Food Chemistry 131 (2012) 500507

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Food Chemistry
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Effect of gelatinisation on slowly digestible starch and resistant starch of

heat-moisture treated and chemically modied canna starches
Juraluck Juansang a, Chureerat Puttanlek b, Vilai Rungsardthong c, Santhanee Puncha-arnon a,
Dudsadee Uttapap a,
a
Division of Biochemical Technology, School of Bioresources and Technology, King Mongkuts University of Technology Thonburi, 49 Soi Tientalay 25, Bangkhuntien-Chaitalay Road,
Takham, Bangkhuntien, Bangkok 10150, Thailand
b
Department of Biotechnology, Faculty of Engineering and Industrial Technology, Silpakorn University, Nakhon Pathom 73000, Thailand
c
Department of Agro-Industrial Technology, Faculty of Applied Science, King Mongkuts University of Technology North Bangkok, 1518 Pibulsongkram Road, Bangsue, Bangkok
10800, Thailand

a r t i c l e

i n f o

Article history:
Received 31 May 2011
Received in revised form 20 July 2011
Accepted 6 September 2011
Available online 16 September 2011
Keywords:
Canna starch
Gelatinisation
Heat-moisture treatment
Modied starch
Octenyl succinylation
Resistant starch
Slowly digestible starch

a b s t r a c t
The effects of gelatinisation on slowly digestible (SDS) and resistant starch (RS) of native and modied
canna starches were investigated. Starch slurries (10% w/w) were gelatinised at 100 C for 5, 10, 20
and 40 min using a rapid visco analyzer (RVA). Signicant change in the degree of gelatinisation (DG) values of all starch samples was observed during the initial 10 min of gelatinisation; after that the DG values
increased gradually with gelatinisation time. The RS contents in all gelatinised starches decreased with
increasing gelatinisation time, while the SDS values uctuated. Chemical modication affected DG values
as well as RS/SDS contents. The RS contents in 10% (w/w) acetylated, hydroxypropylated, octenyl succinylated and cross-linked canna starches gelatinised at 100 C for 40 min were 26.6%, 32.0%, 45.3% and 19.8%,
respectively, which were higher than that of the native starch (12.4%). Canna starch modied by crosslinking had the highest SDS content when gelatinised for 2040 min. Modication of canna starch by
heat-moisture treatment resulted in a lower content of RS for all treated samples. However, the VtHMT25 (canna starch containing moisture content of 25% during heat treatment) when gelatinised for
520 min contained a higher amount of SDS, compared to unmodied starch. The most effective modication method for RS and SDS formation was octenyl succinylation, where the sum of RS and SDS
approached that of Novelose260.
2011 Elsevier Ltd. All rights reserved.

1. Introduction
Starch is the main carbohydrate in human nutrition, and is one
of the most important sources of biological fuel for humans. For
nutritional purposes, starch is generally classied into rapidly
digestible starch (RDS), slowly digestible starch (SDS) and resistant
starch (RS), depending on the rate and extent of its digestion
(Englyst, Kingman, & Cummings, 1992). RDS induces a rapid increase of blood glucose and insulin levels after ingestion. SDS
prolongs the release of glucose, thus preventing hyperglycaemiarelated diseases. RS reduces starch availability for digestion and
produces short-chain fatty acids in the large bowel through fermentation, which is benecial for colon health and protection
against colorectal cancer (Lehmann & Robin, 2007). Consequently,
starch ingredients with high levels of SDS and RS can improve the
nutritional function of foods. RS has been categorised into four

Corresponding author. Tel.: +66 2 470 7754; fax: +66 2 452 3479.
E-mail address: dudsadee.utt@kmutt.ac.th (D. Uttapap).
0308-8146/$ - see front matter 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2011.09.013

types, of which chemically modied starches are classied as RS

type 4 (Eerlingen & Delcour, 1995).
A decrease in the susceptibility of chemically modied starches,
either in native granular form or in a gelatinised state, to enzyme
hydrolysis was shown in a recent report (Lehmann & Robin,
2007). Substitution of starch hydroxyl groups with citryl (Wolf,
Bauer, & Fahey, 1999; Xie & Liu, 2004), acetyl (Chung, Shin, &
Lim, 2008; Hoover & Sosulski, 1985), octenylsuccinyl (Han & BeMiller, 2007; He, Liu, & Zhang, 2008; Heacock, Hertzler, & Wolf,
2004) and hydroxypropyl (Conway & Hood, 1976; Han & BeMiller,
2007) substituents has been found to decrease starch digestibility
by a-amylase. However, studies examining the in vitro digestion of
cross-linked starches have yielded contradictory observations. A
signicant reduction in digestibility of starch cross-linked by a
phosphate bridge was reported by Sang and Seib (2006) and Woo
and Seib (2003); whereas no (or only slight) changes were found
by Chung et al. (2008), stergaard, Bjrck, and Gunnarsson
(1988) and Wolf et al. (1999). The discrepancy in the digestibility
of cross-linked starches might be caused by the inherent properties
of each type of starch, as well as the conditions used for starch

J. Juansang et al. / Food Chemistry 131 (2012) 500507

modication. Although the digestibility of chemically modied

starch has been studied by many researchers, analyses of the nutritional fractions of starch (RDS, SDS and RS) have only recently
drawn more attention.
Heat-moisture treatment (HMT) refers to the exposure of starch
granules to a temperature above the glass transition temperature,
but below the gelatinisation temperature for a certain time period
and at restricted moisture content (below 35%) (Jacobs & Delcour,
1998). It is a physical modication technique that is considered to
be natural and safe, when compared to chemical modications
(Lawal, 2005). Watcharatewinkul, Puttanlek, Rungsardthong, and
Uttapap (2009) and Watcharatewinkul, Uttapap, Puttanlek, and
Rungsardthong (2010) have shown that HMT canna starch displayed equivalent pasting properties to starch cross-linked by sodium trimetaphosphate. When gelatinised, the granules of HMT
canna starches exhibited fewer breakages compared to the native
starch, and the extent of disintegration decreased with increasing
moisture content of starch during HMT. Therefore, it was supposed
that the cooked HMT starches would have less susceptibility to enzyme hydrolysis due to their limited gelatinisation.
Most starch consumed by humans has undergone some form of
processing, which usually involves heating in the presence of moisture, with or without shear, and then cooling. During heat processing, the starch granules swell and gelatinise. As a result, the
molecular order of the starch granule is destroyed and the starch
is easily digested due to increasing accessibility of enzymes to
the starch substrate. Thus, the digestibility of starch depends
strongly on the degree of gelatinisation, which is governed by
the intrinsic properties of starch, as well as, by gelatinisation conditions, such as starch concentration, gelatinisation temperature
and time and heating process. The appearance of gelatinised starch
granules varies from swelled granules, to a combination of starch
fragments and dispersed starch molecules, to completely dispersed
starch molecules, depending on the degree of gelatinisation. Under
identical gelatinisation conditions, starch from the same botanical
source but treated with different modications might display different starch gel morphologies. As a consequence, the key factors
that determine the RDS, SDS and RS contents in cooked starch
are the extent of granule disintegration, which is determined by
the degree of gelatinisation, and the structural change of starch
by each chemical/physical treatment.
In this study, canna starch extracted from rhizomes of the edible canna plant (Canna edulis Ker.) was chosen as a starch base for
modication, since its paste viscosity is quite stable at high temperature and shear force, it has high tendency towards retrogradation (Thitipraphunkul, Uttapap, Piyachomkwan, & Takeda, 2003;
Wandee, Puttanlek, Rungsardthong, & Uttapap, 2011) and also high
resistance to enzyme hydrolysis (Hung & Morita, 2005). Therefore,
it would contain some initial amounts of RS and SDS in the native
form. These fractions were expected to be increased by chemical/
physical modications (acetylation, hydroxypropylation, octenyl
succinylation, crosslinking and HMT). The modied canna starches
were subjected to heat gelatinisation at various conditions, and
RDS, SDS and RS content were analysed. The amounts of these fractions were compared and discussed in relation to the degree of gelatinisation and structural changes of the modied starches. A
commercial RS product (Novelose260) was used as a reference
for comparison.

2. Materials and methods

2.1. Materials
Two varieties of edible canna (Thai-green [TG] and Vietnam
[Vt]) were grown on experimental plots at the Rayong Field Crops

501

Research Center, Rayong, Thailand. Eight-month-old rhizomes

were harvested, and starch was isolated according to a procedure
described by Thitipraphunkul et al. (2003). Chemically modied
canna starch samples were prepared according to the methods described in previous reports, as follows: acetylated canna starch
(TG-AC; DS = 0.06) (Saartrat, Puttanlek, Rungsardthong, & Uttapap,
2005); hydroxypropylated canna starch (TG-HP; DS = 0.02)
(Chuenkamol, Puttanlek, Rungsardthong, & Uttapap, 2007); octenyl
succinylated canna starch (TG-OSA; DS = 0.025) (Kweon, Choi, Kim,
& Lim, 2001); and cross-linked canna starch (TG-CL; 0.2% w/w sodium trimetaphosphate) (Emrat, Uttapap, Puttanlek, & Rungsardthong, 2007). Heat-moisture treated canna starches, named
Vt-HMT18, Vt-HMT22 and Vt-HMT25, were treated at 100 C for
16 h, and had moisture contents of 18%, 22% and 25%, respectively
(Watcharatewinkul et al., 2009). Novelose260 (N260) was provided by the National Starch and Chemical Co. (Bangkok). Alphaamylase (type VI-B from porcine pancreas, A-3173, 28 U/mg) and
amyloglucosidase (EC 3.2.1.3, from Aspergillus niger, 300 U/ml)
were purchased from Sigma Chemical Co. (St. Louis, MO, USA). A
glucose assay kit (liquicolor; Ref. No. 10121) was purchased from
Human GmbH (Wiesbaden, Germany).
2.2. Gelatinisation of starch and light microscopy of starch gel
Starch slurries (10% w/w; 3 g of starch in 27 g of distilled water)
were gelatinised at 100 C for 5, 10, 20 and 40 min using a RVA-3D
rapid visco analyzer (Newport Scientic, Warriewood, Australia)
with a paddle rotated at a xed speed of 160 rpm. The starch slurry
was heated from room temperature to 100 C, at a rate of 3 C/min,
maintained at 100 C for 5 min (or 10, 20, 40 min), and then cooled
to 40 C at the same rate. One set of freshly prepared starch samples was analysed for SDS and RS. Another set of the gelatinised
samples was dried at 40 C for 15 h, and then ground with a mortar
and pestle and passed through a 140-mesh sieve. The ne powder
obtained was analysed for degree of gelatinisation. The gelatinised
starch gels were stained with 0.2% I2/KI and observed under a light
microscope (E200 Eclipse, Nikon Corp., Tokyo, Japan).
2.3. Determination of degree of gelatinisation (DG) by amyloseiodine
complex formation
The degree of gelatinisation (DG) was determined according to
the method of Baks, Ngene, van Soest, Janssen, and Boom (2007),
with modications. Each gelatinised starch sample (0.04 g) was
dissolved in 50 ml of 0.15 M KOH, and then centrifuged at
5000 rpm for 10 min. One ml of the supernatant was taken and
neutralised with 9 ml of 0.17 M HCl. Subsequently, 0.1 ml iodine
reagent (1 g iodine and 4 g potassium iodide in 100 ml water)
was added, to form a blue complex with the dissolved amylose
present in the sample. The absorbance was measured at 25 C
and 600 nm (A1). The same amount of gelatinised starch sample
was dissolved in 50 ml of 0.40 M KOH, and boiled in a water bath
at 95 C for 10 min. The supernatant was neutralised with 0.45 M
HCl. After adding 0.1 ml iodine reagent, the absorbance at 25 C
and 600 nm was measured (A2). The DG was equal to a percentage
ratio of A1 (0.15 M KOH) to A2 (0.40 M KOH).
2.4. Determination of RDS, SDS and RS
RDS, SDS and RS were determined according to the method of
Englyst et al. (1992), with modications. Porcine pancreatic aamylase (0.357 g) was dispersed in 50 ml sodium acetate buffer
(0.1 M, 4 mM CaCl2, pH 5.2), and centrifuged at 5000 rpm for
10 min. The supernatant (4.5 ml) was transferred to a beaker,
and 0.5 ml of amyloglucosidase (15 U/ml) was added to the solution. This enzyme solution was freshly prepared for each digestion.

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J. Juansang et al. / Food Chemistry 131 (2012) 500507

Gelatinised starch samples (100 mg, db), 10 glass beads (0.5 mm

diameter), 5 ml of pH 5.2 sodium acetate buffer and 5 ml of the enzyme solution were added to an Erlenmeyer ask, mixed for 1 min
and then incubated in a shaking water bath (37 C, 130 strokes/
min). After 20 and 120 min of incubation, 0.2 ml aliquots were
added to 4 ml of absolute ethanol, mixed well and centrifuged at
5000 rpm for 10 min. The liberated glucose in the supernatant
was determined using a glucose assay kit. Starch fractions that digested within 20 min, between 20 and 120 min, and that remained
undigested within 120 min, were classied as RDS, SDS and RS,
respectively.
2.5. Statistical analysis
Unless otherwise stated, analyses of starch characteristics and
properties were carried out in duplicate. Experimental data was
analysed using analysis of variance (ANOVA), and expressed as
mean values. A Duncan test was conducted to examine signicant
differences among experimental mean values (a 6 0.05).
3. Results and discussion
3.1. Starch gel morphology
Gel morphologies of chemically and HMT modied canna
starches gelatinised at 100 C for 5 and 20 min are shown in Figs.
1 and 2, respectively. Gels of native canna starch from different
varieties (Thai-green variety in Fig. 1, and Vietnam variety in
Fig. 2) displayed a similar feature. After 5 min gelatinisation, the
native starch granules were broken into small pieces, and the space
surrounding the granules became blue, due to the complexes of
leached amylose and iodine. The extent of disintegration increased,
with an increase in gelatinisation time.
Canna starch modied by hydroxypropylation with a degree of
substitution (DS) of 0.02 was the most fragile, compared to the
other starch samples (Fig. 1, TG-HP). The granules of hydroxypropyl starch were almost completely broken after 20 min gelatinisation, i.e. the granule fragments were hardly observed. Granule
appearances of octenyl succinyl starch (DS = 0.025), after 5 and
20 min, were similar to those of the native starch. However, the
remaining fragments seemed to be larger than those of the native
starch (Fig. 1, TG-OSA). Acetylated starch with DS of 0.06 exhibited
a different feature from the other two substituted starches. Its
granules were obviously less broken, compared to the native and
the other two modied starches (Fig. 1, TG-AC). These morphologies were in accordance with a higher viscosity of hydroxypropyl
canna starch and a lower viscosity of acetylated canna starch, compared to the native starch, as reported by Chuenkamol et al. (2007)
and Saartrat et al. (2005), respectively. On the other hand, starch
granules of cross-linked canna starch were highly swollen, but still
remained in granular form (Fig. 1, TG-CL). This indicated the high
resistance of cross-linked canna starch to heat and shear, which
is a typical characteristic of cross-linked starches.
As shown in Fig. 2, gel morphologies of HMT starches were totally different from that of the native starch. The HMT starches displayed more extensive resistance to heat and shear. The HMT18%
granules were less broken, compared to the native starch, while
the HMT22% and HMT25% granules were swollen but unbroken.
Gel appearance of HMT starch was similar to that of the chemically
cross-linked starch. A decrease in the extent of disintegration with
increasing moisture content of starch samples, during heat treatment, has been reported by Watcharatewinkul et al. (2009). For
the commercial Novelose260 (HMT high amylomaize), very limited
swelling was observed even after 40 min gelatinisation, indicating
that the Novelose260 granules were very resistant to gelatinisation.

3.2. Effect of gelatinisation time on DG values

The effect of gelatinisation time on the DG values of 10% (w/w)
native and modied starches is shown in Fig. 3. The extent of
granule disintegration can be indicated by the DG value, i.e. a
higher DG value indicates a greater extent of granule dispersion.
As shown in the gure, all starch samples displayed a similar pattern: a signicant change in the DG value was observed during
the initial 10 min of gelatinisation, after which the DG value increased gradually with gelatinisation time. However, the DG values and magnitude of changes were different among the starch
samples. The native starches from the two varieties Thai-green
(TG) and Vietnam (Vt) exhibited a very similar prole. The DG
values of native canna starch from Thai-green and Vietnam varieties, gelatinised for 540 min were 68.784.3% and 67.084.6%,
respectively.
Acetylated starch (TG-AC), hydroxypropylated starch (TG-HP)
and octenyl succinylated starch (TG-OSA) had an increase in DG
values as compared to the native starch. The DG values of substituted starches conformed to the following order: TG-HP > TGAC > TG-OSA. After gelatinisation for 40 min, the DG values of
TG-AC, TG-HP and TG-OSA were 92.0%, 94.4% and 87.8%, respectively (Fig. 3A). Except for TG-AC, the DG values obtained were
consistent with the gel morphologies, mentioned in the previous
section. Being less broken after gelatinisation, as revealed in
Fig. 1, it was expected that TG-AC would have a lower DG value
than the native starch. However, it was found that TG-AC had a
slightly higher DG value, implying that the amylose of TG-AC
could be dispersed and reacted with I2 better than the native
starch. The substitution groups might facilitate the penetration
and absorption of water into the starch granules by H-bond disruption, by enlarging the inter-space between starch chains,
and/or because of the hydrophilic nature of the substituent. The
rapid increase in water content inside the granules, as well as
an increase in the plasticising effect of water, rendered the modied starch easy to gelatinise. The diverse effects of different
types of substitution on gelatinisation, as found in this study,
were most probably due to the nature of the substituted groups
(molecular size, hydrophobicity/hydrophilicity, functional groups)
as well as the degree of substitution.
Cross-linked canna starch (TG-CL) showed a decreased DG value
compared to the native starch. Extending the gelatinisation time
from 5 to 40 min resulted in a slight increase of DG value. This indicated that the strength of the additional covalent bonds of the
cross-linked starch enabled the granules to withstand the gelatinisation temperature, and that the cross-linking agents helped
maintain the granular integrity. The DG value of TG-CL gelatinised
for 40 min was 55.9%.
DG values of Vt-HMT18 were much closer to those of the native
starch (Fig. 3B), although their gel appearances were signicantly
different. An increase in moisture content of starch samples during
heat treatment resulted in a lower DG value, i.e. DG values of VtHMT25 < Vt-HMT22 < Vt-HMT18. The DG values of Vt-HMT18,
Vt-HMT22 and Vt-HMT25 gelatinised for 40 min were 86.8%,
74.6% and 69.4%, respectively. The results obtained could be explained in relation to the free amylose dispersed in starch gel, as
reported by Watcharatewinkul et al. (2009). They found that amylose leaching of HMT canna starch tended to be lower when the
moisture content of the starch increased. Therefore, a lesser
amount of amylose dispersed in starch gel resulted in poorer intensity of the blue colour produced by the amyloseiodine complex.
As a consequence, the DG value of heat-treated high-moisture
starch became lower. Novelose260, the commercial HMT highamylose maize starch, had the lowest DG value (13.6%) among
the starches tested.

J. Juansang et al. / Food Chemistry 131 (2012) 500507

503

Fig. 1. Gel morphologies of chemically modied canna starches gelatinised, at 100 C, for 5 min (left) and 20 min (right).

3.3. Effect of gelatinisation time on SDS and RS contents

Changes in the SDS and RS contents of 10% (w/w) native and
modied starches, with gelatinisation time, are shown in Table 1.
In general, RS decreased with an increase in gelatinisation time,
while SDS values uctuated. Native starches from the two varieties
of canna had comparable SDS and RS contents. The initial content

of RS in canna starch was relatively high (88%), but was reduced

extensively upon gelatinisation: to 10.112.4%, after 40 min gelatinisation. Gelatinisation at 100 C, for only 5 min, resulted in a
marked decrease of RS in native canna starch (from 88% to 23
26%), signifying the unstable nature of RS in raw canna starch.
The RS fraction of Novelose260 changed only slightly with
gelatinisation time (from 70.7% in the raw sample to 54.4% after

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J. Juansang et al. / Food Chemistry 131 (2012) 500507

Fig. 2. Gel morphologies of heat-moisture treated canna starches and Novelose 260 gelatinised, at 100 C, for 5 min (left) and 20 min (right).

gelatinisation for 40 min), whereas its SDS contents were quite low
(3.87.4%) throughout the gelatinisation time.
3.4. RS contents
Regardless of the modication methods used, the RS contents of
the modied starches were lower than that of the Novelose260.
Among the modied starches, OSA starch displayed the highest

RS content at all gelatinisation times, and its RS content was not

much different from the Novelose260. All substituted canna
starches had a higher resistance to enzyme hydrolysis, compared
to the native starch. The resistance conformed to the following order: OSA starch > HP starch > AC starch. The results suggested that
the nature of the substituted groups had more inuence on resistance to enzyme hydrolysis than the degree of gelatinisation of
starch granules. The most obvious case was the HP starch: its gran-

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J. Juansang et al. / Food Chemistry 131 (2012) 500507

% Degree of gelatinization

(A)

100

80

60
TG-HP

40

TG-AC
TG-OSA
20

Native TG
TG-CL

0
0

10

20

30

40

Gelatinization time (min)

% Degree of gelatinization

(B)

100

80

60

Vt-HMT18
Native Vt

Vt-HMT22

40

Vt-HMT25
N 260

20

0
0

10

20

30

40

Gelatinization time (min)

Fig. 3. Effect of gelatinisation time on the DG values of 10% (w/w) native and
modied canna starches ((A) chemically modied starches, (B) heat-moisture
treated starches).

ules exhibited much more complete breakage than the native and
acetylated starches, but its RS content was higher. There was also a
tendency that, as the molecular sizes of the substituted group became larger, the modied canna starch became more difcult to
hydrolyse. The results obtained in this study were similar to those

reported in previous literature. Chung et al. (2008) found that the

amounts of RS in gelatinised corn starches were 19.5%, 14.3%,
13.0% and 7.0% in hydroxypropylated, acetylated, oxidised and
cross-linked starches, respectively, whereas the unmodied starch
contained 7.3%. Hoover, Hannouz, and Sosulski (1998) and Wootton and Chaudhry (1981) observed that hydroxypropylation decreased the enzyme susceptibility of pea and wheat starches,
even after the starches were pasted. stergaard et al. (1988) also
found that substitution with acetyl or hydroxypropyl groups reduced the enzyme susceptibility in gelatinised starches. They also
suggested that the hydroxypropyl groups had a greater effect than
the acetyl groups on enzyme resistance, because the bulkier
hydroxypropyl groups provide greater hindrance to attack by enzymes (Hoover & Sosulski, 1986; Wootton & Chaudhry, 1981). It
has also been suggested that the presence of bulky substituents
on C2 of the glucose unit would sterically hinder the proper positioning of the substrate into the active site of a-amylase, and also
effectively restrict enzyme attack on the adjacent glycosidic bonds
of unsubstituted glucose residues (Hoover & Sosulski, 1986; Hoover & Zhou, 2003; stergaard et al., 1988).
Cross-linked canna starch had the highest RS content in its raw
state (94.1%), but when cooked, the RS content was reduced rapidly
with extended gelatinisation time. The remaining RS after gelatinisation for 40 min was 19.8%. Except for few cases, the RS contents
in cross-linked starch were lower than in the substituted starches,
although its starch granules were obviously less broken. Again,
these data conrmed that the susceptibility of starch to enzyme
hydrolysis was inuenced more by the substituted groups in starch
molecules, than by the degree of disintegration of starch granules.
Raw HMT canna starches contained considerably high amounts
of RS (92.294.8%); however when cooked, their RS was reduced
markedly. At the same period of gelatinisation, the RS contents of
all HMT starches were lower than that of the native starch. The
most obvious case was Vt-HMT18; even after only 5 min of gelatinisation, the RS content in Vt-HMT18 was decreased to 3.1%, and it
almost disappeared (0.2%) after 40 min gelatinisation. The result
obtained was contrary to our presupposition that the HMT starch
would have a signicantly higher resistance to enzyme hydrolysis,
due to its limited gelatinisation (in other words, would contain a
higher amount of RS), compared to the native starch. Being kept
in a granular form, with less dispersion of HMT starch granules
after gelatinisation (as shown in Fig. 2), would seemingly obstruct
the accessibility of enzymes to amylose/amylopectin inside the
starch granules. However, the results showed that HMT conversely
facilitated the susceptibility of the treated granules to enzyme
hydrolysis. This was possibly due to the degradation of starch

Table 1
SDS and RS contents of 10% (w/w) native, chemically modied and physically modied canna starches.
%SDS
Gelatinisation time (min)
Chemical modication
TG
TG-AC
TG-HP
TG-OSA
TG-CL
Physical modication
N260
Vt
Vt-HMT18
Vt-HMT22
Vt-HMT25

%RS

10

yz

11.5e
15.7b
y
14.0c
z
23.4a
x
8.6g

5.6f
14.4b
x
13.0c
y
17.6a
x
9.5e

8.8a
2.9h
w
5.1d
w
7.2c
w
3.9f

7.4b
4.4e
y
2.9g
x
2.2i
v
0.6j

5.2h
2.4i
z
10.5f
z
5.5h
z
13.0c

3.8g
3.7g
x
5.6f
z
5.6f
y
11.3d

20

40

10

20

40

2.1f
16.7b
z
16.2b
x
12.5c
z
23.6a

70.7i
88.1h
z
94.4b
z
94.8a
z
92.2e

2.0f
13.7b
y
14.2b
x
12.7c
y
17.9a

6.1e
2.2f
w
0.5g
xy
2.8f
x
9.9d

7.2e
10.3d
w
0.4g
x
1.5f
w
6.9e

Means with different letters (a, b, . . .) in the same column are signicantly different (p 6 0.05).
Means with different letters (x, y, . . .) in the same row are signicantly different (p 6 0.05).

88.2g
91.4f
z
93.4d
z
92.2e
z
94.1c

22.9f
35.9d
y
45.5c
y
54.2b
y
36.6d

x
x

18.1g
32.2d
w
37.6c
w
49.4b
w
25.4e

20.4g
33.8e
x
43.5c
x
51.3b
x
35.3d

62.5a
26.1e
x
3.1h
y
22.6f
y
19.1g

60.1a
24.7f
w
1.6j
x
12.5i
x
17.3h

56.9a
21.6f
y
3.8j
w
5.5i
w
12.8h

12.4f
26.6d
v
32.0c
v
45.3b
v
19.8e

54.4a
10.1g
v
0.2i
v
1.9h
v
10.5g
v

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J. Juansang et al. / Food Chemistry 131 (2012) 500507

molecules by HMT, or because the swelling state of HMT starch

granules might accelerate the sequential catalytic action of the enzyme by increasing the probability of substrateenzyme binding.
As the moisture in starch samples during HMT increased, in most
cases (but not all) the RS content in HMT starch was found to increase (Vt-HMT25 > Vt-HMT22 > Vt-HMT18). This indicated that
the extent of hydrolysis is co-inuenced by many factors, such as
the physical appearance of starch granules, their molecular structure and/or the capability of starchenzyme binding. The effect
of HMT on susceptibility of canna starch to enzyme hydrolysis
was different from those reported previously on other starches.
Brumovsky and Thompson (2001) found that HMT high-amylose
corn starch contained 43.9% resistant starch, compared to 18.4%
in native starch. Gzel and Sayar (2010) reported that RS and
SDS contents in cooked (in a boiling-water bath for 20 min) HMT
bean starches were signicantly higher than those in corresponding native starches. According to Chung, Liu, and Hoover (2009),
when the starch was heat-moisture treated at 120 C for 2 h, the
RS content of corn, pea, and lentil starches increased from 4.6%,
5.2% and 5.3% to 12.3%, 16.4% and 15.7%, respectively, as compared
to gelatinised unmodied starches.
3.5. SDS contents
All starch samples displayed uctuations of SDS content with
gelatinisation time, because it could be transformed from RS or
change into RDS. Native canna starch and Novelose260 contained
less than 10% SDS, whereas acetylated and hydroxypropylated
starches had considerably higher amounts of SDS (1317%), at all
gelatinisation times. The SDS contents of cooked octenyl succinylated and cross-linked starches varied between 12.523.4% and
8.623.6%, respectively. The SDS content of octenyl succinylated
starch was high at the initial period of cooking, but became lower
as the cooking time was prolonged. The opposite was found with
the cross-linked starch, i.e. the SDS content increased with longer
cooking time. When the sum of RS and SDS was taken into consideration, it was found that the RS + SDS of octenyl succinylated
starch was a bit less than, or similar to, that of the Novelose260,
and was somewhat higher when the starch was cooked for a short
period of time (510 min). The RS + SDS of octenyl succinylated
starch after cooking for 5 and 10 min were 77.6% and 68.9%,
whereas those of Novelose260 were 67.7% and 63.9%, respectively.
The HMT starches (Vt-HMT18, Vt-HMT22 and Vt-HMT25) contained much lower contents of SDS (0.613.0%) than the chemically modied starches (2.923.6%). Their SDS decreased with
increasing gelatinisation time (540 min). The SDS of HMT starches
was higher than the native starch after cooking for a short period
(510 min), but became lower when the cooking time was extended to 20 and 40 min except in the case of Vt-HMT25 gelatinised for 20 min, which still contained a higher amount of SDS
compared to the native starch.
3.6. Relationship between DG value and RS content
Relationships between DG value and RS content of 10% native
and modied starches are shown in Fig. 4. RS contents in all samples
decreased with an increase in the degree of gelatinisation. The decrease of RS content in Thai-green canna starches displayed a fairly
linear function of the DG values (R2 = 0.992). This indicated that the
RS content in native starches depended mainly on the level of starch
granule disintegration. In spite of substitution by an acetyl group,
acetylated canna starch also showed a linear relationship of RS content with DG value (R2 = 0.997); however, the slope was much lower
than that of the native starch. This result suggested that the acetyl
group could impede the susceptibility of starch to enzyme hydrolysis. The effect of the substitution group was more clearly observed in

Fig. 4. Relationships between DG value and RS content of 10% native and modied
canna starches ((A) chemically modied starches, (B) heat-moisture treated
starches).

hydroxypropyl starch, which displayed a lower slope than native

and acetylated starches. However, the RS content dropped rapidly
when the DG value was higher than 90%. Some reports have indicated that the amorphous regions (amylose and branch points of
amylopectin) are more accessible to the hydroxypropylating reagent, and thus are modied to a greater extent than the crystalline
regions (amylopectin) of starch granules. It is generally accepted
that amylose molecules are leached out rst at the early stage of gelatinisation. Therefore, at a lower degree of gelatinisation (<90%), the
amylose molecules (which had a higher tendency than the amylopectin to carry the hydroxypropyl groups) would be the majority
in the starch dispersion. Consequently, at this stage the hydroxypropyl starch exhibited a higher resistance to enzyme hydrolysis, compared to the corresponding native starch. Above 90% DG, a greater
extent of starch dispersion, as well as the increased populations of
unmodied starch chains in the dispersion, rendered the starch molecules more easily accessed by the enzyme, thus the RS content
dropped rapidly. Octenyl succinylated starch displayed a trend similar to the hydroxypropyl starch, but its DG values were lower.

J. Juansang et al. / Food Chemistry 131 (2012) 500507

The change in RS content of cross-linked starch with the DG values was slightly higher than that of the native starch at the early
stage of gelatinisation (<50%), but a signicant difference was observed after that. A signicant reduction in the digestibility of starch
cross-linked by a phosphate bridge was also reported by Sang and
Seib (2006) and Woo and Seib (2003). It is still uncertain why the
cross-linked starch displayed higher susceptibility to enzyme
hydrolysis. Typically, cross-linked starch can resist pasting and will
maintain its granular integrity better than native starch upon gelatinisation. It might be possible that the enzyme is able to access
starch molecules more readily when starch granules are in a swollen
state. Additionally, at higher degrees of gelatinisation the phosphate
diester bonds could be destroyed, and the resulting phosphate
monoester might enhance the binding/catalysing capabilities of
the enzyme to the substrate. The decrease in RS content of crosslinked starch was accompanied by an increase of SDS. Within the
gelatinisation period of 540 min, the summation of RS and SDS
contents of cross-linked starch was nearly constant (Table 1), indicating that most of the RS reduced was transformed into SDS.
Cross-linked starch contained the highest amount of SDS (16.5
23.1%) among the starches gelatinised for 2040 min. Also, it had a
tendency to have a higher SDS content with extended gelatinisation
time.
Native Vietnam cv. canna starch showed characteristics similar
to the native Thai-green cultivar. The RS content of all HMT starches
displayed a signicantly higher change with the DG value, compared to the native starch. For Novelose260, the RS content declined
linearly with the DG value, although its DG values were very limited
(less than 20%). Changes in SDS contents of starch samples with the
DG values were in an irregular pattern (Figure not shown). It is difcult to estimate the SDS content, at a particular degree of gelatinisation, because SDS can be increased by transformation of RS into
SDS, and can also be decreased by transformation of SDS into RDS.
4. Conclusions
Octenyl succinylation has shown to be a promising method for
preparation of a high-RS and SDS product from a potential source
of raw material, canna starch. The sum of its RS and SDS was a bit
less than, or similar to, that of the Novelose260, and was somewhat
higher after a short period of cooking. The RS content of octenyl
succinylated canna starch was reduced only slightly by gelatinisation, even at 40 min gelatinisation. The results from this study
showed that heat-moisture treatment was not an efcient means
of producing a high-RS product from canna starch. However,
starches from other sources, or starches treated by using other
HMT conditions, might be examined to conrm this conclusion.
Acknowledgements
We gratefully acknowledge the Thailand Research Fund, Thailand (TRF-MAG Project MRG-WII525S057) and the Higher Education Research Promotion and National Research University
Project of Thailand, Ofce of the Higher Education Commission,
for their nancial supports.
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