Carbohydrate Research 458-459 (2018) 13e18

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Carbohydrate Research
journal homepage: www.elsevier.com/locate/carres

New insight in crosslinking degree determination for crosslinked

starch
Tingting Kou a, b, Qunyu Gao a, b, *
a
College of Light Industry and Food Sciences, South China University of Technology, Guangzhou 510640, China
b
Guangdong Province Key Laboratory of Green Processing of National Products and Product Safety, Guangzhou 510640, China

a r t i c l e i n f o a b s t r a c t

Article history: The crosslinked starch has been studied for many years, but it is difficult to characterize degree of
Received 11 November 2017 substitution on crosslinking of the very high and low crosslinked starches. The available approaches
Received in revised form (including viscosity, settling volume, and P content) all have their limitations, i.e., not applicable in a
17 January 2018
large scope, pollution problem, and can not reflect the internal structure change. Here in this paper
Accepted 30 January 2018
Available online 1 February 2018
starch-iodine (St-I) method was proposed as a new approach to characterize the crosslinking degree. In
this investigation, three starches of A, B, and C crystalline pattern with different amount (from very low
to very high, 0.01, 0.05, 0.1, 0.5, 1, 5, 10%) of crosslinking reagent added were studied. This method is
Keywords:
Crosslinking degree
based on the mechanism that crosslinking reaction take place between amylose/amylopectin, amylo-
Starch-iodine pectin/amylopectin, whereas the amylose dose not crosslink one another. After crosslinked to amylo-
Dose efficiency pectin, the result amylose-amylopectin complex can be considered as a new amylopectin. Results showed
that the St-I method can characterize all the crosslinked starches of the three starches, at low reagent
level (0.01e0.1%), the amylose was found to decrease rapidly, this can also replace the viscosity method,
whereas at high reagent level (1e10%), although the significant differences can still be observed, the
effect was not so obvious as it for the lower crosslinked starches, here we firstly applied dose efficiency to
characterize this phenomenon, which was informative and helpful in determining this modification
process.
© 2018 Elsevier Ltd. All rights reserved.

1. Introduction amylopectin molecules and probably locates primarily in amor-

Starch granules are densely packed with semi-crystalline
structures and composed of two major starch polymers:
branched amylopectin and essentially linear amylose. It is accepted
that amylose is the second most abundant component of starch,
accounting for typically 20e30% of its weight, mainly located in the
amorphous phases [1]. The currently accepted model of amylose
location in starch granules is, therefore, as individual, radially-
orientated chains randomly distributed among the radial amylo-
pectin chains, amylose content increased with increases in granule
size and kernel maturity [1]. In addition, surface gelatinization of
starch granules with concentrated neutral salt solutions reveals
that amylose is more concentrated at the periphery [2]. Report [3]
also states that amylose appears to disperse among the
* Corresponding author. College of Light Industry and Food Sciences, South China
University of Technology, Guangzhou 510640, China.
E-mail address: qygao@scut.edu.cn (Q. Gao).
phous zones of the growth rings, the idea that amylose may destroy
the fine crystalline structure of starch granule, the crystallinity
degree of waxy starch is usually higher than that of the normal,
amylose containing starches. It had been reported that amylose is
located almost exclusively in the centers of the granules, that is the
peripheral regions are amylose-free [4].
Among the ways to modify starches, crosslinking is mostly
applied to improve functional properties, and freeze-thaw and cold
stabilities of starch pastes. [5,6], When a starch is treated with bi- or
multi-functional reagents, crosslinking occurs. Numerous evidence
had shown that amylose molecules do not crosslink to one another,
but do crosslink to amylopectin chains. [7,8], Additionally, in several
studies with crystalline structure of starches found that amorphous
areas are less dense and more susceptible to chemical and enzy-
matic modification than crystalline regions [8e10]. As for the dif-
ficulties for the amylose to crosslink one another and uniqueness of
its location (amorphous and periphery layers), which is the same as
where the crosslinking reaction takes place.
There are many experimental methods found in the literature

https://doi.org/10.1016/j.carres.2018.01.009
0008-6215/© 2018 Elsevier Ltd. All rights reserved.
14 T. Kou, Q. Gao / Carbohydrate Research 458-459 (2018) 13e18
for testing the crosslinking degree of crosslinked starches. They can
be divided into the following three groups based on their under-
lying testing principles: (1) settling volume (swelling power) of the
crosslinked starches [11], (2) pasting properties (peak viscosity,
final viscosity at 95
C, as well as final viscosity) [14], and (3)
phosphorus content (determination of the new element or the new
group that was added). Each method has advantages and disad-
vantages in terms of its requirement for the crosslinking degree, the
crystalline structures of the starches [15]. Ultimately, the benefits of
each parameter must be considered, and the characterization
technique must be chosen to suit the needs of the crosslinked
starches of various crosslinking degrees. In general, settling volume
requires and yields samples in the order of grams, is not suitable
both for the very lower and higher reagent addition for the difficult
to characterize the differences. In fact, pasting properties also
require samples of grams, it can characterize the very lower reagent
added, in this paper, we found that the addition of 0.01% (starch dry
base) can clearly change the pasting characteristics of the native
starches, however, the intermediate and the relatively higher
crosslinking starches, no viscosity can be detected. Phosphorus
content require less samples and are suitable for higher crosslinked
starches, for the very lower crosslinked starches, it is difficult to
distinguish them for the weighing precision, other disadvantages
such as its dangerous analysis and environmental pollution.
In this paper, the amylose content of crosslinked starches of
three typical crystalline structures with various crosslinking de-
grees was investigated, using three conventional methods settling
volume, P content, and paste characteristics to characterize the
crosslinking degree, comparisons were also made. The function and
properties that amylose played during the crosslinking reaction
were disclosed. It should also be noted first that very little amount
of the crosslinking reagent can change physicochemical properties
of the starch a lot, whereas increasing the regent level dose not give
the same effect, we applied dose efficiency to characterize the
crosslinking reaction of different reagent levels.
2. Materials and methods
2.1. Materials
Starches of three different crystalline structures, native maize
starch (A type) was provided by Qinhuangdao Lihua Starch Ltd.
(Hebei, China), native potato starch (B type) was produced by the
Dutch Meelunie Company (Amsterdam Holland), pea starch (C
type) was supplied by Shuangta Food Company (Yantai, China).
2.2. Preparation of the crosslinked starches
Crosslinked starches were prepared using previous reported
method [16] with some modification. Starches were crosslinked at
45
C and pH 11.5 by slurrying the starch (20%, db) with 0.01e10%
99:1 (w/w) mixture of sodium trimetaphosphate (STMP) and so-
dium tripolyphosphate (STPP) for 3 h at various reagent levels (0.01,
0.05, 0.1, 0.5, 1, 5, 10%) with sodium sulfate at 10% (starch basis).
After 3 h, the slurry was adjusted to pH 6.5 with 1M hydrochloric
acid, and the starch was recovered by centrifuging (3,000g, 10min);
washing with water (3X100 mL) and 95% ethanol (1X100 mL); then
drying at 50
C overnight.
2.3. Light microscopy
Starch granule morphology was studied with an Olympus BX
51(Olympus, Tokyo, Japan) polarized microscope equipped with a
CCD camera. Native maize, potato, pea starch and their crosslinked
counterparts (with 10% crosslinking reagent level (starch base)
added) were suspended in distilled water and viewed under
normal and polarized light microscope. A small drop of starch
suspension was placed on the microscope slide and covered with a
coverslip. The starch granule shape and Maltese cross were viewed.
2.4. Settling volume
The settling volume was determined according a previously
reported method [12] with some modification. Starch (2.0 g, db)
was constantly stirred in 40 mL distilled water within a water bath
at 95
C for 30 min. After the slurry was cooled to room tempera-
ture with constant stirring, 20 mL of the slurry was transferred to a
25 mL graduated cylinder. The settling volume of the starch in the
final mixture, which approximately equaled 1.0 g of starch on a dry
basis, was taken after 12 h.
2.5. Determination of the total phosphorus in starch
The total phosphorus includes the bound and residual phos-
phorus was determined according to GB/T 22427.11e2008/ISO
3946: 1982 [18]. Samples of phosphorylated starch
(0.5000 ± 0.0002 g, db) were mineralized with the concentrated
nitric and sulphuric acids. Orthophosphoric acid formed during
mineralization reacted with ammonium molybdate
[(NH4)6Mo7O24$4H2O] giving a complex of molybdenum (VI) ([P(V)
Mo(VI)12O40]3), which afterwards was reduced with ascorbic acid
to a deep blue complex of Mo(V). The absorbance of the blue so-
lution was measured at wavelength 825 nm with a Visible Spec-
trophotomete produced by Lengguang Tech. (Shanghai, China).
2.6. Pasting properties
The pasting profiles were monitored using a Micro Visco-
Amylo-Graph (Brabender, Germany). Maize, potato, and pea
starches (6.00 g, db) were weighed accurately into canisters and
distilled water was added to make a total weight of 100g. With a
700cmg torque, each starch slurry was heated from 30 to 95
C at a
rate of 7.5
C/min maintained at 95
C for 5min, and then cooled to
50
C at the same rate. From the pasting profiles, peak viscosity,
final viscosity at 95
C, and final viscosity were calculated using the
Viscograph software provided with the instrument.
2.7. Colorimetric method for determination of crosslinking degree
This section described the proposed new method for determi-
nation of crosslinking degree and the associated training method-
ology. As one of the ISO method [13], Riceddetermination of
amylose content, has been applied to detect amylose content of
native and modified starches. The ISO approach is based on
amyloseeiodine complexes have a characteristic iodine blue color,
and the color intensity can be used to indicate the concentration of
amylose content.
In this study, a new method, namely St-I method (starch-iodine
method), developed from ISO 6647e1: 2007 [13], was designed to
determine the crosslinking degree efficiently. The architecture of
the method is shown in Fig. 2. It mainly consists of four parts:
chemical gelatinization, color reaction, absorbance measurement
and evaluation of crosslinking degree. It was worth to note that we
applied the strong alkali (sodium hydroxide, NaOH, 1M) as the re-
agent to gelatinize the native and crosslinked starches.
Starches (100 ± 0.5 mg, db) were weighed into a 15 mL centrifuge
tube, then 1 mL ethanol added for sufficient dispersion of starch, it
was preferred to apply a vortex mixer while adding the NaOH (9 mL)
to ensure the homogeneous solution. The above solution was kept
for 10 min at room temperature, after that, the solution was
 T. Kou, Q. Gao / Carbohydrate Research 458-459 (2018) 13e18
transferred into 100 mL volumetric flask and add distilled water to
make 100 mL 5 mL of the above chemical gelatinized starch solution
was transferred into another 100 mL volumetric flask (with 50 mL
distilled water inside), 1 mL acetic acid solution (1M) was added to
neutralization the solution, then 2 mL I2-KI (configured according to
the description of ISO approach) added and made 100 mL for the
color reaction. 30 min later, the absorbance of the native starch (А)
and the crosslinked starches (a) was measured. The blank solution
was also configured according to ISO. The crosslinking degree (CL%)
was determined by the following equation:
CL% ¼ ðA  aÞ=A  100%
Where CL% is the crosslinking degree calculated by the colorimetric
method mentioned above, A is the absorbance of the native starch,
a is the absorbance of the crosslinked starches.
The dose efficiency (DE) was determined by the following
equation:
DE ¼ RL=CL%
Where DE is the dose efficiency, RL is the reagent level, CL% is the
crosslinking degree calculated by equation (1).
3. Results and discussion
3.1. Morphological characteristics
The morphological characteristics of crosslinked starches of
different starches have been portrayed by many papers [14], images
of native starches (images are collected both under normal light
Fig. 1. Light microscope of native and crosslinked starches prepared with the mixture 10% STMP/STPP (200).
15
and polarized light) and crosslinked starches (images taken under
polarized light) are shown in Fig. 1. It can be observed from mi-
crographs that the surface morphology of the starch has not been
changed due to crosslinking reaction. Crosslinking is a reaction that
occurs in magnitude of molecular, where no change in appearance
of the intact granules.
3.2. Calculation of substitution on crosslinking by paste viscosity
Pasting properties is a complicated mechanism, crosslinking
(1) reaction can influence the pasting properties of starches. It has
been hypothesized that crosslinking reaction has two opposite ef-
fects on the paste viscosity by two main mechanisms [15], on the
one hand, crosslinking reduces the loss of solubles (mainly
amylose) from granules and mechanically stiffens them, on the
other hand, crosslinking reaction can inhibit the swelling power of
the granules. For lower regent levels, the former plays the domi-
nant role, the structure of the mass for building the paste is
(2) stronger, as a consequence, the characteristic viscosity is enhanced.
For the higher reagent levels, the later is dominant, because the
starch volume fraction in the aqueous medium is reduced, which
would diminish viscosity. If the crosslinking regent level is high
enough (>1%) no viscosity will be detected (as shown in Table 1, no
viscosity will be shown for the higher crosslinked starches as well).
So it can be hypothesized that the viscosity of the crosslinked starch
paste will increase first and then decrease. That is to say when
applying viscosity to characterize the lower crosslinking degree, we
need to acknowledge the amount of the crosslinking reagent added.
It should be noted first, viscosity cannot be applied for the highly
crosslinked starches as no viscosity could be detected.
16 T. Kou, Q. Gao / Carbohydrate Research 458-459 (2018) 13e18

Fig. 2. The colorimetric results of the three native starch (maize starch, MS; potato starch, PS; and pea starch, PEA) and their crosslinked starches prepared with different levels of
crosslinking with the mixture STMP/STPP.

The properties of crosslinked starches made of identical condi-
tions of different plant origins varied from each other. First and
foremost, the structure of the three native starches are vary
depending on the combination of genes and starch biosynthesis [1],
The three starches with A, B and C crystalline patterns were applied
to prepare crosslinked starches with identical crosslinking reagent
level, although the order of paste viscosity was po-
tato > maize > pea. As the result of the structural differences and
the sample points we taken was not so concentrated, we cannot
point out which was the peak, still, the pea starch, which have a C
crystalline structure, did not followed the hypothesis, all the
crosslinked pea starches have a lower hot paste viscosity than the
native pea starch.
3.3. Calculation of substitution on crosslinking by settling volume
The settling volume profiles showed the extent of swelling of
the starch granules in a slurry; i.e., a higher swelling power resulted

Table 1
Settling volume and pasting properties (peak viscosity, final viscosity at 95
C, and final viscosity) of the three native starch (maize starch, MS; potato starch, PS; and pea starch,
PEA) and their crosslinked starches prepared with different levels of crosslinking with the mixture STMP/STPP.

Samples Peak viscosity (BU) Final viscosity at 95
C (BU) Final viscosity (BU) Settling volume (mL)

0%MS 96 76 144 20.00

0.01%MS 94 83 155 20.00
0.05%MS 104 98 175 20.00
0.1%MS 108 105 181 20.00
0.5%MS ea e e 19.00
1%MS e e e 18.50
5%MS e e e 8.50
10%MS e e e 6.25

0%PS 340 181 289 20.00

0.01%PS 288 246 326 20.00
0.05%PS 388 359 455 20.00
0.1%PS 419 402 534 20.00
0.5%PS 51 51 83 18.50
1%PS e e e 11.25
5%PS e e e 7.00
10%PS e e e 5.50

0%PEA 84 82 122 20.00

0.01%PEA 61 61 81 20.00
0.05%PEA 48 48 66 20.00
0.1%PEA 7 7 7 20.00
0.5%PEA e e e 20.00
1%PEA e e e 10.50
5%PEA e e e 9.50
10%PEA e e e 4.00
a
e: no viscosity can be detected by Brabender.
 T. Kou, Q. Gao / Carbohydrate Research 458-459 (2018) 13e18 17

Table 2
Phosphorus content of the three native starch (maize starch, MS; potato starch, PS; and pea starch, PEA) and their crosslinked starches prepared with different levels of
crosslinking with the mixture STMP/STPP.

Samples P% Samples P% Samples P%

0%MS 0.0109 ± 0.0002 0%PS 0.0299 ± 0 0%PEA 0.0045 ± 0.0001

0.01%MS ea 0.01%PS e 0.01%PEA e
0.05%MS e 0.05%PS e 0.05%PEA e
0.1%MS 0.0042 ± 0.0002 0.1%PS 0.0282 ± 0.0003 0.1%PEA 0.0020 ± 0
0.5%MS 0.0052 ± 0.0002 0.5%PS 0.0308 ± 0.0006 0.5%PEA 0.0089 ± 0.0009
1%MS 0.0104 ± 0.0002 1%PS 0.0347 ± 0.0001 1%PEA 0.0106 ± 0.0004
5%MS 0.0478 ± 0.0004 5%PS 0.0699 ± 0.0004 5%PEA 0.0835 ± 0.0009
10%MS 0.0871 ± 0.0001 10%PS 0.1675 ± 0.0013 10%PEA 0.1709 ± 0.0041
a
e: the ragent level was too small to be detected.

in a higher settling volume [16]. As for the very lower crosslinked

starches, no difference can be detected by their settling volume, for
the gelatinized starch granules were big enough to absorb all the
water. On the other hand, when it comes to the highly crosslinked
starches, with increasing the adding amount of the crosslinking
reagent, the settling volume of the crosslinked starches decreased,
however, the data was not dependent on the amount of the
crosslinking reagent added but in a very close difference. It was also
pointed out that we cannot tell whether the settling volume was
the result of the swelling power or the gelatinization of the starch
granules [11]. Table 1 listed the settling volume of native starches
and crosslinked starches of the three starches, in the meantime,
Table 1 also listed the peak viscosity, final viscosity at 95
C, and
final viscosity.
Apart from the information that Table 1 listed, at 0.01% reagent
level, the peak viscosity of maize starch and potato starch were
lower than the corresponding native starches, whereas the final
viscosity at 95
C and final viscosity were higher, which had not
been reported before, moreover, this trend was more evident for
the potato starch. At 0.5% reagent level, maize starch showed no
viscosity for its resistant to gelatinization, potato starch still
showed a low viscosity that can be detected by the instrument.
While crosslinked pea starches at different reagent levels did not
follow the trends of the other two starches in viscosity, at the same
reagent level, all the pea starches had a lower viscosity than the
other two, and no increase can be detected in viscosity for pea
starch. As a consequence, the characterization of crosslinking de-
gree by viscosity cannot be applied to the lower crosslinking re-
agent level and the pea starches.
3.4. Calculation of substitution on crosslinking by phosphorus
content
30:97  3 25
P% ¼   100%
305:89 100
P% ¼ 7:59%
The required weighting accuracy is ± 0:0002, so the least
amount of the reagent level that can be detected is:
0:0002
m ¼  100%
0:0759
m ¼ 0:264%
This meant that only when the agent level was higher than
0.264%, that the phosphorus may be detected, it is meaningless to
test the P% of the 0.01, 0.05, and 0.1% reagent level. As crosslinked
starch was mostly applied as thickening agent, when the reagent
level added was lower than 0.264%, the P% cannot characterize the
starches.
At 0.1% reagent level, all the crosslinked starches showed a
lower P% compared with their corresponding native starches, even
at 0.5% and 1% reagent levels, the P% of the crosslinked maize
starches were lower than the native maize starch. The P% of the
highly crosslinked starches were in line with the amount of the
reagent added, which was superior to the viscosity and the settling
volume approaches.
3.5. Calculation of substitution on crosslinking by St-I method
The mechanism of crosslinking reaction has been studied for a
long history, it has been suggested that crosslinking reaction can take
place between amylose/amylopectin and amylopectin/amylopectin,
it will not take place between amylose molecules for the relatively

The application of phosphorus content (P%) for crosslinking

degree characterization have become wide-spread for a long time
[17]. As native starches already had some phosphorus, and different
starch varieties have different amount of phosphorus content (see
Table 2). In addition, STMP/STPP was widely accepted as a multi-
functional reagent that can crosslink starch molecules, with the
increase of the reagent level more phosphorus was trapped inside
the starch granules. However, it was reported that the small
amount of the phosphorus in starch is partially removable [18], the
alkali treatment at above time-temperature combination can
reduce the phosphorus content. As shown in Table 2, the 0.01% and
0.05% levels were not taken into account for the combination of the
removable point and the accuracy of the test. The molecular weight
of STMP (Na3(PO3)3) is 305.89, while the molecular weight of
phosphorus (P) is 30.97. The minimum dilution volume is 100 mL,
and the maximum equal-volume is 25 mL, so the P% of the final Fig. 3. The mechanical map of the determination of crosslinking degree by St-I
solution deprived from STMP is: method.
18 T. Kou, Q. Gao / Carbohydrate Research 458-459 (2018) 13e18

Table 3
Crosslinking degree (CL%) maize starch (MS); potato starch (PS); and pea starch (PEA) and their crosslinked starches prepared with different levels of crosslinking with the
mixture STMP/STPP as well as the dose efficiency (DE).

Samples Absorbence CL% DE Samples Absorbence CL% DE Samples Absorbence CL% DE

0%MS 0.378 ± 0.002 0 e 0%PS 0.457 ± 0 0 e 0%PEA 0.635 ± 0.005 0 e

0.01%MS 0.374 ± 0.004 0.93 93.0 0.01%PS 0.419 ± 0.01 8.21 821 0.01%PEA 0.630 ± 0.007 0.79 79.0
0.05%MS 0.362 ± 0.002 4.11 82.2 0.05%PS 0.293 ± 0.02 35.82 716 0.05%PEA 0.576 ± 0.007 9.22 184
0.1%MS 0.348 ± 0.001 7.95 79.5 0.1%PS 0.261 ± 0.05 42.83 428 0.1%PEA 0.528 ± 0.01 16.78 168
0.5%MS 0.300 ± 0.006 20.53 41.1 0.5%PS 0.152 ± 0.003 66.81 134 0.5%PEA 0.355 ± 0.005 44.13 88.3
1%MS 0.285 ± 0.001 24.64 24.6 1%PS 0.099 ± 0.002 78.2 78.2 1%PEA 0.332 ± 0.001 47.68 47.7
5%MS 0.124 ± 0.002 67.15 13.4 5%PS 0.037 ± 0.003 91.89 18.4 5%PEA 0.137 ± 0.002 78.41 15.7
10%MS 0.081 ± 0.003 78.54 7.85 10%PS 0.027 ± 0.001 94.09 9.41 10%PEA 0.073 ± 0.002 88.49 8.85

far distance inside starch granules [19,20]. As shown in Fig. 3, the
amylopectin molecule was big enough to prevent the crosslinking of
two amylose molecules. After crosslinked to the amylopectin, the
new amylose-amylopectin structure can be accepted as a new
amylopectin, as a result, the amylose content decreased.
Amyloseeiodine complexes have a deep blue color, which is a result
of the amylose helix provides a tunnel for iodine molecules to align
[21]. With the increase of the crosslinking reagent levels free
amylose decreased, the intensity of the blue color also decreased,
which can be applied as a new approach to characterize the cross-
linking degree (see Table 3). It was observed that at lower level of
crosslinking reagent (0.01e0.1%) the degree of crosslinking was still
detectable by St-I method used, moreover, the St-I method can also
tell out the differences existed in the crystalline structure of the three
starches, the potato starch, B crystalline, changed much more quickly
than that of maize starch (A crystalline), while pea starch (C crys-
talline, with A outer B inner layers of the starch granules) kept in
between until the very high reagent level (10%).
Additionally, the competitive mechanism of crosslinking of
amylose/amylopectin and amylopectin/amylopectin was also
revealed, as shown in Table 3, the decrease of absorption intensity
at 0.01% reagent level was much higher than that of 10% reagent
level. The dose efficiency (DE) was much more informative, at 0.01%
reagent level, the DE was 93, 821, and 79 for maize, potato, and pea
starch respectively, the potato starch was much more inclined to be
crosslinked. Whereas at 10% reagent level, it was 7.85, 9.41, and
8.85, the reagent added was enough for the crosslinking reaction of
all the crystalline patterns of the three starches. By analyzing the
statistics, we can come a conclusion that in the earlier stages of the
crosslinking reaction, amylose/amylopectin is dominant, whereas
later the amylopectin/amylopectin dominated the reaction, lately,
the amylopectin/amylopectin increased.
4. Conclusion
The St-I method can characterize the crosslinked starches from
very low reagent level (0.01%) to very high level (10%), in addition,
the differences between different reagent levels can also be
detected, e.g., the absorption intensity of 5% is nearly twice the
absorbent of 10%. This method can characterize the crosslinked
starch regardless of its crosslinking reagent level, for the extremely
sensitivity of the crosslinking reaction of amylose/amylopectin,
dose efficiency is informative in characterizing the crosslinking
reaction, very low levels of crosslinking reagent can change the
characteristic viscosity (peak viscosity, final viscosity at 95
C, and
final viscosity) of the starch to a great extent, and very high levels of
crosslinking reagent level did not show many effect.
In addition, during the early stages of the crosslinking reaction,
the amylose/amylopectin dominated the reaction. Starch with B
pattern (potato starch) was found to be more inclined to be cross-
linked, starch with C pattern (pea starch) was found to be difficult to
be crosslinked in the early stages as A pattern (maize starch) did, and
more and more easily crosslinked in the coming stages as B pattern
did, which proves that C pattern was composed of A pattern for the
outside layers and B pattern for the inside layers.
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