Antioxidant activities of some dried fruits consumed in Algeria

Salim Ouchemoukh
a,
*
, Said Hachoud
a
, Hamou Boudraham
a
, Abderrahmane Mokrani
b
,
Hayette Louaileche
b,
*
a
Laboratoire de Biochimie applique, Dpartement de Biologie Physico-Chimique, Facult des Sciences de la Nature et de la Vie, Universit Abderrahmane Mira de Bjaia,
Bjaia, Algeria
b
Laboratoire de Biochimie applique, Dpartement des Sciences Alimentaires, Facult des Sciences de la Nature et de la Vie, Universit Abderrahmane Mira de Bjaia, Bjaia, Algeria
a r t i c l e i n f o
Article history:
Received 12 June 2011
Received in revised form
12 July 2012
Accepted 15 July 2012
Keywords:
Vegetables
Bioactive compounds
Free radicals & chronic diseases
a b s t r a c t
Dried fruits such as prune, apricots, raisins and gs are an important source of different antioxidants
which can inhibit the harmful effects of the free radicals. They receive increasing attention for their
potential role in prevention of human diseases. The aims of our study were the quantication of some
antioxidants (carotenods, total phenolic compounds, anthocyanins, avonods and proanthocyanidins)
using colorimetric assays and the determination of antioxidant activities by three methods. Three
aqueous solvents (distilled water, ethanol and methanol) were used for the extraction of some antiox-
idants. Apricots and gs had the highest concentration of carotenods (10.7 and 10.8 mg bCE/100 g,
respectively). Raisins were the richest fruits in total phenolic concentration (1.18 g GAE/100 g) and
proanthocyanidins (17.53 mg CE/100 g). Also, gs had the highest concentration of avonods (105.6 mg
QE/100 g) and anthocyanins (5.9 mg/100 g). Apricots and raisins possessed a good reducing power, while
Agen prune showed signicant antioxidant activity to the phosphomolybdate. There were signicant
correlations between total phenolic concentration and antiradical activity (r 0.89; p < 0.001) of
ethanolic extract and reducing power (r 0.80; p < 0.01) of aqueous extract.
2012 Elsevier Ltd. All rights reserved.
1. Introduction
In recent times, natural antioxidants have attracted consider-
able interest among nutritionists, food manufacturers and
consumers because of their presumed safety and potential thera-
peutic value (Vijaya Kumar Reddy, Sreeramulu, & Raghunath,
2010). Fruits and vegetables are important dietary sources of
antioxidant polyphenols for humans. Fruits and vegetables
consumption have been shown by multiple epidemiology studies
to reduce the risk of chronic diseases such as cancer and heart
disease (Vinson, Zubik, Bose, Samman, & Proch, 2005). Fruits are
particularly interesting because they are rich in antioxidants and
can be consumed on various occasions and as fresh, dried, juice
and other processed fruits (Patthamakanokporn, Puwastien,
Nitithamyong, Prapaisri, & Sirichakwal, 2008). Antioxidant
activity and in particular polyphenol contents of dried fruits are
expected to be high due to their low moisture content with
increased shelf life (Mariod, Ibrahim, Ismail, & Ismail, 2010). Dried
fruits have been studied by many researchers (Breska et al., 2010;
Coimbra, Nunes, Cunha, & Guin, 2011; Ferreira et al., 2002;
Marelli et al., 2012; Rivero-Cruz, Zhu, Kinghorn, & Wu, 2008). The
drying of fruits is a very ancient practice for food preservation still
in use nowadays. The g is a delicious, nutritive fruit and has
medicinal properties that may reduce the risk of cancer and heart
disease. Fresh and dried gs are especially rich in antioxidant
polyphenols, ber, trace minerals, proteins, sugars and organic
acids. Eight phenols (chlorogenic acid, catechin, epicatechin, rutin,
cyanidin-3-O-rutinoside, luteolin-8-C-glucoside, quercetin-3-O-
glucoside, kaempferol-3-O-glucoside) were identied in fresh and
dried fruits and the predominant phenolic compound was epi-
catechin (Slatnar, Klancar, Stampar, & Veberic, 2011). Raisins are
among solid fruits products having the highest concentration of
total phenolic compounds and the highest level of total antioxi-
dant activity. In raisins, the most abundant phenolic compounds
are usually quercetin, kaempferol and coumaric acid (Williamson
& Carughi, 2010). Meng et al. (2011) reported the presence of 10
phenols (gallic, 3,4 dihydroxybenzoic, caffeic, syringic, ferulic,
salicylic and coumaric acids, catechin, quercetin and rutin) in
Chinese raisins. 3,4-dihydroxybenzoic acid was the most
predominant phenolic compound in these raisins. In this paper,
we report antioxidant concentrations and antioxidant activities of
some dried fruits commonly consumed in Algeria.
* Corresponding authors. Tel./fax: 213 34 21 47 62.
E-mail addresses: ouchemoukhsalim@yahoo.fr (S. Ouchemoukh),
haylouaileche@yahoo.fr (H. Louaileche).
Contents lists available at SciVerse ScienceDirect
LWT - Food Science and Technology
j ournal homepage: www. el sevi er. com/ l ocat e/ l wt
0023-6438/$ e see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.lwt.2012.07.022
LWT - Food Science and Technology 49 (2012) 329e332
2. Material and methods
2.1. Chemicals and reagents
DPPH (2, 2
0
-diphenyl-1-picrylhydrazyl), quercetin, b-carotene,
FolineCiocalteus reagent and hexane was obtained from
SigmaeAldrich (Germany), Alfa Aesar (Germany), Fluka (USA),
Prolabo (USA) and Riedel-de-Han (Germany), respectively. All
other reagents and chemicals were procured from Biochem Che-
mopharma (United kingdom).
2.2. Samples
Some dried fruits, bought in Bejaia (Algeria) market, were used
in the present study: apricots (Prunus armeniaca), white raisins
(Vitis vinifera), black gs (Ficus carica) and prune (Prunus domes-
tica). F. carica belongs to Moraceae family and other species to
Rosaceae family. Two different prunes were used: prune and Agen
prune (region of France). The weight of each sample was approxi-
mately 500 g. The fruits were cleaned and edible portions were cut
into small pieces before extraction.
2.3. Quantication of antioxidants
2.3.1. Carotenods
Carotenods are pigments insoluble in water and soluble in
apolar solvents like hexane. The carotenoids were extracted
according to the method of Sass-Kiss, Kiss, Milotay, Kerek, and
Toth-Markus (2005). A quantity of crushed fruit (0.2 g for apri-
cots and gs: 1 g for prune and raisins) was added to 10 ml of
solvent mixture (hexane, acetone and ethanol, 1:2:1, v/v/v). After
40 min of stirring, the absorbance of upper phase was determined
at 450 nm by UVeVIS spectrophotometer (Shimadzu). b-carotene
was used as the standard and the results were expressed as mg b-
carotene equivalent per 100 g dry weight of fruit (bCE/100 g DW)
using the following equation calibration curve: y 287.63x;
R
2
0.994, where x was the absorbance and y was the b-carotene
equivalent.
2.3.2. Anthocyanins
Anthocyanin concentration was determined according to the
procedure described by Mlo, Lima, Maciel, Caetano, and Leal (2006).
One gramof crushed fruit was mixed with10 ml of ethanolic solution
(ethanol/HCl 1.5 mol/l; 85:15, v/v) and allowed to stand inthe dark at
4

C overnight. After ltration, the absorbance was measured at
530 nm. The concentrations of anthocyanins were obtained by using
a molar extinction coefcient: 38,000 L*mol
1
cm
1
(Ganjewala, Boba, & Raghavendra, 2008).
2.3.3. Phenolic compounds
2.3.3.1. Preparation of the extracts. Three aqueous solvents
(distilled water, ethanol 50 ml/100 ml and methanol 50 ml/100 ml)
were used in order to extract the phenolic compounds. Each sample
(2 g) was mixed with 20 ml of solvent. After stirring for 40 min, the
mixture was centrifuged at 1800 g during 30 min and it was ltered
with lter paper.
2.3.3.2. Total phenolic compounds. The total phenolic compounds
concentration of the samples was measured according to the
method of Nencini et al. (2007). Extract from dried fruits (200 ml)
was mixed with 1 ml of FolineCiocalteus reagent. After 3 min, the
mixture was neutralized with 800 ml of aqueous sodium carbonate
(2 g/100 ml). The blue absorbance was read at 720 nm. The
concentration of total phenolics was expressed as mg of gallic acid
equivalent (GAE)/100 g DW (y 4.2753x; R
2
0.9947).
2.3.3.3. Flavonods. The method of Djeridane et al. (2006) was used
to determine the concentration of avonods. One milliliter of the
extract was mixed with 1 ml of aqueous aluminum chloride
(2 g/100 ml). After incubation at room temperature for 10 min, the
absorbance of the mixture was read at 410 nm. Total avonod
concentration was expressed as mg of quercetin equivalent
(QE)/100 g DW (y 13.03x; R
2
0.9984).
2.3.3.4. Proanthocyanidins. The concentration of proanthocyani-
dins was determined by butanoleHCl assay (Maksimovic, Malencic,
& Kovacevic, 2005). Extract from dried fruits (0.5 ml) was mixed
with 3 ml of butanoleHCl (95:5; v/v) and 0.1 ml of iron sulfate (2 g/
100 ml). The mixture was incubated at 90

C for 1 h. The absorbance

was determined at 530 nm. The results were expressed as mg
cyaniding equivalent (CE)/100 g using a molar extinction coefcient
of cyanidin: 34,700 L*mol
1
cm
1
.
2.4. Antioxidant activities
2.4.1. Antiradical activity
The ability of the extracts to scavenge DPPH radical was deter-
mined according to the method of Al et al. (2009). Methanolic DPPH
solution (100 ml, 6*10
5
mol/l) was added to 1000 ml of extract. After
20 min of incubation at room temperature, the absorbance was read
at 517nm. The percentageof reductionof radical DPPHwas calculated
as follows: Radical-scavenging activity % [(Abs
control
Abs
sample
)/
(Abs
control
)]*100, where Abs
control
is the absorbance of DPPH
radical methanol and Abs
sample
is the absorbance of DPPH
radical sample extract.
2.4.2. Reducing power
The reducing power of the dried fruits tested was estimated
according to the method described by Yildrim, Oktay, and
Bilaloglu (2001). One milliliter of the extract was added to
2.5 ml of phosphate buffer (0.2 mol/l, pH 6.6) and 2.5 ml of
aqueous potassium ferricyanide solution (1 g/100 ml). After stir-
ring, the mixture was incubated at 50

C for 20 min. Then, 2.5 ml of

aqueous trichloroacetic acid solution (10 g/100 ml) were added.
1.25 ml of distilled water and 0.25 ml of aqueous ferric chloride
solution (0.1 g/100 ml) were added to 1.25 ml of the mixture. After
10 min, the absorbance was determined at 700 nm. The results
were expressed as mg ascorbic acid equivalent (AAE)/100 g DW
(y 11.411x; R
2
0.9926).
2.4.3. Phosphomolybdenum method
The total antioxidant capacities of the extracts were evaluated
by the phosphomolybdenum method as described by
Ramalakshim, Rahath, and Jagan (2008). One milliliter of the
phosphomolybdenum solution (sulfuric acid 0.6 mol/l, sodium
phosphate 0.028 mol/l and ammoniummolybdate 0.004 mol/l) was
added to 100 ml of the extract. After incubation at 90

C for 60 min,
the absorbance was determined at 695 nm. The results were
expressed as mg AAE/100 g DW (y 4.8057x; R
2
0.9938).
2.5. Statistical analysis
The averages and the standard deviations are calculated with
Microsoft Ofce Excel 2007. The software STATISTICA 5.5 was
used to compare the different results by the analysis of variance
with one factor (ANOVA). The results were classied by
decreasing order a > b > c > d > e. The values obtained carrying
the same letter do not present any signicant difference at
p < 0.05.
S. Ouchemoukh et al. / LWT - Food Science and Technology 49 (2012) 329e332 330
3. Results and discussion
3.1. Antioxidant concentrations
The solvents used were mixed with water because polyphenols
are polar compounds. The concentrations of carotenods and
anthocyanins are presented in Table 1; total phenolic compounds,
avonods and proanthocyanidins concentrations are given in
Table 2. Antioxidant concentrations of dried fruits presented
differences. The concentration of carotenods of gs and apricots
were the highest. The lowest concentrations were recorded in Agen
prune, prune and raisins. The statistical analysis revealed no
signicant difference (p < 0.05) between apricots and gs and
between the Agen prune, prune and the raisins. The concentrations
of anthocyanins in analyzed fruits presented signicant differences
(p < 0.05). The gs and the Agen prune possessed high concen-
trations of these substances. Nevertheless, the lowest concentra-
tions were recorded for the samples of prune, raisins and apricots.
These results were in agreement with those reported by Vallejo,
marin, and Tomas-Barberan (2012). They obtained high anthocy-
anin concentrations mainly cyanidin-3-rutinoside (16.9e108.9 mg/
100 g) in gs. Ethanolic extract of raisins had the highest concen-
tration of total phenolics and aqueous extract of gs was not rich in
these compounds. The statistical analysis revealed signicant
differences between the fruits (p < 0.05), except for the aqueous
extracts of apricots and the raisins, methanolic extracts of Agen
prune and prune as well as ethanolic apricots and g extracts. Our
result for the concentration of total phenolic compounds of
methanolic extract of raisins was superior to those reported by
Mishra, Dubey, Mishra, and Barik (2010) (0.83 g/100 g). The highest
concentrations of avonods were those of the ethanolic and
aqueous extracts of gs. The aqueous extract of raisins presented
the lowest concentration. Signicant differences were noted
between the analyzed samples (p < 0.05). Higher concentrations
(107e140 mg/100 g) of avonods for gs were obtained also by
Vallejo et al. (2012). The highest concentration of proanthocyani-
dins was observed in raisins ethanolic extract. For each solvent,
apricots presented the lowest concentration of these antioxidants.
The concentrations of proanthocyanidins in aqueous, ethanolic and
methanolic extracts of raisins were high compared to other fruits.
No signicant differences were noted Agen prune, prune, apricots
and gs (aqueous and ethanolic extracts) as well as for Agen prune
and gs (methanolic extracts). The discordance in antioxidant
concentrations of fruits between different studies could be due to
varietal, seasonal, agronomical differences, genomics, moisture
content, method of extraction and standards used (Imeh &
Khokhar, 2002). Also, the difference between extracts is due to
the varying efciency of the solvents used.
3.2. Antioxidant activities
The results of antioxidant activities are given inTable 3. Aqueous
extract of gs had the lowest antiradical activity, whereas ethanolic
extract of raisins had the best DPPHradical scavenging activity. This
can be related to the richness in total phenolic compounds of raisins
fruit compared to the other fruits. All these fruits showed a capacity
to trap radical DPPH and that this property differs signicantly
(p < 0.05) from a fruit to another, except between Agen prune,
prune and raisins (aqueous extracts), prune and raisins (methanolic
extracts) and between apricots and gs (ethanolic extract).
Reducing power is one of the mechanism of antioxidant activity
which measure the conversion of a Fe
3
/ferricyanide complex to
the ferrous form. Aqueous extract of apricots and ethanolic extract
of raisins showed a highest reducing capacity. On the other hand,
aqueous, ethanolic and methanolic extracts of gs showed a weak
capacity to reduce iron (III). Agen prune, apricots and raisins
(methanolic extract) and Agen prune and apricots (ethanolic
extract) did not present a signicant difference in their reducing
power. Mishra et al. (2010) reported that raisins showed a weak
reducing power. However, our results were in discordance with the
reported data. Using the phosphomolybdate, aqueous extract of
Table 1
Concentrations of carotenods and anthocyanins of some dried fruits.
Fruits Carotenods (mg bCE/100 g DW) Anthocyanins (mg/100 g DW)
Agen prune 1.4 0.4
b
4.0 1.0
b
Prune 1.6 0.6
b
2.0 0.0
c
Apricots 10.7 2.6
a
0.5 0.0
d
Raisins 2.2 0.2
b
1.0 0.1
cd
Figs 11.0 1.0
a
5.9 0.2
a
Values are mean standard deviation (n 3). Means followed by the same letter are
not different according to ANOVA (Analysis Of VAriance).
Table 2
Concentrations of antioxidants of some dried fruits.
Fruits Total phenolic
compounds
(g GAE/100 g DW)
Flavonods
(mg QE/100
g DW)
Proanthocyanidins
(mg CE/100 g DW)
Distilled water
Agen prune 0.77 0.02
a
59.7 0.3
c
3.22 0.16
b
Prune 0.73 0.01
ab
70.1 0.6
b
3.15 0.01
b
Apricots 0.65 0.01
b
56.8 0.0
d
2.24 0.02
b
Raisins 0.65 0.07
b
20.8 1.1
e
7.39 0.87
a
Figs 0.47 0.00
c
104.5 2.1
a
2.18 0.08
b
Methanol 50 ml/100 ml
Agen prune 0.74 0.03
b
53.0 0.6
b
3.3 0.2
b
Prune 0.76 0.03
b
46.6 1.0
c
3.0 0.1
bc
Apricots 0.63 0.00
c
31.9 0.8
d
2.2 0.2
c
Raisins 1.03 0.07
a
30.9 0.6
e
13.2 0.1
a
Figs 0.52 0.00
d
79.9 0.8
a
3.4 0.3
b
Ethanol 50 ml/100 ml
Agen prune 0.64 0.02
c
42.0 0.1
c
3.6 0.4
b
Prune 0.74 0.00
b
48.7 0.7
b
2.7 0.0
b
Apricots 0.54 0.00
d
30.6 0.2
d
2.1 0.2
b
Raisins 1.18 0.01
a
28.2 0.0
e
17.5 1.2
a
Figs 0.54 0.00
d
105.6 0.5
a
3.2 0.0
b
Values are mean standard deviation (n 3). Means followed by the same letter are
not different according to ANOVA (Analysis Of VAriance).
Table 3
Antioxidant activities of some dried fruits.
Fruits Antiradical
activity (%)
Reducing power
(mg AAE/100 g DW)
Phosphomolybdenum
method (g AAE/100 g DW)
Distilled water
Agen prune 76.0 7.3
a
599.9 3.7
b
24. 8 0.9
a
Prune 72.6 6.7
ab
548.9 17.3
c
5.4 0.8
b
Apricots 61.2 0.8
b
680.9 2.1
a
22.0 5.0
a
Raisins 78.4 0.6
a
454.5 3.4
d
7.6 0.4
b
Figs 28.1 3.7
c
210.0 1.4
e
22.1 0.5
a
Methanol 50 ml/100 ml
Agen prune 80.7 2.4
b
674.3 11.8
a
14.1 1.2
b
Prune 87.0 0.6
a
623.9 0.4
b
21.5 0.8
a
Apricots 63.3 1.0
c
661.2 1.2
a
16.4 2.1
b
Raisins 89.2 1.5
a
673.7 14.1
a
0.6 0.1
c
Figs 53.0 0.6
d
232.9 6.5
c
14.1 0.4
b
Ethanol 50 ml/100 ml
Agen prune 72.3 1.8
c
516.9 1.0
c
12.9 1.0
c
Prune 81.7 1.6
b
569.2 10.2
b
21.9 1.8
a
Apricots 53.6 0.6
d
485.9 18.9
c
15.4 0.7
bc
Raisins 92.2 0.4
a
679.8 23.5
a
5.9 0.2
d
Figs 55.9 0.7
d
221.7 0.1
d
17.2 2.3
b
Values are mean standard deviation (n 3). Means followed by the same letter are
not different according to ANOVA (Analysis Of VAriance).
S. Ouchemoukh et al. / LWT - Food Science and Technology 49 (2012) 329e332 331
Agen prune had the best antioxidant activity. The statistical analysis
showed signicant differences (p <0.05) between the fruits, except
for the Agen prune, apricots and gs aqueous and methanolic
extracts.
Differences in cultivation region and practices, ripening stage,
harvested conditions and seasons could be the most factors for
variation of the antioxidant activity (Patthamakanokporn et al.,
2008). Moreover, the antioxidant activity depends on the extrac-
tion solvent polarity, the techniques of extraction as well as the
process and the temperature of drying.
3.3. Correlations
There were statistically signicant correlations (Table 4)
between antioxidants and antioxidant activities: DPPH radical
scavenging activity and total phenolic compounds of ethanolic,
methanolic and aqueous extracts, reducing power and total
phenolic compounds of aqueous extracts, antiradical activity and
proanthocyanidins. Moreover, there was a good correlation
between antiradical activity and reducing power of ethanolic
extracts. These results were in agreement with those reported by
Negi, Jayaprakasha, and Jena (2003), Vijaya Kumar Reddy et al.
(2010) & aliskan and Polat (2011). The antioxidant activity of
phenolic constituents may be related to their redox properties,
which allow them to act as reducing agents or hydrogen-atom
donors, their ability to chelate metals, inhibit lipoxygenase and
scavenge free radicals (Mishra et al., 2010).
4. Conclusion
The results obtained showed that the concentrations extracted
by the different solvents were quite close and phenolics from
raisins were poorly extracted by water. Moreover, lowest concen-
trations of proanthocyanidins were consistently found in apricot
and highest proanthocyanidin concentrations were obtained in
raisins, as were total phenolics in the aqueous alcohol extracts. The
antioxidant capacity was related to polyphenols content. The gs
were the richest fruit in carotenods, anthocyanins and avonods.
The aqueous extract of apricot presented the best reducing power
and Agen prune had the highest antioxidant activity with the
phosphomolybdenum method. The raisin extracts showed the
highest reduction of DPPH. Apricots, gs, prune and raisins are an
excellent dietary source of natural antioxidants and can be
considered as foods with remarkable benets for human health.
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Table 4
Correlation matrix between antioxidants and antioxidant activities.
TPC FLA PRO RP AA AAP
TPC 1
FLA 0.52 1
PRO 0.84
***
0.44 1
RP 0.68
***
0.73 0.32 1
AA 0.80
***
0.66 0.56
**
0.74
***
1
AAP 0.41 0.39
*
0.59 0.14 0.36 1
Abbreviations: TPC, total phenolic compounds; FLA, avonods; PRO, proantho-
cyanidins; RP, reducing power; AA, antiradical activity; AAP, antioxidant activity
with phosphomolybdate.
*
p < 0.05,
**
p < 0.01,
***
p < 0.001.
S. Ouchemoukh et al. / LWT - Food Science and Technology 49 (2012) 329e332 332