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Food Hydrocolloids
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Article history:
Received 27 May 2014
Accepted 12 November 2014
Available online 26 November 2014
Starch gels of waxy rice (WR), waxy corn (WC) and waxy potato (WP) were hydrolyzed with pullulanase,
and the products obtained were analyzed for unit chain length distribution and certain physicochemical
properties. Average chain lengths of debranched WR, WC and WP starches were 18.2, 19.2 and 25.6,
respectively. The debranched starches had a greater ability to form complexes with iodine and possessed
higher solubility but lower viscosity compared with their corresponding dispersed native starches. Their
complexing abilities with fatty acids (palmitic and butyric acids) were found to be dependent on the unit
chain length. Among the three debranched starches, debranched waxy potato starch exhibited the
greatest ability to form complexes with iodine and fatty acids, while debranched waxy rice starch had the
highest solubility and lowest viscosity. The debranched starches formed stable gels at high concentrations (10e20%) and formed precipitates at concentrations up to 5%.
2014 Elsevier Ltd. All rights reserved.
Keywords:
Debranched waxy starch
Molecular structure
Solubility
Complexing ability
Viscosity
1. Introduction
Native starches are used in food and non-food applications, in
which starch properties such as viscosity, retrogradation, solubility,
gelation, gel appearance and texture are the main criteria for
choosing an appropriate starch for a certain end-use. In most applications, starch is used in a gelatinized native form, either in
cooked or uncooked products. It is well-known that starch is biosynthesized as granules in higher plants and consists of two types
of a-D-glucose polymers: amylose and amylopectin. Amylose is a
mixture of lightly branched and linear molecules, whereas amylopectin is a much larger molecule with a highly branched structure
consisting of about 95% a-1,4 linkages and 4e5% a-1,6linkages
(Hizukuri, Abe, & Hanashiro, 2006; Tester, Karkalas, & Qi, 2004).
The ratio of amylose to amylopectin in starch varies depending on
the botanical source. Generally, normal starches consist of 20e30%
219
220
Solubility%
2.7. Viscosity
Individual starch samples were suspended in water at concentrations of 1, 5, 10 and 20% (dwb) in a 50 ml centrifuge tube and
then cooked in a boiling water bath for 1 h. The starch solutions
were cooled down by leaving the tubes at room temperature until
the temperature was reduced to 90, 70, 50 and 30 C. At these
temperatures, the viscosity of the starch solution was determined
by a viscometer (model LVDV-IP; Brookeld Engineering Laboratories, Middleborough, MA) equipped with a suitable spindle (nos.
61e64) that rotated at a xed speed of 60 rpm.
Fig. 1. HPSEC chromatograms of waxy rice, waxy corn and waxy potato starches after
debranching with pullulanase (molar basis).
Average chain
length
c
18.2 0.1
19.2 0.4b
25.6 0.2a
Distribution (%)
DP 6e12
a
47.8 0.1
42.4 0.4b
31.0 0.6c
Table 2
Iodine staining indexes of native (N) and debranched (D) starches, with and without
fatty acid (FA) complexation.
Samples
DP 13e24
c
36.4 0.1
40.3 0.2b
41.5 0.5a
DP 25e36
c
6.8 0.1
7.7 0.1b
9.8 0.1a
DP 37
Waxy rice
NWRS
9.1 0.1
9.6 0.3b
17.7 0.2a
Means with different letters in the same column are signicantly different
(p 0.05).
221
Waxy corn
DWRS
Without FA complexation
ISI460
0.196aB
0.282aA
ISI570
0.089aB
0.490aA
With butyric acid complexation
ISI460
0.116bB
0.267aA
D460
0.080
0.015
ISI570
0.058bB
0.447bA
D570
0.031
0.043
With palmitic acid complexation
0.068cB
0.218bA
ISI460
D460
0.128
0.064
0.028cB
0.290cA
ISI570
D570
0.061
0.200
Waxy potato
NWCS
DWCS
NWPS
DWPS
0.237aB
0.173aB
0.315aA
0.527aA
0.403aB
0.322aB
0.451aA
0.846aA
0.142bB
0.095
0.089bB
0.084
0.283bA
0.032
0.490bA
0.037
0.364bB
0.039
0.298bB
0.024
0.440aA
0.011
0.795bA
0.051
0.103cB
0.134
0.067bB
0.106
0.252cA
0.063
0.307cA
0.220
0.298cB
0.105
0.195cB
0.127
0.377bA
0.074
0.542cA
0.304
Capital letters (A, B) indicate a signicant difference (p 0.05) between native and
debranched starches of the same starch.
Small letters (a, b) indicate a signicant difference (p 0.05) among samples at the
same wavelength in the same column.
D difference in ISI values of free starch and starcheFA complex.
association with the unit chain length distributions (Fig. 1), it could
be estimated that wavelengths of less than approximately 530 nm
reected the light absorption of iodine and short starch chains (<DP
30), while those between 530 and 645 nm reected the light absorption of iodine and longer starch chains (around DP 35e300).
The iodine staining indexes (ISIs) herein were thus expressed as the
absorbance at 460 nm (representative of short chains) and at
570 nm (representative of longer chains); the values are summarized in Table 2. DBSs exhibited higher ISI values than their corresponding intact starch molecules (NWSs). The higher ISI of the DBSs
as compared with the intact amylopectin molecules is thought to be
associated with inner and outer branch chains in the cluster
structure of amylopectins. Embedding of the inner branch chains in
organized clusters makes it difcult for them to form complexes
with iodine. Hydrolysis of the branch points with pullulanase
released these chains from the ordered structure, enabling them to
have a higher possibility to form complexes. Among the three waxy
starches, DWPS had the highest ISI, followed by DWCS and DWRS.
These ISI values had a positive correlation with the average chain
length of the DBS, suggesting that longer branch chains were more
effective in forming complexes.
The complex-forming ability of DBS with fatty acids (FA) was
determined by measuring the free DBSs remaining in the solution
using iodine staining because the procedure is fast and simple. The
method was also able to reveal the chain lengths of DBS that
preferentially form inclusion complexes with FA according to the
decrease in the absorbance at different wavelengths
(Yotsawimonwat et al., 2008). Palmitic acid (PA, 16:0) and butyric
acid (BA, 4:0) were used as representatives of long-chain and shortchain fatty acids, respectively. It should be noted that the starcheFA
complexes measured by this procedure represented those that
were more stable than the complex between iodine and starch. The
222
Fig. 3. Solubility values of native (N) and debranched (D) waxy starches in water at different starch concentrations and temperatures.
Fig. 4. Solubility values of 5% (w/w) debranched waxy starches in 0, 0.2 and 2 M NaCl solutions at different temperatures.
Starch
concentration (%)
DWRS
1
5
1
5
1
5
DWCS
DWPS
Solubility (%)
0h
100.8
99.9
99.5
95.6
100.2
99.3
D (%)
2h
98.3
96.6
91.5
86.8
84.3
77.1
0.8
2.3A
1.8A
4.4A
1.9A
1.2A
AB
5.8
1.1AB
7.5B
1.3B
7.4B
2.8B
2.5
3.3
8.0
8.8
15.9
22.2
Capital letters (A, B) indicate a signicant difference (p 0.05) between DBS solutions at 95 C and those after leaving at room temperature for 2 h for the same starch
and starch concentration.
D percentage difference of solubility values at initial time and after leaving at
room temperature for 2 h.
in
accordance
with
the
empirical
equation
of
MarkeHouwinkeSakurada, in which the viscosity of polymers ()
is a function of the molecular weight (M) [ kMa, where k and a
are constants related to the degree of molecular expansion]. It was
also observed that the difference was greater at high DBS concentration (10e20%) and low temperature (50 and 30 C). Therefore,
not only the chain length of the DBSs, but their ability to retrograde
into a network structure might also affect the viscosity of the DBS
solutions. The superior re-association ability of DWPS compared
with DWCS and DWRS resulted in a signicant difference in their
viscosity values, especially at high DBS concentration and low
temperature.
3.5. Gelling properties
It is known that when starch is subjected to gelatinization,
either by heating in excess water or suspending in some chemical
solutions, the starch granules will be destroyed to some extent,
depending on gelatinizing conditions. The dissolved starch molecules, especially amylose, can re-associate by H-bonding (between
hydroxyl groups or via a water bridge), resulting in a precipitate or
gel (Biliaderis, 2009). The ability of starch to retrograde depends on
many factors, such as starch composition (amylose content, protein
content, lipid content), structure of starch molecules (amylose/
amylopectin structure), storage conditions (e.g. temperature, time),
starch concentration, etc. Generally, retrogradation of highTable 4
Viscosities of native (N) and debranched (D) waxy starches at different starch concentrations and temperatures.
Starch Starch Viscosity (centipoise, cP)
sample conc.
90 C
70 C
(% w/v)
NWRS
DWRS
NWCS
DWCS
NWPS
DWPS
1
5
1
5
10
20
1
5
1
5
10
20
1
5
1
5
10
20
11.0
2696.7
1.7
2.0
4.1
5.5
13.9
3433.3
1.8
1.9
3.0
7.8
325.7
4266.7
1.6
1.8
3.9
26.8
50 C
30 C
0.4
14.6 0.6
16.8 1.7
20.1 1.6
66.6 2843.3 41.6 3163.3 80.8 3216.7 40.4
0.1
2.0 0.1
2.1 0.1
2.1 0.1
0.1
2.2 0.1
2.5 0.1
3.0 0.1
0.2
5.5 0.4
6.5 0.2
7.6 0.2
0.1
13.1 0.1
28.4 0.3
45.5 0.7
0.4
21.1 1.2
21.4 0.7
22.9 1.2
72.3 3623.3 76.4 4256.7 65.1 4450.0 75.5
0.1
1.8 0.1
2.0 0.1
2.4 0.1
0.1
2.1 0.1
2.3 0.1
2.8 0.1
0.1
3.8 0.2
6.5 0.3
8.3 0.3
0.1
17.7 0.1
30.5 0.9
42.1 1.8
4.3
337.0 4.4
353.5 3.8
380.7 4.5
66.6 4553.3 60.3 4676.7 95.0 4936.7 76.4
0.1
1.7 0.1
1.9 0.1
2.1 0.1
0.1
2.5 0.1
4.8 0.6
12.2 0.8
0.1
10.7 0.4
25.9 0.8
39.3 1.1
0.4
39.0 0.7
85.8 1.8
135.8 2.5
223
224
Fig. 5. Appearances of cooked native (N) and debranched (D) starches at different starch concentrations after storing at 30 C (left column) and 4 C (right column) for 24 h.
225
Table 5
Turbidity, gel consistency and sediment volume of cooked native (N) and debranched (D) starches at different starch concentrations after storing at 30 C and 4 C for 24 h.
Starch sample
Turbidity (%)
30 C
NWRS
DWRS
NWCS
DWCS
NWPS
DWPS
5
1
5
10
20
5
1
5
10
20
5
1
5
10
20
69.4
17.8
99.4
100.0
100.0
89.1
66.4
99.9
100.0
100.0
22.5
99.5
99.9
100.0
100.0
4 C
0.3
1.8
0.2
0.1
0.1
0.3
1.9
0.1
0.1
0.1
1.3
0.1
0.1
0.1
0.1
70.4
99.2
100.0
100.0
100.0
88.7
99.7
100.0
100.0
100.0
36.3
99.8
100.0
100.0
100.0
and DWPS was unexpected, since it had the shortest average chain
length. Therefore, besides chain length, other factors e such as the
remaining a-1,6 linkages, phosphate ester in glucose residues, and
conformation of the debranched chains e might also be involved in
the efciency of gel formation. It is worth noting that DWS gels
appeared as particle gels (gure not shown) rather than as a
continuous network in the case of native starch gels. Therefore, the
gel of DWS would have signicantly different characteristics from
the gel of native starch.
3.6. Potential applications of DBSs
In view of the potential industrial applications of DBSs, information on each attribute derived from this study indicated the
following potential uses.
3.6.1. Complexing ability
The results indicated a signicant increase in complex formation
with iodine and fatty acids when starches were debranched.
Debranched potato starch displayed the highest ability to form
complexes. Therefore, DBSs could be an alternative source of carriers for drug delivery and encapsulating agents for active compounds such as avonoids, vitamins and antioxidants. Similar to
cyclodextrins, the complex-forming mechanism of DBSs with other
guest molecules is molecular inclusion; thus, the guest molecules
must have the appropriate size and conformation to t into the
cavities of the DBSs. DBSs can increase the dispersion of highly
insoluble compounds, control the release of compounds, and protect sensitive compounds against the environment.
3.6.2. Solubility and viscosity
The DBSs displayed much higher solubility and extremely low
viscosity compared with native starches; therefore, they might be
suitable for use as an ingredient in products that need a highly
dispersed bulking agent ller/carrier. The low viscosity of DBSs also
facilitates product processing, such as pumping through a pipeline,
mixing by agitation, or atomizing in a spray dryer. The DBSs from
rice starch would be the rst choice if a DBS that can be solubilized
at low temperature is required.
3.6.3. Gelling properties
The DBSs formed gels at high concentration and formed precipitates at low concentration. DBS gels are non-sticky, smooth, and
have a glossy appearance; therefore, they can be used as a gelling
agent in food, cosmetics and pharmaceutical products. Due to the
textural appearance of these gels, they have also received attention
0.2
0.4
0.1
0.1
0.1
0.4
0.1
0.1
0.1
0.1
1.1
0.1
0.1
0.1
0.1
30 C
30 C
29.7
144.7
141.7
20.0
20.7
17.3
149.3
90.0
24.0
20.3
19.0
156.3
60.0
26.3
20.3
4 C
1.5
2.1
4.5
1.0
2.1
0.6
7.6
8.0
1.0
1.2
1.0
3.8
2.0
1.5
0.6
26.3
150.7
18.0
19.7
19.7
20.0
150.7
27.3
23.7
20.7
18.3
114.8
29.0
25.0
19.7
2.5
2.5
1.0
0.6
1.2
1.0
4.0
2.1
0.6
0.6
0.6
4.0
1.0
1.0
0.6
100.0
0.4
3.3
42.1
52.9
100.0
0.1
15.4
26.7
45.4
100.0
4.2
20.3
36.7
62.9
4 C
0.1
0.7
1.2
2.6
0.7
0.1
0.1
1.2
0.7
0.7
0.1
1.4
0.6
1.9
1.9
100.0
3.8
34.3
74.2
76.2
100.0
5.2
22.4
31.2
52.8
100.0
5.0
21.3
39.6
66.7
0.1
2.1
1.5
1.4
1.3
0.1
2.5
1.4
1.3
1.9
0.1
2.5
1.2
2.6
1.4
226
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