty fish muscle final product was named WGDF. The WGDF was mixed
during frozen storage (Pazos, Gallardo, Torres, & Medina, into the MFM. Concentrations of 0, 2, and 4% WGDF (0-
2005a) and ability to preserve endogenous antioxidant WGDF, 2-WGDF, and 4-WGDF, respectively) were
systems (like vitamin E) in fish muscle (Pazos, González, added to MFM based on final weight. They constitute
Gallardo, Torres, & Medina, 2005b). In addition, the the restructured fish products (RFP). We also prepared
authors (Sánchez-Alonso, Jiménez-Escrig, Saura-Calixto, another sample with 2% WGDF, which was vacuum-
& Borderı́as, 2007; Sánchez-Alonso & Borderı́as, 2007) packed in plastic bags (2-WGDF-vac). All the WGDF
have reported high levels of inhibition of lipid oxidation in samples were stored at 20 1C for 6 months. For sample
minced horse mackerel during frozen storage when red preparation, restructured fish products (RFPs), fish muscle
grape antioxidant DF was added. White grape pomace was mixed in a mixing machine model RM-20 (Mainca,
could therefore be a potential source of antioxidants; it Granollers, Spain), then WGDF was dispersed in cold
may also have technological applications as a natural food water (as per the formulation) and added to the MFM.
additive, and possibly nutritional benefits due to its Samples were mixed for 6 minutes. Moisture was adjusted
composition, in the design of healthy (functional) foods. in all samples to the same as in the original muscle
Lipid oxidation is a highly complex process involving (75.52%). The samples were placed on 21.5  15  3.5 cm3
numerous reactions which produce a variety of chemical aluminium trays and then frozen in a Benjamin SMC 4-65
and physical changes. These reactions appear to follow model horizontal plate freezer (Sabroe, Aarhus, Denmark),
known pathways, but they frequently occur simultaneously which cooled the thermal core to 20 1C. The samples were
and in competition. Numerous methods have been devised then sliced (15  1.5  3.5 cm3), packed in plastic bags
for their assessment, but there is none that permits (Cryovacs BB4L, oxygen permeability of 30 cm3/m2/24 h
simultaneous monitoring of all oxidative events or that is at 23 1C, 0% HR and 1 bar) and stored at 20 1C. The
equally applicable either at all stages of the process or to all RFPs containing WGDF were analysed at the beginning of
fats, all foods and all processing conditions. In specific the experiment, immediately after preparation of samples,
systems and under certain conditions, a single test can and then every month for 6 months. The pH values in
monitor only a few changes, so that for numerous purposes samples with WGDF added were the same as in the
it is best to use a combination of several tests. For proper original MFM (6.170.05) at the beginning and throughout
monitoring of oxidation, at least two methods have to be the study.
used to determine oxidation compounds at different stages
of deterioration (Frankel, 1998). 2.2. Chemicals
This paper characterizes a DF concentrate (called white
grape antioxidant dietary fibre (WGDF)), which was All the chemicals used were analytical grade and were
obtained from by-products of the white wine industry, in obtained from Panreac Quı́mica S.A. (Barcelona, Spain),
order to determine DF, total polyphenols and antioxidant Sigma-Aldrich Co. or Merck (Darmstadt, Germany).
properties. The main objective was to study the effect of Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-car-
addition of this DF concentrate to minced horse mackerel boxylic acid), a water-soluble analogue of vitamin E, was
(Trachurus trachurus) muscle on the prevention of lipid from Aldrich Co. (Madrid, Spain). 2,20 -azinobis(3-ethyl-
oxidation during six months’ frozen storage at 20 1C. A benzothiazolin-6-sulfonate) (ABTS) and 2,4,6-tri(2-pyri-
different methodology was assayed in order to assess the dyl)-s-triazine (TPTZ) were from Fluka Chemicals
multifunctional antioxidant activity of WGDF on minced (Madrid, Spain).
fish muscle (MFM) during frozen storage.
2.3. Chemical analysis of WGDF
2. Materials and methods
2.3.1. Dietary Fibre
2.1. Preparation of fish and samples The AOAC enzymatic-gravimetric method (Prosky, Asp,
Schweizer, Devries, & Furda, 1988) was modified in our
Minced muscle was prepared from ice-stored horse laboratory: dialysis against water was used instead of
mackerel (Trachurus trachurus) fillets. Individuals were ethanol precipitation of soluble dietary fibre (sDF) (Mañas
filleted without removing the skin by a local seafood & Saura-Calixto, 1995) for DF analysis. After enzymatic
company and transported to the pilot plant. The muscle hydrolysis of digestible components, insoluble DF (iDF)
was extracted using a Baader model 694 de-boning and sDF fractions were separated and chemically hydro-
machine (Lübeck, Germany) equipped with a drum with lysed. iDF fractions were hydrolysed with 12 M sulphuric
3 mm holes. White grape pomace (peels and seeds) acid (30 1C, 1 h) and then diluted to 1 M sulphuric acid
ARTICLE IN PRESS
44 I. Sánchez-Alonso et al. / LWT 41 (2008) 42–50

(100 1C, 90 min). The remaining residues were gravimetri- 2.4.2. Indirect methods
cally quantified as Klason lignin (KL) after drying at Extracts from RFP were obtained by sequential extrac-
105 1C to constant weight. SDF dialysates were hydrolysed tions with methanol/water (50:50, v/v) plus HCl to attain a
with 1 M sulphuric acid (100 1C, 90 min). Constituent final pH of 2.0, and acetone/water (70/30, v/v) at room
neutral sugars (NSs) and uronic acids (UAs) were temperature for 60 min in each case. The supernatants were
quantified in the hydrolysates. NSs were quantified by combined and the RFP extracts were used for total
Gas Chromatography (Jiménez-Escrig, Rincon, Pulido, & reduction power (TRP) by FRAP (ferric-reducing antiox-
Saura-Calixto, 2001). UAs were quantified spectrophoto- idant power) assay and free radical scavenging activity
metrically by the Scott method (Scott, 1979) using (FRSA) by ABTS assay.
galacturonic acid as standard. iDF was calculated as
(NS+UA+KL), and sDF as (NS+UA). 2.4.2.1. FRAP assay. The TRP by FRAP assay of
acetone–methanol–aqueous extracts was estimated accord-
2.3.2. Extraction of polyphenols ing to the procedure described by Benzie and Strain (1996),
One gram of ground freeze-dried WGDF sample was with some modifications introduced in our laboratory
placed in a test tube; 40 mL methanol/water (50:50) was (Sánchez-González, Jiménez-Escrig, & Saura-Calixto,
added, plus HCl to obtain a final pH 2.0. The solution was 2005). Briefly, 900 mL of FRAP reagent, freshly prepared
thoroughly shaken at room temperature for 1 h then and warmed at 37 1C, was mixed with 90 mL distilled water
centrifuged at 2500  g for 10 min, and the supernatant and either 30 mL of test sample or standard or appropriate
was recovered. Forty microlitres of acetone/water (70:30) reagent blank. The FRAP reagent contained 2.5 mL of a
was added to the residue, and shaking and centrifugation 10 mmol/L TPTZ solution in 40 mmol/L HCl, plus 2.5 ml
were repeated. The two extracts were mixed together. This of 20 mmol/L FeCl3  6H2O, plus 25 mL 0.3 mol/L acetate
procedure has been used by our group before and is buffer pH 3.6. Readings at the absorption maximum
described elsewhere (Jiménez-Escrig et al., 2001). Extrac- (595 nm) were taken every 15 s using a Beckman DU-640
tions were performed in triplicate and used to calculate the spectrophotometer (Beckman Instruments Inc., Fullerton,
total phenolics content and the antioxidant capacity. The CA, USA) equipped with a thermostatized auto-cell-
total phenolics in the extracts was determined spectro- holder. Temperature was maintained at 37 1C. The read-
photometrically according to the Folin-Ciocalteu proce- ings at 30 min were selected for calculation of TRP values.
dure (Singleton, Orthofer, & Lamuela-Raventós, 1999) Methanolic solutions of known Trolox concentrations were
using gallic acid as standard (concentration range 5–25 mg used for calibration, and the results were expressed as mmol
per 100 mL), and the results were expressed as gallic acid of Trolox eqivalent (TE)/g dry matter (dm) of RFP.
equivalents.
2.4.2.2. ABTS assay. The ABTS-FRSA was estimated
2.4. Methods for determining antioxidant activity following the procedure described elsewhere (Sánchez-
González et al., 2005). Briefly, ABTS radical cation
2.4.1. Direct methods based on the kinetics of lipid (ABTS+) was produced by reacting 7 mmol/L ABTS
peroxidation stock solution with 2.45 mmol/L potassium persulphate
Measurement of conjugated diene and triene hydroper- and allowing the mixture to stand in the dark at room
oxides. Lipids were extracted from mackerel muscle (Bligh temperature for 12–16 h before use. The ABTS+d solution
& Dyer, 1959) and the lipid content was determined (2 days stable) was diluted with 5 mM phosphate-buffered
gravimetrically in triplicate (Herbes & Allen, 1983). saline (pH 7.4) to an absorbance of 0.7070.02 at 658 nm.
Conjugated hydroperoxides were measured from fish oil After addition of 0.1 mL of sample or Trolox standard to
samples dissolved in hexane, and absorbance was measured 3.9 mL of diluted ABTS+ solution, absorbance readings
at 234 and 268 nm. Concentrations of hydroperoxides were were taken every 20 s using a Beckman DU-640 spectro-
calculated as mmol of hydroperoxides per kg of oil as photometer (Beckman Instruments Inc., Fullerton, CA,
described by Frankel, Huang, Kanner, and Bruce-German USA). The reaction was monitored for 6 min. Absorbance
(1994). versus time was plotted and the area below the curve
The thiobarbituric acid index (TBA-i) was determined (0–6 min) was calculated. Methanolic solutions of known
according to Vynke (1970) on a 5% trichloracetic acid Trolox were used for calibration, and the results were
extract of the restructured fish muscle. Results were expressed as mmol of TE/g dm of RFP.
expressed as mg malondialdehyde per kg of dry sample. The spectrophotometer used was a UV/VIS Perkin-
Elmer Lambda 15.
2.4.1.1. Antioxidant effectiveness. Antioxidant effective-
ness was calculated as per cent inhibition of oxidation (%I) 2.5. Measurement of colour
as described by Frankel (1998): %I ¼ (cs/c)*100, where c
is the increment of 0-WGDF in the experiment and s the Colour measurements consisted of determining redness
increment of sample with added WGDF at the same time. (a*) and yellowness (b*) using a CIELab scale (Young &
High levels of %I indicate greater antioxidant effectiveness. Whittle, 1985; Park, 1995) in raw samples. Measurements
ARTICLE IN PRESS
I. Sánchez-Alonso et al. / LWT 41 (2008) 42–50 45

were done in a HunterLab model D25-9 colorimeter (D45/ (Larrauri, Ruperez, & Saura-Calixto, 1996; Sánchez-
21) (Hunter Associates Laboratory Inc., Reston, VA, Alonso et al., 2007). In the preparation, of white wines,
USA), and measurements were standardized with respect the pomace is removed before fermentation, thus leaving
to the white calibration plate. little time for the solubilization of extractable polyphenols
(Larrauri et al., 1996); however, the phenolic composition
2.6. Statistical analysis of wine pomace depends on cultivar differences, regional
variations in climate and winemaking techniques (duration
One- and two-way ANOVA was performed using of pomace contact, cell disruption during crushing and
Statgraphics 2.1 (STSC Inc., Rockville, MD). The differ- pressing) (Sánchez-Moreno, Cao, Ou, & Prior, 2003). The
ence in means was analysed using a Tukey HSD test high polyphenol levels and the antioxidant capacity of this
(po0.05). WGDF suggest that it may have a potential use as a
natural antioxidant in food systems.
3. Results and discussion During lipid oxidation, antioxidants act in various ways,
binding metal ions, scavenging radicals and decomposing
Table 1 shows the composition of the WGDF. The chief peroxides. In food-related systems, antioxidant activity
characteristics of this natural product are that it is rich in means chain-breaking inhibition of lipid peroxidation. The
both DF and total polyphenolic compounds. The WGDF most frequently measured products are conjugated hydro-
had more than 76% of total DF (dm basis). It is worth peroxides (dienes and trienes) for primary oxidation and
noting that the sDF content of this WGDF was greater in volatile compounds (TBA-i) for secondary. By continuous
relative terms than the iDF content (sDF/iDF ratio of monitoring of these compounds during frozen storage, the
1:2.3); this ratio is higher than others reported by process of lipid damage can be observed in close detail.
Figuerola, Hurtado, Estévez, Chiffelle, and Asenjo (2005) Less conjugated hydroperoxides, dienes and trienes were
for DF concentrates from apple pomace and citrus peel, formed from PUFAs in samples with added WGDF (2%
but is comparable to others reported for orange DF and 4%) than in the control sample over a period of 6
concentrates (Grigelmo-Miguel & Martı́n-Belloso, 1999) months at 20 1C. Initial values, immediately after
and red grape antioxidant DF (Sánchez-Alonso, Jiménez- preparation of samples, were slightly lower in samples
Escrig, Saura-Calixto, & Borderı́as, 2007). Most consumers with WGDF added than in the control. During the first
need to increase their DF intake. The amount added to the month of storage, the control sample showed a significant
mince could contribute to achieve the recommended intake increase (po0.05) in the development of conjugated dienes
of DF, which is 14 g per 1000 calories (Dietary Guidelines (Fig. 1); sample 2-WGDF showed a smaller increase, and
for Americans, 2005). Apart from that, some suggestions of samples 2-WGDF-vac and 4-WGDF did not change
the health benefits of DF in the intestinal track could been significantly. The highest values occurred in sample with-
found (Jiménez-Escrig & Sánchez-Muniz, 2000; Goñi, out WGDF in the last period of storage (between 120
Jiménez-Escrig, Gudiel, & Saura-Calixto, 2005). and 180 days). Samples with 2% (2-WGDF) and 4%
WGDF contains high amounts of extractable polyphe- (4-WGDF) added WGDF showed no significant changes
nols associated with the DF matrix (Table 1) and had until the last months of storage, when they started to
relatively high antioxidant activity as measured by FRAP increase; vacuum-packed sample with 2% WGDF
TRP and ABTS (radical scavenging capacity) methods. (2-WGDF-vac) showed no significant increases during
These values fall within the usual ranges described for frozen storage and presented the lowest values for
antioxidant DF obtained from white grape pomace development of conjugated dienes throughout the study
originating from wine production (Saura-Calixto, 1998).
This WGDF had a higher extractable polyphenol value
45
than others reported for red grape antioxidant DF
40
mmol hydrp/kg oil

35
Table 1
Proximate composition of WGDF (grams per 100 g of dm)a 30

Protein 7.3170.2 25
Fat 5.5070.1 20
Extractable polyphenols 7.8570.3
Insoluble dietary fibre 53.3671.3 15
Soluble dietary fibre 23.0170.1 0 30 60 90 120 150 180
Ash 3.5070.2 Days
FRAP 466743 0-WGDF 2-WGDF 2-WGDF-vac 4-WGDF
ABTS 284724
Fig. 1. Formation of conjugated dienes in samples of minced fish muscle
Ferric reducing antioxidant power (FRAP) and ABTS values expressed as with added white grape dietary fibre during frozen storage at 20 1C. 0-
mmol of Trolox equivalents per gram of WGDF (dm). dm: dry matter. WGDF: control without WGDF; 2-WGDF, 4-WGDF: 2% and 4% of
a
Values are mean7standard deviation of three replicate determinations. WGDF added; 2-WGDF-vac: 2% WGDF added and vacuum-packed.
ARTICLE IN PRESS
46 I. Sánchez-Alonso et al. / LWT 41 (2008) 42–50

(Fig. 1). Table 2 shows the percentage inhibition of These kinds of products are chain initiators, and their
development of conjugated dienes attained by WGDF presence in a lipid system accelerates the process and
concentrate in MFM at 60 and 120 days of storage. At reduces the time taken by inhibitor consumption (Yanish-
both times the percentage inhibition of formation of lieva, 1973). Antioxidants should therefore be added to
conjugated dienes was significant in samples containing foodstuffs as early as possible to achieve maximum
WGDF. The addition of 4% WGDF to MFM inhibited protection against oxidation.
the development of conjugated dienes more significantly Secondary lipid oxidation products as reflected by the
than when 2% WGDF was added. Between samples with TBA-i presented no significant differences between samples
2% added WGDF and different packing (2-WGDF and 2- with added WGDF and without it at the beginning of
WGDF-vac), there was a significant difference in the effect storage (Fig. 2). Levels attained in control sample
of the packing on the effectiveness of the WGDF in (0-WGDF) kept at 20 1C were higher and increased
inhibiting lipid oxidation; the percentage inhibition of continuously with storage time, reaching a maximum value
diene development was higher in vacuum packed sample, at 150 days of storage. Samples with WGDF presented
and it was significantly higher when compared to sample lower values throughout the storage period. Samples with
with 4% WGDF added. 4% WGDF (4-WGDF) and vacuum-packed samples with
In the case of conjugated trienes, the evolution of 2% (2-WGDF-vac) did not increase significantly during
samples with WGDF added did not reach maximum storage at 20 1C. Values in sample with 2% WGDF
during frozen storage; only the control sample showed a (2-WGDF) increased significantly in the last stage of
significant increase at 180 days of storage. In this case the storage (between 150 and 180 days). The rate of inhibition
percentage of inhibition (180 days) did not differ signifi- at 90 days of storage (Table 2) was significantly different
cantly between samples with different amounts of WGDF for the two levels of DF concentrate: 66.8% in sample with
(26.0% in samples with 2% and 23.4% in samples with 4% 2% and 76.8% in sample with 4% WGDF; sample 2-
added WGDF), but vacuum-packed sample with 2% WGDF-vac was not significantly different from those
WGDF showed a significantly higher rate of inhibition (74.6%). At 150 days of storage the rates of inhibition of
(45.0%) of conjugated triene formation. Values recorded in lipid oxidation remained high, with no significant differ-
the course of the experiment in the development of ence between sample with 4% WGDF and vacuum-packed
conjugated hydroperoxides were in the same range as sample with 2% (81.9% and 85.2% of inhibition). The
those recorded in a previous study when red grape fibre antioxidant capacity of white grape fibre was effective up
was added to MFM (Sánchez-Alonso & Borderı́as, 2007), until 180 days of storage (experimental values in samples
but the rate of inhibition of lipid oxidation when WGDF with WGDF were lower than in the control in the last
was added is significantly higher than with red grape month of analyses). In a previous study (Sánchez-Alonso &
antioxidant DF (Sánchez-Alonso et al., 2007). This could Borderı́as, 2007), the values of i-TBA in frozen horse
be related to the fact that the red grape antioxidant DF mackerel with added red grape fibre at 180 days of storage
contained less extractable polyphenols (around 5%). In at 20 1C were similar to the control sample, but in this
addition, the initial concentration of primary oxidation study white grape fibre was effective at this time (23.0%
products (hydroperoxides) from lipids in food systems can i-TBA inhibition in samples with 2% WGDF, 87.1% in
substantially reduce the efficiency of the antioxidant added samples with 2% WGDF-vac and 68.9% in samples with
to the system that is being stabilized (Yanishlieva, 1973). 4% WGDF). The antioxidant effect for the two types of
packing in samples with same amount of added fibre was
Table 2
Per cent inhibitiona of formation of conjugated dienes and trienes, and
3.5
thiobarbituric index (TBA-i) in minced fish samples (means7DS)b with
white grape fibre added 3.0
mg MDA/Kg sample

Sample Conjugated dienes Conjugated TBA-i 2.5


trienes 2.0
Day 60 Day 120 Day 180 Day 90 Day 150
1.5
0-WGDF 0.073.3a 0.071.0a 0.072.5a 0.072.9a 0.072.1a
2-WGDF 25.470.1b 13.671.5b 26.073.3b 66.873.7b 64.771.2b
1.0
2-WGDF- 50.871.9c 88.971.8c 45.072.5c 74.673.4bc 85.272.8c 0.5
vac
4-WGDF 43.770.4 d
69.371.7 d
23.473.9b
76.872.9 c
81.970.5 c 0.0
0 30 60 90 120 150 180
a Days
Percentage of inhibition ¼ (cs/c)*100, where c is the oxidation
product formed in control sample and s the oxidation product formed 0-WGDF 2-WGDF 2-WGDF-vac 4-WGDF
in sample with added white grape dietary fibre (WGDF) at the same time.
a
0-WGDF: control without WGDF; 2-WGDF, 4-WGDF: 2% and 4% of Fig. 2. Formation of aldehydes in samples of MFM with added white
WGDF added; 2-WGDF-vac: 2% WGDF added vacuum-packed. grape dietary fibre during frozen storage at 20 1C. 0-WGDF: control
b
Different letters in the same column indicate significant differences without WGDF; 2-WGDF, 4-WGDF: 2% and 4% of WGDF added; 2-
(*po0.05). WGDF-vac: 2% WGDF added and vacuum-packed.
ARTICLE IN PRESS
I. Sánchez-Alonso et al. / LWT 41 (2008) 42–50 47

significantly different after 90 days of frozen storage; the 70


effect in samples 4-WGDF and 2-WGDF-vac was similar
60
up until the last month of storage.

micro Mol TE/g d.m.


The effectiveness of antioxidants cannot be evaluated 50
using only one assay protocol. The antioxidant activity of
vegetables includes several multifunctional mechanisms 40
(Prior, Wu, & Schaich, 2005). Thus, to evaluate this 30
potential antioxidant activity it is necessary to assess
several antioxidant assays that include different antiox- 20
idant mechanisms. A useful way of viewing the interactions
10
among various antioxidants is to take into account
oxidation-reduction potentials. The FRAP assay is based 0
0 30 60 90 120 150 180
on a single electron transfer reaction between an oxidant
Days
and the antioxidants, i.e., WGDF antioxidants are oxidized
0-WGDF 2-WGDF 2-WGDF-vac 4-WGDF
by the oxidant Fe (III). As a result, a single electron is
transferred from the antioxidant molecule (phenolic Fig. 4. Radical scavenging capacity (ABTS method) of minced fish muscle
hydroxyl groups) to the oxidant. The standard redox with added white grape dietary fibre during frozen storage at 20 1C. 0-
potential of Fe(III)/Fe(II) is 0.77 V; any compound with WGDF: control without WGDF; 2-WGDF, 4-WGDF: 2% and 4% of
WGDF added; 2-WGDF-vac: 2% WGDF added and vacuum-packed.
lower redox potential can theoretically reduce Fe(III) to
Fe(II) (Ou, Huang, Hampsch-Woodill, Flanagan, &
Deemer, 2002). Another mechanism commonly used to difference favourable to the samples with added WGDF
measure the prevention of oxidation in foods is FRSA was maintained in both assays. This indicates the effective
(FRSA of free radicals involved in lipid oxidation). protection that WGDF exerts on the MFM during frozen
Therefore, to evaluate the protection of MFM by WGDF, storage. However, it should be stressed that this depletion
two different multifunctional systems were chosen: FRSA was very evident during the first 2 months in the case of the
and TRP, measured by ABTS and FRAP assays respec- samples with added WGDF (ranging from 35% to 47%),
tively. whereas in the case of the vacuum packed sample it was
The results obtained by TRP (Fig. 3), and FRSA (Fig. 4) around 21%. At the end of the storage period this feature
procedures are discussed below. Prior to storage, signifi- was maintained: the 2-WGDF and the 4-WGDF samples
cant differences in the TRP and FRSA values were found presented twice as much depletion of the mentioned
between the 0-WGDF sample and the samples with antioxidant values than the 2-WGDF-vac sample. At the
WGDF added. In the case of TRP these differences were end of storage at 20 1C, there was a significant difference
166% (2-WGDF), 161% (2-WGDF-vac), and 267% in averaged contents between samples containing WGDF
(4-WGDF) higher than the 0-WGDF sample. The same and sample without. In both assays, 2-WGDF-vac and 4-
pattern was observed in the case of FRSA: 170% WGDF samples gave the best antioxidant values among
(2-WGDF), 144% (2-WGDF-vac), and 388% (4-WGDF). tested samples.
During the whole 6-month storage period a significant Previous papers (Larrauri et al., 1996; Larrauri, Sán-
chez-Moreno, & Saura-Calixto, 1998; Lu & Foo, 1999;
Kammerer et al., 2004) have shown that WGDFs have a
80 rich qualitative and quantitative polyphenolic profile.
70 These data are often correlated with the antioxidant
activity of WGDF extracts. Thus, the antioxidant protec-
micro Mol TE/g d.m.

60
tion of the MFM by WGDF demonstrated in our work
50 may be a result of its polyphenol content. The grape
40 polyphenols in white grape fibre have considerable anti-
30
oxidant potential and can be divided into two groups: on
the one hand nonextractable poplyphenols (polymeric
20
proanthocyanidins and high molecular weight hydrolysable
10 tannins), and on the other hand extractable polyphenols
0 (mainly flavonoid derivatives: flavonols (quercetin) and
0 30 60 90 120 150 180 flavan-3-ols (catechin and epicatechin, monomers and
Days compounds of different polymeric degrees, and glycosilated
0-WGDF 2-WGDF 2-WGDF-vac 4-WGDF forms)), and phenolic acids (gallic acid), both of which
show some promise in the fields of nutrition and health.
Fig. 3. Profile of antioxidant activity by FRAP assay in RFPs during
storage at 20 1C for different amounts of added white grape dietary fibre.
The ability of flavonoids, one of the most abundant groups
0-WGDF: control without WGDF; 2-WGDF, 4-WGDF: 2% and 4% of of polyphenols in white grape fibre, to inhibit lipid
WGDF added; 2-WGDF-vac: 2% WGDF added and vacuum-packed. oxidation is well documented, both for natural lipid
ARTICLE IN PRESS
48 I. Sánchez-Alonso et al. / LWT 41 (2008) 42–50

products and for model lipids (Mora, Paya, Rios, & Table 3
Alcaraz, 1990; Terao, Piskula, & Yao, 1994; Bors, Heller, Surface colour measurements of samplesa
Michael, & Stettmaier, 1996). Flavonoids act as antiox- Sample a* b*
idants by donating electrons (scavenging radicals), and
stopping radical chains. Thus, lipid oxidation can be Day 0 Day 180 Day 0 Day 180
prevented at the outset by free radical scavengers and
0-WGDF 7.970.6 3.770.5 10.270.6 13.270.3
singlet oxygen quenchers, and the propagation chain 2-WGDF 7.170.1 6.870.1 12.970.3 13.870.3
reaction can be broken by peroxy-radical scavengers 2-WGDF-vac 7.170.1 6.770.3 13.170.3 13.970.2
(Torel, Cillard & Cillard, 1986). Besides free radical 4-WGDF 7.070.1 6.870.2 13.970.2 14.270.3
scavenging activities, grape flavonoids may also delay lipid a
For sample description see Table 2 footnote.
oxidation via inactivation of metallic promotion of lipid
oxidation by chelating metals (Pazos, Alonso, Fernández-
Bolaños, Torres, & Medina, 2006). This is especially (a* from 7.9 to 3.75 at 180 days). This decrease of redness
important in fish minces where blood is homogenized with in raw control sample may be related to the development of
muscle because haemoglobin is one of the strongest lipid oxidation in this sample. Yellowness values (b*) did
endogenous catalysts of fish muscle lipid oxidation not change significantly in sample with added fibre but did
(Richards, Modra, & Li, 2002). pH values are crucial in increase significantly in sample without fibre during frozen
haemoglobin-catalized lipid oxidation and is particularly storage; this could be related to losses of redness, since
pro-oxidative in nature since it falls below pH 6.5. The oxygenated haeme proteins confer a bright colour while
native lower post-mortem pH of pelagic fish, such as oxidized pigments are brown (met-haeme proteins).
mackerel (when compared to other fish species) may play a
role in further deoxygenation of their anodic haemoglobin
4. Conclusions
(Richards and Hultin, 2003). In our study, as we have
mentioned above, the pH values in samples were 6.170.05.
The WGDF by-product contained high concentrations
Richards and Hultin (2000) showed a lag phase decrease in
of DF and associated polyphenols with significant anti-
the development of rancidity and thiobarbituric acid
oxidant capacity as determined by the ABTS and FRAP
reactive substances (po0.01) coupled with autoxidation
methods. These properties suggest a wide range of possible
of haeme protein when fish muscle pH was reduced from
applications for WGDF as a food ingredient. The
7.2 to 6.0.
evaluation of antioxidant capacity of WGDF added to
Pazos et al. (2005a) reported that an optimal combina-
frozen minced muscle over 6 months of storage was similar
tion of degree of procyanidin polymerization and percen-
for all the methods followed. The addition of WGDF
tage of galloylation, as in white grape fibre, may help
considerably inhibited the development of oxidation in
account for the high antioxidant efficacy of grape
minced horse mackerel muscle during storage at 20 1C.
polyphenols in frozen fish muscle. It has been suggested
These results indicate that WGDF could be used as a
(Pazos et al., 2005b) that the preservation of endogenous
natural ingredient to prevent oxidation in minced fish
antioxidants by grape pomace polyphenols could be a
during frozen storage and could be used as functional
useful measure for increasing the oxidative stability of fatty
ingredient in the design of healthy (functional) foods.
fish. These results are consistent with our own findings
regarding high white grape fibre antioxidant capacity
during frozen storage (20 1C) in a matrix made of minced Acknowledgements
muscle. We found intense antiradical activity in the white
grape fibre concentrate (Table 2), and the results show that This work has been supported by the Spanish Ministerio
WGDF possesses notable antioxidant properties and is de Educación y Ciencia under Projects AGL2002-04104-
able to significantly inhibit the development of rancidity in C04-03 and AGL2002-04104-C04-01, and by the EU under
frozen fish muscle stored at 20 1C. Integrated Project SEAFOODplus (Ref. FP6/506359). The
Surface colour was evaluated to detect whether changes authors wish to thank the Spanish Ministerio de Educación
in the oxidation development could be related to changes in y Ciencia for Ms. Sánchez-Alonso’s predoctoral fellowship.
the surface colour of samples. Decreases in the value a*
provide an indirect means of monitoring Hb-mediated lipid References
oxidation in fish muscle—for instance in species like horse
mackerel (Trachurus trachurus) in whose muscle large Ackman, R. G. (1999). In R. G. Ackman (Ed.), Marine biogenic lipids, fats
amounts of haemoglobin, a well-known activator of lipid and oils. Boca Raton, FL: CRC Press.
oxidation, coexist with lipids—during lipid oxidation Benzie, I. F. F., & Strain, J. J. (1996). The ferric reducing ability of plasma
(Undeland, Ekstrand, & Lingnert, 1998). During frozen (FRAP) as a measure of ‘‘antioxidant power’’: The FRAP assay.
Analytical Biochemistry, 239, 70–76.
storage, there was no change in a* (shift towards red) Bligh, E. G., & Dyer, W. J. (1959). A rapid method of total lipid extraction
values of raw samples with WGDF (Table 3), while the and purification. Canadian Journal of Biochemistry and Physiology, 37,
values in sample without fibre decreased significantly 911–917.
ARTICLE IN PRESS
I. Sánchez-Alonso et al. / LWT 41 (2008) 42–50 49

Bors, W., Heller, W., Michael, C., & Stettmaier, K. (1996). Flavonoids employing oxygen radical absorbance capacity (ORAC) and ferric
and polyphenols: chemistry and biology. In E. Cadenas, & L. Parker reducing antioxidant power (FRAP) assays: a comparative study.
(Eds.), Handbook of Antioxidants (pp. 409–466). New York: Marcel Journal of Agricultural and Food Chemistry, 50, 3122–3128.
Dekker. Park, J. W. (1995). Surimi gel colors as affected by moisture content and
Dietary Guidelines for Americans (2005) in http://www.usda.gov. physical conditions. Journal of Food Science, 60, 15–18.
(Accessed December 2006). Pazos, M., Alonso, A., Fernández-Bolaños, J., Torres, J. L., & Medina, I.
Escribano-Bailón, M. T., Guerra, M. T., Rivas-Gonzalo, J. C., & Santos- (2006). Physicochemical properties of natural phenolics from grapes
Buelga, C. (1995). Proanthocyanidins in skins from different grape and olive oil byproducts and their antioxidant activity in frozen horse
varieties. Zeitschrift für Lebensmittel Untersuchung und Forschung, 200, mackerel fillets. Journal of Agricultural and Food Chemistry, 54,
201–224. 366–373.
Frankel, E. N. (1998). Foods. In Lipid Oxidation (p. 202). Great Britain: Pazos, M., Gallardo, J. M., Torres, J. L., & Medina, I. (2005a). Activity of
The Oily Press (Chapter 10). grape polyphenol as inhibitors of the oxidation of fish lipids and frozen
Frankel, E. N., Huang, S. W., Kanner, J., & Bruce-German, J. B. (1994). fish muscle. Food Chemistry, 92, 547–557.
Interfacial phenomena in the evaluation of antioxidants: bulk oils Pazos, M., González, M. J., Gallardo, J. M., Torres, J. L., & Medina, I.
versus emulsions. Journal of Agricultural and Food Chemistry, 42, (2005b). Preservation of the endogenous antioxidant system of fish
1054–1059. muscle by grape polyphenols during frozen storage. European Food
Frankel, E. N., Satué-Gracia, T., Meyer, A. S., & German, J. B. (2002). Research and Technology, 220, 514–519.
Oxidative stability of fish and algae oils containing long-chain Prior, R. L., Wu, X., & Schaich, K. (2005). Standardized methods for the
polyunsaturated fatty acids in bulk and in oil-in-water emulsions. determination of antioxidant capacity and phenolics in foods and
Journal of Agricultural and Food Chemistry, 50, 2094–2099. dietary supplements. Journal of Agricultural and Food Chemistry, 53,
Figuerola, F., Hurtado, M. L., Estévez, A. M., Chiffelle, I., & Asenjo, F. 4290–4302.
(2005). Fibre concentrates from apple pomace and citrus peel as Prosky, L., Asp, N. G., Schweizer, T. F., Devries, J. W., & Furda, I. J.
potential fibre sources for food enrichment. Food Chemistry, 91, (1988). Determination of insoluble, soluble and total dietary fiber in
395–401. foods and food products: interlaboratory study. Journal Association of
González-Paramás, A. M., Esteban-Ruano, S., Santos-Buelga, C., Official Analytical Chemists, 71, 1017–1023.
Pascual-Teresa, S., & Rivas-Gonzalo, J. C. (2004). Flavanol content Richards, M. P., & Hultin, H. O. (2000). Effects of pH on lipid oxidation
and antioxidant activity in winery byproducts. Journal of Agricultural using trout hemolysate as a catalyst: A possible role for deoxyhe-
and Food Chemistry, 52, 234–238. moglobin. Journal of Agricultural and Food Chemistry, 48, 3141–3147.
Goñi, I., Jiménez-Escrig, A., Gudiel, M., & Saura-Calixto, F. (2005). Richards, M. P., & Hultin, H. O. (2002). Contributions of blood and
Artichoke (Cynara scolymus L) modifies bacterial enzymatic activities blood components to lipid oxidation in fish muscle. Journal of
and antioxidant status in rat cecum. Nutrition Research, 25, 607–615. Agriculture and Food Chemistry, 50, 555–564.
Grigelmo-Miguel, N., & Martı́n-Belloso, O. (1999). Characterization of Richards, M. P., & Hultin, H. O. (2003). Effects of added hemolysate from
dietary fiber from orange juice extraction. Food Research International, mackerel, herring, and rainbow trout on lipid oxidation of washed cod
31, 355–361. muscle. Fisheries Science, 69, 1298–1300.
Herbes, S., & Allen, C. (1983). Lipid quantification of freshwater Richards, M. P., Modra, A. M., & Li, R. (2002). Role of deoxyhemoglo-
invertebrates: meted modification for microquantification. Canadian bin in lipid oxidation of washed cod muscle mediated by trout, poultry
Journal Fisheries and Aquatic Sciences, 14, 1315–1317. and beef hemoglobins. Meat Science, 62, 157–163.
Jiménez-Escrig, A., Rincon, M., Pulido, R., & Saura-Calixto, F. (2001). Sánchez-Alonso, I., & Borderı́as A.J. (2007). Technological effect of red
Guava fruit (Psidium guajava L.) as a new source of antioxidant grape antioxidant dietary fibre added to minced fish muscle.
dietary fiber. Journal of Agricultural and Food Chemistry, 49, International Journal of Food Science and Technology, in press,
5489–5493. doi:10.1111/j.1365-2621.2007.01554x.
Jiménez-Escrig, A., & Sánchez-Muniz, F. J. (2000). Dietary fibre from Sánchez-Alonso, I., Jiménez-Escrig, A., Saura-Calixto, F., & Borderı́as, A.
edible seaweeds: chemical structure, physicochemical properties and J. (2007). Effect of grape antioxidant dietary fibre on the prevention of
effects on cholesterol metabolism. Nutrition Research, 20, 585–598. lipid oxidation in minced fish: evaluation by different methodologies.
Kammerer, D., Claus, A., Carle, R., & Schieber, A. (2004). Polyphenol Food Chemistry, 101, 372–378.
screening of pomace from red and white grape varieties (Vitis vinifera Sánchez-González, I., Jiménez-Escrig, A., & Saura-Calixto, F. (2005). In
L.) by HPLC-DAD-MS/MS. Journal of Agricultural and Food vitro antioxidant activity of coffees brewed using different procedures
Chemistry, 52, 4360–4367. (Italian, espresso and filter). Food Chemistry, 90, 133–139.
Larrauri, J. A., Ruperez, P., & Saura-Calixto, F. (1996). Antioxidant Sánchez-Moreno, C., Cao, G., Ou, B., & Prior, R. L. (2003). Anthocyanin
activity of wine pomace. American Journal of Enology and Viticulture, and proanthocyanidin content in selected white and red wines. oxygen
47, 369–372. radical absorbance capacity comparison with nontraditional wines
Larrauri, J. A., Sánchez-Moreno, C., & Saura-Calixto, F. (1998). Effect of obtained from Highbush Blueberry. Journal of Agricultural and Food
temperature on the free radical scavenging capacity of extracts from Chemistry, 51, 4889–4896.
red and white grape pomace peels. Journal of Agricultural and Food Saura-Calixto, F. (1998). Antioxidant dietary fiber product: A new
Chemistry, 46, 2694–2697. concept and a potential food ingredient. Journal of Agricultural and
Lu, Y., & Foo, L. Y. (1999). The polyphenol constituents of grape Food Chemistry, 48, 4303–4306.
pomace. Food Chemistry, 65, 1–8. Saura-Calixto, F., & Larrauri, J., inventors; CSIC. (1997). Concentrado
Mañas, E., & Saura-Calixto, F. (1995). Dietary fibre analysis: methodo- de fibra dietética antioxidante natural de uva y su procedimiento de
logical error sources. European Journal of Clinical Nutrition, 49, obtención. Patente Española 9702397.
S158–S162. Schieber, A., Stintzing, F. C., & Carle, R. (2001). By-products of plant
Mazza, G. (1995). Anthocyanins in grapes and grapes products. Critical food processing as a source of functional compounds—recent
Reviews in Food Science and Nutrition, 35, 341–371. developments. Trends in Food Science and Technology, 12, 401–413.
Mora, A., Paya, M., Rios, J. L., & Alcaraz, M. J. (1990). Structure–- Scott, R. W. (1979). Colorimetric determination of hexauronic acids in
activity relationship of polymethoxyflavonoids and other flavonoids as plant materials. Analytical Chemistry, 51, 936–941.
inhibitors of non-enzymic lipid peroxidation. Biochemical Pharmacol- Singleton, V. L., Orthofer, R., & Lamuela-Raventós, R. M. (1999).
ogy, 40, 393–397. Analysis of total phenols and other oxidation substrates and
Ou, B., Huang, D., Hampsch-Woodill, M., Flanagan, J. A., & Deemer, E. antioxidants by means of Folin-Ciocalteau reagent. Methods in
K. (2002). Analysis of antioxidant activities of common vegetables Enzymology, 299, 152–178.
ARTICLE IN PRESS
50 I. Sánchez-Alonso et al. / LWT 41 (2008) 42–50

Teissedre, P. L., Frankel, E. N., Waterhouse, A. L., Peteg, H., & German, Vynke, W. (1970). Direct determination of the thiobarbituric acid value in
B. (1996). Inhibition of in vitro human LDL oxidation by phenolic trichloracetic acid extracts of fish as measure of oxidative rancidity.
antioxidants from grapes and wines. Journal of the Science of Food and Fette Seifen Anstrichmittel, 72, 1084–1087.
Agriculture, 70, 55–61. Yanishlieva, N. (1973). Uber einige Eigentümlichkeiten in der Kinetik zu
Terao, J., Piskula, M., & Yao, Q. (1994). Protective effect of epicatechin, Beginn der Autoxydation von Estern ungesättigter Fettsäuren, 6 Mitt
epicatechingallate, and quercetin on lipid peroxidation in phospholi- Hydroperoxide und Inhibierung. Nahrung, 17, 323–334.
pids bilayers. Archives of Biochemistry and Biophysics, 308, 278–284. Yildrim, H. K., Akcay, Y. D., Guvenc, U., Altindisli, A., & Sozmen, E. Y.
Torel, J., Cillard, J., & Cillard, P. (1986). Antioxidant activity of (2005). Antioxidant activities of organic grape, pomace, juice, must,
flavonoids and reactivity with peroxy radical. Phytochemistry, 25, wine and their correlation with phenolic content. International Journal
383–385. of Food Science and Technology, 40, 133–142.
Undeland, I., Ekstrand, B., & Lingnert, H. (1998). Lipid oxidation in Yilmaz, Y., & Toledo, R. T. (2004). Major flavonoids in grape seeds and
minced herring (Clupea harengus) during frozen storage: influence of skins: Antioxidant capacity of catechin, epicatechin and gallic acid.
washing and pre-cooking. Journal of Agricultural and Food Chemistry, Journal of Agricultural and Food Chemistry, 52, 255–260.
46, 2319–2328. Young, K. W., & Whittle, J. (1985). Colour measurement of fish minces
Valiente, C., Arrigoni, C., Esteban, R. M., & Amado, R. (1995). Grape using Hunter L, a, b values. Journal of the Science of Food and
pomace as a potential food fiber. Journal of Food Science, 60, 818–820. Agriculture, 36, 383–392.

Vous aimerez peut-être aussi