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Carbohydrate Polymers 150 (2016) 299307

Contents lists available at ScienceDirect

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

Effect of high-pressure treatment on the structural and rheological


properties of resistant corn starch/locust bean gum mixtures
Raza Hussain, Hamed Vatankhah, Ajaypal Singh, Hosahalli S. Ramaswamy
Department of Food Science and Agricultural Chemistry, Macdonald Campus, McGill University, 21, 111 Lakeshore Ste. Anne de Bellevue, Qubec, H9X 3V9,
Canada

a r t i c l e

i n f o

Article history:
Received 20 November 2015
Received in revised form 9 May 2016
Accepted 12 May 2016
Available online 16 May 2016
Keywords:
Resistant cornstarch
Rheology
DSC
Locust bean gum
CLSM
High pressure

a b s t r a c t
In this study, effects of a 30 min high pressure (HP) treatment (200600 MPa) at room temperature on the
rheological, thermal and morphological properties of resistant corn starch (RS) (5% w/w) and locust bean
gum (LBG) (0.25, 0.50 and 1.0% w/v) dispersions were evaluated. Results showed that the storage modulus
(G ), loss modulus (G  ), and apparent viscosity values of starch/gum (RS/LBG) mixtures were enhanced
with an increase pressure level, and demonstrated a bi-phasic behavior. HP treated RS/LBG samples were
predominantly either solid like (G > G  ) or viscous (G  > G ), depending on the pressure level and LBG
concentrations. Differential scanning calorimetry (DSC) analysis of the pressurized mixtures showed a
major effect on gelatinization temperatures (To , Tp ,), and it was observed that RS/LBG mixtures gelatinized completely at 400 MPa with a 30 min holding time. Confocal laser scanning microscopy (CLSM)
images conrmed that at 600 MPa, RS/LBG mixtures retained granular structures and their complete
disintegration was not observed even at the endpoint of the gelatinization.
2016 Elsevier Ltd. All rights reserved.

1. Introduction
Starch/hydrocolloid mixtures are frequently used in the food
industry as they impart better quality and stability to the nal
products. Starches, in their native form, often do not exhibit properties prerequisite for a specic application. (Zhu, 2015). Native
starches are often chemically modied and/or blended with other
components such as hydrocolloids (e.g. guar gum, corn ber gum,
hydroxypropylmethylcellulose (HPMC), xanthan gum) to impart
desirable and amending properties for improving product texture
and rheology (Qiu et al., 2015; Rosell, Yokoyama, & Shoemaker,
2011), enhancing cold-storage and freeze-thaw stability (BeMiller,
2011), moisture retention (Kohajdov & Karovicov, 2009), syneresis (Yamazaki et al., 2013), and resistance to various physical
changes during processing (heat, shear, and acidic medium), retrogradation and breakdown.
Locust bean gum (LBG), also known as Carob gum is a natural
hydrocolloid, which has been used in numerous industrial applications (food, cosmetic, textile, pharmaceutical, paint, milling and
construction industries) because of its ability to produce highly viscous solutions at relatively low concentrations (0.02%) to stabilize

Corresponding author.
E-mail address: hosahalli.ramaswamy@mcgill.ca (H.S. Ramaswamy).
http://dx.doi.org/10.1016/j.carbpol.2016.05.039
0144-8617/ 2016 Elsevier Ltd. All rights reserved.

emulsions and as a fat replacement (Barak & Mudgil, 2014). In food


industry, LBG alone or in combination with other hydrocolloids (e.g.
carrageenan, xanthan gum) is widely used in beverages (Dogan,
Toker, & Goksel, 2011), bakery products (Kohajdov, Karovicov, &
Schmidt, 2009), noodles (Silva, Birkenhake, Scholten, Sagis, & van
der Linden, 2013), ice cream (Cropper, Kocaoglu-Vurma, Tharp, &
Harper, 2013), low-fat yoghurt (nal, Metin, & Isikli, 2003) and as
edible coatings (Martins et al., 2012).
Corn starch is also used in many food applications for its
desirable rheological properties (Amini, Razavi, & Mortazavi,
2015). Further, starch and its derivatives can be classied on
the basis of their digestibility as rapidly digested starch (RDS),
slowly digested starch (SDS) and resistant starch (RS) (Englyst,
Kingman, & Cummings, 1992). RS is classied into ve groups
on the basis of its structure and properties: RS 1 physically
inaccessible starch; RS II native granular starch; RS III retrograded starch; RS IV chemically-modied starch and RS V
amylose-lipid complex (Vamadevan & Bertoft, 2015). RS has been
primarily used as a functional ingredient to improve the quality
and dietary ber content (ztrk & Kksel, 2014) of low-moisture
foods such as bakery products (bread and mufns), pasta products,
and in breakfast cereals and snacks (Fuentes-Zaragoza, RiquelmeNavarrete, Snchez-Zapata, & Prez-lvarez, 2010). Additionally,
being a dietary bre, it plays several physiological functions against
colonic carcinogenesis and reducing the risk of diabetes, obesity,

300

R. Hussain et al. / Carbohydrate Polymers 150 (2016) 299307

high cholesterol and other chronic diseases (Choi, Lee, Cho, Choi, &
Moon, 2010).
Several studies have reported that the inuence of hydrocolloids
on corn starch is dependent on the structure and concentration
of the hydrocolloid (Funami et al., 2008; Qiu et al., 2015). The
interaction between starch and hydrocolloid is also reported to
be dependent on the starch source, differences in granule size
(Amini et al., 2015) and crystalline microstructure (Techawipharat,
Suphantharika, & BeMiller, 2008). In order to take advantage of
starch/gum interactions and enhance their applications in food
formulations, it is important to assess the inuence of different
physical and chemical processes, such as heat, high pressure (HP)
and/or enzymatic treatments, on their functional and physicochemical properties (Li, Bai, Mousaa, Zhang, & Shen, 2012). The
majority of earlier reports are focused mainly on the nutritional,
functional and digestibility characteristics of resistant starches,
while the use of resistant starches in food formulations often
involve noticeable presence of other hydrocolloids (gums). Therefore, understanding the interactions between RS and gums are of
considerable interest.
High pressure-processing (HPP) is a non-thermal processing
technology used in the food industry for preparing products
with high functional (Ramaswamy, Singh, & Sharma, 2015), nutritional (Ramaswamy, Chen, & Rattan, 2015) and textural (Alvarez,
Ramaswamy, & Ismail, 2008) qualities. Over the last two decades,
the inuence of HP treatment (i.e. pressure level, treatment time,
starch source, concentration, temperature, etc.) on starch gelatinization in general (Ahmed, Ramaswamy, Ayad, Alli, & Alvarez,
2007; Ahmed, Varshney, & Ramaswamy, 2009; Liu, Selomulyo, &
Zhou, 2008; Oh, Hemar, Anema, Wong, & Neil Pinder, 2008; Oh,
Pinder, Hemar, Anema, & Wong, 2008; Stolt, Oinonen, & Autio,
2000) or more specically on gelatinization of corn starch (Ahmed,
Singh, Ramaswamy, Pandey, & Raghavan, 2014; Baszczak et al.,
2007; Buckow, Heinz, & Knorr, 2007) has been widely investigated. The above studies have been focused on understanding the
mechanism of pressure-induced gelatinization and its inuence on
physicochemical and structural properties of starches. HP treatment has been shown to inuence mostly the non-covalent bonds
in starches, so this treatment can cause major structural, textural,
sensory and nutritional damages (Balny, 2002).
In a recent review covering over fty studies (BeMiller, 2011), a
few have focused on the effect of LBG on different starches but none
have included HPP. To the best of our knowledge, there has been no
study on the effect of HP treatment on starch/LBG mixtures to date.
With this background, the aim of this experimental study was to
evaluate the effect of HP treatment on the rheological, calorimetric
(gelatinization) and morphological properties of starch-LBG mixtures. From an industrial point of view, this research is signicant
because of the dearth of basic data on the dynamics of interactions
between LBG and RS treated with HP which will be useful in the
development of RS-gum based food products.

2. Materials and methods


2.1. Materials
Commercial grade chemically modied resistant starch with
average particle size 1015 m (Hi Maize 260 corn starch termed
as RS) (National Starch and Chemical Co., NJ, USA) was used in the
study. The proximate composition of the starch was: moisture content, 10.8 0.1%; ash content, < 0.4%; fat content, <0.8%; and protein
content, 0.8% (all on dry basis), as provided by the manufacturer. In
order to verify moisture content of the starch sample, RS samples
were dried (in duplicate) in an oven at 130 C to constant weight.
The evaluated moisture content was within 1% of the supplier

data. Laboratory food-grade LBG (galactomannan polysaccharide


100 K0220) was from Sigma Chemicals (St. Louis, USA).
2.2. Preparation of starch and starch/LBG dispersions
Aqueous 5% dispersion of RS was prepared by dispersing 5 g
of starch to 95 g deionized water at room temperature. LBG
powder was dispersed in deionized water at 0.25, 0.50 and 1%
(w/w). Both dispersions were prepared with constant stirring
overnight on a magnetic stirrer (400 rpm) to achieve complete
hydration. Starch/gum dispersion mixtures were prepared by gradually adding 5% (w/w) RS powder into fully-hydrated gum solutions
to achieve samples with the required concentrations of starch/LBG
dispersions. The nal dispersions were stirred for 2 h, and immediately prepared for HP treatments.
2.3. High pressure treatment
Prepared starch/gum dispersion samples were sealed (approx.
20 mL) in low density polyethylene bags (Whirl Pak , Nasco, Fort
Atkinson, WI, USA) and treated by HP at 200, 400 or 600 MPa
for 30 min in a 5 L static HP unit (ACIP 6500/5/12VB; ACB Pressure Systems, Nantes, France) equipped with temperature and
pressure regulators. The pressurization and depressurization rates
were maintained at 4.4 MPa/s and 26 MPa/s, respectively. The initial temperature of pressure transmission medium (water) was
around 18 C, which increased to 2330 C up on pressurization to
200600 MPa due to adiabatic compression. The adiabatic temperature rise is expected to be about 3 C per 100 MPa increase. The nal
processing temperature became steady between 22 and 25 C during the treatment time because the pressure vessel was operated at
room temperature. Each experiment was carried out in triplicate.
2.4. Rheological measurements
2.4.1. Dynamic rheology measurements
Oscillatory rheology measurements of HP treated starch/LBG
dispersions were made using a controlled-stress rheometer (AR
2000, TA Instruments, New Castle, DE, USA). A cone plate geometry was used with a diameter of 60 mm and a cone angle of 2 .
The gap between the base plate and geometry was set at 64 m.
The sample temperature (25 C) was controlled by a Peltier system
and monitored by platinum resistance thermometer sensors (accuracy 0.1 C), which were positioned both in the upper and lower
plates. The sample perimeter was covered with a thin layer of hightemperature-resistant silicone oil to prevent sample dehydration.
Small-amplitude oscillatory strain sweep experiments (0.00110%)
were carried out, and elastic (G ) and viscous (G ) shear moduli, at
a constant frequency of 0.1 Hz were measured to determine the
limit of the linear viscoelastic region (LVR) (Ahmed et al., 2014).
The LVR determination was carried out for the entire range (25 Pa
depending on gel stiffness), and measurements were carried out
accordingly. Frequency sweep tests (0.0125 Hz) were carried out
in the linear regime, at constant strain (0.05%) at 25 C (Ahmed
et al., 2014). All the rheological measurements were carried out
in triplicate and rheological parameters (G , G ) were obtained
directly from the manufacturer supplied computer software (Rheology Advantage software, TA Instruments, New Castle, DE, USA).
2.4.2. Steady ow measurements
Steady ow measurements (shear stress, shear rate, apparent
viscosity) made over a shear rate range from 0.1 to 100 s1 at 25 C
in triplicate. Flow curves of HP treated dispersions at 25 C was
modeled using Rheology Advantage software, TA Instruments, New
Castle, DE, USA. The Power Law model, represented by the following

R. Hussain et al. / Carbohydrate Polymers 150 (2016) 299307

301

Table 1
Storage modulus (G ), loss modulus (G  ) at lower (10 rad/s) and higher (50 rad/s) frequencies and apparent viscosity (at 10 s1 ), consistency index (Pa sn ), ow behavior index
for starch (RS), and RS-LBG dispersions with different concentration at 25 C (without and with high pressure processing).
Samples

G at 10 rad/s

G  at 10 rad/s

G at 50 rad/s

G  at 50 rad/s

 (Pa s) at 10 s1

K (Pa sn )

n ()

R2

5% RS
5% RS/0.25% LBG
5% RS/0.50% LBG
5% RS/1.0% LBG
200 HP/5% RS
400 HP/5% RS
600 HP/5% RS
200/5% RS/0.25% LBG
400/5% RS/0.25% LBG
600/5% RS/0.25% LBG
200/5% RS/0.5% LBG
400/5% RS/0.5% LBG
600/5% RS/0.5% LBG
200/5% RS/1.0% LBG
400/5% RS/1.0% LBG
600/5% RS/1.0% LBG

0.03
0.04
2.81
4.40
0.03
0.04
0.03
0.05
0.04
0.04
0.14
0.17
0.16
6.20
17.65
23.12

0.04
0.09
5.36
6.75
0.07
0.08
0.06
0.08
0.08
0.08
0.52
0.57
0.61
8.34
15.35
19.18

0.92
1.16
11.1
15.7
1.62
1.70
1.56
0.68
1.15
1.12
1.60
1.84
1.74
19.1
35.7
48.47

0.37
0.61
15.12
18.35
0.62
0.74
0.60
0.59
0.74
0.73
2.55
2.71
2.78
20.3
26.9
34.64

0.005
0.020
0.060
0.080
0.012
0.010
0.012
0.014
0.017
0.014
0.053
0.051
0.064
0.530
0.740
0.880

4.12
27.1
21.5
25.3
7.23
8.35
7.02
33.7
35.2
38.5
36.2
42.4
44.3
45.8
54.6
58.3

0.72
0.68
0.66
0.66
0.60
0.58
0.57
0.65
0.62
0.59
0.57
0.56
0.54
0.52
0.50
0.50

0.99
0.95
0.98
0.99
0.96
0.98
0.99
0.93
0.96
0.99
0.98
0.99
0.98
0.98
0.99
0.97

Values are average of three determinations (n = 3); k = consistency index (Pa sn ), n = ow behavior index (dimensionless). R2 values are for n and k.

equation, was used to describe the pseudoplastic behavior of test


samples (Marcotte, Hoshahili, & Ramaswamy, 2001).
(1)
is the shear stress (N/m2 ),

ysis software (Minitab Inc. USA). Statistically signicant effects


were further analyzed, and the means were compared using spearmens multiple range test. Statistical signicance was determined
at p < 0.05.

is the shear rate (s1 ), K (Pa sn )

where 
is the consistency coefcient and n is the ow behavior index (for
Newtonian uids, n is equal to 1 while it is lower than 1 for pseudoplastic uids).
2.5. Differential scanning calorimetry (DSC)
The gelatinization behavior of starch alone and starch/LBG dispersions (before and after HP treatments) were evaluated using a
differential scanning calorimeter (DSC) (TA Q 2000, TA Instruments,
New Castle, DE, USA). Prior to sample measurements, the DSC
was calibrated using indium standard. The samples (68 mg) were
heated from 20 to 150 C at 10 C/min heating rate. An empty pan
was used as a reference and dry nitrogen at a ow rate of 50 mL/min,
was used as the purge gas. Thermal transitions of treated samples
were used to gather information of the gelatinization behavior of
test samples. After treatment at higher pressure levels (400 MPa),
the RS/LBG dispersions were fully gelatinized, so demonstrated
no gelatinization peaks. Gelatinization peaks in the nominal gelatinization temperature range (7090 C) were also not observed for
some starch/LBG samples.
2.6. Confocal laser scanning microscopy (CLSM)
Morphological characteristics of starch/LBG dispersions were
observed using confocal scanning microscope (Nikon Instruments,
Nikon Eclipse TE 800-E, Radiance 2100, BIO-RAD, Japan). A few
drops of the starch dispersions with or without LBG were transferred to a 2-cavity slide (Electron Microscopy Sciences, Hateld,
PA, USA), sealed with nail enamel and observed within 15 min.
A laser with an excitation wavelength of 488 nm and a 40 airimmersion objective was used. Only transmission detectors were
used. Digital image les were acquired at 1024 1024-pixel resolution using 1 3 zoom factors. At least 4 images were obtained
per sample for each replicate, and representative micrographs were
selected.
2.7. Statistical analysis
All experiments were carried out in triplicate and the results
were reported as mean. Data were analyzed by Minitab 16 anal-

3. Results and discussion


3.1. Dynamic rheological properties of starch-gum mixtures
before HP treatments
Fig. 1 shows typical changes in dynamic mechanical spectra
(G and G ) as function of oscillation frequency () for 5% RS and
RS/LBG mixtures at 25 C and the computed values are reported in
Table 1. For RS, G values predominated over G values at lower frequencies (0.110 rad/s), while G exceeded G at high frequencies
(20100 rad/s). The frequency dependent variations in the oscillatory parameters of RS have been recognized to be linked to the
source and concentrations (Ahmed et al., 2014; Lee, Yao, Bae, Lee,
& Lee, 2012). The viscoelastic properties of RS were modied with
the addition of LBG at concentration levels tested (0.251%). For
example, RS/LBG mixture with the lowest concentration of LBG
(0.25%) showed a liquid like behavior (G > G ) at higher frequencies
(25100 rad/s).
However, at higher concentration of LBG (0.50%1%), the G
and G values of the mixtures were much higher than those of
RS alone and/or with 0.25% LBG and the rising curves of these
moduli were almost parallel (Fig. 1). The increase in the values for
the dynamic moduli (G and G ) can be attributed to the dominant viscoelastic properties of added LBG, which is concentrated
within the continuous phase in RS/LBG mixtures as also conrmed by Alloncle and Doublier (1991). Addition of LBG seems to
have a synergistic effect on the rheological properties of RS/LBG
mixtures. In addition, the G values were much higher than the
G at all values (0.1100 rad/s), indicating the dominant viscous ow behavior of RS/LBG mixtures (G > G ) (Table 1). This
result is in agreement with the ndings of Qiu et al. (2015) who
reported that the G values in maize-starch corn-ber gum blends
increased due to the effective concentration of polymers in their
respective micro phases. Generally, this overall shift of starch/gum
gel network from an elastic to a more viscous nature can be
attributed to the mechanisms of phase separation (Closs, CondePetit, Roberts, Tolstoguzov, & Escher, 1999) and thermodynamic
incompatibility between like and unlike polysaccharides (BeMiller,
2011).

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R. Hussain et al. / Carbohydrate Polymers 150 (2016) 299307

Fig. 1. Rheological properties: dynamic mechanical spectra (closed symbols, storage modulus, G ; open symbols, loss modulus, G  ) measured at 0.00110% strain of 5% RS
(w/w) alone and in the presence of LBG (0.25, 0.50 and 1% LBG) (RS and LBG stands for resistant corn starch and locust bean gum respectively). All measurements were made
at 25 C.

3.2. Dynamic rheological properties of starch-gum mixtures after


HP treatments
HP treatment (200600 MPa) of RS slightly increased the G and
G values as compared to the control, followed by a subsequent
decrease at 600 MPa (Table 1). Similar resistance of high amylose
corn and potato starches by HP treatments has been reported earlier
(Ahmed et al., 2014; Oh, Pinder et al., 2008). As shown in Fig. 2, the
magnitudes of G and G of RS/LBG dispersions remained parallel
with an increasing frequency (), showing an expected trend i.e.
the greater the applied pressure and the LBG concentration, the
higher the curves of elastic (G ) and viscous (G ) modulus.
Apparently, the highest viscoelastic moduli curves were
obtained for samples after 600 HP treatment with the highest LBG
concentration (LBG 1%), followed by treatment at 400 and 200 MPa.
Two distinct behaviors were observed based on the pressure levels
and the LBG concentrations. HP treated RS/LBG mixtures with lower
LBG concentration (0.25%) showed a solid like behavior (G > G ) at
higher angular frequencies (20 100 rad/s). RS/LBG mixtures with
0.5% LBG concentration showed a predominant viscous behavior
(G > G ). Moreover, HP treated RS/LBG mixtures displayed lowering
of G values at 0.25 and 0.50% LBG, compared to untreated samples
(Fig. 2, Table 1). This indicated that HP had no signicant effect on
the viscoelastic behavior of the RS/LBG mixtures at these LBG concentrations. However, mixtures HP treated at 400 and 600 MPa with
1% LBG exhibited solid like behavior (G > G ), unlike the untreated
samples. This dramatic shift of HP treated mixtures from elastic
(G > G ) to more viscous (G > G ) may be due to the intensied and
synergistic effect of HP treatment and concentration of the added
LBG (1%).
Although RS granules have a compact outer layer that offers
resistance to pressure (Baszczak, Valverde, & Fornal, 2005), intensive HP treatments may disintegrate the non-covalent bonds and
thus weaken the gel structures (Pei-Ling, Xiao-Song, & Qun, 2010).
Therefore, LBG at variable concentrations (0.251%) appears to play
an important role in inuencing the viscoelastic properties of the
RS/LBG mixtures at different pressure levels. Another explanation
for these differences in rheological behavior (elastic or viscous) at
different LBG concentrations (0.255% and 0.51%) could be the
structural arrangement and intramolecular interactions of the LBG
(Dionisio & Grenha, 2012) accompanied by starch gum intermolecular interaction and a possible phase separation (Kim and
Yoo, 2011). Although, the magnitude of pressure and the concen-

tration of LBG were the main parameters considered in this study,


other factors including type and source of the starch, its swelling
behavior, its modication/manufacturing history, amylose content,
amylose to amylopectin ratio, water-starch-gum interactions, pressure holding time may also inuence the rheological properties
of the starch hydrocolloid systems (Ahmed et al., 2014; Barak
and Mudgil, 2014; Oh, Pinder et al., 2008; Qiu et al., 2015; Stute,
Klingler, Boguslawski, Eshtiaghi, & Knorr, 1996). Further investigations of inter- and intra-molecular starch gum interactions at
other HP processing conditions and other concentrations of gum
may be useful to fully explore the effect of these parameters on the
viscoelastic behavior.

3.3. Steady shear ow behavior of starch-gum mixtures before HP


treatments
The steady shear ow behavior of RS and RS/LBG mixtures (HP
treated and untreated) are presented as apparent viscosity determined at shear rate of 10 s1 , and the ow behavior index (n)
and consistency index (k) data are included in Table 1. The shear
stress () versus shear rate ( ) data at 25 C were well tted with
power law model (Eq. (1)) with high determination coefcients
(R2 = 0.930.99) as shown in Table 1. The ow behavior index (n)
which indicates the extent of shear-thinning behavior or pseudoplasticity as it deviates from 1, was in the range of 0.500.72
(Table 1). The n values did not change much with inclusion of LBG
in the concentration range, 0.251%, indicating low effect of LBG on
ow behavior of starch.
The low degree of pseudoplasticity of LBG can be explained in
terms of its structure. According to Sajjan and Rao (1987), LBG consists of a mannan backbone which tends to coil with the formation
of intramolecular hydrogen bonds. Therefore, it interacts less with
linear amylose molecules because of the lower number of hydroxyl
groups available to form intermolecular hydrogen bonding, and
hence pseudoplasticity decreases. In contrast, K, which represents
the consistency of the sample, a measure sample viscosity and resistance to ow, increased with an increasing concentration of LBG.
These results are in good agreement with those found in corn starch
and rice starch-LBG mixtures (Kim and Yoo, 2011; Yoo, Kim, & Yoo,
2005).

R. Hussain et al. / Carbohydrate Polymers 150 (2016) 299307

303

Fig. 2. Rheological properties: dynamic mechanical spectra (closed symbols, storage modulus, G ; open symbols, loss modulus, G  ) measured at 0.00110% strain of HP
treated RS LBG (5% w/w) dispersions in the presence of LBG (0.25, 0.50 and 1% LBG) (RS and LBG stands for resistant corn starch and locust bean gum respectively). All
measurements were made at 25 C.

3.4. Steady shear ow behavior of starch-gum mixtures after HP


treatments
The n value for RS samples decreased with an increase in pressure level, suggesting that the pseudoplastic behavior of starch
dispesrsion was strengthened by HP treatment. Besides, all HP
treated samples showed decreasing n and increasing K values, compared to the untreated samples, and these values depended on
pressure level and LBG concentration (Table 1). As lower values
of n indicate more pronounced pseudoplasticity, it can be assumed
that HP treatment tends to induce higher pseudoplasticity.
Rheograms of HP treated RS/LBG mixtures with different LBG
concentrations (0, 0.25, 0.50 and 1%) at 25 C as a function of
shear rate (1100 s1 ) indicated the intensication non-Newtonian
(pseudoplastic) nature of the mixture (Fig. 3). The apparent viscosity and consistency coefcient of HP treated mixtures increased
with the incorporation of LBG, a fact attributed to synergism
(Alloncle, Lefebvre, Llamas, & Doublier, 1989). Among all samples, HP treated RS/LBG mixtures at the maximum pressure level
(600 MPa) and highest LBG concentration (1%) had the highest
apparent viscosity and highest shear thinning behavior (Table 1).
As stated by Alloncle and Doublier (1991), starch dispersions may be considered as composite materials comprising of
swollen granules (amylopectin) dispersed in a continuous biopolymer matrix (amylose). Alloncle et al. (1989) reported that in
galactomannan (e.g. locust bean gum) and corn starch mixture dispersions, gums are located within the continuous phase (amylose).
Consequently, if the volume of this phase is reduced, it will cause
an obvious increase in the concentration of gum in the continuous phase, thereby resulting in higher viscosity. The shear thinning
behavior of HP treated mixtures could be due to the alteration
in macromolecular organization of the mixtures at variable shear
rates and applied pressures (Oh, Pinder et al., 2008).
The low viscosity of RS compared to other native starches (corn,
rice, potato, wheat, and tapioca) may result from the modied
structure that is more shear resistant than native starches (Jun,
Lee, Lee, & Lee, 2014). The reasons for low viscosity of HP treated
RS/LBG mixtures are not fully known, but one explanation could be
that higher amylose content of RS somehow, stabilizes the structure under pressure. It could be concluded that the rheological
behavior of RS/LBG mixtures (HP treated and untreated) was less
pressure-dependent but greater gum concentration-dependent.

Table 2
DSC measurements for gelatinization properties of resistant corn starch only (RS)
and RS LBG mixtures (with and without high pressure processing).
Samples

5% RS
5% RS/0.25% LBG
5% RS/0.50% LBG
5% RS/1% LBG
200 HP/5% RS
400 HP/5% RS
600 HP/5% RS
200 HP (5%RS/0.25% LBG)
400 HP (5%RS/0.25% LBG)
600 HP (5%RS/0.25% LBG)
200 HP (5%RS/0.50% LBG)
400 HP (5%RS/0.50% LBG)
600 HP (5%RS/0.50% LBG)
200 HP (5%RS/1% LBG)
400 HP (5%RS/1% LBG)
600 HP (5%RS/1% LBG)

Gelatinization Activity
To ( C)

Tp ( C)

56
70
80
80
37
34
32
63
ND
ND
80
ND
ND
80
ND
ND

81
91
85
85
85
74
68
83
ND
ND
85
ND
ND
85
ND
ND

*
Values are average of three determinations (n = 3); ND: Not detected, RS-LBG
(Resistant starch-locust bean gum dispersions), To (onset temperature), Tp (peak
gelatinization).

3.5. Gelatinization properties of starch-gum mixtures before HP


treatments
The gelatinization behavior of RS-LBG mixtures at different
concentrations of LBG (0.251%) as obtained from the DSC thermograms is shown in Table 2. For RS dispersions (control), melting
peak (Tp ) was recorded at 80.5 C, which is referred as gelatinization
temperature of resistant corn starch (Ahmed et al., 2014). Deviation
from this as inuenced by the addition of LBG and HP treatment is recorded for correlating with rheological and structural
aspects. RS dispersions with the added LBG (0.251% concentrations) showed a different thermal behavior than the control. The
gelatinization temperature range for the RS/LBG mixtures was
predominantly increased beyond the nominal gelatinization temperature range (7090 C). This increase is attributed to RS-LBG
interactions, which produced more stable structures, requiring a
higher temperature for disorganization (Aguirre-Cruz, MndezMontealvo, Solorza-Feria, & Bello-Prez, 2005). Increasing the LBG
concentration (0.25%) reduced the crystallinity of RS suggesting
an interference with gelatinization of RS/LBG mixtures. It can be
presumed that LBG has entrapped RS granules and thus reduced

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R. Hussain et al. / Carbohydrate Polymers 150 (2016) 299307

Fig. 3. Effect of HP treatment on the apparent viscosities of HP treated RS LBG (5% w/w) dispersions in the presence of LBG (0.25, 0.50 and 1% LBG) at 25 C (RS and LBG
stands for resistant corn starch and locust bean gum respectively).

their water absorption activity which is a critical factor for initiating


gelatinization. These results correspond well with some previous
studies on starch-gum interactions (Alamri, Mohamed, & Hussain,
2013; Ferrero, Martino, & Zaritzky, 1996).

3.6. Gelatinization properties of starch-gum mixtures after HP


treatments
HP treatments (200600 MPa) resulted in partial gelatinization
of the RS resulting from partial destruction of the crystalline structure. To values of RS were signicantly decreased from 56 to 32 C,
with no effect on the Tp (Table 2). The gelatinization behavior of HP
treated dispersions could be highly inuenced by starch concentration and amylose contents, pressurization level, temperature and
holding time (Pei-Ling et al., 2010). According to a report on the
classication of starches (based on the pressure resistance) carried
out by X-ray diffraction method, the native corn starches belong to
B type that are partially gelatinized at 600 MPa (Stute et al., 1996).
Nevertheless, the resistant starches contain higher amylose content than the native starches (Jiang, Lio, Blanco, Campbell, & Jane,
2010). Accordingly, it can be speculated that amylose and displaced
amylopectin in starch may develop a pressure stable arrangement
(Oh, Hemar et al., 2008). A similar trend was reported for rice (Oh,
Pinder et al., 2008), potato (Baszczak et al., 2005), barley (Stolt et al.,
2000) and corn starches (Liu et al., 2008) studied under various
high-pressure levels.
HP treatment (200 MPa) of RS/LBG mixtures at 0.25% LBG
concentration showed a signicant decrease in gelatinization temperatures (To , Tp ) as compared to the same concentration of LBG
for the untreated, although this effect was not seen at higher
concentrations (0.501%) (Table 2). The pressure-induced weak
interactions of RS and LBG could be the reason of the decreased
gelatinization parameters. However, no gelatinization peaks were
detected in test samples after treatment at higher-pressure levels
(400600 MPa) indicating the complete gelatinization of RS/LBG
mixtures by HP treatment. Recently, Ma et al. (2012) studied the
effects of high pressure processing on the gelatinization properties
of salt-soluble meat protein gels containing locust bean gum (0.2%)
and reported a decrease in thermal transition temperatures treated
at a pressure of 300 MPa but no peak observed after a treatment at
400 MPa.

Further investigations are required to understand the possible


molecular interactions in starch hydrocolloid systems to fully
explain the mechanism of the inuence of HP treatment on the thermal transition temperatures. It remains unclear if further changes
can occur when the magnitude of pressure and/or holding time are
increased beyond the levels used in this study.
3.7. Morphological descriptions of starch-gum mixtures before
HP treatments
Fig. 4 shows CLSM micrographs of resistant corn starch (5% w/w)
with and without LBG (0.251%), as well as before and after HP
treatment obtained from the confocal scanning microscope. The
RS granules (without gum addition) appeared to exhibit a continuous phase, are angular shaped, well-separated, and fully swollen
due to complete hydration (A). The different microstructures
are included as presented in an earlier study for the concomitant process of starch/gum network formation and starch-water,
and/or starch/starch interactions (Ferrero et al., 1996). RS/LBG
mixtures with the lowest added concentration of LBG (0.25%) represented a weak gel like network in which starch particles tend
to become closer (B). In some sense, at higher concentration of LBG
(0.501.0%), there was indication of formation of clumpy structures
cross-linked by dense aggregated junctions, in addition to the small
evenly dispersed granules (C, D). Although these clumps seem to
be coarser and denser than those of individual starch (A) and of
lower LBG concentration (B), their individual size was considerably reduced. It is well documented that mixing of two polymers
(e.g. starch and gum) could result in phase separation and development of two liquid phases, each phase being enriched with one
of the two polymers (Flory, 1976).
In our particular case, mixing of resistant starch (nongelatinized) into hydrated gum solution was carried out at room
temperature (cold mixing) so it can be expected that no chemical reaction took place and the interactions were intermolecular.
Furthermore, hydrocolloids, such as locust bean gum, are capable of enwrapping the starch granules and physically stabilizing
the particles by acting as a protective layer (BeMiller, 2011). Thus,
several factors such as a possible phase separation and thermodynamic incompatibility of two polymers, high water binding ability
of LBG, and the exclusion and immobilization of water molecules
by the hydrocolloid might contribute to the formation of dense

R. Hussain et al. / Carbohydrate Polymers 150 (2016) 299307

305

Fig. 4. CLSM images of 5% resistant corn starch (w/w) containing 0 (A), 0.25% (B), 0.50% (C), (D) 1% LBG (w/v) with HP treatment at 200 MPa (BD 200 A), 400 MPa (BD 400 A)
and 600 MPa (BD 600 A).

entanglements of RS-LBG dispersions, which are dependent on


LBG concentration.
3.8. Morphological descriptions of starch-gum mixtures after HP
treatments
HP treatment (200 MPa) of RS/LBG mixtures at different concentrations of LBG (0.251%) showed that the majority of the
mixtures kept their integrity (B 200 A to D 200 A), leading to
relatively homogeneous distribution of dispersed phase with limited swelling (C 200 A) and congealed structures (D 200 A).
In general, treatment at a lower pressure not only improved the
starch-gum distribution, but also induced a homogeneity, which
was conrmed by the appearance of single endothermic peak during DSC observations. After being treated at higher pressure levels
(400600 MPa), the RS/LBG granules seem to be stressed and entangled, evident from the loss in shape, optical birefringence, increased
swelling and gelatinization (B D 400 A and B D 600 A). It shows
that this range of pressure was sufcient to distort crystalline
regions of starch and to weaken starch/gum interactions, resulting
in the gelatinization of RS/LBG mixtures (Table 2), but not enough to
disrupt/disintegrate the entire granular structures. Initially, RS/LBG
system collapsed and became sponge-shaped and HP treatment
led to even stronger deformation (C 400 A, D 400 A and C 600 A,
D 600 A). Nevertheless, some studies have shown that after HP
treatments (100% degree of gelatinization), majority of the starch
granules retained their integrity (Pei-Ling et al., 2010; Stolt et al.,
2000; Vallons & Arendt, 2009).
It appears that LBG provide the essential molecular strength
through intermolecular starch-gum interactions. Therefore, it
could be speculated that treatment at a higher pressure is needed
(>600 MPa) to bring out further molecular changes in the morphological characteristics of RS/LBG mixtures though the micrographs
of pressure-induced gelatinization could be different from the
head-induced one. Many small starch particles still retained their
granular structure (see the arrow pointed, B 400 A, C 400 A
and B 600 A), suggesting that small particles were more resistant
to high pressure (Wang et al., 2008). Invariably, these morpho-

logical illustrations provide merely complementary evidence and


complete structural interpretation cannot be ruled out unless it is
complemented with other structural and physical techniques (e.g.
rheological and thermal measurements) (Hussain, Gaiani, & Scher,
2012).
3.9. Relationship between rheological, thermal and
morphological characteristics
Indeed, the strong increase in dynamic moduli (G , G ) of HP
treated starch-hydrocolloid mixtures, correlates well with their
thermal and morphological properties. Two distinctive behavior
were observed based on the pressure level (200600 MPa) and
the LBG concentration (0.251%). The RS/LBG gel network shifted
from an elastic to more viscous nature (G > G ), indicating the
mechanisms of phase separation and thermodynamic incompatibility between the polysaccharides. This phenomenon is also
evident from the micrographs, showing the weaker and more
homogeneous gels at variable pressure and LBG concentration. The
results of higher values of G discussed in Section 3.3 should also
explain why the gelatinization temperatures (To , Tp ) of HP treated
(200 MPa) mixtures decreased compared to the untreated samples.
Concomitant results obtained with CLSM are also associated with
DSC measurements. HP treatment with different concentrations of
LBG not only improved the distribution and homogeneity of RS/LBG
mixtures (Fig. 4C 200 A and D 200 A), but also induced the gelatinization, which was conrmed by the DSC observations (Table 2).
4. Conclusions
It is concluded that characteristics of resistant corn starch/LBG
mixtures treated with high pressure (400600 MPa) were highly
correlated with most of the rheological and structural properties of starch/LBG mixtures. Steady-ow and dynamic rheological
parameters were more dependent on the concentration of gum
rather than the magnitude of pressure treatment, showing a synergistic effect. The extent of pressure-induced gelatinization of
various RS/LBG mixtures was monitored by DSC and it was per-

306

R. Hussain et al. / Carbohydrate Polymers 150 (2016) 299307

ceived that gelatinization parameters are controlled by pressure


and LBG concentration in the starch/gum mixture. The interaction
between starch and hydrocolloid produces a signicant modication of DSC parameters with two distinct behaviors (before and
after HP treatment), and complete gelatinization of mixtures was
observed, being ascertained from their microstructure. Overall, the
HP treatment contributed extensively to the stability of starch-gum
mixtures and their physical and structural properties based on the
magnitude of pressure and the concentration of LBG. These results
suggest that HP treatment offers promise in modifying the structure
and texture of processed resistant starch and gum based products
(e.g. dough used for baking, sauces, beverages and meat products).
Considering the food processing industry, the proper adjustment of pressure levels and resistant starch/gum combinations,
might be useful to slow down the gelatinization process, hence
preserving properties such as retention of avor, color, and aroma,
inactivation of enzymes and microorganisms and impart physical properties, which will enable better control of processing.
As this is the very rst report in this vein, further investigation
should be done for a better understanding of the mechanism
of pressure-induced gelatinization. Particularly, pressure-induced
macromolecular interactions between starch and gum during gelatinization should be investigated at molecular levels with Fourier
transfer infrared (FTIR) and particle size analyzer. The effect of different starches (native corn, waxy corn etc.) and gums with vast
range of concentrations should be explored. The effect of higher
pressures (>600 MPa) combined with various holding times and
temperatures are also important factors that remained unexplored.
A future investigation will compare the effect of these parameters on temperature-induced gelatinization and pressure-induced
gelatinization.
Acknowledgements
The authors are gratefully indebted to Natural Sciences and
Engineering Research Council of Canada (NSERC) for the Post Doc.
fellowship of the rst author. The authors also wish to thank Dr.
Pramod Pandey, Ali. R. Teherian and Dr. Jaun. Pablo. C. Montes for
helpful discussions to improve the quality of this paper.
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