Journal of Engineering Science and Technology

Special Issue on SOMCHE 2014 & RSCE 2014 Conference, January (2015) 22 - 29
© School of Engineering, Taylor’s University

EXTRACTION AND CHARACTERISATION OF PECTIN

FROM SWEET POTATO (IPOMOEA BATATAS) PULP

DAYANG NORULFAIRUZ ABANG ZAIDEL1,*, NURUL NADIA ZAINUDIN1,

YANTI MASLINA MOHD JUSOH1, IDA IDAYU MUHAMAD2
1
Faculty of Chemical Engineering, Universiti Teknologi Malaysia,81310 UTM Johor
Bahru, Johor, Malaysia
2
IJN-UTM Cardiovascular Engineering Centre, Universiti Teknologi Malaysia, 81310
UTM Johor Bahru, Johor, Malaysia
*Corresponding Author: dayang@cheme.utm.my

Abstract
Sweet potato (Ipomoea batatas) pulp contains pectin, which acts as thickening
and gelling agents in food application. This study investigates the yield and
profile of the galacturonic acid (GalA) and rheological properties of the sweet
potato pectin produced using acid extraction method. In this work, pectins were
extracted using hydrochloric acid at different concentrations (0.05 M, 0.1 M,
0.15 M, 0.2 M, and 0.25 M) at pH 1.5 and temperature 90 °C for 1 hour.
Hydrolysis of residual starch in the cell wall of sweet potato using heat stable α-
amylase and amyloglucosidase was employed prior to pectin extraction to
eliminate starch contamination. Pectins were characterised for yield, GalA
profile, degree of esterification (DE) and its rheological properties. Profile of
GalA and DE were determined by using Fourier Transfer Infrared Spectroscopy
(FTIR). Rheological properties of pectin were performed by addition of calcium
ions (Ca2+) to investigate the viscosity changes due to gelation of pectin. The
yield of pectin obtained was in the range of 5.0 – 6.0% (w/w) and the DE was
affected by the acid concentrations used during extraction. For its rheological
properties, viscosity of pectin increases as Ca2+ was added into the pectin
solution indicating the gelation of pectin by the cross-linking formation
between calcium ion and non-esterified GalA in pectic polysaccharides.
Keywords: Degree of esterification, Pectin, Polysaccharides, Rheology, Sweet potato.

1. Introduction
Sweet potatoes are mainly used to produce starch and starchy foods, which
generates a considerable quantity of residues. The majorities of these residues are
22
 Extraction and Characterisation of Pectin from Sweet Potato . . . . 23

directly thrown out and consequently pollute the environment. The peels are
mostly waste materials resulting from the sweet potato processing industry and
are normally discarded. These discarded peels may cause an environmental
problem, particularly water pollution [1]. Thus, in addition to being fed to
animals, the peels can be used in the production of pectin, which would then
increase the potential return for the sweet potato processing industry.
Sweet potato contains pectin, which has many important functions in plants.
Commercially, pectin has wide applications in both the pharmaceutical and food
industries, where it acts as thickening and gelling agents, regulates the thickness
and mouth-feel of fruit drink powder when the powder is dissolved in cold water,
and prevents the formation of cheesy milk layer in gelled milk dessert [1]. It is
reported that sweet potato residues, a good resource of pectin production, contain
about 15% pectin on dry matter basis [2]. Therefore, extractions of pectin from
the residues not only lower the pollution but also reduce the waste of resources. In
addition, pectin has proven to have beneficial effects on human health [3-5].
Extraction of pectins can lead to large variations in the chemical structure of
the final material. The ease of pectin extraction ispartly affected by the degree of
esterification [6]. Pectins are industrially extracted from citrus peels and apple
pomace by hot acidified water. Extraction conditions of pH 1.5 to 3.0 and
temperatures of 60 to 100 °C for 0.5 to 6 hours were varied to give a material that
has the desired gelling capacity and degree of methylation [7].
This study was conducted to investigate the extraction parameters for sweet
potato pectin such as concentration of solvent by using acid extraction method to
yield highest production of pectin. The highest pectin yields were obtained by hot
acid extraction, as reported previously [7, 8]. In this study, chemical properties of
sweet potato pectin, such as galacturonic acid content, degree of esterification,
molecular weight and rheological properties were investigated.

2. Materials and Methods

2.1. Materials
Sweet potato (Red Sweet Potato) was obtained from local supermarket in Taman
Universiti, Skudai. The cell wall materials had been prepared from sweet potato
pulp as described previously [9]. Commercial citrus pectin was purchased from
Sigma (USA) in order to compare its physicochemical properties with sweet
potato. Heat-stable α-amylase (Termamyl 120 type LS) and amyloglucosidase
(EC 3.2.1.3 from Aspergillusniger, 30-60 units per mg protein) for enzymatic
hydrolysis and all reagents were obtained from Sigma (St. Louis, MO).

2.2. Sample preparation

Sweet potatoes (5 kg) were washed with tap water and peeled. Pulp were then
sliced into small pieces and macerated in a Waring blender (Scientific Industries,
New York). The slurry was filtered through double layers of cheesecloth to
separate the residue from the starch milk. The residue was rinsed thoroughly with
running water until the water ran clear, and was then dried in a forced-air dryer
(Whisper 500, Philip, Germany) at 45 °C for 18 hours. The resultant dried starch

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24 D. N. A. Zaidel et al.

residue (500 g) was ground and passed through a 60-mesh (250 µm) sieve before
used for enzymatic hydrolysis.
Ground dried sweet potato starch residue (10 g) was suspended in distilled
water (200 mL) and boiled for 5 minutes. The suspension was kept at 80 °C,
and 0.5 mL of heat-stable α-amylase (Termamyl 120 type LS from Sigma
Chemical Co, St. Louis, USA) was added, and then incubated for 30 minutes to
hydrolyse the residual starch. The mixture was centrifuged at 3000 rpm for 10
minutes, supernatant was discarded and digestion of the residue was repeated
with 0.5 mL amyloglucosidase (EC 3.2.1.3 from Aspergillusniger, 30-60 units
per mg protein, Sigma Chemical Co, St. Louis, USA ), then incubated at 55 °C
for 30 minutes at pH 4.5. The mixture was filtered using two layers of
cheesecloth. The residue was washed with distilled water, methanol and
acetone, successively, and air-dried for 48 hours.

2.3. Extraction of pectin

Pectin was extracted according to previous method with slight modification [10].
Hydrochloric (HCl) was used as extracting solvents in this experiment. Samples
(10 g) of dried ground cell wall materials were dispersed in 250 mL 0.05 M HCl.
The dispersion was stirred and kept at 90 °C for 1 hour. After incubation, the
suspensions were centrifuged at 10 °C for 15 minutes at 10000 rpm (Kubota
Model, Germany). The liquid fraction containing extracted pectin materials was
neutralised with 32% NaOH, then the same volume of 95% ethanol was added
and the mixture was stirred for 5 minutes and then stored at 4 °C for 12 hours.
The mixture was then centrifuged at 10000 rpm for 15 minutes and the pectin
residue washed successively with 70, 80, 90% ethanol [11]. Finally the extracted
pectin was dried in a freeze dryer (model HETO FD 4.0, United Kingdom) for 18
hours, ground and stored in a desiccator until further analysis. The extracted
pectin was analysed for moisture content, pectin yield, detection of starch in
pectin, galacturonic acid, degree of esterification and the molecular weight. The
procedure as described above was repeated by adjusting the concentration of the
acid to 0.1 M, 0.15 M, 0.2 M, and 0.25 M HCl. The experiment was duplicated
for each concentration and 10 samples were extracted.

2.4. Analysis of pectin

2.4.1. Yield of pectin
Pectin yield was calculated as the ratio of the weight of dried pectin obtained to
dried cell wall materials [12].

2.4.2. Determination of the degree of esterification

The degree of esterification (DE) of pectin was determined by Fourier transform
infrared (FTIR) spectrometry (Perkin Elmer, USA) and enzymatically using
pectate lyase [13]. For the enzymatic determination of DE, pectin samples were
solubilised (2 mg/mL) in 50 mM Tris–HCl buffer, pH 8, and the pectin solution
were mixed with 790 µL of 50 mM Tris–HCl buffer, 1 mM CaCl2, pH 8, and 10

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 Extraction and Characterisation of Pectin from Sweet Potato . . . . 25

µL of enzyme (0.01 U in 50 mM Tris–HCl buffer, 1 mM CaCl2, pH 8). The

reaction and blanks were conducted at 40 °C for 30 minutes and monitored at 235 nm.
The amount of product was calculated using
235 = 4600 M/cm. The degree of
esterification was calculated from the calibration curve of the pectin standards.
All measurements were performed in triplicate.

2.4.3. Rheological properties of pectin

Brookfield digital viscometer, model DV-II + Pro, with an attached UL adapter
was used to describe the rheological characteristics of the blend solutions in this
work. The viscosity was determined in 20 mL of the sample and the shearing
time was 15 seconds. For the storage time measurements, solutions were kept at
room temperature in glass bottles in a dark place until analysis. Each
measurement was recorded as an average value of five readings when a constant
shear rate was applied.

3. Results and Discussion

3.1. Yield of pectin
The yield of pectin extracted from this study was shown in Table 1. The result
shows that the yield of pectin decreased as the acid concentrations increased. This
indicates that the conditions of extraction significantly affected pectin yields. It is
expected that the highest acid concentration will provide the most efficient
procedure. This indicates that pectins in sweet potato cell walls are mainly calcium-
bound low methoxyl pectin. From previous research, the yields of pectin extracted
from cell wall materials of sweet potato using various conditions were between
7.2% and 29.3% of dry cell wall materials [14]. These pectins are not extractable
with mild acid or alkaline [15]. A high yield of pectin obtained from extractions at
pH 1 of acid extraction was reported previously [16]. It has been reported that,
despite the presence of starch, lowering pH increased the level of galacturonic acid
detected, with pH 1 resulting in the highest galacturonic acid content of pectin
extracted [14]. This was attributed to the lower pH, leading to greater loosening of
the cell wall matrix so that pectin could be extracted more easily.

Table 1. Yield of pectin (%) as extracted by different acid concentrations.

Acid Concentration Pectin Yield (%)
0.05 M HCl 74.43
0.10 M HCl 60.73
0.15M HCl 46.12
0.20 M HCl 20.38
0.25 M HCl 18.05

3.2. Determination of the degree of esterification

Although degree of esterification depends on the original sources and treatment,
pectin extracted under acidic conditions usually contains around 60% methyl ester
groups [16]. Lowering the pH to 1 also increased the degree of esterification of
the pectin from no peak to 38.8% in the Beauregard variety sweet potato. This

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26 D. N. A. Zaidel et al.

may be due to starch degradation at pH 1, leading to a proportional increase in

galacturonic acid or pectin released from the residue [16].
Figure 1 shows the infrared spectra of the sweet potato pectin from extraction of
various acid concentrations. Among the absorption bands common to all spectra at
about 3410 cm-1 due to O-H stretching vibration, while absorption at 1640 cm-1 was
due to C=O stretching vibration of salt-form carboxyl group. The intensity of the
absorption at 1740 cm-1 declined with the decrease in the degree of esterification.
The degree of esterification was calculated from absorbance intensities for
1630 and 1745 cm-1 band. The result of FTIR spectroscopy in this study proved
that the sweet potato pectin has a very low degree of methyl-esterification. The
carbohydrates show high absorbance between 1200 and 950 cm-1, which
constitutes the ‘fingerprint’ region, specific for each polysaccharide. Structural
features arising from particular conformations around the glycosidic bond of the
pectin were observed in the 990-1100 cm-1 range.

Fig. 1. Infrared spectra of the sweet potato pectin

from extraction at various acid concentrations.

3.3. Rheological properties of pectin

The rheological characteristics of pectin solutions and Ca2+ pectin gels partially
depend on the concentration regime of the pectin solutions [17]. In all cases,
viscosity was almost independent of shear rate similar to the result reported for
apple pectin [18]. Low methoxyl pectin needs Ca2+ to form gel meanwhile high
methoxyl pectin can form gel itself by presence of sugar and water.
The gelation properties of pectin have been explained by the formation of
junction zones called “egg boxes” between two pectin chains in the presence of
divalent ions such as calcium. Gelation of pectin confines sugar, water and others

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 Extraction and Characterisation of Pectin from Sweet Potato . . . . 27

in a 3-dimensional network formed through junction zones. The gel formation and
gel properties of pectin are markedly affected by the DE, molecular weight of
pectin, pH, temperature, and concentration of calcium.
Therefore in this study, the effect of extrinsic parameter, such as concentration
of calcium, on the breaking pressure of the pectin gel was evaluated. Since pectin
extracted by using 0.05 M HCl gave the highest percentage yield, it has been used
to check on pectin viscosity.
Figure 2 shows the changes in breaking pressure with increasing calcium:
pectin ratio. Calcium concentration is an important factor affecting the breaking
pressure of the gel. As the calcium concentration increased from 10 to 25 mg/g-
pectin, the breaking pressure increased linearly. The maximum breaking pressure
was obtained at 25 mg/g-pectin and the value was 6.45×104 N/m2. As the
concentration exceeded 25 mg/g-pectin, the gels were short and brittle, and the
breaking pressure value dropped sharply. In LM-pectin, junction zones of suitable
length are believed to be required for producing stable gels. There is also a
tendency to decrease the amount of liquid that can held in the gel network as the
junction zones become longer in the presence of excess calcium. Therefore,
calcium concentration is considered to be very important in gel formation. In this
experiment, the breaking pressure of the gel dropped sharply as the calcium
content exceeded 25 mg/g-pectin due to the reason described above.

7000
Breaking pressure of gel (N/m2)

6000

5000

4000

3000

2000

1000

0
0 20 40 60
Concentration of Ca2+ (mg/mg-pectin)

Fig. 2. Effect of Calcium (Ca2+) on breaking pressure of pectin.

4. Conclusions
In this work, an investigation to find new parameter for pectin extraction from
sweet potato pulp with highest percentage yield was carried out. Yield, degree of
esterification and rheological properties such as their stiffness and stress
relaxation was determined. Pectin could be prepared from sweet potato pulp by
hydrochloric acid extraction method with yield of 74.43% from 0.05 M HCl. The
maximum breaking pressure was obtained at 25 mg/g-pectin and the value was
6.45×104 N/m2.

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28 D. N. A. Zaidel et al.

Acknowledgment
The authors would like to acknowledge Universiti Teknologi Malaysia and
Ministry of Higher Education Malaysia for the financial support under
Fundamental Research Grant Scheme (R.J130000.7844.4F447) and Research
University Grant (Q.J130000.2544.06H39).

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