Carbohydrate Polymers 140 (2016) 59–65

Contents lists available at ScienceDirect

Carbohydrate Polymers
journal homepage: www.elsevier.com/locate/carbpol

Optimization of microwave assisted extraction of pectin from sour

orange peel and its physicochemical properties
Seyed Saeid Hosseini, Faramarz Khodaiyan ∗ , Mohammad Saeid Yarmand
Bioprocessing and Biodetection Laboratory, Department of Food Science and Engineering, University of Tehran, Karaj 31587-77871, Iran

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

Article history: Microwave assisted extraction technique was used to extract pectin from sour orange peel. Box–Behnken
Received 9 October 2015 design was used to study the effect of irradiation time, microwave power and pH on the yield and degree
Received in revised form of esterification (DE) of pectin. The results showed that the optimum conditions for the highest yield of
17 November 2015
pectin (29.1%) were obtained at pH of 1.50, microwave power of 700 W, and irradiation time of 3 min. DE
Accepted 20 December 2015
values of pectin ranged from 1.7% to 37.5%, indicating that the obtained pectin was low in methoxyl. Under
Available online 23 December 2015
optimal conditions, the galacturonic acid content and emulsifying activity were 71.0 ± 0.8% and 40.7%,
respectively. In addition, the emulsion stability value ranged from 72.1% to 83.4%. Viscosity measurement
Keywords:
Sour orange
revealed that the solutions of pectin at low concentrations showed nearly Newtonian flow behavior, and
Pectin as the concentration increased, pseudoplastic flow became dominant.
Optimization © 2015 Elsevier Ltd. All rights reserved.
Degree of esterification

1. Introduction physicochemical properties and thus different applications (Chan

Pectin is a complex mixture of polysaccharides in cell walls of all
land plants, which is composed mainly of d-galacturonic acid and
neutral sugars, such as l-rhamnose, l-arabinose, and d-galactose
(Axelos, Thibault, & Lefebvre, 1989; Basanta, Ponce, Rojas, & Stortz,
2012; Pagan & Ibarz, 1999). The majority of the structure of pectin
contains a backbone of ˛(1 → 4) linked to d-galacturonic acid units
that are partially esterified with methanol or acetic acid at the car-
boxylic acid (Xu et al., 2014; Yapo, Robert, Etienne, Wathelet, &
Paquot, 2007).
The degree of esterification (DE; is defined as the percentage
of carboxyl groups esterified with methanol) of the pectin is an
important parameter for the definition of their application (Santos,
Espeleta, Branco, & de Assis, 2013; Seixas et al., 2014). Depend-
ing on DE, pectin is divided into two major groups: high methoxyl
pectin (HMP), with DE higher than 50% and low methoxyl pectin
(LMP), with DE lower than 50% (Thakur, Singh, Handa, & Rao, 1997;
Yapo et al., 2007). HMP form gels when heated in solutions with
a low pH (2–3.5) and at high concentrations of sugar (55–75%);
while LMP can gel in a wide pH range (2–6) with or without a
little amount of sugar in the presence of divalent ions such as cal-
cium (Ca2+ ) (Lofgren & Hermansson, 2007; Lopes da Silva & Rao,
2006; Wai, Alkarkhi, & Easa, 2010). HMP and LMP have different
∗ Corresponding author.
E-mail address: khodaiyan@ut.ac.ir (F. Khodaiyan).
& Choo, 2013). Pectin is used as a gelling agent, stabilizer, emulsi-
fier and thickener in the food industry for the production of jams
and jellies (Liu, Cao, Huang, Cai, & Yao, 2010; Thakur et al., 1997). In
addition, LMP may be used as a gelling agent in low calorie products
for diabetics and overweight patients (Gardner, Schwartz, Krista, &
Merimee, 1984; Wai et al., 2010).
The majority of sources for commercial pectin production are
citrus peels, apple pomace and sugar beet pulp (May, 1990;
Mesbahi, Jamalian, & Farahnaky, 2005). Also, pectin from non-
commercial sources such as cocoa husks (Chan & Choo, 2013),
mulberry branch bark (Liu et al., 2010), peach pomace (Pagan &
Ibarz, 1999), sisal waste (Santos et al., 2013), watermelon rind
(Maran, Sivakumar, Thirugnanasambandham, & Sridhar, 2014), soy
hull (Kalapathy & Proctor, 2001), pumpkin (Ptichkina, Markina, &
Rumyantseva, 2008) and banana peel (Qiu et al., 2010) have been
studied.
Citrus aurantium is the scientific name for a plant that is com-
monly called sour orange, bitter orange or seville orange (Haaz et al.,
2006). This fruit is one of the citrus fruits that exist in large quan-
tity in northern and southern Iran (Khormaei, Nasernejad, Edrisi,
& Eslamzadeh, 2007). Due to its sour and bitter taste, it is used
less often as edible fruit, but the fruit is more widely used as a
medicinal or dietary supplement (He, Lian, Lin, & Bernart, 1997).
Sour orange peel is a by-product of fruit juice factories, and it is
largely used as animal feed. The use of this waste as a new source for
the production of high value-added products such as pectin can be
valuable.

http://dx.doi.org/10.1016/j.carbpol.2015.12.051
0144-8617/© 2015 Elsevier Ltd. All rights reserved.
60
9.0
8.0
Yield of SOPP (%)
7.0
6.0
5.0
4.0
0.0 5.0 10.0 15.0 20.0 25.0 30.0
liquid-solid ratio (v/w)
Fig. 1. Effects of different LSR on extraction yield of SOPP with irradiation time of
2 min, microwave power of 500 W, pH of 2.25.
In the traditional extraction process, pectin is extracted using
hot water acidified with mineral acids (Xu et al., 2014). The use
of such strong acids in the extraction step leads to corrosion of
equipment and devices as well as damage to the environment.
In addition, the process is time consuming and leads to pectin
degradation (Seixas et al., 2014). The need for replacement of
this extraction method with newer and better methods could
be a potential means of minimizing the stated adverse effects.
Microwave assisted extraction is an interesting alternative to tra-
ditional extraction methods, and it has many advantages such
as shorter processing time, less solvent, higher extraction yield,
better quality products with lower production cost (Bagherian,
Ashtiani, Fouladitajar, & Mohtashamy, 2011; Kratchanova, Pavlova,
& Panchev, 2004).
The aim of this paper is to develop a microwave assisted extrac-
tion of sour orange peel pectin (SOPP) as a new source of pectin
and to obtain optimum conditions for maximum extraction yield
using Box–Behnken experimental design, and to study the physico-
chemical properties of the pectin extracted from sour orange fruit
peel.
2. Materials and methods
2.1. Raw materials and reagents
Sour orange (C. aurantium L.) peels were obtained from a fruit
orchard in Fasa, Fars, Iran. The peels were removed, finely cut
with a sharp knife. The cut fragments were placed on stainless
trays, and dried in a hot air oven at 50 ◦ C until it attained a con-
stant weight. The dried peels were pulverized and passed through
a 40-mesh sieve to obtain powdered sample. The powder was
stored in dark bags and kept in a dry environment prior to the
experimental analysis. Citric acid, sodium hydroxide, hydrochlo-
ric acid, phenolphthalein reagent, sulfuric acid, sodium tetraborate
and sodium azide were purchased from Merck Chemical Co. (Darm-
stadt, Germany). Sigma Chemical Co. (St. Louis, MO, USA) supplied
meta-hydroxydiphenyl reagent and standard citrus pectin (poly-d-
galacturonic acid methyl ester).
2.2. Pectin extraction
Microwave assisted extraction of SOPP was performed accord-
ing to the methods described by Li, Jia, Wei, and Liu (2012) with
minor modifications. The dried peels powder (liquid–solid ratio
(LSR) of 15 (v/w), Fig. 1) was poured into citric acid aqueous solu-
tion adjusted to the desired pH values (1.5, 2.25, 3), and thereafter
stirred. The solution was then extracted with three powers of 300,
500, 700 W for three times, 1, 2, 3 min. After extraction, the mixture
S.S. Hosseini et al. / Carbohydrate Polymers 140 (2016) 59–65
was allowed to cool down to room temperature (25 ◦ C), filtered
using filter paper, centrifuged (10 000 × g for 15 min) and the super-
natant was precipitated with ethyl alcohol (96%). The coagulated
pectin mass was washed with ethanol (96%) for three times to
remove the monosaccharides and disaccharides (Minkov, Minchev,
& Paev, 1996). In the next step, the wet pectin was dried at 40 ◦ C
in the hot air oven until a constant weight was obtained (approxi-
mately 7 h). The pectin extraction yield was calculated as follows:
Weight of dried pectin (g)
% Yield = × 100 (1)
Weight of dried powder (g)
35.0 40.0 45.0
2.3. Determination of degree of esterification
The degree of esterification (DE) of the SOPP was determined by
the titrimetric method (USP 26 NF 21, 2003) with a slight modifica-
tion. Briefly, dried pectin sample (100 mg) was moistened with 2 ml
ethanol and dissolved in 20 ml of distilled water free of carbon diox-
ide at 40 ◦ C. After complete dissolution of the sample, five drops of
phenolphthalein reagent were added and the sample was titrated
with 0.1 M NaOH (V1 ). Thereafter, 10 ml of 0.5 M NaOH was added
and the sample was stirred, after which the sample was allowed to
stand for 20 min for hydrolysis. In the next step, 10 ml of 0.5 M HCl
was added and the sample was shaken until the pink color disap-
peared. Finally, five drops of phenolphthalein reagent were added
and the mixture was titrated with 0.1 M NaOH until a pale pink
color that persisted after vigorous shaking was obtained (V2 ). The
DE of the pectin (%) was calculated as follows:
V2
% DE = × 100 (2)
V1 + V2
2.4. Galacturonic acid content
The galacturonic acid (GA) content was determined by the
meta-hydroxydiphenyl method using a UV–Vis spectrophotometer
according to Blumenkrantz and Asboe-Hansen (1973) with minor
modifications. To 1 ml of the sample (200 ␮g/ml), 6 ml of H2 SO4
containing 0.0125 M sodium tetraborate was added. The tubes were
cooled in a water-ice bath. The mixture was agitated with a vortex
mixer and the tubes heated in a boiler for 5 min. After cooling in
a water-ice bath, 20 ␮l of the meta-hydroxydiphenyl reagent was
added. The tubes were shaken for 5 min and the absorbance was
read at 520 nm. The galacturonic acid content was determined by
comparison with a standard curve obtained from standard galac-
turonic acid solutions (0–250 ␮g/ml).
2.5. Emulsifying properties
Emulsifying activity (EA) and emulsion stability (ES) were
assessed using the methods described by Dalev and Simeonova
(1995) with minor modifications. In a nutshell, oil-in-water (O/W)
emulsions were prepared by adding 5 ml of sunflower oil to 5 ml
pectin solutions (0.5%, w/w) containing 0.02% sodium azide as a
bactericide. The pre-emulsions were spun vigorously in a homog-
enizer at 10 000 × g for 3 min. In the next step, the emulsions were
centrifuged in graduated tubes at 3000 × g for 5 min. The EA was
calculated as follows:
EPV
EA (%) = × 100 (3)
Tv
where Tv was the total volume of the system and EPV was the
volume of the emulsion phase.
 S.S. Hosseini et al. / Carbohydrate Polymers 140 (2016) 59–65 61

Table 1 Table 3
Independent variables and their levels used in the Box–Behnken design. Analysis of variance (ANOVA) for the yield and degree of esterification of SOPP.

Factors Coded symbols Levels Source Sum of squares DF Mean square F-Value p-Value

−1 0 1 (A) Yield
Regression 761.0 9 84.6 293.6 <0.0001
Irradiation time (min) X1 1 2 3 Linear 580.5 3 193.5 671.9 <0.0001
Microwave power (W) X2 300 500 700 Square 168.7 3 56.2 195.3 <0.0001
pH X3 1.5 2.25 3 Interaction 11.7 3 3.9 13.5 0.008
Residual error 1.4 5 0.3
Lack-of-fit 0.9 3 0.3 1.1 0.522
Pure error 0.6 2 0.3
Table 2 Total 762.4 14
Experimental and predicted values for the yield and degree of esterification of SOPP
from the Box–Behnken experimental design. R2 0.998
Adj R2 0.995
Run Independent variables Measured responses Predicted responses Pred R2 0.980

X1 (min) X2 (W) X3 Yield (%) DE (%) Yield (%) DE (%)

(B) DE
1 1 300 2.25 6.2 9.4 5.8 9.0 Regression 2308.4 9 256.5 76.7 <0.0001
2 3 300 2.25 8.2 17.9 8.0 17.8 Linear 1742.7 3 580.9 173.7 <0.0001
3 1 700 2.25 7.6 11.1 7.8 11.2 Square 505.8 3 168.6 50.4 <0.0001
4 3 700 2.25 11.4 18.5 11.8 18.8 Interaction 59.8 3 19.9 6.0 0.042
5 1 500 1.50 20.0 2.9 20.0 1.9 Residual error 16.7 5 3.3
6 3 500 1.50 25.2 3.9 25.0 2.6 Lack-of-fit 10.5 3 3.5 1.1 0.499
7 1 500 3.00 5.2 21.4 5.4 22.7 Pure error 6.2 2 3.1
8 3 500 3.00 6.6 37.5 6.6 38.5 Total 2325.1 14
9 2 300 1.50 20.2 1.7 20.6 3.0
10 2 700 1.50 26.4 2.0 26.2 2.9 R2 0.993
11 2 300 3.00 6.6 30.6 6.8 29.6 Adj R2 0.980
12 2 700 3.00 7.4 34.2 7.0 32.9 Pred R2 0.921
13 2 500 2.25 8.0 30.0 7.6 30.2
14 2 500 2.25 7.8 32.0 7.6 30.2
15 2 500 2.25 7.0 28.5 7.6 30.2
variables are presented in Table 1. All computation and graphics
in this study were conducted using the statistical software Design
Expert 7.0 and Excel.
Similar EA were prepared to study the ES after 1 and 30 days
of storage at 4 ◦ C and room temperature. Then, the samples were
centrifuged at 3000 × g for 5 min. The ES was calculated as follows: 3. Results and discussion
VEr
ES (%) = × 100 (4) 3.1. Effect of different LSR on extraction yield of SOPP
VEi
where VEr was the remaining emulsified layer volume and VEi was The effect of different LSR (5, 10, 15, 20, 25, 30, 35, 40 and 45
the initial emulsified layer volume. (v/w)) on the extraction yield of SOPP was presented in Fig. 1,
when the other factors were fixed (irradiation time of 2 min,
2.6. Viscosity measurement microwave power of 500 W, pH of 2.25). The extraction yields
of the SOPP increased significantly from 6.7% to 9.2% with an
The viscosity of the pectin solutions was measured at different increase in LSR from 5 to 15 (v/w). However, the extraction yields
concentrations of SOPP (0.1%, 0.5%, 1.0%, 1.5% and 2.0% (w/v)) by a decreased with further increase in LSR. The solvent can increase
rotational programmable viscometer (LVDV-II Pro, Brookfield Engi- the contact surface area between the plant matrix and the solvent,
neering Inc., USA) using a LV spindle. About 25 ml of each sample which results in an increase in the extraction yield of pectin up
was cast into the cylinder of the viscometer and the shear rate was to LSR of 15 (v/w) (Maran et al., 2014). However, the higher LSR
programmed between 2.01 and 98.12 s−1 at an interval 2 s. The flow decreased the microwave adsorption of solid material (because the
behavior of dilute solutions of SOPP was analyzed at 25 ◦ C. solvent absorbed more energy), leading to a decrease in pectin
mass transfer rate and reduction in pectin extraction yield (Li
2.7. FT-IR spectroscopy et al., 2010; Maran, Sivakumar, Thirugnanasambandham, & Sridhar,
2013). Therefore, in this study, the LSR of 15 (v/w) was used for all
Fourier transform infrared (FT-IR) spectroscopy of the SOPP experiments.
was characterized by means of a Perkin Elmer FT-IR spectrom-
eter (Perkin Elmer Co., MA, USA) using the KBr disk method in
4000–450 cm−1 range with a resolution of 4 cm−1 . 3.2. Statistical analysis

2.8. Experimental design Box–Behnken design is a collection of mathematical and statis-

The one Factor at the Time (where only one factor (liquid–solid
ratio (LSR)) was changed at one time while the other factors were
kept fixed (irradiation time of 2 min, microwave power of 500 W,
pH of 2.25)) was used to estimate the best LSR for the extraction of
SOPP. A Box–Behnken response surface experimental design was
used to optimize and investigate the effect of independent vari-
ables (time (X1 ), microwave power (X2 ), pH (X3 )) on the extraction
yield and degree of esterification. The levels of the three retained
tical techniques for empirical model building. The advantage of this
method is the simultaneous investigation of individual and inter-
active effects of process variables on the response from a lower
number of experiments in comparison with other experimental
designs (Maran et al., 2014). The experimental and predicted val-
ues for the yield and DE of SOPP at different extracted conditions
are shown in Table 2.
Factorial Box–Behnken design of type 33 was used to evalu-
ate the effect of irradiation time, microwave power and pH on the
62 S.S. Hosseini et al. / Carbohydrate Polymers 140 (2016) 59–65

Fig. 2. Effect of irradiation time, microwave power and pH on the extraction yield (a, b, c) and DE (d, e, f) of the SOPP.

yield and DE of SOPP. The quadratic models for yield and DE were
expressed as follows:
Yield (%) = 7.60 + 1.55X1 + 1.45X2 − 8.25X3 − 0.07X12 + 0.82X22
+ 6.72X32 + 0.45X1 X2 − 0.95X1 X3 − 1.35X2 X3
DE (%) = 30.17 + 4.11X1 + 0.78X2 + 14.15X3 − 8.32X12 − 7.64X22
− 5.42X32 − 0.28X1 X2 + 3.76X1 X3 + 0.84X2 X3
where Xi is a coded independent factor (X1 = irradiation time,
X2 = microwave power, X3 = pH).
The results of the second order response surface models in the
form of analysis of variance (ANOVA) are summarized in Table 3.
(5) The results showed that the p-values of the developed model for
yield and DE were below 0.0001, which indicated that the fitness
of the model was highly significant (p < 0.0001). The lack-of-fit tests
were insignificant (p = 0.522 and p = 0.499 for yield and DE, respec-
(6) tively), which indicated that the models could be used to predict
 S.S. Hosseini et al. / Carbohydrate Polymers 140 (2016) 59–65 63

the responses. The values of R2 for yield and DE were 0.998 and Table 4
Emulsifying activity and emulsion stability of oil/0.5% (w/w) pectin solutions.
0.993, respectively. This indicated that the models explained 99.8%
and 99.3% of the total variation and only 0.2% and 0.7% of the total Emulsion activity (%) Emulsion stability (%)
variation was unexplained (Wai et al., 2010). In addition, the values Storage time 1 day 30 day
of the adjusted determination coefficient (adjusted R2 = 0.995 and
Temperature (◦ C) 23 4 23 4 23
0.980 for yield and DE, respectively) were high to advocate a high
Pectina 40.7 83.4 72.3 83.3 72.1
significance of the model (Khodaiyan, Razavi, & Mousavi, 2008).
a
Pectin extracted under optimal extraction conditions.

3.3. The effect of extraction condition on the SOPP yield

14.0
0.10%
The yield of SOPP, which ranged from 5.2% to 26.4% (w/w) 12.0 0.50%
(Table 2), depended on the irradiation time, microwave power and

pH (Fig. 2(a–c)). Microwave power was an important factor that
influenced the extraction efficiency. The results showed that the
extraction of pectin increased with an increase in microwave power
from 300 to 700 W (Fig. 2(a, c)). The increasing microwave irra-
diation energy can enhance the penetration of extraction solvent
into the pectin source matrix, and are delivered to the materials
efficiently through molecular interaction with the electromagnetic
field and offer a quick transfer of energy to the extraction solvent
and matrix, allowing the components from the dissolution to be
extracted (Yan et al., 2010).
Irradiation time is also an important factor that influences the
yield of pectin. A longer irradiation time presents a positive effect
on the yield of pectin (Fig. 2(a, b)). It has been reported that a longer
irradiation time favors the production of pectin (Bagherian et al.,
2011; Li et al., 2012). This might be due to the time requirement
for the exposure of the SOPP to the release medium where the
solvent penetrated into the raw materials, dissolved the SOPP and
subsequently diffused out from the raw solid materials (Samavati,
2013).pH was one of the key factors that influenced the extraction
yield of pectin and it is necessary to select a proper pH to assure
maximum extraction of SOPP. From Fig. 2(b, c) it is evident that the
yield increased with decreasing pH values. The extraction solvent
with high acidity had the ability to mix with the insoluble pectin
and favor the hydrolysis of the insoluble pectin constituents into
soluble pectin, thus increasing the extraction of pectin from plant
materials (Ma et al., 2013; Maran et al., 2013).
3.4. Optimization of microwave extraction conditions
Based on the above-mentioned results and discussion, an opti-
mization study is required to find the optimal conditions for yield of
SOPP. The optimum extraction conditions were irradiation time of
3 min, microwave power of 700 W and pH of 1.5 and maximum
yield of SOPP was 29.1%. The result was obtained under opti-
mized experimental conditions in triplicate and the mean values
(28.8 ± 0.3%) obtained from real experiments, demonstrated the
validation of the optimized conditions.
3.5. The effect of extraction condition on DE of SOPP
As shown in Table 2, the DE of SOPP ranged from 1.7% to 37.5%. In
addition, pectin samples extracted under optimal extraction condi-
tions (irradiation time of 3 min, microwave power of 700 W and pH
of 1.5) had DE of 1.5 ± 0.2. These results showed that the DE was not
related to extraction yield because the highest yield was obtained
under optimal extraction conditions, whereas the DE was quite
low at these conditions. Based on the above-mentioned results, the
SOPP extracted in this study showed a DE value lower than 50% and
thus can be classified as LMP.
The effects of the irradiation time, microwave power and pH on
the degree of esterification (DE) of the extracted pectin are pre-
sented in Fig. 2(d–f). The results showed that the DE of the SOPP
decreased with a large increase in irradiation time and microwave
power and a decrease in pH values. Generally, pectin obtained
Viscosity (Pa.S)
10.0 1.00%
8.0 1.50%
2.00%
6.0
4.0
2.0
0.0
0.0 20.0 40.0 60.0 80.0 100.0
Shear rate (1/S)
Fig. 3. The flow behavior of a dilute solution of SOPP at different concentrations.
under very harsh extraction conditions (high power, long irradia-
tion interval and low pH) have a low degree of esterification because
these harsh conditions could increase deesterification of polygalac-
turonic chains (Mort, Qiu, & Maness, 1993; Wai et al., 2010).
3.6. Total galacturonic acid content
All pectins are rich in galacturonic acid and according to the
provisions of FAO and EU, ‘pectin’ must contain at least 65% galac-
turonic acid (Willats, Knox, & Mikkelsen, 2006). In this study, the
galacturonic acid content was obtained under optimal extraction
conditions (irradiation time of 3 min, microwave power of 700 W
and pH of 1.5). The results showed that the galacturonic acid
content of SOPP was 71.0 ± 0.8% under the above-mentioned con-
ditions. Galacturonic acid amount in this research was close to
the data of fresh orange pectin and sugar beet pectin reported by
Kratchanova, Pavlova, Panchev, and Kratchanov (1996) and Ma et al.
(2013), respectively.
3.7. Emulsifying properties
In this experiment, the EA and ES of the pectin extracted at
irradiation time of 3 min, microwave power of 700 W and pH
of 1.5 (optimum extraction conditions) were studied with emul-
sions prepared with 0.5% (w/w) pectin solutions (Table 4). After
centrifuging the emulsions, oil phase, emulsified layer phase and
aqueous phase were observed on the top, middle and bottom,
respectively. Table 4 shows that the emulsifying activity values of
the emulsions prepared with the pectin extracted under optimal
extraction conditions were 40.7%. This value was nearly similar
to the data of the pectin extracted from sugar beet pulp under
acidic conditions (Yapo et al., 2007). Moreover, the emulsions were
83.4% stable at 4 ◦ C and 72.3% at 23 ◦ C after 1 day of storage. After
30 days of storage, the emulsions stability was 83.3% at 4 ◦ C and
72.1% 23 ◦ C, indicating the high stability levels of the emulsions
at two different storage temperatures. However, emulsions were
slightly steadier when they were stored at 4 ◦ C. In comparison with
other data published, these values were higher than that obtained
for pectin extracted from sugar beet under acidic conditions (Ma
et al., 2013). From these results, it could be inferred that under cer-
tain extraction conditions, microwave extracted pectin from sour
orange peel could be a suitable emulsifier.
64 S.S. Hosseini et al. / Carbohydrate Polymers 140 (2016) 59–65
Fig. 4. FTIR spectra of SOPP extracted under optimal extraction conditions and citrus pectin.
3.8. Viscosity measurement
In this stage, the flow behavior of a dilute solution of SOPP
obtained under optimal extraction conditions was studied by a vis-
cometer at 25 ◦ C. The results showed that the viscosity increased
with an increase in the concentration of pectin solutions. In addi-
tion, the viscosity of SOPP depended on the shear rate at different
concentrations, such that at low concentrations (≤1.0%, w/v),
all samples showed nearly Newtonian flow behavior (viscosity
remained constant with an increase or decrease in the shear rate),
whereas as the concentration increased, pseudoplastic flow (vis-
cosity decreased with increasing shear rate) became dominant
(Fig. 3). This is in accordance with the study of Chen, Zhang, Sun, and
Wei (2014) who studied the extraction of Abelmoschus esculentus
pectin. They stated that pectin of this product at low concentra-
tions showed Newtonian behavior, whereas as the concentration
increased, the behavior changed from Newtonian to pseudoplas-
tic flow. The molecular origin of pseudoplastic behavior has been
attributed to the fact that applied shear stress can cause disentan-
glement of biopolymers, alignment of biopolymers with the shear
field, or disruption of weak physical interactions holding biopoly-
mers together (McClements, 2004, chap. 4).
3.9. FTIR spectra of SOPP
Fig. 4 shows the FTIR spectra of SOPP obtained under optimal
extraction conditions and a citrus pectin pattern. The absorption
peak between 3300 and 3500 cm−1 was due to OH stretching. The
peak between 2850 and 3000 cm−1 was attributed to CH vibra-
tional modes, including CH, CH2 , and CH3 stretching and bending
vibrations (Chen et al., 2014). The peak at 1744 cm−1 was due to
the stretching vibration of the CO group of the carboxylic acid
methyl ester. Carboxylate groups showed two peaks at 1633 and
1440 cm−1 corresponding to the asymmetric and weaker symmet-
ric stretching of the carboxylate group, respectively (Manrique &
Lajolo, 2002). The effect of the displacement of the wavelengths
was due to the physical state of the sample and the areas where the
polymer COO− groups of galacturonic acid were inserted (Santos
et al., 2013). The absorption patterns between 800 and 1200 cm−1
are collectively referred to as the “finger print” region that is unique
to a compound and thus its interpretation can be a difficult matter
(Chen et al., 2014).
4. Conclusion
Pectin is a natural additive that is widely used in the food
industry. In the present study, microwave assisted extraction of
pectin from sour orange peel was investigated under different
extraction conditions. The Box–Behnken design was carried out to
study the effect of irradiation time, microwave power and pH on
the yield and DE of pectin. The results showed that the highest
yield of sour orange peel pectin extracted at the optimal condi-
tions (irradiation time of 3 min, microwave power of 700 W, pH of
1.50) was 29.1%, which was confirmed through validation experi-
ments (28.8 ± 0.3%). In addition, the pectin extracted under optimal
extraction conditions had a suitable purity (galacturonic acid con-
tent was 71.0 ± 0.8%) and is classed as low methoxyl pectin. Also,
the results indicated the high stability levels of the emulsions at two
different storage temperatures (4 and 23 ◦ C). Viscosity measure-
ment revealed that the solutions of pectin at low concentrations
showed nearly Newtonian flow behavior, whereas at higher con-
centrations, pseudoplastic flow behavior became dominant.
References
Axelos, M. A. V., Thibault, J., & Lefebvre, J. (1989). Structure of citrus pectins and
viscometric study of their solution properties. International Journal of Biological
Macromolecules, 11(3), 186–191.
Bagherian, H., Ashtiani, F. Z., Fouladitajar, A., & Mohtashamy, M. (2011).
Comparisons between conventional, microwave- and ultrasound-assisted
methods for extraction of pectin from grapefruit. Chemical Engineering and
Processing: Process Intensification, 50(11), 1237–1243.
Basanta, M. F., Ponce, N. M. A., Rojas, A. M., & Stortz, C. A. (2012). Effect of
extraction time and temperature on the characteristics of loosely bound
pectins from Japanese plum. Carbohydrate Polymers, 89(1), 230–235.
Blumenkrantz, N., & Asboe-Hansen, G. (1973). New method for quantitative
determination of uronic acids. Analytical Biochemistry, 54(2), 484–489.
Chan, S., & Choo, W. (2013). Effect of extraction conditions on the yield and
chemical properties of pectin from cocoa husks. Food Chemistry, 141(4),
3752–3758.
Chen, Y., Zhang, J. G., Sun, H. J., & Wei, Z. J. (2014). Pectin from Abelmoschus
esculentus: Optimization of extraction and rheological properties. International
Journal of Biological Macromolecules, 70, 498–505.
 S.S. Hosseini et al. / Carbohydrate Polymers 140 (2016) 59–65
Dalev, P. G., & Simeonova, L. S. (1995). Emulsifying properties of protein–pectin
complexes and their use in oil-containing foodstuffs. Journal of the Science of
Food and Agriculture, 68(2), 203–206.
Gardner, D. F., Schwartz, L., Krista, M., & Merimee, T. J. (1984). Dietary pectin and
glycemic control in diabetes. Diabetes Care, 7(2), 143–146.
Haaz, S., Fontaine, K. R., Cutter, G., Limdi, N., Perumean-Chaney, S., & Allison, D. B.
(2006). Citrus aurantium and synephrine alkaloids in the treatment of
overweight and obesity: An update. Obesity Reviews, 7(1), 79–88.
He, X., Lian, L., Lin, L., & Bernart, M. W. (1997). High-performance liquid
chromatography–electrospray mass spectrometry in phytochemical analysis
of sour orange (Citrus aurantium L.). Journal of Chromatography A, 791(1),
127–134.
Kalapathy, U., & Proctor, A. (2001). Effect of acid extraction and alcohol
precipitation conditions on the yield and purity of soy hull pectin. Food
Chemistry, 73(4), 393–396.
Khodaiyan, F., Razavi, S. H., & Mousavi, S. M. (2008). Optimization of canthaxanthin
production by Dietzia natronolimnaea HS-1 from cheese whey using statistical
experimental methods. Biochemical Engineering Journal, 40(3), 415–422.
Khormaei, M., Nasernejad, B., Edrisi, M., & Eslamzadeh, T. (2007). Copper
biosorption from aqueous solutions by sour orange residue. Journal of
Hazardous Materials, 149(2), 269–274.
Kratchanova, M., Pavlova, E., & Panchev, I. (2004). The effect of microwave heating
of fresh orange peels on the fruit tissue and quality of extracted pectin.
Carbohydrate Polymers, 56(2), 181–185.
Kratchanova, M., Pavlova, E., Panchev, I., & Kratchanov, C. (1996). Influence of
microwave pretreatment of fresh orange peels on pectin extraction. Progress in
Biotechnology, 14, 941–946.
Li, D., Jia, X., Wei, Z., & Liu, Z. (2012). Box–Behnken experimental design for
investigation of microwave-assisted extracted sugar beet pulp pectin.
Carbohydrate Polymers, 88(1), 342–346.
Li, J., Zu, Y. G., Fu, Y. J., Yang, Y. C., Li, S. M., Li, Z. N., et al. (2010). Optimization of
microwave-assisted extraction of triterpene saponins from defatted residue of
yellow horn (Xanthoceras sorbifolia Bunge.) kernel and evaluation of its
antioxidant activity. Innovative Food Science & Emerging Technologies, 11(4),
637–643.
Liu, L., Cao, J., Huang, J., Cai, Y., & Yao, J. (2010). Extraction of pectins with different
degrees of esterification from mulberry branch bark. Bioresource Technology,
101(9), 3268–3273.
Lofgren, C., & Hermansson, A. M. (2007). Synergistic rheological behaviour of
mixed HM/LM pectin gels. Food Hydrocolloids, 21(3), 480–486.
Lopes da Silva, J. A., & Rao, M. A. (2006). Pectins: Structure, functionality, and uses.
In A. M. Stephen, G. O. Philips, & P. A. Williams (Eds.), Food polysaccharides and
their applications (pp. 353–411). Boca Raton: CRC Press.
Ma, S., Yu, S., Zheng, X., Wang, X., Bao, Q. D., & Guo, X. (2013). Extraction,
characterization and spontaneous emulsifying properties of pectin from sugar
beet pulp. Carbohydrate Polymers, 98(1), 750–753.
Manrique, G. D., & Lajolo, F. M. (2002). FT-IR spectroscopy as a tool for measuring
degree of methyl esterification in pectins isolated from ripening papaya fruit.
Postharvest Biology and Technology, 25(1), 99–107.
Maran, J. P., Sivakumar, V., Thirugnanasambandham, K., & Sridhar, R. (2013).
Optimization of microwave assisted extraction of pectin from orange peel.
Carbohydrate Polymers, 97(2), 703–709.
65
Maran, J. P., Sivakumar, V., Thirugnanasambandham, K., & Sridhar, R. (2014).
Microwave assisted extraction of pectin from waste Citrullus lanatus fruit
rinds. Carbohydrate Polymers, 101, 786–791.
May, C. D. (1990). Industrial pectins: Sources, production and applications.
Carbohydrate Polymers, 12(1), 79–99.
McClements, D. J. (2004). Food emulsions: Principles, practices, and techniques (2nd
ed.). Boca Raton: CRC Press.
Mesbahi, G., Jamalian, J., & Farahnaky, A. (2005). A comparative study on functional
properties of beet and citrus pectins in food systems. Food Hydrocolloids, 19(4),
731–738.
Minkov, S., Minchev, A., & Paev, K. (1996). Modelling of the hydrolysis and
extraction of apple pectin. Journal of Food Engineering, 29(1), 107–113.
Mort, A. J., Qiu, F., & Maness, N. O. (1993). Determination of the pattern of methyl
esterification in pectin. Distribution of contiguous nonesterified residues.
Carbohydrate Research, 247, 21–35.
Pagan, J., & Ibarz, A. (1999). Extraction and rheological properties of pectin from
fresh peach pomace. Journal of Food Engineering, 39(2), 193–201.
Ptichkina, N. M., Markina, O. A., & Rumyantseva, G. N. (2008). Pectin extraction
from pumpkin with the aid of microbial enzymes. Food Hydrocolloids, 22(1),
192–195.
Qiu, L., Zhao, G., Wu, H., Jiang, L., Li, X., & Liu, J. (2010). Investigation of combined
effects of independent variables on extraction of pectin from banana peel
using response surface methodology. Carbohydrate Polymers, 80(2),
326–331.
Samavati, V. (2013). Polysaccharide extraction from Abelmoschus esculentus:
Optimization by response surface methodology. Carbohydrate Polymers, 95(1),
588–597.
Santos, J. D. G., Espeleta, A. F., Branco, A., & de Assis, S. A. (2013). Aqueous
extraction of pectin from sisal waste. Carbohydrate Polymers, 92(2),
1997–2001.
Seixas, F. L., Fukuda, D. L., Turbiani, F. R. B., Garcia, P. S., Carmen, L. de O., Jagadevan,
S., et al. (2014). Extraction of pectin from passion fruit peel (Passiflora edulis f.
flavicarpa) by microwave-induced heating. Food Hydrocolloids, 38, 186–192.
Thakur, B. R., Singh, R. K., Handa, A. K., & Rao, M. A. (1997). Chemistry and uses of
pectin – A review. Critical Reviews in Food Science & Nutrition, 37(1), 47–73.
USP NF 21. (2003). The United States pharmacopeia – The national formulary. pp.
1401–1402. Rockville, MD: United States Pharmacopeial Convention.
Wai, W. W., Alkarkhi, A. F. M., & Easa, A. M. (2010). Effect of extraction conditions
on yield and degree of esterification of durian rind pectin: An experimental
design. Food and Bioproducts Processing, 88(2), 209–214.
Willats, W. G. T., Knox, J. P., & Mikkelsen, J. D. (2006). Pectin: New insights into an
old polymer are starting to gel. Trends in Food Science & Technology, 17(3),
97–104.
Xu, Y., Zhang, L., Bailina, Y., Ge, Z., Ding, T., Ye, X., et al. (2014). Effects of ultrasound
and/or heating on the extraction of pectin from grapefruit peel. Journal of Food
Engineering, 126, 72–81.
Yan, M. M., Liu, W., Fu, Y. J., Zu, Y. G., Chen, C. Y., & Luo, M. (2010). Optimisation of
the microwave-assisted extraction process for four main astragalosides in
Radix Astragali. Food Chemistry, 119(4), 1663–1670.
Yapo, B. M., Robert, C., Etienne, I., Wathelet, B., & Paquot, M. (2007). Effect of
extraction conditions on the yield, purity and surface properties of sugar beet
pulp pectin extracts. Food Chemistry, 100(4), 1356–1364.