International Journal of Biological Macromolecules 92 (2016) 441–450

Contents lists available at ScienceDirect

International Journal of Biological Macromolecules

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

Extraction and characterization of three polysaccharides extracted

from Opuntia ficus indica cladodes
Nadia Bayar a , Mouna Kriaa a , Radhouane Kammoun a,b,∗
a
Laboratory of Microrganisms and Biomolecules, Center of Biotechnology of Sfax, University of Sfax, Road Sidi Mansour Km 6, B.P 1177 Sfax 3018, Tunisia
b
Biotechnology High School of Sfax (ISBS) Soukra Km 3; P.O. Box 261, Sfax 3000, Tunisia

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

Article history: The chemical extraction and the characterization of polysaccharides from mucilage (MC), pectin (PC)
Received 11 March 2016 and total pectic mucilage fraction (TFC) of Opuntia ficus indica cladodes as well as the evaluation of their
Received in revised form 11 July 2016 antioxidant activities was investigated. The FTIR spectroscopic analysis revealed the presence of carboxyl
Accepted 13 July 2016
and hydroxyl groups corresponding to polysaccharides. Uronic acid and the total sugar contents of PC
Available online 15 July 2016
were higher than those of TFC and MC whereas ash content of MC was considerably more important. In
addition, the findings showed that all the samples had little protein content and low average molecular
Keywords:
weight compared to the results mentioned in literature. Furthermore, MC reached not only the highest
Opuntia ficus indica cladodes
Polysaccharides
water (WHC) and oil holding (OHC) capacities (7.81 g/g and 1.34 g/g, respectively) but also the highest
Functional properties antioxidant properties (DPPH and ABTS scavenging activities, ␤-carotene bleaching inhibition activity
and reducing power). However, PC had the strongest emulsifying and foaming properties. As for TFC,
it had low WHC, OHC and emulsifying properties whereas it had higher foaming properties than MC
and greater antioxidant properties compared to PC. These outcomes can encourage the use of PC as a
surfactant and MC and TFC as natural antioxidants in food and pharmaceutical industries.
© 2016 Elsevier B.V. All rights reserved.

1. Introduction
Opuntia ficus indica (OFI) is a higher plant which belongs to the
Cactaceae family and the order Centrospermae. This plant grows,
essentially, in the tropical and subtropical regions. It is one of the
fewest plants that can adapt to difficult conditions [1]. OFI also
grows throughout Tunisia in areas characterized by water stress
and low soil quality. Fresh fruits have been exclusively used as
human food while cladodes have been exploited as livestock forage,
essentially, during the dry season. In Tunisia, cactus has also been
used to prevent soil degradation and control desertification [2].
The cultivated area of OFI in Tunisia was estimated to 600,000 ha
which was similar to the area cultivated in Brazil. The chemical
composition of the cactus forage was varied with the cultivar, the
development stage, the fertilization, the plant population and the
cladode order [3]. The main compounds presented were pectin,
mucilage and minerals [4].
∗ Corresponding author at: Laboratory of Microrganisms and Biomolecules, Center
of Biotechnology of Sfax, University of Sfax, Road Sidi Mansour Km 6, B.P 1177, Sfax
3018, Tunisia.
E-mail address: radhouan.kammoun@cbs.rnrt.tn (R. Kammoun).
The vegetative parts of OFI plants which were traditionally used
to protect people’s health now display an important effect on food
and drug industry because they contain pectic mucilage which is a
complex polysaccharide [4]. Several researchers have studied this
whole fraction and found out that we could extract two water sol-
uble polymers which were the mucilage and pectin.
The mucilage is the result of the polymerization of more
monosaccharides whose majority is associated with uronic acids.
This complex hydrocolloid which is naturally abundant in certain
plant seeds and cactus plants is a viscous substance responsible for
retaining water [5,6]. The mucilage gums were used in food and
medicine because of their functional properties such as gelling,
thickening and emulsifying and their ability to prevent and treat
certain diseases [7]. Pectin is located in the primary cell wall and in
the middle lamella of higher plant tissues [8]. It designs a large and
complex family of polysaccharides which is essentially composed
of ␣-d-(1 → 4) galacturonic acid units (homogalacturonans) inter-
rupted by l-rhamnose residues with side-chains of neutral sugars
especially d-galactose and l-arabinose (rhamnogalacturonans I and
II) [9,10]. This carbohydrate is a natural food additive (E440) widely
applied in the food industry for its different properties such as the
gelling agent, the thickener, the stabilizer and the emulsifier. There-
fore, global demand for pectin is in access of 30,000 t per year, with
a growth of 4–5% per year [11].

http://dx.doi.org/10.1016/j.ijbiomac.2016.07.042
0141-8130/© 2016 Elsevier B.V. All rights reserved.
442 N. Bayar et al. / International Journal of Biological Macromolecules 92 (2016) 441–450
This study aims to isolate and characterize pectin from mucilage
and the whole fraction of pectic mucilage presented in Opuntia ficus
indica cladodes and to investigate and compare their functional
properties and antioxidant activities in order to assess their poten-
tial uses as new food hydrocolloids in food and pharmaceuticals.
2. Materials and experimental methodology
2.1. Materials
Cladodes were harvested from the region of Siliana in North
West Tunisia. They were washed to remove their impurities and
then cut into small pieces. After that, they were dried in an oven
with a flow of air at 60 ◦ C for 24 h. The dried pieces were grounded
and stored at room temperature until use. The raw materials were
selected due to their abundance in Tunisia.
2.2. Extraction
The extraction of the total pectic mucilage fraction was carried
out by the conventional method described by Bagherian et al. [12]
with slight modifications. Dried cladodes were suspended in dis-
tilled water with a solid/liquid ratio of 1:15 (w/v). The suspension
was then adjusted to pH 2.8 with hydrochloric acid before being
heated at 90 ◦ C in water bath for 2 h. Subsequently, the mixture was
filtered in synthetic cloth and centrifuged at 4500 rpm for 15 min
in order to remove solid residues. The filtrate was concentrated
using a rotary evaporator. Next, the concentrated solution was pre-
cipitated with two volumes of isopropanol and kept overnight at
4 ◦ C. Afterward, the precipitate was separated by centrifugation at
4500 rpm for 15 min and the wet polysaccharides were washed
twice with absolute ethanol and dried in a laboratory oven at 50 ◦ C
for 24 h. The obtained fraction was designed by TFC. The mucilage
was extracted from dried cladodes by distilled water according to
the method described by Habibi et al. [13]: 90 min, at 25 ◦ C, with
stirring at 250 rpm and solid/liquid ratio of 1:10 (w/v). Then, the
mixture was centrifuged for 15 min at 4500 rpm. The filtrate was
precipitated with two volumes of isopropanol overnight at 4 ◦ C.
Afterward, the precipitate was washed twice with absolute ethanol
and dried at 50 ◦ C for 24 h. This fraction was named MC.
The residue was used to extract pectic fraction with the same
procedure applied to the TFC extraction. Pectic fraction was
designed by PC.
The polysaccharide yields were expressed by the ratio of the
dry weight of the polysaccharide extracted to the dry weight of the
samples in% (w/w).
2.3. Chemical and structural analyses
The moisture and ash content of the cladodes extracts were
recorded according to the AOAC standard methods 930.15 and
942.05 [14], respectively.
The protein content was determined using the Bradford method
[15].
Total carbohydrates were estimated according to the phenol-
sulfuric method [16].
The uronic acid content was determined using the method
described by Filisetti-Cozzi and Carpita [17]. In fact, 0.2 ml of pectin
solution was mixed with 1.2 ml of sulfuric acid containing up to
120 mM sodium tetraborate. The samples were heated at 100 ◦ C
in boiling water for 20 min. The tubes were cooled in an ice bath
and 100 ␮l of carbazole reagent (1% in ethanol) were added. Then,
the tubes were heated at 100 ◦ C for 20 min and left to cool at room
temperature. The absorbance took place at 525 nm. The uronic acid
content was estimated according to the standard curve obtained
with galacturonic acid solutions (25–200 ␮g/ml).
The average molecular weight (Mw) was determined using a gel
filtration high pressure chromatography equipped with a refrac-
tive index detector using Zorbax PSM 300 column (6.2 * 250) with
a mobile phase of bi-distilled water, a flow rate of 0.8 ml/min and
30 ◦ C. 100 ␮l of polysaccharide solution (0.2%) was injected after
a filtration through a 0.2 ␮m membrane. The average molecular
weight (Mw) of the three polysaccharides was evaluated by the
comparison of their retention times with the calibration curve
traced using the retention times of different polysaccharides hav-
ing known molecular weights which were lactose (0.342 kDa),
␣-cyclodextrin (1.135 kDa), dextran (40 kDa) and dextran (60 kDa).
The structural characteristics of the cladodes extracts were
investigated by the FTIR spectra which were recorded using
a Bruker Vector 22 instrument (Germany) in the region of
400–4000 cm−1 at a resolution of 4 cm−1 and then processed by
Bruker OPUS software.
All analysis was performed in triplicate.
2.4. Functional properties
2.4.1. Water holding capacity (WHC)
The water holding capacity of the extracted fractions was deter-
mined through the method described by Lin et al. [18] and improved
by Sila et al. [19]. Each sample (0.5 g) was placed into a centrifuge
tube to be weighed (tube + sample). Then, 50 ml of distilled water
were added and the suspensions were held at room temperature for
1 h with stirring for 5 s every 15 min. After 20 min of centrifugation
using a refrigerated centrifuge (Hettich Zentrifugen, ROTINA 380R,
Germany) at 5000g, the upper phase was eliminated and the tube
was tilted to a 45◦ angle on a filter paper; then drained for 30 min.
The WHC was recorded according to the following formula:
the weight of the content of the tube after drainage
WHC =
the weight of the dried sample
2.4.2. Oil holding capacity (OHC)
The samples (1.0 g) were added to a 10 ml of vegetable oil and
stirred for 5 h. After that, the suspension was centrifuged for 15 min
at 5000g. The supernatant was eliminated, the solids were weighed
and the OHC was expressed as the gram of oil held per gram of
sample [20,21].
2.4.3. Foaming properties
The foaming properties which included Foam capacity (FC)
and Foam stability (FS) were evaluated according to the method
described by Shahidi et al. [22] with some modifications. 10 ml
of each sample at 2% (w/v) of concentration were placed into a
conic tube and homogenized using the vortex for 3 min at room
temperature. Foam capacity was expressed as the percentage of
volume increasing just after homogenization and Foam stability
was expressed as the volume of foam remaining after 30 min. FC
and FS were calculated using the following equations:
VT − V0
FC (%) = × 100
V0
Vt − V0
FS (%) = × 100
V0
2.4.4. Emulsifying properties
Emulsion capacity (EC) and emulsion stability (ES) were carried
out according to the method of Cui and Chang [23] with slight modi-
fications. 10 ml of each sample at the concentration of 2% (w/v) were
mixed with 5 ml of corn oil and homogenized using the vortex for
3 min at room temperature. Then, the emulsions were centrifuged
 N. Bayar et al. / International Journal of Biological Macromolecules 92 (2016) 441–450
at 4600g for 5 min using a refrigerated centrifuge (Hettich Zen-
trifugen, ROTINA 380R, Germany). EC was calculated as follows:
Vf
EC (%) = × 100
Vi
Where Vf is the emulsion volume and Vi is the total volume
The emulsions were then incubated for 30 min at 80 ◦ C and cen-
trifuged at 8000g for 5 min. The emulsion stability ES was calculated
according to the following equation:
Vt
ES (%) = × 100
Vf
Where Vt is the emulsion volume after incubation and centrifuga-
tion.
2.5. In vitro antioxidants activities
2.5.1. DPPH• radical scavenging activity
The DPPH• radical scavenging activity of the three cladode
extracts was assessed using the method described by Bersuder
et al. [24]. 500 ␮l of each sample at different concentrations
(1–20 mg/ml) was mixed with 375 ␮l of 99% ethanol and 125 ␮l
of DPPH solution (0.02% (w/v) in ethanol) at different concentra-
tions. The mixture was stirred and then incubated in the absence
of light at room temperature for 1 h. The scavenging capacity was
determined by monitoring the decrease in absorbance at 517 nm.
The DPPH radical had a maximum absorbance at 517 nm which
disappeared upon reduction by an antiradical compound. BHA was
used as positive control. The DPPH• radical scavenging activity was
calculated as follows:
Acontrol − Asample
DPPH • radical scavenging activity (%) = × 100
Acontrol
Where Acontrol is the absorbance of the control reaction which con-
tains all reagents except the sample and Asample is the absorbance
of the sample with the DPPH solution. All experiments were done
in triplicate for each pectin solution.
2.5.2. ABTS+ radical scavenging activity
The ABTS+ radical scavenging activity of MC, PC and TFC
was determined according to the method detailed by Braca
et al. [25] with some modifications. The ABTS radical cation
was the result of the reaction between 7 mM ABTS (2,2 -azino-
bis (3-ethylbenzothiazoline-6-sulphonic acid) reagent in H2 O and
2,45 mM potassium persulfate stored in obscurity at room tem-
perature for 12 h. The mixture was then diluted to the absorbance
of 0.70 ± 0.02 at 734 nm by ethanol. For this test, different con-
centrations (1–10 mg/ml) of pectin solutions were used and 0.5 ml
samples were mixed with 1 ml ABTS radical. After 6 min incubation
at room temperature in obscurity, the absorbance of the mixture
was measured at 734 nm. Trolox was used as positive control. The
ABTS+ radical scavenging activity was calculated with the same
formula of the DPPH• radical scavenging activity.
2.5.3. ˇ-Carotene bleaching inhibition activity
The abilities of the carbohydrates to prevent the bleaching of ␤-
Carotene were determined as described by Koleva et al. [26]. 0.5 g
of ␤-Carotene, 25 ␮l of linoleic acid and 200 ␮l of Tween 40 were
dissolved in 1 ml of chloroform which was then completely evap-
orated under vacuum in rotary evaporator at 40 ◦ C and 100 ml of
bi-distilled water were added. The resulting mixture was vigor-
ously stirred until obtaining an emulsion. The aliquots (2.5 ml) of
the emulsion which had to be freshly prepared were added to 0.5 ml
of each sample at different concentrations (1–20 mg/ml) and fol-
lowed by incubation for 2 h in dark and at 50 ◦ C. The absorbance of
443
each sample was measured at 470 nm. A control contained 0.5 ml
of distilled water instead of sample. BHA was used as a reference
antioxidant. The ␤-Carotene bleaching inhibition activity was eval-
uated by the formula:
 
(A0 − At )
ˇ − Carotene bleaching inhibition activity = 1 − × 100
(A 0 − A t )
Where:
A0 and A’0 were the absorbance, respectively, of the sample and
the control at time zero, At and A’t were the absorbance, respec-
tively, of the sample and the control after incubation for 2 h.
2.5.4. Reducing power assay
The reducing power assay was determined with reference to the
detailed method of Zhu et al. [27] with some modifications. 0.5 ml
of each sample at different concentrations (1–20 mg/ml) was mixed
with 1.25 ml of 0.2 M phosphate buffer (pH 6.6) and 1.25 ml of 1%
(w/v) potassium ferricyanide solution. The mixture was then incu-
bated at 50 ◦ C for 30 min. After incubation, 1.25 ml of 10% (w/v) TCA
was added and 1 ml of the resulted mixture was centrifuged for
10 min at 10000 rpm. The supernatant solution from each sample
mixture was then mixed with 1 ml of distilled water and 0.25 ml
of 0.1% (w/v) ferric chloride. After 10 min of reaction time, the
absorbance was measured at 700 nm. BHA was used as a positive
control. Higher absorbance of reaction samples indicated higher
reducing power.
2.6. Statistical analysis
All tests were carried out in triplicate and expressed as
mean ± standard deviation (SD). Data were analyzed by one-way
analysis of variance (ANOVA) using the SPSS 21.0 statistical soft-
ware program and deviations were significant at P <0.05.
3. Results and discussion
3.1. Extraction yields
The effect of the acid process on the yield extract of the three
water-soluble hydrocolloids was investigated. The obtained results
(Table 1) showed that the extraction yield of MC (10.24% of dry
weight) was more important than that of PC (6.13% of dry weight).
Indeed, the TFC extraction yield was lower than the sum of MC
and PC yields. This could be due to the extraction conditions. As
compared to the study of Càrdenas et al. [28], the PC extraction
yield was relatively similar to that afforded by alkaline process (6%
of dry weight). Indeed, MC yield was greater than that from the
skin of prickly pear fruits which represented 4.1% of dry weight
[13]. Kaewmanee et al. [7] reported the research of Barbary et al.
[29] who used the extraction of the flaxseed mucilage at 25 ◦ C from
1 h to 8 h. It is about 3% to 5.2%.The concentration of mucilage and
pectin in Opuntia spp. was influenced by the cultivar, cultivation
site, climatic conditions and cladode order [30,31].
Moreover, TFC yield was higher than that for the nopal pads
in which the yield reached 11.7% of dry weight [31]. Furthermore,
the results were conform to those found by Goldstein et al. [32]
and reported by Sepùlveda et al. [31] who mentioned that the pec-
tic mucilage fraction yield ranged from 9% to 19% in dry weight
in relation to the climatic conditions at the moment of cladodes
collection.
3.2. Moisture and ash
The moisture of the three OFI cladode extracts fluctuates
between 9.67% and 10.66% without any significant difference
444 N. Bayar et al. / International Journal of Biological Macromolecules 92 (2016) 441–450
Table 1
Yield, chemicals analysis and average molecular weight of cladodes extracts.
Sample Yield (% of Moisture Ash content
dry weight) (%) (%dry weight)
TFC 13.12 ± 2.19a 10.66 ± 0.73a 13.78 ± 1.15a
PC 6.13 ± 0.60a 9.67 ± 1.44a 5.08 ± 0.13b
MC 10.24 ± 0.69a 10.25 ± 2.62a 35.52 ± 0.41c
The underline represents the average molecular weights which correspond to the major peaks.
a,b,c
Values within a column followed by different letters are significantly different at P < 0.05 using Duncan’s multiple range test.
(p < 0.05) (Table 1). This relatively high moisture may be due to
the conditions of analysis and drying.
As shown in Table 1, the three polysaccharide fractions present
high ash content which can be related to the high ash content of
the starting raw materials and the absence of the step of dialysis
in the extraction process. Moreover, MC has the highest ash con-
tent compared to PC and TFC (p < 0.05). This result can be related
to the extraction temperature. In fact, MC was extracted at 25 ◦ C
versus 90 ◦ C for PC and TFC. Wang et al. [33] demonstrated that
ash content decreased with the increase of extraction temperature.
This content (35.52%) was much higher than that of the mucilage
of OFI cladodes from Marocco which reached 19.6% [34,31] but it
was approximately similar to those for the mucilage in Opuntia spp.
which ranged from 34.9% to 39.1% [31].
PC exhibited lower ash content than TFC although they were
extracted under the same conditions. This could be explained by the
fact that PC was extracted after the extraction of MC. So, the highest
ash proportion was extracted with mucilage. It was also lower than
that of potato pulp pectin which was extracted by alkaline process
(10.2%) and commercial citrus pectin (6.6%) [35,28].
3.3. Protein
In our study, the protein content of the three polysaccharides
extracted from OFI cladodes were evaluated (Table 1). The results
showed low protein content in all fractions. The lowest value being
0.29% was found in the pectin fraction, while the highest being
0.83% was found in the mucilage fraction. The contamination of the
polysaccharides with water soluble protein and its content were
related to the extraction conditions. In fact, Jouki et al. [36] demon-
strated that the protein content increased when the temperature
extraction of queen seed mucilage increased from 25 ◦ C to 55 ◦ C. In
that way, we could explain the low content of protein in the three
fractions by the fact that mucilage was extracted at room temper-
ature whereas pectin and the total fraction were extracted at high
temperature (90 ◦ C). These results were lower than those for the
Parkia speciosa pod pectin and the commercial citrus pectin which
reached 1.1% at pH 7 and 2.0%, respectively [21]. Gan et al. [21]
proved that protein content was also related to the extraction pH
of polysaccharides. By this means, we could explain the difference
in the protein content between mucilage and pectic fractions see-
ing that the mucilage was extracted at the pH of approximately 5.3
but pectin was extracted at pH 2.8. Conversely, the protein content
is high when the pH of extraction was near the isoelectric point.
The protein content of TFC was much lower than that found by
Sepùlveda et al. [31] (6.7% to 7.5%). This difference could be related
to the extraction conditions, the starting raw materials and the
method used for protein quantification (Bradford test in our study
against Kjeldhal in the reported study).
Protein content Total sugars content Uronic acid content Average molecular
(% dry weight) (% dry weight) (% dry weight) weight (kDa)
0.32 ± 0.01a 85.31 ± 1.77a 60.66 ± 7.02a 93.56 ± 2.86a
80.00 ± 1.30
49.62 ± 9.28
0.47 ± 0.02b 93.81 ± 0.48b 79.82 ± 0.25b 110.70 ± 3.37
78.40 ± 5.49
58.57 ± 4.31b
0.92 ± 0.03c 63.45 ± 1.75c 11.64 ± 0.51c 52.78 ± 4.77b
3.4. Total sugar and uronic acid
The total sugar content was determined using the phenol-
sulfuric method and the results were presented in Table 1. PC had
a significantly higher content of carbohydrates than MC and TFC
(p < 0.05) which was mainly composed of acidic sugar as uronic
acid.
Uronic acid content was performed using the carbazole test. The
results mentioned in Table 1 show that the three fractions have a
considerably different uronic acid content (p < 0.05). PC was rich in
uronic acid with a percentage of 79.82%. This content was beyond
the recommended limits (65%) by the European Union and the FAO
[37]. Furthermore, it was higher than that of the Opuntia ficus indica
peel pectin (64.5%) [13]. The uronic acid content of TFC was slightly
lower than that for the prickly pear peel pectin (64.30%) [38].
Cladodes mucilage is composed of 11.64% of uronic acid which
was significantly lower than that of PC and TFC. This content was
equivalent to that of the mucilage of OFI (8–12.7%) [39–41], but it
was lower than that of the fruit of OFI which was 23.4% [42].
3.5. Average molecular weight
The molecular weight distribution of the three fractions (Fig. 1)
was investigated by the gel filtration high performance liquid chro-
matography coupled with the refractive index detector (RID). MC
showed a major peak with an average molecular weight of approx-
imately 52.8 kDa (Table 1) whereas the molecular weight profiles
of both PC and TFC samples contained each two major peaks which
were determined to be around 58.6 and 110.7 kDa for PC and 49.6
and 93.6 kDa for TFC. In addition, other minor peaks were essen-
tially observed for PC and TFC. Yet, the most pronounced peaks
were determined to be around 78.4 kDa and 79.1 kDa, respectively.
This large distribution of molecular weight of PC and TFC could be
caused by the acid hydrolysis at time of extraction. It could also be
due to the degradation of the molecules by the high temperature
used for their dissolution.
3.6. Fourier transform infrared spectroscopy
FTIR spectrum is one of the most used techniques to analyze
the structure of polymers and to determine their major functional
groups. The infrared spectra of the three fractions extracted from
cladodes are shown in Fig. 2. The region of 3270–3286 cm−1 rep-
resents the stretching of hydroxyl groups. Peaks at 2918.56 cm−1
for PC and 2919.89 cm−1 for FTC correspond to the symmetric
stretching vibrations of the CH group of the methyl ester of galac-
turonic acid [43]. This band reflects the binding site for ions
which can interact with water and facilitate the gelling capac-
ity of the fractions [44]. Peak at 1253.27 for MC indicate the
presence of o-acetyl groups [45]. The area between 800 and
1200 cm−1 is the region for carbohydrates fingerprint and it allows
the identification of some functional groups characteristics of
 N. Bayar et al. / International Journal of Biological Macromolecules 92 (2016) 441–450 445

Fig. 1. Gel filtration chromatographs of MC (A), PC (B) and TFC (C).

polysaccharides such as stretching (C O C), bending (O H) and
deforming (CH3 ) vibrations [46]. The bands at 851.41 cm−1 and
841.55 cm−1 for PC and MC, respectively, indicated that the
major linkages were ␣-glycosidic [47]. Peaks at 1651.06 cm−1 ,
1621.24 cm−1 and 1567.89 cm−1 for TFC, PC and MC, respectively,
were attributed to the stretching vibration of free carboxylic
groups. The vibration at 1731.33 cm−1 for TFC was assigned to the
esterified groups which allowed the estimation of the degree of
methylation of pectin and the peak at 1409.65 cm−1 for MC which
referred to the presence of free carboxylic acids [37]. The absorp-
tion peaks at 1146.70 and 1012.91 cm−1 for TFC and the peaks at
1148.43 and 1015.53 cm−1 for PC were attributed to the C O H
bonds of primary alcoholic and d-arabinose, respectively [48,44].
3.7. Functional properties
3.7.1. Water holding capacity
Water holding capacity is a functional property that indicates
the capability of samples to include water. Mucilage and pectin
showed important water holding capacities as shown in Table 2
which were, respectively, 7.81 g/g and 5.64 g/g; but which were
lower than that displayed by the carraghenane (9.79 ± 0.06 g/g).
446 N. Bayar et al. / International Journal of Biological Macromolecules 92 (2016) 441–450

Fig. 2. FTIR spectra of MC (A), PC (B) and TFC (C).

Table 2 They were also lower than that reported by Sciarini et al. [49] for
Water and oil holding capacities of cladodes extracts (g water or oil/g wet sample).
galactomannans (15.20 g H2 O/g sample). WHC of MC was higher
MC PC TFC than that for the cactus fiber isolate which was 7.1 g/g [50,39].
WHC 7,81 ± 1,12a 5,64 ± 0,30b 2,24 ± 0,22c The total pectic mucilage fraction had lower WHC than MC and
OHC 1,34 ± 0,05a 1,24 ± 0,13ab 0,62 ± 0,04b PC. We can conclude that the interaction between mucilage and
a,b,ab,c
Values within a column followed by different letters are significantly different
pectin has a negative effect on WHC. The greatest WHC of MC and
at P < 0.05 using Duncan’s multiple range test. PC could be due to the high concentration of free hydroxyl groups
 N. Bayar et al. / International Journal of Biological Macromolecules 92 (2016) 441–450
Fig. 3. (A) Foam capacity (FC), (B) Foam stability (FS), (C) Emulsion capacity (EC) and (D) Emulsion stability (ES).
in their structures [51] and high content of calcium [52,31]. This
finding implied the possibility of its use as a potential ingredient in
many food products to avoid syneresis.
In addition to the source and the chemical structure, water hold-
ing capacity of dietary fibers such as pectin and mucilage is related
to the ionic strength and ion form present in solution, pH, temper-
ature, particle size and porosity of samples [53].
3.7.2. Oil holding capacity
OHC was determined for the three fractions extracted from
OFI cladodes. Table 2 revealed that MC gave a higher OHC value
(1.34 g/g) than PC (1.24 g/g) and TFC (0.62 g/g). However, these
finding were lower than those for the Parkia speciosa pod pectin
and commercial citrus pectin [21] in which OHC values (at pH
7) were 3.9 g/g and 2.1 g/g, respectively. They were, also, lower
than the OHC value displayed by cactus fiber isolate which was
4.5 g/g [50]. Elleuch et al. [53] suggested that OHC could be related
to the hydrophilic character and to the overall charge density of
constituents.
3.7.3. Foaming properties
Foam is the dispersion of gas bubbles in water. So, it requires
the presence of surface active compounds which can adsorb at the
water-air interface. The foam capacity (FC) and the foam stability
(FS) of the three fractions extracted from cladodes (TFC, PC and
447
MC) at 2% of concentration are shown in Fig. 2A and B. The anal-
ysis of the data indicated that TFC and PC had high FC and FS. In
contrast, MC was able to form foam. PC had higher FC and lower
FS than those given by TFC. This can be explained by the fact that
TFC is a mixture of pectin and mucilage. Furthermore, mucilage is
more hydrophilic than pectin so it has a low tendency to adsorb
at the air-water interface which reduces the foaming capacity but
increases the viscosity and the stability of foam [54,7]. Generally,
gums such as mucilage are not known as surfactants; however, they
can increase the viscosity of solution and prevent gas-bubbles coa-
lescence in food preparation [54]. Consequently this fraction causes
an increase in the foaming stability of TFC compared to PC. Sàenz
et al. [39] studied the effect of the addition of cladodes mucilage
dispersions (0.5% and 0.8%) to eggs foams and found a decrease in
syneresis and an increase in foam volume.
3.7.4. Emulsifying properties
Emulsion is a mixture of two normally immiscible liquids. Emul-
sifying properties include two parameters: emulsion capacity (EC)
and emulsion stability (ES). These properties were one of the most
considerable functionalities of pectin. In this study, the different
emulsions were prepared with 2% (w/v) polysaccharides-water
solutions. The results are presented in Fig. 2(C and D). EC of PC
was 35% which was slightly higher than that of sugar beet pectin
extracted by hydrochloric acid (33.5%) [55] and was similar to the
448 N. Bayar et al. / International Journal of Biological Macromolecules 92 (2016) 441–450
Fig. 4. Antioxidants activities: (A) DPPH radical scavenging activity; (B) ABTS radical scavenging activity; (C) ␤-carotene bleaching inhibition activity; (D) Reducing power.
EC of citrus pectin (35%) [56]. Moreover, PC emulsion was 90.45%
stable after 30 min of incubation at 80 ◦ C. Ma et al. [55] indicated
that emulsion is more stable at low temperature which confirmed
the greater ability of PC to stabilize oil-in-water emulsion.
Cladodes mucilage displayed the lowest emulsion capacity
(15%). Nonetheless, MC showed great ES after 30 min of incuba-
tion at 80 ◦ C (65.28%), but, it was lower than that of quince seeds
mucilage which was more than 80% [36] and than that of flax
mucilage which exceeded 80% [7]. In fact, mucilage stabilized oil-
in-water emulsion by forming a layer around each oil droplet and
their hydrophilic character delayed the coalescence of globules
[51]. This stability could be due to the low extraction temperature
of mucilage (less than 45 ◦ C) and the contamination of extract frac-
tion by other constituents such as ferulic acid, proteins and minerals
[57,36].
EC displayed by TFC was 16.67% which was lower than that of
PC and higher than that of MC. Besides, TFC was able to stabilize
emulsion less than PC but slightly higher than MC.
The functional properties of polysaccharides were largely
related to their structure. Indeed, the presence of protein and fer-
ulic acid was responsible for the increase of emulsion capacity. In
fact, they decreased the interfacial tension between water and oil
droplet due to their hydrophobicity [58,59]. In addition, the pres-
ence of arabinose and galactose in the lateral chains (which link the
main chains to the ferulic acid and protein) had a positive effect on
the emulsion properties [58,59]. Concerning the emulsion stability,
it decreased with the decrease of molecular weight; but increased
with the increase of the concentration of galacturonic acid which
formed, with the side chain, a network resulting in the prevention
of oil droplets coalescence [60]. Methyl and acetyl groups, in the
main chain, had a positive effect on emulsion stability [59].
3.8. In vitro antioxidants activities
Antioxidants had different mechanisms of action. So, it was
necessary to combine tests in order to determine in vitro the antiox-
idant ability of compounds [61].
In this paper, we used four methods (DPPH, ABTS, reducing
power and ␤-carotene) to test the antioxidant activities in three
polysaccharides fractions extracted from OFI cladodes.
3.8.1. DPPH radical scavenging activity
DPPH• radical scavenging activity is one of the different tests
used for estimating the antioxidant activity of natural products. Its
action mechanism was based on the donation of hydrogen which
resulted in the reduction of DPPH on DPPH-H and the inhibition
of the propagation of oxidizing reaction. DPPH• radical scaveng-
ing activity of MC, PC and TFC increased in a dose-dependent
manner as presented in Fig. 3A. MC and TFC had a good antiox-
idant capacity and the highest DPPH• radical scavenging activity
 N. Bayar et al. / International Journal of Biological Macromolecules 92 (2016) 441–450
was 100% and 93.87%, respectively, at 20 mg/ml; however, it was
33.87% for PC. The IC50 of MC and TFC were, respectively, 4.6 mg/ml
and 5.6 mg/ml. At the some concentration level, PC had a lower
DPPH• radical scavenging activity than MC and TFC. These results
could be explained by the diffusion of the antioxidants compounds
into the mucilage fraction. On the other hand, the antioxidants
compounds were thermo-sensitive. So, the further increase of tem-
perature extraction decreased the antioxidant activities [36]. DPPH
radical scavenging activity of 2% of TFC was lower than that found
by Chaouch et al. [62] with pectic mucilage fraction extracted by
ultrasound (10 min, 100 W) at the same concentration level (30.79%
against 37.33%). The DPPH• radical scavenging activity of the three
fractions was lower than that displayed by BHA at all concentra-
tions tested. The results mentioned above implied that MC and
TFC can replace synthetics antioxidants in the food preparation as
natural antioxidants (Fig. 4).
3.8.2. ABTS radical scavenging activity
The ABTS+ radical scavenging activity was also studied in order
to further verify and compare the antioxidant activity of MC, PC
and TFC. As shown in Fig. 3B, the three fractions had high antiox-
idant activity in a dose-dependent manner which exceeded 60%
at 10 mg/ml of concentration. In fact, MC showed more important
ABTS+ radical scavenging activity than those displayed by PC and
TFC and the highest values were 100% for MC against 72.79% and
68.08% for TFC and PC, respectively. The result displayed by PC was
lower than those of the pectin extracted by subcritical water from
apple pomace (99.65%) and citrus peels (98.49%) at the concen-
tration of 4.5 mg/ml [33]. The cladodes extracts had lower ABTS+
radical scavenging activities than the trolox. They can be consid-
ered as a good natural antioxidant to scavenge radical such as DPPH
and ABTS by donating hydrogen atoms or electrons.
3.8.3. ˇ-Carotene bleaching inhibition activity
The ␤-Carotene bleaching inhibition activity of MC, PC and TFC
was investigated. The mechanism of this test was based on generat-
ing peroxyl radicals by the linoleic acid. The free radicals produced
could react with the ␤-carotene chromophore resulting in a bleach-
ing effect but, this reaction can be inhibited by the free radicals
scavenger [19].
As shown in Fig. 3C, the antioxidant activities of the three
polysaccharides increased with the concentration. The higher val-
ues were 94.89% for MC, 69.91% for PC and 73.47% for TFC at
20 mg/ml. However, these results were lower than those displayed
by BHA. The IC50 of MC, TFC and PC were 2.8 mg/ml, 6.8 mg/ml
and 11 mg/ml, respectively. This data implied that all fractions pre-
sented a good antioxidant activity.
According to Zeng et al. [47], the antioxidant activities of the
three cladodes extracts could be caused by the hydroxyl and
carboxylate groups which transferred electron with radicals. Fur-
thermore, these activities could be enhanced by the low average
molecular weight [62]. Mengome et al. [63] demonstrated that
the homogalacturonic chain and the methylation degree were not
involved in the ability of pectin to scavenge free radicals. However,
they proposed that this activity may be related to the presence of
other molecules which were linked to the carbohydrates through
an ester function.
3.8.4. Reducing power
The reducing power of compounds is related to the presence
of reductones which can donate a hydrogen atom to free radicals
in order to convert them to stable products and break the oxidiz-
ing chain reaction. The reductones can also prevent the peroxide
formation by reacting with some precursors of peroxide [64,19].
According to the results presented in Fig. 3D MC, PC
and TFC exhibited reducing powers which increased with the
449
concentration. As the previous tests, MC had stronger reducing
power than TFC and PC. However, the three samples had antioxi-
dant activities much lower than BHA used as a reference. Mengone
et al. [63] proved that the Fe3+ reducing activity was not due to the
general structure of polysaccharides but rather to specific domains
or motifs. So, the fractionation of molecules should be carried out
in order to define these motifs. They also suggested that the scav-
enging and the reducing activity of pectin didn’t have the same
target.
4. Conclusion
This paper was undertaken to isolate three polysaccharides (MC,
PC and TFC) from the OFI cladodes using chemical methods. We
tried to compare their physicochemical, functional and antioxidant
properties. The three fractions had relatively high moisture and
low protein content. Contrary to PC, MC was rich with ash but had
low uronic acid content. PC had stronger foaming and emulsify-
ing properties and lower water and oil holding capacities than MC.
Furthermore, these carbohydrates, essentially the mucilage frac-
tion, showed greater capacities to scavenge radicals such us DPPH
and ABTS and also to inhibit the bleaching of ␤-carotene. However,
they had relatively low reducing power. These findings implied
that cladodes extracts could be applied as a surfactant or a nat-
ural antioxidant in several food and drug formulations. In addition,
we demonstrated that the extraction of mucilage and pectin can be
a novelty process to produce fractions with specific activities useful
for determinant applications.
Acknowledgements
« This research and innovation work was carried out under the
MOBIDOC project funded by the European Union under the PASRI
program administered by the ANPR». The authors would like to
express their sincere gratitude to Mrs Salma KARRAY for her con-
structive proofreading and valuable language polishing services.
References
[1] H. Han, P. Felker, Field validation of water use efficiency of CAM plant Opuntia
ellisiana in south Texas, J. Arid Environ. 36 (1997) 133–148.
[2] H.N. Le Houérou, Cacti (Opuntia spp.) as a fodder crop for marginal lands in
the Mediterranean basin. Proc. 4th IC on Cactus pear and cochineal, Acta Hort.
581 (2002) 21–46.
[3] J.C. Dubeux, Use of Cactus for Livestock Feeding, UFRPE, Brazil, 2011.
[4] F.C. Stintzing, R. Carle, Cactus stems (Opuntia spp.): a review on their
chemistry, technology and uses, Mol. Nutr. Food Res. 49 (2005) 175–194.
[5] U.M. Deogade, V.N. Deshmukh, D.M. Sakarkar, Natural gums and mucilage’s in
NDDS: Applications and recent approaches, Int. J. PharmTech. Res. 4 (2012)
799–814.
[6] C.M.O. Simões, E.P. Schenkel, G. Gosmann, J.C.P. Mello, L.A. Mentz, P.R.
Petrovick, Farmacognosia: da Planta ao Medicamento, 6th ed., UFRGS Editora,
Florianópolis: Editora da UFSC, Porto Alegre, 2007.
[7] T. Kaewmanee, L. Bagnasco, S. Benjakul, S. Lanteri, C.F. Morelli, G. Speranza,
M.E. Cosulich, Characterisation of mucilages extracted from seven Italian
cultivars of flax, Food Chem. 148 (2014) 60–69.
[8] S. Wang, F. Chen, J. Wu, Z. Wang, X. Liao, X. Hu, Optimization of pectin
extraction assisted by microwave from apple pomace using response surface
methodology, J. Food Eng. 78 (2007) 693–700.
[9] B.L. Ridley, M.A. O’Neill, D.A. Mohnen, Pectins Structure, biosynthesis, and
oligogalacturonide-related signaling, Phytochemistry 57 (2001) 929–967.
[10] D. Mohnen, Pectin structure and biosynthesis, Curr. Opin. Plant Biol. 11 (2008)
266–277.
[11] L. Partos, A First Pectin Ingredient Enters Cargill’s Portfolio., 2006, http://
www.dairyreporter.com/news/ng.asp?id=61305, 2006 (accessed 22.07.06).
[12] H. Bagherian, F.Z. Ashtiani, A. Fouladitajar, M. Mohtashamy, Comparisons
between conventional, microwave and ultrasound-assisted methods for
extraction of pectin from grapefruit, Chem. Eng. Process. 50 (2011)
1237–1243.
[13] Y. Habibi, A. Heyraud, M. Mahrouz, M.R. Vignon, Structural features of pectic
polysaccharides from the skin of Opuntia ficus-indica prickly pear fruits,
Carbohyd. Res. 339 (2004) 1119–1127.
[14] AOAC (Association of Official Analytical Chemists), Method of Analysi
Gaithersburg, AOAC International, Gaithersburg, MD, 2000.
450 N. Bayar et al. / International Journal of Biological Macromolecules 92 (2016) 441–450
[15] M.M. Bradford, A rapid and sensitive method for the quantitation of
microgram quantities of protein utilizing the principle of protein-dye binding,
Anal. Biochem. 72 (1976) 248–254.
[16] M. Dubois, K.A. Gilles, J.K. Hamilton, P.A. Rebers, F. Smith, Colorimetric
method for determination of sugars and related substances, Anal. Chem. 28
(1956) 350–356.
[17] T.M.C.C. Filisetti-Cozzi, N.C. Carpita, Measurement of uronic acids without
interference from neutral sugars, Anal. Biochem. 197 (1991) 157–162.
[18] M.H.Y. Lin, E.S. Humbert, F.W. Sosulki, Certain functional properties of
sunflower meal products, J. Food Sci. 39 (1974) 368–370.
[19] A. Sila, N. Bayar, I. Ghazela, A. Bougatef, R. Ellouz-Ghorbel, S.
Ellouz-Chaabouni, Water-soluble polysaccharides from agro-industrial
by-products: functional and biological properties, Int. J. Biol. Macromol. 69
(2014) 236–243.
[20] D. Betancur-Ancona, G. Peraza-Mercado, Y. Moguel-Ordonez, S.
Fuertes-Blanco, Physicochemical characterization of lima bean (Phaseolus
lunatus) and Jack bean (Canavalia ensiformis) fibrous residues, Food Chem. 84
(2004) 287–295.
[21] C.Y. Gan, N. Hj Abdul Manaf, A.A. Latiff, Physico-chemical properties of alcohol
precipitate pectin-like polysaccharides from Parkia speciosa pod, Food
Hydrocolloid. 24 (2010) 471–478.
[22] F. Shahidi, X.Q. Han, J. Synowiecki, Production and characteristics of protein
hydrolysates from capelin, Food Chem. 53 (1995) 285–293.
[23] S.W. Cui, Y.H. Chang, Emulsifying and structural properties of pectin
enzymatically extracted from pumpkin, LWT−Food Sci. Technol. 58 (2014)
396–403.
[24] P. Bersuder, M. Hole, G. j. Smith, Antioxidants from a heated histidine-glucose
model system I: investigation of the antioxidant role of histidine and isolation
of antioxidants by high performance liquid chromatography, J. Am. Oil Chem.
Soc. 75 (1998) 181–187.
[25] A. Braca, N. De Tommasi, L. Di Bari, C. Pizza, M. Politi, I. Morelli, Antioxidant
principles from Bauhinia tarapotensis, J. Nat. Prod. 64 (2001) 892–895.
[26] I.I. Koleva, T.A. Van Beek, J.P.H. Linssen, A. de Groot, L.N. Evstatieva, Screening
of plant extracts for antioxidant activity: a comparative study on three testing
methods, Phytochem. Anal. 13 (2002) 8–17.
[27] K. Zhu, H. Zhou, H. Qian, Antioxidant and free radical-scavenging activities of
wheat germ protein hydrolysates (WGPH) prepared with alcalase, Process
Biochem. 41 (2006) 1296–1302.
[28] A. Càrdenas, F.M. Goycoolea, M. Rinaudo, On the gelling behaviour of ‘nopal’
(Opuntia ficus indica) low methoxyl pectin, Carbohydr. Polym. 73 (2008)
212–222.
[29] O.M. Barbary, S.A. Al-Sohaimy, M.A. El-Saadani, A.M.A. Zeitoun, Extraction,
composition and physicochemical properties of flaxseed mucilage, J. Adv.
Agric. Res. 14 (2009) 605–620.
[30] J.H. Kim, H.J. Lee, Y. Park, K.S. Ra, K.S. Shin, K.W. Yu, H.J. Suh, Mucilage removal
from cactus cladodes (Opuntia humifusa Raf.) by enzymatic treatment to
improve extraction efficiency and radical scavenging activity, LWT−Food Sci.
Technol. 51 (2013) 337–342.
[31] E. Sepùlveda, C. Sàenz, E. Aliaga, C. Aceituno, Extraction and characterization
of mucilage in Opuntia spp, J. Arid. Environ. 68 (2007) 534–545.
[32] G. Goldstein, J.L. Andrade, P. Nobel, Differences in water relations parameters
for the chlorenchyma and parenchyma of Opuntia ficus indica under wet
Versus dry conditions, Aust. J. Plant Physiol. 18 (1991) 95–107.
[33] X. Wang, Q. Chen, X. Lü, Pectin extracted from apple pomace and citrus peel
by subcritical water, Food Hydrocolloid. 38 (2014) 129–137.
[34] M.E. Malainnine, A. Dufresne, D. Dupeyre, M. Mahrouz, R. Vuong, M. Vignon,
Structure and morphology of cladodes and spines of Opuntia ficus indica.
Cellulose extraction and characterisation, Carbohydr. Polym. 51 (2003) 77–83.
[35] T. Turquois, M. Rinaudo, F.R. Taravel, A. Heyraud, Extraction of highly gelling
pectic substances from sugar beet pulp and potato pulp: influence of extrinsic
parameters on their gelling properties, Food Hydrocolloid. 13 (1999) 255–262.
[36] M. Jouki, S.A. Mortazavi, F.T. Yazdi, A. Koocheki, Optimization of extraction,
antioxidant activity and functional properties of quince seed mucilage by
RSM, Int. J. Biol. Macromol. 66 (2014) 113–124.
[37] A.L. Lira-Ortiz, F. Reséndiz-Vega, E. Ríos-Leal, J.C. Contreras-Esquivel, N.
Chavarría-Hernández, A. Vargas-Torres, A.I. Rodríguez-Hernández, Pectins
from waste of prickly pear fruits (Opuntia albicarpa Scheinvar ‘Reyna’):
chemical and rheological properties, Food Hydrocolloid. 37 (2014) 93–99.
[38] E. Forni, M. Penci, A. Polesello, A preliminary characterization of some pectin
from quince fruit (Cydonia oblonga Mill.) and prickly pear (Opuntia ficus indica)
peel, Carbohydr. Polym. (1994) 231–234.
[39] C. Sàenz, E. Sepùlveda, B. Matsuhiro, Opuntia spp. mucilage’s: a functional
component with industrial perspectives, J. Arid. Environ. 57 (2004) 275–290.
[40] S. Trachtenberg, A.M. Mayer, Composition and properties of Opuntia ficus
indica mucilage, Phytochemistry 20 (1981) 2665–2668.
[41] P. Nobel, J. Cavelier, J.L. Andrade, Mucilage in cacti: its apoplastic capacitance,
associated solutes, and influence on tissue water relations, J. Exp. Bot. 43
(1992) 641–648.
[42] B. Matsuhiro, L.E. Lillo, C. Sáenz, C.C. Urzúa, O. Zárate, Chemical
characterization of the mucilage from fruits of Opuntia ficus indica,
Carbohydr. Polym. 63 (2006) 263–267.
[43] D. Gopi, K. Kanimozhi, L. Kavitha, Opuntia ficus indica peel derived pectin
mediated hydroxyapatite nanoparticles: synthesis spectral characterization,
biological and antimicrobial activities, Spectrochim. Acta Part A: Mol. Biomol.
Spectrosc. 141 (2015) 135–143.
[44] S.M.A. Razavi, S.W. Cui, Q. Guo, H. Ding, Some physicochemical properties of
sage (Salvia macrosiphon) seed gum, Food Hydrocolloid. 35 (2014) 453–462.
[45] P. Khonkar, Composition and partial structure characterization of Tremella
polysaccharides, Mycobiology 37 (2009) 286–294.
[46] Y. Jiang, Y. Du, X. Zhu, H. Xiong, M.W. Woo, J. Hu, Physicochemical and
comparative properties of pectins extracted from Akebia trifoliata var.
australis peel, Carbohydr. Polym. 87 (2012) 1663–1669.
[47] H. Zeng, S. Miao, Y. Zhang, S. Lin, Y. Jian, Y. Tian, B. Zheng, Isolation,
preliminary structural characterization and hypolipidemic effect of
polysaccharide fractions from Fortunella margarita (Lour.) Swingle, Food
Hydrocolloid. 52 (2016) 126–136.
[48] M. Cerná, A.S. Barros, A. Nunes, S.M. Rocha, I. Delgadillo, J. Copıkova, M.A.
Coim-bra, Use of FT-IR spectroscopy as a tool for the analysis of
polysaccharide food additives, Carbohydr. Polym. 51 (2003) 383–389.
[49] L.S. Sciarini, F. Maldonado, P.D. Ribotta, G.T. Pérez, A.E. León, Chemical
composition and functional properties of Gleditsia triacanthos gum, Food
Hydrocolloid. 23 (2009) 306–313.
[50] J.L. Rosado, M. Dîaz, Physicochemical properties related to gastrointestinal
effects of six dietary fibers, Rev. Invest. Clin. 47 (1995) 283–289.
[51] V.D. Prajapati, G.K. Jani, N.G. Moradiya, N.P. Randeria, Pharmaceutical
applications of various natural gums, mucilages and their modified forms,
Carbohydr. Polym. 92 (2013) 1685–1699.
[52] S.H. Trachtenberg, A. Mayer, Mucilage cells, calcium oxalate crystals and
soluble calcium in Opuntia ficus indica, Ann. Bot. 50 (1982) 549–557.
[53] M. Elleuch, D. Bedigian, O. Roiseux, S. Besbes, C. Blecker, H. Attia, Dietary fibre
and fibre-rich by-products of food processing: characterisation, technological
functionality and commercial applications: a review, Food Chem. 124 (2011)
411–421.
[54] E.A. Makri, G.I. Doxastakis, Surface tension of Phaseolus vulgaris and
coccineus proteins and effect of polysaccharides on their foaming properties,
Food Chem. 101 (2007) 37–48.
[55] S. Ma, S.J. Yu, X.L. Zheng, X.X. Wang, Q.D. Bao, X.M. Guo, Extraction,
characterization and spontaneous emulsifying properties of pectin from sugar
beet pulp, Carbohydr. Polym. 98 (2013) 750–753.
[56] R.H. Liang, L.H. Wang, J. Chen, W. Liu, C.M. Liu, Alkylated pectin: synthesis
characterization, viscosity and emulsifying properties, Food Hydrocolloid. 50
(2015) 65–73.
[57] C. Lv, Y. Wang, L.J. Wang, D. Li, B. Adhikari, Optimization of production yield
and functional properties of pectin extracted from sugar beet pulp,
Carbohydr. Polym. 95 (2013) 233–240.
[58] P.A. Williams, C. Sayers, C. Viebke, C. Senan, J. Mazoyer, P. Boulenguer,
Elucidation of the emulsification properties of sugar beet pectin, J. Agric. Food.
Chem. 53 (2005) 3592–3597.
[59] H.M. Chen, X. Fu, Z.G. Luo, Effect of molecular structure on emulsifying
properties of sugar beet pulp pectin, Food Hydrocolloid. 54 (2016) 99–106.
[60] T. Funami, M. Nakauma, S. Ishihara, R. Tanaka, T. Inoue, G.O. Phillips,
Structural modifications of sugar beet pectin and the relationship of structure
to functionality, Food Hydrocolloid. 25 (2011) 221–229.
[61] Q.Y. Zafra-Rojas, N. Cruz-Cansino, E. Ramírez-Moreno, L. Delgado-Olivares, J.
Villanueva- Sánchez, E. Alanís-García, Effects of ultrasound treatment in
purple cactus pear (Opuntia ficus-indica) juice, Ultrason. Sonochem. 20 (2013)
1283–1288.
[62] M.A. Chaouch, J. Hafsa, C. Rihouey, D. Le Cerf, H. Majdoub, Depolymerization
of polysaccharides from Opuntia ficus indica: antioxidant and antiglycated
activities, Int. J. Biol. Macromol. 79 (2015) 779–786.
[63] L.E. Mengome, A. Voxeur, J.P. Akue, P. Lerouge, Screening of antioxidant
activities of polysaccharides extracts from endemic plants in Gabon, Bioact.
Carbohydr. Diet. Fibre. 3 (2014) 77–88.
[64] G. Hikmet, A. Burhan, D. Gokhan, E. Selim, Y. Ismet, Antioxidant, free radical
scavenging and metal chelating characteristics of propolis, Am. J. Biochem.
Biotechnol. 1 (2005) 27–31.