LWT - Food Science and Technology 42 (2009) 312–318

Contents lists available at ScienceDirect

LWT - Food Science and Technology

journal homepage: www.elsevier.com/locate/lwt

Grape wine lees improves the rheological and adds

antioxidant properties to ice cream
Jean-Yu Hwang a, Yung-Shin Shyu b, Cheng-Kuang Hsu c, *
a
Department of Food Science and Technology, Chung-Hwa University of Medical Technology, 89, Wenhwa 1st Street, Rende Shiang, Tainan 717, Taiwan, ROC
b
Department of Baking Technology and Management, National Kaohsiung Hospitality College, 1, Sung-Ho Road, Hsiao-Kang, Kaohsiung 812, Taiwan, ROC
c
Department of Health and Nutrition Biotechnology, Asia University, 500, Lioufeng Road, Wufeng Shiang, Taichung 413, Taiwan, ROC

a r t i c l e i n f o a b s t r a c t

Article history: The effect of adding grape wine lees (GWL) (50, 100 and 150 g kg1, wet weight basis) on the rheological
Received 18 September 2007 and antioxidant properties of ice cream was evaluated to determine the potential of using GWL as
Received in revised form 8 March 2008 a value-added ingredient in ice cream. Black queen grape was selected because it was the only species for
Accepted 11 March 2008
red winemaking in Taiwan. The addition of GWL decreased the specific gravity, pH, melting rate, firm-
ness, lightness and the amount of freezable water in ice cream, but increased the viscosity, yellowness
Keywords: and fat destabilization in a dose dependent circumstance. However, the ice cream with high GWL
Grape wine lees
contents (100 and 150 g kg1) also showed unpleasant effects, such as the decrease of overrun and the
Ice cream
Antioxidant
increase of particle size of fat globule. The level of 50 g kg1 was applicable because it improved the
Rheological property functionality without the disadvantages in the overrun and particle size. Furthermore, GWL significantly
increased the DPPH radical scavenging activity and reducing power of ice cream, and also its inhibitory
effect toward the oxidation of human erythrocyte membrane. It appeared that the antioxidants in GWL
were quite stable to the process of ice cream making.
Ó 2008 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.

1. Introduction a health ingredient in ice cream, we also evaluated the effect of

Grape wine lees (GWL) is the sediment in the bottom of the
barrel, about 100 g kg1 of GWL can be obtained from the pro-
cessing of grape wine. It contains yeast lees and grape stalks, grape
pomace, grape peel and grape seed, so it is considered as a waste-
product (Salmon, Fornairon-Bonnefond, Mazauric, & Moutounet,
2000). It has been shown that the extract form grape peel could
inhibit the oxidation of human LDL and moderate atherosclerosis
(Celotti, Ferrarini, Zironi, & Conte, 1996; Fitzpatrick, Coffey, &
Jantzen, 1997; Negro, Tommasi, & Miceli, 2003). Negro et al. (2003)
demonstrated that GWL exhibited strong antioxidant activity due
to its high phenolic contents, and proanthocyanidines in grape seed
also had strong antioxidant ability. Goni, Martin, and Saura-Calixto
(2005) found that the microbes in human intestine could
decompose 950 g kg1 total phenolic compounds, 300 g kg1 fiber
and 600–700 g kg1 proteins in the GWL. The beneficial fermen-
tation product, short-chain fatty acid, indicated that GWL could be
utilized by the microbes in human intestine (Toping & Clifton,
2001).
In this study, we used GWL to add the antioxidant properties to
ice cream. In order to determine the potential of using GWL as
* Corresponding author. Tel.: þ886 4 23323456x5160; fax: þ886 4 23337010.
E-mail address: ckhsu@asia.edu.tw (C.-K. Hsu).
adding GWL on the rheological properties of ice cream. Black queen
GWL was selected because it is the only species available for red
winemaking in Taiwan.
2. Materials and methods
2.1. Materials
The ingredients of ice cream mix included granular sugar (Tai-
wan Sugar Corporation, Tainan, Taiwan), skim milk powder (Hunter
OAK industries Co., Queensland, Australia) and cream (Bunge
Canada Ltd., Toronto, Canada). Carboxy methyl cellulose (CMC) was
obtained from Nippon paper industries Co., Tokyo, Japan. Gallic
acid, 1,1-diphenyl-2-picrylhydrazyl hydrate (DPPH), ascorbic acid
and thiobarbituric acid were purchased from Sigma Co. (St. Louis,
MO). Hydrochloric acid, sodium carbonate, potassium chloride,
sodium hydroxide, potassium ferricyanide were obtained from
Wako Co. (Tokyo, Japan). Sodium dihydrogen phosphate, disodium
hydrogen phosphate and potassium dihydrogen phosphate were
purchased from Nacalai Co. (Kyoto, Japan). Trichloroacetic acid, iron
chloride hexahydrate and Folin–Ciocalteu’s phenol reagent and
iron sulfate were obtained from Merck Co. (Darmstadt, Germany).
The black queen GWL was obtained from EPL Fruits Wine Co.,
Kaohsiung, Taiwan. The moisture and solid contents of the GWL
were 893 and 107 g kg1, respectively.

0023-6438/$34.00 Ó 2008 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.
doi:10.1016/j.lwt.2008.03.008
 J.-Y. Hwang et al. / LWT - Food Science and Technology 42 (2009) 312–318
2.2. Preparation of GWL
For the determination of antioxidant activities of black queen
GWL, the lees was homogenized with distilled water (the weight
ratio was 1:3) at 3000g, 5  C for 3 min under vacuum (Stephan UMC
5, Stephan Food Service Equipment, Hameln, Germany). The paste
(containing 0.17 g GWL) served as the sample for the measure-
ments of total anthocyanin content, total phenolic content, DPPH
radical scavenging activity, reducing power and inhibitory ability
toward oxidative damage on human erythrocyte membrane.
2.3. Preparation of ice cream with GWL
Ice cream mix was blended using the ingredients in the
following order: distilled water (620 g kg1), cream (120 g kg1),
skim milk powder (110 g kg1) and granular sugar (150 g kg1). The
blending process was conducted at 50  C. The mixture was filtered
with cheese cloth and then was sterilized at 80  2  C for 20 min.
For the ice cream with the addition of GWL, the lees in the range of
50, 100 and 150 g kg1 (wet weight basis) was pre-homogenized
with 1/4 of the amount of distilled water required for the ice cream
mix at 3000g, 5  C for 3 min under vacuum. The paste was heated
for 1 min in a water bath with boiling water. After cooling down the
temperature to 25  C, the paste was added to 2 kg ice cream mix,
and then the mixture was homogenized in vacuum for 5 min. After
homogenization, the mixture was refrigerated at 4  C for 18 h, and
then it was semi-frozen in an ice cream freezer (model SM-500,
Shiman Enterprise Co., Ltd., Yungho, Taiwan). The semi-frozen ice
cream was packaged into a 270 ml container and was hardened and
stored at temperatures of 25  C. For the control group (without
the addition of GWL), additional 4 g kg1 CMC was added as
a stabilizer.
2.4. Determinations of specific weight, viscosity, pH, overrun and
meltdown test
Frozen ice cream samples were thawed out in a refrigerator at
4  C for 24 h, and then were set at room temperature for the sample
temperature to reach 20  C. The melted ice cream was put into
a cylinder, and the volume and weight were recorded to calculate
the specific weight of ice cream samples. The viscosity of the
melted ice cream samples was determined using a Brookfield
viscometer with an RV spindle set (spindle No. 2). For the pH
measurement, about 10 g ice cream samples were dissolved in
90 ml distilled water. After homogenization, the pH was measured
using a pH meter. To determine the overrun, ice cream mix was put
in a container (270 ml) and weighted. Once the ice cream mix was
made into frozen ice cream, the same volume of frozen ice cream
was cut and put in the container. The overrun ¼ (WA  WB)/
WB  100%, where WA is the weight of ice cream mix; WB is the
weight of ice cream had the same volume as the mix. Meltdown
tests were conducted based on Martinou-Voulasiki and Zerfiridis
(1990) with slight modification. Approximately 140 g hardened ice
cream samples were put in a refrigerator at the temperature of
20  C for 16 h, and then the samples were suspended on a wire
mesh and allowed to melt at room temperature. The material that
melted and drained through the wire mesh was collected and
weighted every 10 min for 130 min.
2.5. Determination of fat destabilization and particle size
The fat destabilization index was determined by a method based
on the procedure of Goff and Jordan (1989) with slight modifica-
tion. The melted ice cream and ice cream mix (3 g each) were
diluted 1:600 in two steps with distilled water, and then the
absorbance of each diluent was measured at 540 nm using distilled
313
water as a blank. Fat destabilization index (%) ¼ [(A0  Ab)/
A0]  100%, where A0: value of the diluent of ice cream mix; Ab:
value of the diluent of melted ice cream.
The measurement of the particle size of fat globule was carried
out by using a commercial dynamic light scattering (DLS) spec-
trometer (Malvern CGS-3 with an ALV/LSE-5003 Multiple-tau
digital correlator, Malvern, England). The light source was a JDS-
Uniphase solid-state He–Ne laser (output power: 22 mW; wave-
length: 632.8 nm). DLS is a highly effective tool for probing the
particle size and particle size distribution of mix ice cream samples
in solution states. The temperature dependence of the particle size
was measured using the step scattering measurement for 10 min
with 90 scattering angle at room temperature (20  C).
2.6. Texture profile analysis (TPA) of ice cream
The firmness of the ice cream sample was measured by a texture
analyzer (TA-XT2 Texture Analyzer, Stable Micro Systems Co., Ltd.,
Surrey, United Kingdom) with a cylinder probe (diameter 50 mm).
The samples were cut into cubes with the lengths of 2  2  2 cm
and were compressed at a speed of 5 mm/s.
2.7. Color measurement
The color of ice cream sample was measured using a colorimeter
(Model TC-1, Tokyo Denshoku Co., Ltd., Japan). The L, a, and b values
were recorded, with L denoting lightness on a 0–100 scale from
black to white; a, red (þ) or green (); and b, yellow (þ) or blue ().
2.8. Differential scanning calorimetry (DSC) of ice cream
The thermal properties of ice cream samples were determined
by a DSC (Modulated DSC 2910, TA instrument, New Castle, USA)
according to the method reported by Goff, Caldwell, and Stanle
(1993). About 5 mg ice cream sample was sealed in an aluminum
sample pan and scanned from 30 to 30  C with a heating rate of
5  C/min. An empty pan was used as the reference. The flow rates of
nitrogen gas for cooling and heating were 110 and 40 cc/min,
respectively. The onset temperatures (T0), peak temperatures (Tp)
and enthalpies (DH ) of the transitions of ice-formation and ice-
melting were recorded.
2.9. Determination of total anthocyanin content
After melting 50 g of ice cream sample, 2 ml of the melted
sample was mixed with 2 ml methanol (containing 100 g kg1 HCl)
and then centrifuged at 3000g for 10 min (Picinelli, Bakker, & Bridle,
1994). The supernatant (1 ml) was added to 9 ml 1 mol equiv/l HCl.
The absorbance was measured at 520 nm. The total anthocyanin
content was calculated as A  101 18.89 (mg/l), where A: the
absorbance at 520 nm. For the sample of GWL, 2 ml of its pre-
homogenized paste was used.
2.10. Determination of total phenolic content
Twenty-five milliliters 500 ml l1 methanol solution was used to
extract 50 g of ice cream sample for 12 h. For grape a higher amount
of polyphenols and anthocyanins (antioxidants) is expected in
methanol extract than in water extract. However, the use of
500 ml l1 methanol solution can somehow extract both methanol
and water extractable polyphenols. Therefore, 500 ml l1 methanol
solution was used instead of pure methanol solution. The mixture
was filtered with Whatman No. 4 paper, and then 0.2 ml of the
sample was added with 1.8 ml distilled water in a 10 ml tube. After
adding 1 ml Folin–Ciocalteu’s phenol reagent and 2 ml Na2CO3
solution (20% w/v), the mixture was kept at 25  C for 20 min and
314 J.-Y. Hwang et al. / LWT - Food Science and Technology 42 (2009) 312–318
then the absorbance at 735 nm was measured (Julkunen-Titto,
1985). Gallic acid (0, 0.1, 0.25, 0.5, 0.75, and 1 mg/ml) was used as
the standard for the calibration curve, and the total phenolic
content in the sample was expressed as mg gallic acid equivalent
per ml tested sample. For the sample of GWL, 50 g of its pre-
homogenized paste was used.
2.11. Assay for DPPH radical scavenging activity
Briefly, ice cream sample (50 g) was extracted with 25 ml
500 ml l1 methanol solution for 12 h. The mixture was filtered
with Whatman No. 4 paper, and then 0.3 ml of the sample was
added with 1.2 ml methanol and 1.5 ml of 0.5 mmol/l DPPH (in
methanol). The solution was kept at room temperature for 90 min
and the absorption at 517 nm was measured (Blois, 1958). The DPPH
radical scavenging effect was calculated as follows: scavenging
effect (%) ¼ [(A0  (A Ab))/A0]  100%, where A0: value of DPPH
without sample; A: value of sample and DPPH; Ab: value of sample
without DPPH. For the sample of GWL, 50 g of its pre-homogenized
paste was used.
2.12. Measurement of reducing power
Fifty grams of ice cream sample was extracted with 25 ml
500 ml l1 methanol solution for 12 h. The mixture was filtered
with Whatman No. 4 paper, and then 0.1 ml of the sample was
mixed with 0.5 ml 0.2 mol/l phosphate buffer (pH 6.6) and 0.5 ml
10 g kg1 potassium ferricyanide, and then incubated at 50  C for
20 min. After cooling down the temperature, 0.5 ml of 100 g kg1
trichloroacetic acid was added to the mixture to stop the reaction,
and then the mixture was centrifuged at 7000g for 30 min. The
supernatant (1 ml) was mixed with 1 ml distilled water and 0.2 ml
1 g kg1 FeCl3, and then the absorbance was measured at 700 nm
(Oyaizu, 1986). The reducing powers of the tested samples
increased with the absorbance values. For the sample of GWL, 50 g
of its pre-homogenized paste was used.
2.13. The inhibitory ability toward oxidation of human erythrocyte
membrane
Human blood (100 ml) obtained from health subjects (age 20)
was diluted with equal volume of 10 mmol/l phosphate/152 mmol/l
KCl buffer (pH 7.4), and then the diluent was centrifuged at 1500g
for 20 min (Liao & Yin, 2000; Tsuda, Ohshima, Kawakishi, & Osawa,
1994). The erythrocyte was put in 10 mmol/l phosphate buffer (pH
7.4) and set at 4  C refrigerator for 24 h, and then the mixture was
centrifuged at 20,000g for 40 min. The sediment (broken erythro-
cyte membrane) was collected for subsequent tests.
Fifty grams of melted ice cream sample was extracted with
25 ml 500 ml l1 methanol solution for 12 h. The mixture was
filtered with Whatman No. 4 paper, and then 0.1 ml of the sample
was mixed with 0.5 ml erythrocyte membrane (protein concen-
tration was adjusted to 1 mg/ml), and then 0.1 ml 5 mmol/l FeSO4,
0.1 ml 10 mmol/l KCl, 0.1 ml 0.05 mmol/l ascorbic acid and 0.1 ml
PBS buffer were added to a final volume of 1 ml. The mixture was
incubated at 37  C for 24 h. After incubation, 0.5 ml 2.0 mol/l TCA/
1.7 mol/l HCl and 0.5 ml 10 g kg1 thiobarbituric acid (TBA) (dis-
solved in 0.05 mol equiv/l NaOH) were added to the mixture, and
then the final mixture was heated at 100  C for 30 min. After
cooling down the temperature of the mixture, it was centrifuged at
7000g for 10 min. The absorbance of the supernatant was measured
at 532 nm. The inhibitory ability of the tested samples toward
oxidation of human erythrocyte membrane increased with the
decrease of the absorbance values. The inhibitory ability was
represented as percentage of inhibition (%) ¼ [1  (AS/A0)]  100%,
where AS: the absorbance value with the sample; A0: the
absorbance value without sample. For the sample of GWL, 50 g of
its pre-homogenized paste was used.
2.14. Statistical analysis
One-way analysis of variance (one-way ANOVA) was conducted
using a package (SAS Institute Inc., Cary, NC). A significance level of
5% was adopted for all comparisons. Duncan’s multiple range test
was used to determine the significant difference between different
treatments.
3. Results and discussion
3.1. Effect of adding GWL on the specific gravity, pH, overrun,
viscosity and meltdown of ice cream
Physico-chemical properties of ice cream with the addition of
GWL are shown in Table 1. The addition of GWL slightly decreased
the specific gravity of ice cream; however, the differences among
50, 100 and 150 g kg1 GWL groups were negligible. The pH value of
ice cream decreased with the increase of GWL concentration. The
decline of pH was because there were various acid compounds,
such as tartaric acid and malic acid, in grapes. Furthermore, the
oxidation of ethanol remained in GWL could also contribute to
lower pH. Although GWL significantly decreased the overrun of ice
cream at 100 and 150 g kg1 levels, GWL at 50 g kg1 level did not
cause significant change in the overrun of ice cream. The viscosity
of ice cream was raised significantly by the addition of GWL. The
increase of the viscosity was about 0.9, 1.9 and 3.2 folds with the
addition of GWL at 50, 100 and 150 g kg1, respectively. Since the
viscosity of ice cream increased with GWL, it was possible that less
air was incorporated in the ice cream mix with GWL during batch
freezing, which resulted in lower overrun than for the ice cream
without GWL. Martinou-Voulasiki and Zerfiridis (1990) found that
the use of xanthan gum or guar gum increased the viscosity of
yogurt ice cream; a significant decrease in the overrun was
observed as the concentration of xanthan gum or guar gum
increased to level of 2–3 g kg1. Besides overrun, the melting rate is
also an important quality index for ice cream products. It was found
that the ice cream without GWL (the control) and the ice cream
with 4 g kg1 CMC were completely melted after 80 min, but ice
cream with GWL slowed down the melting rate significantly (Fig. 1).
About 32% reduction in the melting rate was found at 50 g kg1
GWL. More than 80% reduction in the melting rate was found at 100
and 150 g kg1 GWL; however, the difference in the reduction
between 100 and 150 g kg1 GWL was negligible. It has been
reported that the addition of hydrocolloid stabilizers or poly-
saccharides to ice cream not only increased the viscosity, but also
reduced the melting rate (Goff & Sahagian, 1996; Segall & Goff,
2002). Therefore, it was suspected that GWL contained some
components having the ability to absorb water, thus it could
increase the viscosity and reduce the melting rate.
Table 1
Effect of the addition of grape wine lees on the physico-chemical properties of ice
cream
Specific Viscosity pH Overrun
gravity (mPa s)
0 g kg1 GWL 1.120  0.00a 330  0.00e 7.12  0.01a 53.9  1.8b
50 g kg1 GWL 1.096  0.03b 640  28.28c 6.79  0.02b 51.9  2.5b
100 g kg1 GWL 1.084  0.03c 955  91.92b 6.56  0.23bc 41.6  1.3c
150 g kg1 GWL 1.071  0.01d 1390  42.43a 6.32  0.11c 35.3  2.1d
4 g kg1 CMC 1.120  0.00a 470  14.14d 7.14  0.00a 60.2  4.6a
GWL: grape wine lees; CMC: carboxy methyl cellulose.
The values are mean  standard deviation.
a,b,c,d,e
The values in the same column followed by different superscripts were sig-
nificantly different ( p < 0.05).
 J.-Y. Hwang et al. / LWT - Food Science and Technology 42 (2009) 312–318 315

160 Table 3
Effect of the addition of grape wine lees on the texture profile of ice creams

Firmness (g) Cohesiveness

140
0 g kg1 GWL 291  35a 0.25  0.01a
50 g kg1 GWL 173  29b 0.23  0.01a
100 g kg1 GWL 142  19bc 0.23  0.03a
120 150 g kg1 GWL 113  1c 0.16  0.02b
4 g kg1 CMC 251  11a 0.29  0.04a

100 GWL: grape wine lees; CMC: carboxy methyl cellulose.

The values are mean  standard deviation.
Weight (g)

a,b,c
The values in the same column followed by different superscripts were signifi-
cantly different ( p < 0.05).
80

reductions in the firmness were found at 50, 100 and 150 g kg1
60 GWL, respectively. However, the cohesiveness was less affected by
GWL. Our data showed that the cohesiveness did not change at 50
and 100 g kg1 GWL. For 150 g kg1 GWL, about 36% decline in the
40
cohesiveness was found. Table 4 shows the color properties of ice
cream influenced by the addition of GWL. As expected, the addition
20 of GWL increased the redness and yellowness of ice creams, but
decreased the lightness.

0 3.4. The thermal properties of ice cream with GWL

0 20 40 60 80 100 120 140
Time (mins)
The thermal properties associated with ice crystal-melting of ice
Fig. 1. Effect of the addition of GWL on ice cream melting. The values are mean and the cream with and without GWL were determined by differential
error bar represents standard deviation (A: control; -: 50 g kg1 GWL; :: 100 g kg1 scanning calorimetry (DSC). Fig. 2 shows the typical DSC curves for
GWL; B:150 g kg1 GWL; J: 4 g kg1 CMC).
the ice crystal-melting curves. The enthalpy for the ice crystal-
melting of 0 g kg1 GWL, 50 g kg1 GWL, 100 g kg1 GWL,
3.2. The fat structure of ice cream with GWL 150 g kg1 GWL and 4 g kg1 CMC was 7.16, 6.73, 6.25, 4.43 and
9.76 J/g, respectively. Addition of GWL decreased the enthalpy
Table 2 shows the fat destabilization index and particle size of values associated with the ice-melting transition. The decrease in
fat globule in ice cream containing GWL. Both GWL and 4 g kg1 enthalpy value might have two possible reasons: the decrease of
CMC significantly increase the fat destabilization index of ice cream. the final moisture content and the decrease of freezable water in
About 0.6-, 1.4- and 2.2-fold increase in the index were obtained the sample. However, the addition of GWL (800 g kg1 moisture
with the addition of 50, 100 and 150 g kg1 GWL, respectively. It has content) increased the final moisture content; therefore, the
been stated that fat destabilization in ice cream is partial moisture content was not the factor for the reduction in the
coalescence of fat globules due to the destabilizing effect during enthalpy. Vittadini and Vodovotz (2003) reported that the enthalpy
freezing conditions (Goff, 1997). Since the use of GWL increased the of ice-melting transition had a positive correlation with the amount
fat destabilization index, it was reasonable to consider that GWL of freezable water in the tested sample. Our data indicated that the
could enhance the coalescence of fat globules. In general, ice cream addition of GWL significantly decreased the amount of freezable
products with high fat destabilization index and low particle size of water in ice cream and that GWL had the ability to hold water and
fat globule were considered to be better quality. GWL increased the thus increases the melting rate and freezable water in ice cream.
fat destabilization index. A 2.3- and 3-fold increase in the particle
size was observed at 100 and 150 g kg1 GWL, respectively, while 3.5. The total anthocyanin and total phenolic contents of ice cream
no significant increase in the particle size was found at 50 g kg1 with GWL
GWL (Table 2).
It has also been shown that waste by-products obtained from
3.3. The texture and color properties of ice cream with GWL grape processing exhibited strong antioxidant activity (Jette et al.,
2000; Lu & Foo, 1999). Torel, Cillard, and Cillard (1986) reported
Effect of GWL on the texture profile of ice cream is shown in that anthocyanins were the main antioxidants in grape peel, while
Table 3. The firmness of ice cream decreased with the increase of proanthocyanidin and resveratrol were the major antioxidants in
GWL concentrations. Compared to the control, about 41, 52 and 61% grape seed (Jayaprakasha, Selvi, & Sakariah, 2003; Negro et al.,
2003). The total anthocyanin contents of the ice creams after
Table 2
Effect of the addition of grape wine lees on the fat structure of ice cream Table 4
Effect of the addition of grape wine lees on the color properties of ice creams
Fat destabilization Particle size of
index (%) fat globule (nm) L a b
0 g kg1 GWL 10.3  1.6d 198  15c 0 g kg1 GWL 96.4  0.7a 0.2  0.1d 2.3  0.3e
50 g kg1 GWL 16.4  2.1cd 221  19c 50 g kg1 GWL 68.3  1.4b 2.3  0.0c 8.0  0.2d
100 g kg1 GWL 24.6  1.0bc 448  76b 100 g kg1 GWL 67.8  0.7b 4.3  0.1b 10.7  0.3b
150 g kg1 GWL 32.5  8.0b 587  95a 150 g kg1 GWL 59.4  0.5c 5.4  0.2a 9.6  0.3c
4 g kg1 CMC 49.5  4.4a 197  14c 4 g kg1 CMC 96.2  0.4a 3.4  0.0e 11.5  0.2a

GWL: grape wine lees; CMC: carboxy methyl cellulose. GWL: grape wine lees; CMC: carboxy methyl cellulose.
The values are mean  standard deviation. The values are mean  standard deviation.
a,b,c,d a,b,c,d,e
The values in the same column followed by different superscripts were sig- The values in the same column followed by different superscripts were sig-
nificantly different ( p < 0.05). nificantly different ( p < 0.05).
316 J.-Y. Hwang et al. / LWT - Food Science and Technology 42 (2009) 312–318

0 The total phenolic contents of the ice creams with the addition
of GWL in the range of 0–150 g kg1 are shown in Fig. 4. It was
found that the total phenolic contents of ice cream and GWL were
1.52 and 7.51 mg/ml, respectively. The addition of GWL resulted in
-0.05
the increase of total phenolic content. When adding 150 g kg1
GWL, the total phenolic content reached 3.58 mg/ml. We estimated
the amount of total phenolic content in ice cream with 150 g kg1
-0.1 GWL was 2.42 mg/ml (0.85  1.52 mg/ml þ 0.15  7.51 mg/ml).
Thus, like the total anthocyanin content, the total phenolic content
Heat flow (W/g)

also did not decrease due to the processing of ice cream. When
converting the unit to mg/g of GWL, we obtained the value of total
-0.15
phenolic content of 45 mg/g GWL, about 5% of GWL. As expected,
our data indicated that the GWL still contained great amount of
phenolic compounds. Our results indicated that both anthocyanins
-0.2 and phenolic compounds in the GWL were quite stable during the
process of ice cream making. Therefore, the addition of GWL could
increase the antioxidant activity of ice cream.

-0.25 3.6. The antioxidant activity of ice cream with GWL

DPPH is a stable free radical that would be scavenged by a pro-

-0.3
-25 -20 -15 -10
Temperature (ºC)
Fig. 2. Effect of the addition of grape wine lees on the ice crystal-melting of ice creams
measured by differential scanning calorimetry (DSC) (A: control; -: 50 g kg1 GWL;
:: 100 g kg1 GWL; B:150 g kg1 GWL; J: 4 g kg1 CMC). Enthalpy (DH ) for the ice
crystal-melting of 0 g kg1 GWL, 50 g kg1 GWL, 100 g kg1 GWL, 150 g kg1 GWL and
4 g kg1 CMC was 7.16, 6.73, 6.25, 4.43 and 9.76 J/g, respectively.
addition of GWL in the range of 0–150 g kg1 are shown in Fig. 3.
For the ice cream without the addition of GWL or with the addition
of 4 g kg1 CMC, the total anthocyanin content was not detectable.
With the addition of GWL, the total anthocyanin contents in the ice
creams increased with the amount of added GWL. The total
anthocyanin content in the GWL was 568.6 mg/l, while the content
in the ice cream with 150 g kg1 GWL was 120.2 mg/l (21% of the
value of GWL). This result indicated that the addition of GWL
increased the anthocyanin content in the lees-containing ice cream,
and also the anthocyanin content did not decrease due to the
processing of ice cream. Therefore, GWL could contribute antho-
cyanins to enhance the antioxidant activity of ice cream.
ton-donating substance, for example, an antioxidant (Espin, Rivas,
-5 0 5 & Wichers, 2000). Based on this principle, the antioxidant activity
of a substance can be expressed as its ability in scavenging the
DPPH free radical. Fig. 5 shows the effect of GWL on the DPPH
radical scavenging activity of ice cream. The DPPH radical scav-
enging activity of GWL (0.17 g) was equivalent to 1 mg butylated
hydroxyanisole (BHA). Adding GWL significantly increased the
DPPH radical scavenging activity; however, the amount of addition
did not differ in the range of 50–150 g kg1.
The reducing powers of ice creams with the addition of GWL are
shown in Fig. 6. The reducing power of GWL (0.17 g) was equivalent
to 0.5 mg BHA. No reducing power was detectable in the ice cream
without GWL or with 4 g kg1 CMC. Addition of GWL, however,
increased the reducing power of ice cream increased with the
amount of GWL. When compared to the profiles of total anthocy-
anins and phenolic contents, it was noted that the profile of
reducing power is more similar to that of total anthocyanin content
than total phenolic content. Therefore, the reducing power
appeared to be influenced by anthocyanins in GWL.
9

700 a
8
Total polyphenolic contents (mg/ml)

600 a 7

6
Total anthocyanins (mg/L)

500

5
400
4 b
300 bc
3 c

200
b 2 d
c
100 d
1 e
ND ND
0 0
0 g kg-1 50 g kg-1 100 g kg-1 150 g kg-1 4 g kg-1 Wine lees 0 g kg-1 50 g kg-1 100 g kg-1 150 g kg-1 4 g kg-1 Wine lees
GWL GWL GWL GWL CMC GWL GWL GWL GWL CMC

Fig. 3. The total anthocyanin contents of ice creams containing grape wine lees in the Fig. 4. The total phenolic content of ice creams containing grape wine lees in the range
range of 0–150 g kg1 (ND: value was not detected). of 0–150 g kg1.
 J.-Y. Hwang et al. / LWT - Food Science and Technology 42 (2009) 312–318 317

100 a 80
a a

90
70

80
b b a
60
70
b
50 b
60 b

50 40

40 c
30

30
c 20
20
c
c 10 c
10

0 0
0 g kg-1 50 g kg-1 100 g kg-1 150 g kg-1 4 g kg-1 Wine BHA 0 g kg-1 50 g kg-1 100 g kg-1 150 g kg-1 4 g kg-1 Wine BHA
GWL GWL GWL GWL CMC lees 1 mg GWL GWL GWL GWL CMC lees 1 mg

Fig. 5. The DPPH scavenging activity of ice creams containing grape wine lees in the Fig. 7. The inhibitory effect of ice creams containing grape wine lees in the range of 0–
range of 0–150 g kg1. 150 g kg1 toward the oxidation of human erythrocyte membrane.

GWL (0.17 g) also exhibited an inhibitory effect toward the
oxidation of human erythrocyte membrane, equivalent to the
inhibition of 1 mg BHA (Fig. 7). Adding 50 g kg1 GWL did not show
significant increase in the inhibitory effect, but at both 100 and
150 g kg1 levels the inhibition increased significantly. Ohnishi
et al. (1994) stated that phenolic compounds such as caffeic acid,
chlorogenic, 3,5-dicaffeoylquinic and protocatechuic acid could
inhibit hydrogen peroxide-induced oxidation of erythrocyte
membrane. Therefore, it was suggested that phenolic compounds
in the GWL contributed to the increase of the inhibition toward
human erythrocyte membrane.
Our results demonstrated that the addition of GWL significantly
increased the antioxidant activities of ice cream, including DPPH
radical scavenging activity, reducing power and the inhibition to
the oxidation of human erythrocyte membrane. Our data also
indicated that the addition of GWL significantly increased the total
anthocyanins and phenolic contents. It is well known that antho-
cyanins and polyphenolic compounds had significant antioxidant
activity, and these compounds were quite abundant in grape.
Increase in the antioxidant activity in ice cream containing GWL
was due to the antioxidant activity contributed by GWL. Also,
anthocyanins and polyphenolic compounds might be the active
compounds contributed to the antioxidant activity because they
were quite stable during the process of ice cream making.
4. Conclusions

The addition of GWL increased the melting rate, fat

2.5
destabilization and decreased the freezable water of ice cream.
However, ice cream with high GWL contents also showed
a unpleasant effects, such as the decrease of overrun and the increase
of particle size of fat globule. These unwanted effects were minor at
2 a
low GWL concentration. For example, no significant decrease of the
overrun and increase of the particle size were found at 50 g kg1
level. GWL contains a significant amount of phenolic compounds
Absorbance at 700 nm

and may be used to produce value-added food products. In this

1.5
study, we demonstrated that the addition of GWL could signifi-
cantly increase the antioxidant activity of ice cream, and the anti-
oxidant in the GWL appeared to be quite stable during the process
b of ice cream making. Therefore, GWL has the potential to be used as
1 a value-added ingredient in ice cream industry to enhance the
antioxidant activity.

c References
0.5
Blois, M. S. (1958). Antioxidant determinations by the use of stable free radical.
Nature, 26, 1199–1200.
d Celotti, E., Ferrarini, R., Zironi, R., & Conte, L. S. (1996). Resveratrol content of some
ND ND wines obtained from dried Valpolicella grapes: Recioto and Amarone. Journal of
0 Chromatography, 730, 47–52.
-1 -1 -1 -1
0 g kg 50 g kg 100 g kg 150 g kg 4 g kg-1 Wine BHA Espin, J. C., Rivas, C. S., & Wichers, H. J. (2000). Characterization of the free radical
GWL GWL GWL GWL CMC lees 0.5 mg scavenger capacity of vegetable oils and oil fractions using 2,2-diphenyl-1-
picrylhydrazyl radical. Journal of Agricultural and Food Chemistry, 48, 648–656.
Fig. 6. The reducing activity of ice creams containing grape wine lees in the range of Fitzpatrick, D. F., Coffey, R. G., & Jantzen, P. T. (1997). Endothelium-dependent
0–150 g kg1 (ND: value was not detected). vasorelaxing activity of wine, grape and other plant products. In Wine:
318 J.-Y. Hwang et al. / LWT - Food Science and Technology 42 (2009) 312–318
Nutritional and therapeutic benefits (pp. 136–158). Washington: American
Chemical society.
Goff, H. D. (1997). Colloidal aspects of ice cream – a review. International Dairy
Journal, 7, 363–373.
Goff, H. D., Caldwell, K. B., & Stanle, D. W. (1993). The influence of polysaccharides
on the glass transition in frozen sucrose solutions and ice cream. Journal of
Dairy Science, 76, 1268–1277.
Goff, H. D., & Jordan, W. K. (1989). Action of emulsifiers in promoting fat
destabilization during the manufacture of ice cream. Journal of Dairy Science, 72,
18–29.
Goff, H. D., & Sahagian, M. E. (1996). Glass transitions in aqueous carbohydrate
solutions and their relevance to frozen food stability. [Special issue]. Thermo-
chimica Acta, 280–281, 449–464.
Goni, I., Martin, N., & Saura-Calixto, F. (2005). In vitro digestibility and intestinal
fermentation of grape seed and peel. Food Chemistry, 90, 281–286.
Jayaprakasha, G. K., Selvi, T., & Sakariah, K. K. (2003). Antibacterial and antioxidant
activities of grape (Vitis vinifera) seed extracts. Food Research International, 36,
117–122.
Jette, F., Young, L. O., Dragsted, B. D., Sorem, T., Lauridsen, M. H., & Brittmarie, S.
(2000). The effect of grape-skin extract on oxidative status. British Journal of
Nutrition, 84, 505–513.
Julkunen-Titto, R. (1985). Phenolic constituents in the leaves of northern willows:
methods for the analysis of certain phenolics. Journal of Agricultural and Food
Chemistry, 33, 213–217.
Liao, K. L., & Yin, M. C. (2000). Individual and combined antioxidant effects of seven
phenolic agents in human erythrocyte membrane ghosts and phosphatidyl-
choline liposome systems: importance of the partition coefficient. Journal of
Agricultural and Food Chemistry, 48, 2266–2270.
Lu, Y., & Foo, L. Y. (1999). The polyphenol constituents of grape pomace. Food
Chemistry, 65, 1–8.
Martinou-Voulasiki, I. S., & Zerfiridis, G. K. (1990). Effect of some stabilizers on
texture and sensory characteristics of yogurt ice cream from sheep’s milk.
Journal of Food Science, 55, 703–707.
Negro, C., Tommasi, L., & Miceli, A. (2003). Phenolic compounds and antioxidant
activity from red grape marc extracts. Bioresource Technology, 87, 41–44.
Ohnishi, M., Morishita, H., Iwahashi, H., Toda, S., Shirataki, Y., & Kimura, M., et al.
(1994). Inhibitory effects of chlorogenic acids on linoleic acid peroxidation and
haemolysis. Phytochemistry, 36, 579–583.
Oyaizu, M. (1986). Studies on products of the browning reaction. Antioxidative
activities of browning reaction products prepared from glucosamine. Japanese
Journal of Nutrition, 44, 307–315.
Picinelli, A., Bakker, J., & Bridle, P. (1994). Model wine solutions: effect of sulphur
dioxide on colour and composition during aging. Vitis, 33, 31–35.
Salmon, J. M., Fornairon-Bonnefond, C., Mazauric, J. P., & Moutounet, M. (2000).
Oxygen consumption by wine lees: impact on lees integrity during wine ageing.
Food Chemistry, 71, 519–528.
Segall, K. I., & Goff, H. D. (2002). A modified ice cream processing routine that
promotes fat destabilization in the absence of added emulsifier. International
Dairy Journal, 12, 1013–1018.
Toping, D. L., & Clifton, P. M. (2001). Short chain fatty acids and human colonic
function: roles of resistant starch and nonstarch polysaccharides. Physical
Review, 81, 1031–1064.
Torel, J., Cillard, J., & Cillard, P. (1986). Antioxidant activity of flavonoids and
reactivity with peroxy radicals. Phytochemistry, 25, 383–385.
Tsuda, T., Ohshima, K., Kawakishi, S., & Osawa, T. (1994). Antioxidative pigments
isolated from the seeds of Phaseolus vulgaris L. Journal of Agricultural and Food
Chemistry, 42, 248–251.
Vittadini, E., & Vodovotz, Y. (2003). Changes in the physicochemical properties of
wheat- and soy-containing breads during storage as studied by thermal anal-
yses. Journal of Food Science, 68, 2022–2027.