Food Structure 11 (2017) 16–23

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Food Structure
journal homepage: www.elsevier.com/locate/foostr

Influence of pH, pectin and Ca concentration on gelation properties of

low-methoxyl pectin extracted from Cyclea barbata Miers
Oni Yuliartia,* , Amanda Low Sok Hoonb , Sin Yee Chongb
a
School of Chemical and Life Sciences, Singapore Polytechnic, 500 Dover Road, Singapore, Singapore
b
Massey University, Singapore Campus, Block T1A, 500 Dover Road, Singapore

A R T I C L E I N F O A B S T R A C T S

Article history:
Received 23 March 2016 Pectin isolated from Cyclea barbata Miers leaf (green jelly leaf) forms a gel at a low pH (
3) in the
Received in revised form 21 September 2016 absence of co-solutes and divalent ions at room temperature. These gelling properties are important and
Accepted 21 October 2016 particularly useful in the development of low-sugar food systems. In this study, rheological
Available online 8 November 2016 measurements were employed to determine the effect of varying different levels of pH, pectin, urea
and divalent ions (Ca2+) concentration towards the gelation properties of the isolated pectin. The gelation
Keywords: time and gel physical properties were investigated by determining the gel-sol transition time and the
Green jelly leaf mechanical spectra (G’ and G”) using oscillatory measurements. This study showed that acid-induced
Gelation
gelation of the extracted pectin was mediated by hydrogen bonding and gelation only occurred at pH 3.
Low-methoxyl pectin
The results obtained from oscillatory measurements showed that gel strength of green jelly leaf (GJL)
Acid
Calcium pectin was affected by varying pectin concentration where gel strength increased with increasing pectin
concentration. However, when compared with a pectin with a lesser charge density (Genu pectin, DE
31%), the gel strength of GJL pectin at different concentrations were lower. Presumably the abrupt
formation (due to highly negative charge) of cross-linking junction of GJL pectin molecules had led to un-
homogenous gel network and resulting in weaker gels.
© 2016 Elsevier Ltd. All rights reserved.

1. Introduction formation and protein stabilisation. Currently apple pomace and

citrus peels are the two main sources of commercial pectins (May,
Pectin is a complex polysaccharide of the cell walls of growing 1990). Based on their degree of esterification (DE), pectins are
plants, and mainly consists of partly methylesterified a-1,4-linked grouped into high methoxyl pectin (HMP, DE > 50%) and low-
d-galacturonic acid (Mohnen, 2008). Pectins are categorised into methoxyl pectin (LMP, DE < 50%) (Rolin & De Vries, 1990). In
three main polymer groups (Ridley, O'Neill, & Mohnen, 2001): general, HMP forms a gel in an acidic environment (typically
homogalacturonan, rhamnogalacturonan I and substituted gal- pH
3) and in the presence of low molecular weight co-solutes,
acturonan. Homogalacturonan has a backbone with a linear chain such as sucrose, at concentrations of
65% (Evageliou, Richardson,
of a-(1,4)-linked D-galacturonosyl acid, where Rhamnogalactur- & Morris, 2000). In contrast, LMP forms a gel in the presence of
onan I has a backbone of a-1,2-linked L-rhamnosyl and a-1, 4- ionic calcium. However, LMP can also gel in the absence of Ca2+ by
linked D-galacturonosyl acid residues, with neutral sugars such as lowering the pH to below 3.3 (Gilsenan, Richardson, & Morri,
arabinose and galactose as the side chains. 2003). The ability of LMP to gel at acidic condition is related to the
Pectin is a water soluble polysaccharide (WSP) that is progressive reduction of the charge density of pectin with
deliberately added to food systems to enhance their functional decreasing pH (Capel , Nicolai, Durand, Boulenguer, & Langendorff,
properties. The most significant functional properties of WSPs are 2006) which in turn induces aggregation and eventually gelation.
their water binding capacity and their viscosity enhancement. Vincent, Mansel, Kramer, Kroy, and Williams (2013) describes
WSPs are widely used as stabilisers and viscosifiers to control the that the acid-induced gelation of LMP is attributed by hydrogen
texture of semi-solid foods (Wang & Cui, 2005). Thus, pectin, in bonding between protonated carboxylic acid groups on the
general, has three functional properties: viscosity build-up, gel galacturonic acid units of pectin backbone and also between the
hydroxyl groups of neighbouring molecules. This gelation mecha-
nism of LMP depends on intrinsic and extrinsic factors of pectin
* Corresponding author. such as pectin concentration, divalent ions concentration, pH,
E-mail address: [email protected] (O. Yuliarti).

http://dx.doi.org/10.1016/j.foostr.2016.10.005
2213-3291/© 2016 Elsevier Ltd. All rights reserved.
 O. Yuliarti et al. / Food Structure 11 (2017) 16–23 17

temperature and soluble-solid type and concentration (Axelos & couple plasma spectrophotometry (ICP, Shimadzu ICPE-9000,
Thibault, 1991). Singapore) based on the AOAC method (AOAC, 1990). Neutral
Cyclea barbata Miers leaf is a climbing shrub grown in many sugar composition of the GJL polysaccharide fraction was
parts of Asia. Traditionally, the water extract from the leaves is determined by gas liquid chromatography (GLC, BPX-70 column),
consumed in the form of a gel or infusion and is used to treat in the form of alditol acetate derivatives as described by Englyst,
digestive disorders and fever in some parts of Asia such as Thailand Quigley, and Hudson (1994) using an Englyst Kit for NSP
and Indonesia. It is found that the leaves of Cyclea barbata Miers determination (Englyst Carbohydrate Ltd, UK). The GLC column
form a gel instantly, without addition of other ingredients, when temperature was 220  C, and the injector and detector were
blended with water at room temperature (Kooiman, 1969; maintained at 180  C. The carrier gas (hydrogen) flow rate was
Arkarapanthu, Chavasit, Sungpuag, & Phuphathanaphong, 2005). 8 mL/min. The GalA content was determined through colorimetric
These authors indicated that pectin is the polymer responsible for method as method described by Scott (1979) using an UV-160A
the gelation to occur. spectrophotometer (Shimadzu, Douglas Scientific, Singapore).
As the spontaneous gelling of the pectic polysaccharide can be The recovered GJL purified pectin was analysed for DE by using
useful in food gel systems that require no heat, additional infrared spectroscopy (Spectrum Two IR, Perkin Elmer U.S.A)
ingredients or pH reduction, study on gelation rheological according to the method described by Videcoq, Garnier, Robert,
properties of pectin extracted from Cyclea barbata Miers need to and Bonnin (2011). Briefly, four commercial pectin films with
be carried out. The aim of this study is to investigate the effect of different DE (Sigma P9135, Sigma P8471, Danisco AMD 783 and
varying pH values, pectin concentration, and divalent ion (Ca2+) Genu Pectin CP Kelco) were prepared in duplicate by drying the
concentration on rheological properties of gelation of pectin pectin solutions (0.5% w/w) at 50  C on petri dishes for
18 h. From
isolated from Cyclea barbata Miers leaf. the spectra of Fourier Transform Infra-Red (FT-IR), the absorptions
bands at 1740 and 1624 cm1 are assigned to C¼O stretching
2. Materials and methods vibrations of esterified carboxyl groups and carboxylic acid
respectively. An equation of prediction was obtained from the
2.1. Materials standard curve with a coefficient of determination R2 = 0. 9942.
This analysis was operated using the Spectrum AssureID software
The Cyclea barbata Miers leaf was obtained from a plantation (AssureID ES version, Perkin Elmer U.S.A). The spectrum obtained
area in West Lombok, Indonesia. The leaves were sorted, washed, was then compared against the library of spectra from the database
packaged in a container and freeze (–20  0.4  C). The pH of the fresh (Sadtler Basic Monomers & Polymers, V1, 2012, Perkin Elmer U.S.
leaf water extract was 6.9  0.2. The purified pectin has a A).
galacturonic acid (GalA) content of 36% w/w, DE 10% and The size exclusion chromatography coupled to a multi-angle
average-weight molecular weight (Mw) of
4.4  105 g/mol. With laser light scattering (SEC-MALLS) was used to determine the
regards to the minerals content, it consists of calcium (
0.57% w/w) weight-average molecular weight (Mw) according to the method
and zinc (
0.54% w/w). Commercial pectin used in this study was described by Yuliarti et al. (2015). The eluent used was 0.1 M
Genu1 Pectin (DE 31%) obtained from Cp Kelco (Singapore). All sodium chloride and 0.02% w/v sodium azide in Milli-Q water. The
chemicals were purchased from Sigma Aldrich (Singapore), unless SEC-MALLS system used in this study comprised of a HPLC
otherwise specified. quaternary pump (Agilent Technologies 1200 series, U.S.A)
connected in series to a Shodex SB-6 guard column, Shodex SB-
2.2. Extraction of cyclea barbata miers leaf pectin 805 size exclusion column (Shodex, Tokyo, Japan), DAWNTM 8+
MALLS detectors (Wyatt Technology, U.S.A), ViscoStarTMII viscom-
Sodium citrate, 1% w/v (Danisco, Singapore) in deionised water eter (Wyatt Technology, U.S.A) and a DRI detector (Agilent
(DI) was used to extract the pectin from Cyclea barbata Miers leaves Technologies, U.S.A). The software used to operate the HPLC was
in order to prevent cation-mediated gelation during the extraction Agilent Chem Station LC1200 (Agilent Technologies 1200 series, U.
process. The extractionwas carried out by blending (Waring Blender S.A) and the light scattering and viscometric data were analysed
7011 HS, U.S.A) the leaves with the chelator-solution at a ratio of using Astra 6.1 (Wyatt Technology, U.S.A).
1:25 (w/v) at low speed for 30 s. The mixture was filtered through
three layers of cheesecloths to obtain a clear water extract. The 2.4. Rheological measurements
residue was re-dispersed in the chelator-solution and blended again
to obtain a second extract. The two filtrate samples were combined. Pectin stock solution (0.5% w/v) of GJL and commercial LMP
The filtrate was stirred for 1 h at room temperature for the chelating (Genu1 Pectin) was prepared by hydrating the samples in Milli-Q
process to take place. The filtrate was then centrifuged at 4000g for water for 4 h (
25  C) to achieve complete dissolution. The pH of
15 mins at 20  C (Biofuge Stratos Haraeus, Buckinghamshire, the GJL and Genu1 Pectin solutions was found to be 6.08 and 4.05
England) in order to remove any insoluble matter. Ethanol (80% respectively. The pH of Genu1 Pectin was adjusted similar to GJL
v/v, VWR, Singapore) was used to precipitate the pectin at the ratio pectin by addition of 0.1 M NaOH. Preliminary experiment was
of 1:4 (v/v). The recovered precipitated pectin was rinsed twice with conducted to determine the relation between pH and amount
80% (v/v) ethanol, followed by drying in an oven (EU53JOUAN SA, Glucono-delta-lactone (GDL) required using 0.2% w/w pectin
France) at 50  C  0.5  C for 12 h. The crude pectin extract was concentration at room temperature. GDL was used as an acidulant
further purified by hydrating the pectin in DI water (0.5% w/v) for in this study as GDL slowly hydrolyses in aqueous solution to form
2 h. The solubilised crude pectin was then centrifuged at 14,000g for gluconic acid, which gives a gradual decrease in pectin solution pH.
40 mins at 20  C to remove any insoluble components. The collected Small and large deformation oscillatory was performed using a
supernatant was precipitated and dried as described above to Paar Physica MCR 301 rheometer (Anton-Paar, GmbH, Germany)
recover the green jelly leaf (GJL) purified pectin. equipped with a cup and bob measuring system (C-CC27/T200
Cup, B-CC27/Q1 Bob).
2.3. Characterisation of isolated purified pectin The effect of pH on the gelation properties of GJL pectin was
studied using 0.2% (w/v) pectin solution and 15%, 7%, 0.7% and
The mineral compositions of the GJL and the extracted 0.075% (w/w) of GDL crystals to achieve the pectin solution final pH
polysaccharide fractions were determined using an inductively of 2, 3, 4 and 5, respectively. To prevent the formation of gel due to
18 O. Yuliarti et al. / Food Structure 11 (2017) 16–23

addition of GDL before the measurement, pectin solutions were concentration of calcium phosphate monobasic monohydrate at
cooled to 5  C in an ice water bath during addition of GDL. Slow pH 3. The calcium concentration is expressed as relative to the
dissociation of GDL to gluconic acid occurs at lower temperature. amount of unesterified galacturonic acid residues in the pectin
The mixture was stirred under constant stirring for 1 min and the backbone (COO) (Ström et al., 2007), and in this study the
temperature of pectin solutions were maintained at 5  C. stoichiometric ratio (R = 2 * [Ca2+]/[COO]) gave the R value of 1.5,
Approximately 20 mL of the pectin solution was loaded into a 3.0, 4.5 and 6.0. Calcium phosphate was added to the pectin
pre-equilibrated (5  C) rheometer. The temperature of the sample solution (5  C) followed by addition of GDL. The mixture was
was raised from 5  C to 20  C at 1  C/min to allow gradual stirred for 1 min and the temperature was maintained at 5  C. As
dissociation of GDL in pectin solution and the transition of sol the pH of pectin solution decreases with slow dissociation of GDL,
to gel to take place. Temperature sweep (5  C–20  C) at constant Ca2+ will be released slowly into solution (Lootens et al., 2003). GDL
strain, 1% (within the linear viscoelastic region) and frequency of was used to decrease the pH of pectin solution as it dissociates
1 Hz was conducted to capture the changes in G’ and G” of GJL slowly in solution ensuring a more uniform distribution of H+ in
pectin gels. As the temperature of the sample reached 20  C, time solution. The release of H+ due to GDL hydrolysis causes the
sweep measurement was carried out for 140 mins at 20  C. calcium phosphate to dissociate forming calcium ions. The Ca2+
Frequency sweep was conducted from 0.1 rad/s to 100 rad/s then crosslink the protonated carboxylic acid groups on the
angular frequency at constant 1% strain and 20  C following the galacturonic acid units. In this study, the stoichiometric amount of
time sweep. GDL used was 0. 5, 10, 15 and 20 mM.
A frequency sweep test (0.1–100 rad/s, 1% strain, pH 3) was The frequency sweep was conducted at 20  C, from 0.1 to
performed at 20  C to investigate the frequency dependence of the 100 rad/s angular frequency at constant 1% strain following the
moduli of the GJL pectin gels. The pectin concentration was varied method as described above. Small deformation oscillatory time
to i.e. 0.1, 0.2, 0.3, 0.4 and 0.5% w/v. Similar to the method as sweep (1% strain, 1 Hz) was carried out to observe the gel network
described above, the pectin solution temperature was raised from development of GJL pectin when urea (1 mol/L) was added. The
5  C to 20  C at 1  C/min to allow the gradual dissociation of GDL in pectin concentration and pH used was 0.2% w/v and 3 respectively.
pectin solution and the gelation to occur. The gel was allowed to set Similar to the method as described above, the pectin solution
for 140 mins at 20  C before frequency sweep measurement was temperature was raised from 5  C to 20  C at 1  C/min to allow the
carried out. gradual dissociation of GDL in pectin solution and the gelation to
The effect of addition of calcium ions (Ca2+) on the gelation occur. To prevent the evaporation during measurement a mineral
properties GJL pectin (0.2% w/v) was studied by varying the oil was placed on the surface of the sample for all measurements.

Fig. 1. Temperature sweep responses of GJL pectin gel (0.2% w/w) at different pH (1 Hz and 1% strain). Filled and unfilled symbols represent storage modulus (G’) and loss
modulus (G”), respectively.
 O. Yuliarti et al. / Food Structure 11 (2017) 16–23 19

3. Results and discussions chain interactions occur, promoting association and gelation of the
system. At a lesser acidic condition (pH 3), less dissociation of GDL
3.1. Effect of pH occurred and this resulted in a higher number of charge density of
pectin molecules ( . . . COO) which led to stronger electrostatic
The effect of pH on the gelation of Cyclea barbata Miers leaf repulsion between chains. As shown (Fig. 1b), it seems that more
pectin in the absence of calcium ions (Ca2+) was studied using a energy was required to promote the association between pectin
fixed pectin concentration (0.2% w/w) at different levels of pH i.e. 2, chains, hence the gel formed at higher temperature. In the case of
3, 4 and 5. In this study, the acidification of GJL pectin solution by pH 4 and pH 5, there was no cross-over between the storage
GDL was carried out at 5  C in order to prevent the spontaneous modulus G’ and the loss modulus G” of the pectin solution
gelation by rapid dissociation of GDL at higher temperature. observed; rather a continuous decrease in both moduli with
Acidification at higher temperature approximately >14  C and at increasing temperature, with G” dominating over the entire
lower pH value would lead to a rapid gel formation. Fig. 1a–d show temperature range (Fig. 1c and d).
the changes in the storage modulus G’ and the loss modulus G” Fig. 2a–d shows the dependence of gelation time of GJL pectin
obtained when pectin solutions temperature was increased from 5 on the pH. As observed, the gelation only occurred at pH 2 and 3
to 20  C at different pH values. and there was no gel formed at above pH 3. At pH 2, the gel formed
As observed in Fig. 1a for pH 2, initially, the viscous character of eairlier (
8 mins) than at pH 3 (
17 mins). This implies that with
the system dominated at lower temperature (G” > G’). As the decreasing the pH to below 4, the charge density of pectin
gelation temperature (
14  C) was reached, the elastic modulus molecules decreased, and this consequently decreased the electro-
crossed over the loss modulus. A similar trend was observed for pH satic repulsion between pectin molecules and therefore gel was
3, however the development of storage modulus occurred at higher formed. Fig. 3a and b show the illustrative mechanical spectra
temperature (
20  C). This finding could suggest that, the gelation recorded after 150 mins of storage at 20  C for two different pHs (2
of GJL pectin was strongly promoted by the reduction of pH. As and 3). The pH 4 and 5 were excluded since there was no gel formed
dissociation of GDL increased at higher temperature (pH was observed. The mechanical response at pH 3.0 (Fig. 3a) was typically
decreased with addition of H+), the progressive association of GalA gel-like with storage modulus G’ was greater than G” (G’ > G”) over
units of pectin molecules ( . . . COO) and H+ occurred, forming most of the accessible frequency range (0.1–100 rad/s). When
larger un-dissociated form of carboxylic acid ( . . . COOH), decreas- further reduction in pH to 2, the gel-like character became more
ing the charge density until the critical temperature was reached. pronounced with less variation over frequency range was observed
Rascon-Chu et al. (2009) reported that the repulsive forces and the increase in the storage (G’) and loss (G”) modulus became
between carboxyl groups of pectin molecules are weaker when more obvious. Ström et al. (2007) reported that the modulus of the
there are fewer charges on the pectin chains, hence more chain– acid-pectin gelation network increases and influenced by the

Fig. 2. Time sweep of GJL pectin (0.2% w/w) at different pH (1 Hz and 1% strain). Filled and unfilled symbols represent storage modulus (G’) and loss modulus (G”),
respectively.
20 O. Yuliarti et al. / Food Structure 11 (2017) 16–23

Fig. 3. Mechanical spectra (1% strain) recorded after 150 mins storage at 20  C (a) pH 2; (b) pH 3.

formation of junction zones, where according to Gilsenan, of both pectins were strongly dependent on pectin concentrations
Richardson, and Morris (2000) this network is governed solely where G’ increased with increasing pectin concentration (Fig. 4c).
by hydrogen bonds between the un-protonated and protonated This could be attributed to an increase in the number of cross-
carboxyl group of the pectin molecules. The significant change in linking junction zones between the pectin chains. As the
both moduli in higher acidic condition (pH 2), could suggest that concentration of pectin increases, the number of carboxylic acid
the interactions between the pectin chains was more intensive, groups ( . . . COOH) of pectin molecules in the system (at pH 3)
due to less repulsion and led to a higher number of binding sites increases, providing less repulsive forces and hydrogen bonding to
that formed the gel network in a given polymer concentration. form, thereby strengthen the gelation network. An increase in the
gel strength with increasing pectin concentration is widely reported
3.2. Effect of pectin concentration in the literature (Rascon-Chu et al., 2009; da Silva & Rao, 2006; Rao,
van Buren, & Cooley, 1993). When the pH is remained constant, an
The mechanical spectra of GJL pectin gel for different pectin increase in the number of pectin molecules that are able to interact
concentration (0.1, 0.2, 0.3, 0.4 and 0.5% w/v) at constant pH (pH 3) is to form the junction zone network leads to stronger gels.
shown in Fig. 4a. For a comparison, commercial LMP Genu pectin (DE Interestingly, the change in elastic modulus (G’) of Genu pectin
31%) was used in this study (Fig. 4b). As shown, the magnitudes of G’ was more apparent than GJL pectin. The elastic modulus values for
 O. Yuliarti et al. / Food Structure 11 (2017) 16–23 21

Fig. 4. Effect of pectin concentration (% w/w) on the changes of modulus (G’, G”) prepared with different pectins (a) GJL; (b) Commercial Genu pectin (pH 3); (c) storage
modulus of GJL (*) and commercial pectin () as function of pectin concentration (pH 3, 1rad/s).

pectin concentration of 0.1 and 0.5% w/w were 1.34 Pa and 156 Pa gelation thereby strengthening the gel network. However, the
(GJL pectin) respectively and 3.93 and 383 Pa for commercial Genu increase in gel strength was not observed from the critical limit R
pectin. Both of the samples showed an approximately increase in value of 3.0. Beyond this critical limit (R > 3.0), the gel strength
the moduli by a factor of 10 over the investigated range. This remained low as the excess Ca2+ could bind to other anionic sites
indicates that pectin with a lesser charge density (Genu pectin, DE along the pectin molecules and contribute to ionic repulsion
31%) exhibited stronger gel strength than higher charge density (Lootens et al., 2003).
pectin (GJL pectin, 10% DE), regardless the pectin molecular weight.
Noted that GJL pectin was higher in molecular weight (
4.4  105 3.4. Effect of addition urea
g/mol) than Genu pectin (
1.6  105 g/mol). Presumably the abrupt
formation (due to highly negative charge) of cross-linking junction The effect of addition urea on acid-induced gelation of Cyclea
of GJL pectin molecules had led to un-homogenous gel network barbata Miers leaf pectin was studied by addition of 1 mol/L urea to
formation and resulting in weaker gels. 0.2% (w/v) pectin solution at a constant pH 3 and temperature of
20  C. Fig. 6 shows the time sweep of pectin solution without and
3.3. Effect of calcium concentration with the presence of urea for duration of 90 mins. The cross-over of
storage modulus G’ and the loss modulus G” at 17 mins for pectin
The effect of addition of divalent ions (Ca2+) on gelation solution indicated the occurrence of a sol-gel transition. On the
strength of Cyclea barbata Miers leaf pectin was studied using 0.2% other hand, no sol-gel transition was observed for pectin solution
(w/v) pectin solution at pH 3. Fig. 5 shows the elastic modulus G’ of at pH 3 when 1 mol/L urea was included. This implies that addition
pectin solutions at different R values (0, 1.5, 3.0, 4.5 and 6.0) at the of urea had inhibited the formation of hydrogen bonds of pectin
frequency of 1 rad/s and temperature of 20  C. The addition of molecules, hence no gelation was formed. Urea forms more
divalent ions at R values of 1.5 and 3.0 had increased the gel energetically favourable hydrogen bonds with carboxylic acid
strength of pectin as divalent ions provide another means of groups and hydroxyl groups than hydrogen bonding stabilizing the
formation of junction zones via the ‘egg-box’ model. This ionic pectin network junction zones. Thus, as when urea was added,
bonding between Ca2+ and free carboxylic pectin molecules there is less hydrogen bonding between protonated carboxylic acid
( . . . COO) supplements the hydrogen bonding in acid-induced groups ( . . . COOH) on the galacturonic acid units of pectin
22 O. Yuliarti et al. / Food Structure 11 (2017) 16–23

Fig. 5. Effect of calcium phosphate solution concentrations on the storage modulus (G’) of GJL pectin gels (0.2% w/v, pH 3).

backbone and between the hydroxyl groups ( . . . OH) of neighbour- 4. Conclusions

ing molecules stabilizing the pectin network junction zones.
Therefore, the resulting gel strength deceased up to the point In this study, rheological measurements were employed to
where no gel formation was observed. The results of this study is in determine the effect of varying different levels of pH, pectin and
agreement with literature data reported by Vincent et al. (2013), divalent ions (Ca2+) concentration towards the gelation properties
where hydrogen bonding are responsible in the formation of of pectin isolated from Cyclea barbata Miers leaf. The gelation time
cross-link in acid-pectin gel network. Therefore, it can be deduced and gel physical properties were investigated by determining the
that hydrogen bonds are the primary bonds which stabilize the gel-sol transition time and the mechanical spectra (G’ and G”) using
pectin network in acid-induced gelation of Cyclea barbata Miers oscillatory measurements. This study showed that acid-induced
leaf pectin. gelation of the extracted pectin was mediated by hydrogen

Fig. 6. Effect of addition urea (1 mol/L) on the gelation time of GJL pectin gel (0.2% w/v, pH 3) with constant frequency (1 Hz) and strain (1%).
 O. Yuliarti et al. / Food Structure 11 (2017) 16–23 23

bonding and gelation only occurred at pH  3. The results obtained Gilsenan, P. M., Richardson, R. K., & Morris, E. R. (2003). Associative and segregative
from oscillatory measurements showed that gel strength was interactions between gelatin and low-methoxy pectin: Part 1. Associative
interactions in the absence of Ca 2+. Food Hydrocolloids, 17(6), 723–737.
affected by varying pH, pectin concentration and divalent ions (Ca2 Kooiman, P. (1969). Cold water-extractable pectin in cell walls of plant leaves.
+
) levels. Gel strength increased with decreasing pH, increasing Journal of the Science of Food and Agriculture, 20(1), 18–20.
pectin concentration, and a similar increasing pattern was Lootens, D., Capel, F., Durand, D., Nicolai, T., Boulenguer, P., & Langendorff, V. (2003).
Influence of pH, Ca concentration, temperature and amidation on the gelation of
observed with increasing Ca2+ concentration levels, however it low-methoxyl pectin. Food Hydrocolloids, 17(3), 237–244.
was only up to a critical limit R value of 3.0. The ability to form soft May, C. D. (1990). Industrial pectins–Sources, production and applications.
gel via acid-induced gelation without the need for co-solutes Carbohydrate Polymers, 12(1), 79–99.
Mohnen, D. (2008). Pectin structure and biosynthesis. Current Opinion in Plant
enables Cyclea barbata Miers leaf pectin to be used in low pH food Biology, 11(3), 266–277.
systems such as low-caloric fruit jams or beverages. Rao, M. A., van Buren, J. P., & Cooley, H. J. (1993). Rheological changes during gelation
of high-methoxyl pectin fructose dispersions-effect of temperature and aging.
Journal of Food Science, 58(1), 173–176.
Acknowledgements
Rascon-Chu, A., Martinez-Lopez, A. L., Carvajal-Millan, E., de Leon-Renova, N. E. P.,
Marquez-Escalante, J. A., Romo-Chacon, A., et al. (2009). Pectin from low quality
Authors acknowledge Or Cheng Cheng for the help during the ‘Golden Delicious' apples: Composition and gelling capability. Food Chemistry,
research and Utari for providing the green jelly leaf. 116(1), 101–103.
Ridley, B. L., O'Neill, M. A., & Mohnen, D. (2001). Pectins: Structure, biosynthesis, and
oligogalacturonide-related signaling. Phytochemistry, 57(6), 929–967.
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