Food Chemistry 133 (2012) 1196–1205

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Food Chemistry
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Synthesis, characterisation and in vitro digestibility of carboxymethyl potato

starch rapidly prepared with microwave-assistance
Jia Liu a, Jian Ming a, Weijin Li d, Guohua Zhao a,b,c,⇑
a
College of Food Science, Southwest University, Tiansheng Road 1, Chongqing 400715, PR China
b
Key Laboratory of Food Processing and Technology of Chongqing, Chongqing 400715, PR China
c
Chongqing Sweet Potato Engineering and Technology Centre, Chongqing 400715, PR China
d
College of Food Science, Sichuan Agriculture University, Yaan 625000, PR China

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

Article history: Carboxymethyl potato starch was synthesised with the aid of microwave. Optimal degree of substitution
Available online 18 May 2011 (DS) of 0.32 was obtained at 45 °C in 25 min using aqueous ethanol media with water/solvent of 0.15 at
200 W. The molar ratio of sodium hydroxide and monochloroacetic acid to anhydroglucose unit for opti-
Keywords: mal DS were 2.5 and 1.0, respectively. FT-IR spectrometry revealed the carboxymethyl starch to show
Potato new bands at m = 1614 cm1 and m = 1429 cm1. Wide angle X-ray diffractometry and DSC revealed a
Carboxymethyl starch remarkable reduction in starch crystallinity after carboxymethylation, which was consistent with
Microwave-assisted
destroyed surface observed in SEM. The digestibility of carboxymethyl starch (CMS) was lower than that
In vitro digestibility
Resistant starch
of native starch. With similar DS, there was no difference in digestibility of carboxymethyl starch pre-
Paste properties pared with and without microwave. As DS increased from 0.05 to 0.32, the amount of resistant starch
in microwave-assisted carboxymethyl starch was elevated from 14.6% to 20.0%, which was much higher
than that of native starch (10.8%).
Ó 2011 Elsevier Ltd. All rights reserved.

1. Introduction they are easily-prepared, non-toxic and renewable, they play an

China is the largest producer of potatoes in the world with an
annual yield of approx. 70 million tons. In China, potatoes are used
for the production of starch and other starchy foods. The physical
properties of native starches and the colloidal solutions produced
by heating their suspensions limit their commercial applications
(Bhattacharyya, Singhal, & Kulkarni, 1995a). For improving some
specific properties to suit industrial process, native starch can be
chemically modified (Tijsen, Kolk, Stamhuis, & Beenackers, 2001a,
2001b), for instance, by hydroxypropylation (Wootton & Chaudhry,
1981), carboxymethylation (Tijsen, Kolk, Stamhuis, & Beenackers,
1999), cross-linking (Woo & Seib, 2002) among many others. Since
Abbreviations: AUC, area under the hydrolysis curve; CT, conventional modifi-
cation method; CMS, carboxymethyl starch; DS, degree of substitution; DSC,
differential scanning calorimetry; FG, free glucose; FT-IR, Fourier transform
infrared; GI, estimated glycemic index; H-CMS, carboxymethyl starch in hydrogen
form; HI, hydrolysis index; MAT, microwave-assisted; MCA, monochloroacetic acid;
NPS, native potato starch; RDS, rapidly digestible starch; RS, resistant starch; SDS,
slowly digestible starch; SEM, scanning electron micrographs; TS, total starch; XRD,
X-ray diffractometry.
⇑ Corresponding author at: College of Food Science, Southwest University,
Tiansheng Road 1, Chongqing 400715, PR China. Tel.: +86 23 68 25 03 74; fax:
+86 68 25 19 47.
E-mail address: zhaogh@swu.edu.cn (G. Zhao).
important role in industries such as food, pharmaceuticals, textile,
and paper (Bhattacharyya, Singhal, & Kulkarni, 1995b; Khalil,
Hashem, & Hebeish, 2006). Among these starch derivatives, car-
boxymethyl starch (CMS) as a kind of etherified starch has at-
tracted a lot of attention in both research and industry (Lawal,
Lechner, Hartmann, & Kulicke, 2007). The presence of functional
group (CH2 COO ) yields starch with many unique properties, such
as low gelatinization temperature, excellent flexibility, improved
paste storage stability and clarity (Qiu & He, 1999). Due to its un-
ique properties, CMS with degree of substitution (DS) 6 0.4 can be
used as stabilizer, thickening and water retaining agent to improve
the quality of food (Björck, Gunnarsson, & Östergård, 1988; Lawal
et al., 2007; Stojanovic, Jeremic, & Jovanovic, 2000; Östergård,
Björck, & Gunnarsson, 1988).
The resistant starch (RS) is thought to be desirable in human
health, as it has functional properties similar to fermentable die-
tary fibres (Noda et al., 2008). In the colon, RS increases faecal bulk,
lowers colonic pH and the portion fermented by the intestinal
microflora produces a range of short chain fatty acids, especially
butyrate which is thought to provide a degree of protection against
bowel cancer. Chemically modified starch belongs to RS4 and is re-
ported to reduce digestibility (Chung, Shin, & Lim, 2008). Wootton
and Chaudhry (1981) observed that the digestibility of the
gelatinized wheat starches decreased as the degree of

0308-8146/$ - see front matter Ó 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2011.05.061
 J. Liu et al. / Food Chemistry 133 (2012) 1196–1205 1197

Table 1
Degree of substitution (DS) of carboxymethyl potato starch and reaction conditions used for carboxymethylation of potato starch.

Treatment CodeA Microwave Power (W) Temperature (°C) Time (min) H2O/solvent nNaOH/nAGUB nMCA/nAGUC DS
CT-1 – 45 150 0.15 2.5 1.0 0.30 ± 0.01
CT-2 – 45 30 0.15 2.5 1.0 0.08 ± 0.01
MAT-1 100 45 25 0.15 2.5 1.0 0.26 ± 0.01
MAT-2 100 45 45 0.15 2.5 1.0 0.31 ± 0.02
MAT-3 200 45 25 0.15 2.5 1.0 0.32 ± 0.01
MAT-4 200 25 25 0.15 2.0 1.0 0.02 ± 0.01
MAT-5 200 35 25 0.15 2.0 1.0 0.14 ± 0.01
MAT-6 200 50 25 0.15 2.0 1.0 0.28 ± 0.01
MAT-7 200 50 25 0.15 2.5 1.0 0.34 ± 0.01
MAT-8 200 45 1 0.15 2.5 1.0 0.06 ± 0.02
MAT-9 200 45 15 0.15 2.5 1.0 0.27 ± 0.01
MAT-10 200 45 30 0.15 2.5 1.0 0.33 ± 0.01
MAT-11 200 45 25 0.05 2.5 1.0 0.09 ± 0.01
MAT-12 200 45 25 0.25 2.5 1.0 0.22 ± 0.01
MAT-13 200 45 25 0.15 2.5 1.4 0.21 ± 0.01
MAT-14 200 45 25 0.15 2.5 0.6 0.13 ± 0.01
MAT-15 200 45 25 0.15 3.5 1.0 0.31 ± 0.01
MAT-16 200 45 25 0.15 3.0 1.0 0.35 ± 0.01
MAT-17 200 45 25 0.15 2.0 1.0 0.27 ± 0.01
MAT-18 300 45 25 0.15 2.5 1.0 0.32 ± 0.01
A
CT, carboxymethyl starch prepared without microwave-assisted; MAT, carboxymethyl starch prepared with microwave-assisted.
B
nNaOH/nAGU, the addition of sodium hydroxide in reaction solution expressed as the ratio of sodium hydroxide moles to anhydroglucose unit moles.
C
nMCA/nAGU, the addition of monochloroacetic acid in reaction solution expressed as the ratio of monochloroacetic acid moles to anhydroglucose unit moles.

hydroxypropylation increased. Östergård et al. (1988), Woo and
seib (2002) and Chung et al. (2008) found that starch substituted
with acetyl or hydroxypropyl groups or oxidised would reduce
the ultimate level of enzymatic hydrolysis of starches in the gela-
tinized starches. Jyothi, Rajasekharan, Moorthy, and Sreekumar
(2005) showed the in vitro digestibility of cassava starch succinates
prepared by microwave-assistance to be lower than native starch.
Microwave radiation is an efficient source of thermal energy,
providing fast and uniform heating, and is becoming a standard
technique in various fields of chemical synthesis including modi-
fication of starch. Microwave dielectric heating dissipates heat in-
side the medium and raises the energy of the molecules rapidly.
Consequently, under microwave dielectric heating, more mole-
cules become energised, and this usually results in higher reac-
tion rates under microwave dielectric heating (Li et al., 2001).
Many studies have reported the microwave-assisted chemical
modifications of starch, such as methyl starch (Singh & Tiwari,
2008), starch acetates (Shogren & Biswas, 2006), starch carbamate
ester and phosphate ester (Lewandowicz et al., 2000). However,
reports on microwave-assisted carboxymethylation of starch are
scant.
The objective of the present works was to investigate micro-
wave-assisted (MAT) carboxymethylation of starch in a solvent
system and its comparison with conventional modification method
(CT) as control. The effects of microwave reaction conditions were
optimised so as to develop a new process to prepare CMS rapidly.
In addition, the digestibility of CMS is not fully understood, and the
starch nutritional fraction of CMS has not been discussed before. A
secondary objective was to evaluate the in vitro digestibility of
MAT-CMS and CT-CMS.
2. Material and methods
2.1. Materials
Native potato starch (NPS) was kindly supplied by the JingTian
Agriculture Development Co. Ltd. (Chongqin, China). NaOH, meth-
anol, ethanol, acetone, monochloroacetic acid (MCA) were of ana-
lytical grade and purchased from Kelong Reagent Chemical Co.
(Chengdu, China). Porcine pancreatic a-amylase (No. 3176,
30 kU/g, 1 U will liberate 1.0 mg of maltose from starch in
3 min at pH 6.9 and 20 °C) and amyloglucosidase (No. 9913,
320 U/mL, 1 U will liberate 1.0 mg of glucose from starch in
3 min at pH 4.2 and 60 °C) were purchased from the Sigma (St.
Louis, MO, USA).
2.2. Preparation of CMS
CMS was prepared by a substitution reaction in aqueous etha-
nol under alkaline condition using MCA as carboxymethyl group
providing regent.
(1) CMS prepared by conventional modification method (CT) was
conducted as per the scheme suggested by Hebeish and Khalil
(1988). NPS (13.6 g, dry weight), ethanol (85 mL), water
(15 mL) and NaOH (4.20 g) were added to a three-necked
flask installed with an agitator and mixed for 30 min at
30 °C. MCA (7.93 g) and NaOH (4.19 g) were then added to
the flask. The reaction mixture was warmed at 45 °C and con-
tinually agitated for 30 and 150 min to respectively obtain
two control samples having significantly different DS.
(2) The synthesis of CMS with microwave-assistance was also
carried out in two steps. In the first step, the alkalization
of starch was carried out in a 250 mL round-bottom flask,
equipped with the motor-driven stirrer. NaOH (2.73–
4.33 g) was added to water (6.5–32.5 mL) in the flask and
the mixture was stirred at 250 rpm to reach complete disso-
lution of NaOH. Alkalization took place when NPS (10.4 g,
dry weight) and ethanol (97.5–123.5 mL) were added to
the flask and the temperature of water bath was raised to
35 °C for 0.5 h. In the second step, monochloroacetic acid
(3.51–8.19 g) and sodium hydroxide (2.73–4.33 g) were
added to the mixture. Then, the reactor was placed at the
centre of the microwave apparatus (MAS-II, max output,
1 kW; frequency, 2.45 GHz; Chemical Science and Technol-
ogy Co. Ltd., Shanghai, China). The mixture was stirred with
magnetic stirrer bar at 1100 rpm for homogeneous heating.
The reaction conditions are presented in Table 1.
At the end of the reaction, the product was washed several
times with 85% ethanol until the silver nitrate test for chloride of
the filtrate was negative. The slurry obtained was suspended in
acetone, stirred for 20 min, and dried in an oven at 40 °C for 48 h.
1198 J. Liu et al. / Food Chemistry 133 (2012) 1196–1205
2.3. Determination of degree of substitution
Titrimetry was used for the determination of the DS (Li et al.,
2001). CMS (5 g) was dispersed in acetone (150 mL) and 5 M HCl
(15 mL) was added to the dispersion and stirred for 30 min. During
this process, the sodium form CMS was converted to the H-CMS
(carboxymethyl starch in hydrogen form). H-CMS was washed sev-
eral times with 80% (v/v) methanol until the solution became neu-
tral with pH test. The neutral dispersion was filtered again,
suspended in acetone, and it was stirred for another 15 min, fol-
lowing which it was filtered and dried for 24 h in a desiccator over
silica gel. Two grams of H-CMS were dissolved in 1% (w/v) NaCl
solution and it was titrated with 1 M NaOH. The DS was deter-
mined as follows:
 
nNaOH  M0 mP  F
DS ¼ mc ¼ mP 
mc  nNaOH  M R 100
where M0 is molar mass of the anhydroglucose unit (162 g/mol);
MR is molar mass of carboxymethyl residue (58 g/mol); nNaOH is
the quantity of sodium hydroxide used (mol); mp is the weight of
polymer taken (g); mc is the corrected weight of polymer (g), and
F is the moisture content of the sample (g/100 g).
2.4. Fourier transform infrared (FT-IR) spectroscopy
The IR spectra of NPS, MAT-CMS and CT-CMS were obtained
with a Spectrum Scanner (Perkin–Elmer, model 2000, USA) in the
frequency range 500–4000 cm1. The starch sample was blended
with KBr powder and pressed into tablets before measurement.
2.5. Scanning electron microscopy (SEM)
The structure was observed using a scanning electron micro-
scope (SEM). NPS, MAT-CMS and CT-CMS were placed in an oven
at 45 °C for 5 days for dehydration. The dehydrated samples were
then coated with gold powder to avoid charging under the electron
beam. The granule surface and shape of starch were observed using
Hitachi S-4800 scanning electron microscope (Tokyo, Japan).
2.6. X-ray diffraction (XRD)
The X-ray diffraction pattern of NPS, MAT-CMS and CT-CMS
were recorded with an X’Pert pro X-ray diffractometer equipped
with X’celerator as detector (Panalytical, Kassel, Germany). The dif-
fractograms were registered at Bragg angle (2h) = 5–30° at a scan
rate of 5°/min, while step with = 0.02°.
2.7. Differential scanning calorimetry (DSC)
The thermal analyses of NPS, MAT-CMS and CT-CMS were car-
ried out using a differential scanning calorimeter (SDTQ600, TA
Instruments, USA). A total of 10 mg samples sealed in aluminium
pans with 5.0 lL of deionized water was scanned from 30 to
120 °C at a rate of 10 °C/min while sealed empty pans were used
as reference (Lawal, Lechner, & Kulicke, 2008a). The onset temper-
ature (T0), peak temperature (Tp), conclusion temperature (Tc) and
the enthalpy of gelatinization were determined.
2.8. In vitro starch digestibility
The in vitro starch digestibility was determined by following the
procedure of Englyst, Kingman, and Cummings (1992) and Miao,
Zhang, Mu, and Jiang (2010) with a slight modification. Amyloglu-
cosidase (1 mL) was added to deionized water (2 mL). Porcine pan-
creatic a-amylase (3.21 g) was dispersed in water (24.7 mL) and
centrifuged for 10 min at 2500g, and 18.7 mL of supernatant was
collected. This supernatant was mixed with 1 mL of diluted amylo-
glucosidase for making the enzyme solution. The solution was
freshly prepared for the digestion analysis.Two hundred milligram
samples and 10 mL deionized water were added to the tube. The
tubes were then heated in a boiling water bath for 30 min. The
pH of each sample was adjusted to around 5 using 2.0 M HCl solu-
tion. Seven glass balls (10 mm in diameter), 0.5 M sodium acetate
solution (5 mL) and the enzyme solution (10 mL) were then added
to the tube containing the starch gels, followed by incubation in a
water bath at 37 °C with agitation of 120 rpm. Aliquots (0.5 mL)
were taken at 0, 20, 40, 60, 80, 100, 120 and 180 min, and mixed
with 4 mL of 80% ethanol, and the reducing sugar of the hydroly-
sates was determined using 3,5-dinitrosalicylic acid (Bernfeld,
1955). A glucose standard curve was prepared and the amount of
hydrolysis calculated as the proportion of starch converted to
glucose.
The total starch (TS) content in the starch samples and the
amount of starch fractions based on digestibility was calculated
by using the procedure of Englyst et al. (1992). The values of differ-
ent starch fractions of RDS, SDS and RS were calculated using the
following formulas: RDS (g/100 g) = [(G20  FG)  0.9/TS]  100,
SDS (g/100 g) = [(G120  G20)  0.9/TS]  100, and RS (g/100 g) =
[[TS  (RDS + SDS)]/TS]  100, where G20 is the amount of glucose
released after 20 min hydrolysis (g); G120 is the amount of glucose
released after 120 min hydrolysis (g), FG is the amount of free glu-
cose in original sample (g) and TS is the amount of starch in origi-
nal sample (g).
2.9. Estimation of glycemic index (GI)
A non-linear model established by Goni, Garcia-Alonso, and
Saura-Calixto (1997) was applied to describe the kinetics of starch
hydrolysis. The first order equation has the form: C ¼ C 1 ð1  ekt Þ,
where C corresponds to the percentage of starch hydrolysed at
time t, C1 is the equilibrium percentage of starch hydrolysed after
180 min, k is the kinetic constant and t is the hydrolysis time (min).
The parameters C1 and k were estimated for each treatment based
on the data obtained from the in vitro hydrolysis procedure. Param-
eter estimation was carried out using the ORIGIN version 8.0.
The area under the hydrolysis curve (AUC) was calculated
using the equation AUC = C1(tf  t0)  (C1/k)[1  exp[k(tf  t0)]],
where C1 corresponds to the equilibrium percentage of starch
hydrolysed after 180 min, tf is the final time (180 min), t0 is the
initial time (0 min) and k is the kinetic constant. A hydrolysis in-
dex (HI) was obtained by dividing the area under the hydrolysis
curve of each sample by the corresponding area of a reference
sample (white bread) (Granfeldt, Björck, Drews, & Tovar, 1992).
Previous research has shown HI to be a good predictor of glyce-
mic response (Goni et al., 1997). The glycemic indices (GI) of
the samples were calculated according to the equation proposed
by Goni et al. (1997): GI = 39.71 + 0.549HI.
2.10. Freeze–thaw stability
Freeze–thaw stability of NPS, MAT-CMS and CT-CMS pastes was
measured at a concentration of 2% (w/v) in distilled water. NPS
paste was prepared by gelatinizing aqueous NPS suspension in
boiling water bath for 30 min with constant mild agitation and
cooled to 25 °C. CMS pastes were obtained by dispersing CMS pow-
der in the stirring water, mixed exhaustively, and the suspensions
were allowed to stand overnight at room temperature (25 °C) for
complete swelling. Twenty grams accurate weight of the starch
paste was added into each of the pre-weighed 50 mL centrifuge
tubes, then frozen at 18 °C in a freezer for 18 h. All tubes were re-
moved from the freezer and thawed at room temperature (25 °C)
 J. Liu et al. / Food Chemistry 133 (2012) 1196–1205
for 6 h. Three tubes from each thawing cycle of these samples were
centrifuged at 4000 rpm for 30 min. The clear liquid was decanted,
and the residue was weighed. The percentage of syneresis (%) was
then calculated as the ratio of the weight of the liquid decanted to
the total weight of the paste before centrifugation and multiplied
by 100. The remaining tubes were then put back to the freezer
for further freeze–thaw cycling. Six freeze–thaw cycles were per-
formed for this study.
2.11. Paste clarity
The NPS, MAT-CMS, and CT-CMS pastes were prepared as men-
tioned above and their clarity determined using starch aqueous
dispersions 1% (w/v), as described by Craig, Maningat, Seib, and
Hoseney (1989). The pastes were stored in a freezer at 4 °C. Ten
millilitres of totally mixed paste was removed from the freezer
and equilibrated at temperature (25 °C) for 1 h prior to paste clar-
ity determination. The paste clarity was evaluated with a spectro-
photometer (722-P, Xianke Co., Shanghai) at 640 nm as a function
of storage time. Paste clarity was expressed as percent transmit-
tance (%T) of the paste.
2.12. Paste Viscosity
Viscosity was measured using a rotational rheometer (Brook-
field Engineering Laboratories, MA, USA; model LVDV-II+PRO) in
2% (w/v) aqueous starch paste prepared as mentioned above.
Depending on the range of viscosity to be determined, spindles
of No. S61 (MAT-CMS, DS 0.05; CT-CMS, DS 0.08) and S62 (MAT-
CMS, DS 0.32; CT-CMS, DS 0.30; NPS) were used. Viscosity as a
function of shear rate varied between 1.5 and 100 s1 was mea-
sured at a controlled temperature of 25 °C. Viscosity as a function
of temperature varied from 25 to 85 °C was measured with fixed
shear rates of 30 s1 for thin samples (MAT-CMS, DS 0.05, CT-
CMS, DS 0.08) and 3 s1 for thick samples (MAT-CMS, DS 0.32,
CT-CMS, DS 0.30, NPS). Viscosity in mPa s was recorded.
2.13. Statistical analysis
All measurements were replicated in triplicate. Data were sub-
jected to analysis of variance (ANOVA), and comparison of means
was carried out by Tukey’s HSD test. Differences were considered
to be significant at p < 0.05 (Miao et al., 2010). Statistical compu-
tations and analyses were conducted using SPSS16.0.
3. Results and discussion
3.1. Carboxymethylation of potato starch with microwave-assistance
The DS and synthesis parameters of the CMS subjected to car-
boxymethylation of starch with and without microwave-assistance
are presented in Table 1. The DS of CMS under similar reaction con-
ditions in this study was lower than those reported by Ornanong,
Jakkapan, and Reinhard (2006) and Lee et al. (2010). Stojanovic,
Jeremic, Jovanovic, and Lechner (2005) found that direct titration
gave lower DS, while the Cu salt precipitation method gave higher
DS values than the back titration method, which was found to be
the most accurate. Moreover, the CMS obtained from corn starch
is reported to have a higher DS than that from potato starch under
similar experimental conditions. Thus, the lower DS observed in
the present study may be due to different methods of determina-
tion of DS of CMS and different origin of starch.
1199
3.1.1. Effect of microwave power
At a power above 400 W, it was found that the starch was com-
pletely gelatinized and formed a hard mass. Thus, to prevent gela-
tinization of potato starch in the reaction medium and extend the
duration of the carboxymethylation, the carboxymethylation of
potato starch under microwave radiation was conducted at low
power (from 100 to 300 W). An increase in DS (0.32) was observed
when the power was raised to 200 W, beyond which no further in-
crease in DS was observed. Starch modified under microwave radi-
ation (25 min) could produce product with similar DS in shorter
reaction time than starch modified conventionally (150 min).
Through promoting the accessibility of reagent to starch molecular
and raising the energy of the molecules rapidly, power is a key fac-
tor to accelerate the carboxymethylation of starch. To achieve the
same DS, microwave-assistance consumed much less time than
conventional method. However, carboxymethylation under
100 W consumed more time than 200 and 300 W to acquire the
product with satisfied DS. In other words, reaction time exerted
more acute influence on DS of product from microwave-assistance
than that from conventional method, which was implied by the
difference in DS of CT-1, MAT-1 and MAT-2. As the reaction time
of microwave-assistance was extended to 45 min, the CMS ob-
tained had a DS similar to that prepared with conventional method
for 150 min.
3.1.2. Effect of molar ratios of MCA to starch
An increase in the DS was observed when the amount of MCA
added to the reaction mixture was increased from 0.6 to 1.0 in
terms of nMCA/nAGU (Table 1). With an increase in the MCA concen-
tration, the contact between starch molecules and etherifying
agent increased. However, the DS decreased above nMCA/nAGU of 1
and a maximal DS of 0.32 was recorded within the range evaluated.
This could reasonably be attributed to the increasing ratio of MCA
will lead to usage of NaOH, while under the same reaction condi-
tion, the amount of NaOH is changeless. So the more MCA is, the
less NaOH can react with AGU, which leads to lower DS (Wang,
Pan, Hu, Miao, & Xu, 2010).
3.1.3. Effect of molar ratios of NaOH to starch
As observed, the DS of the CMS increased with an increase in
NaOH concentration and attained a maximum DS of 0.35 at
nNaOH/nAGU of 3.0 (Table 1). However, gelatinization occurred under
this condition and the recovery of product was difficult. Therefore,
the optimal nNaOH/nAGU is reasonably adjusted to 2.5. During the
carboxymethylation process, the NaOH provides the alkaline envi-
ronment for the reaction as well as serving as the swelling agent to
facilitate diffusion and penetration of the etherifying agent to the
starch granular structure (Lawal et al., 2007). However, as the
amount of NaOH increased, the starch gelatinized easily and
the contact between starch and MCA was inhibited in the reaction
mixture. Moreover, further increase in sodium NaOH concentration
diverted the MCA towards the side reaction. This may explain the
reduction in the DS at higher nNaOH/nAGU (3.5).
3.1.4. Effect of water content in the reaction medium
Water is a good absorber of MR and plays the important role of
aiding dissociation, diffusion and adsorption of etherifying agent. It
is probable that any reaction containing water would be greatly
accelerated by microwave heating. As observed in the present
study, carboxymethylation of starch was not very significant and
CMS with a rather low DS (0.09) was obtained at a volume ratio
of water to solvent of 0.05. Further increase in volume ratio of
water to solvent to 0.25 reduced the DS, and showed agglomera-
tion and starch gelatinization. It is reasonable to say that the crit-
ical volume ratio of water to solvent is around 0.15.
1200 J. Liu et al. / Food Chemistry 133 (2012) 1196–1205
3.1.5. Effect of temperature
The DS increased with an increase in temperature from 25 to
50 °C. Generally, higher temperature increases the solubility and
diffusion of the etherifying reagents and swelling of starch mole-
cules. The proportion of molecules with higher energy than the
activation energy is reported to increase with an increase in tem-
perature which consequently increases the rate of reaction (Lawal,
Lechner, & Kulicke, 2008b). The highest DS (0.34) was obtained
when the carboxymethylation was performed at 50 °C. However,
gelatinization occurred at 50 °C, and the recovery of product was
Fig. 1. The infrared spectra of native potato starch (a) and carboxymethyl potato
starch prepared with microwave-assisted method (b, with a DS of 0.32) and
conventional method (c, with a DS of 0.30).
Fig. 2. Scanning electron micrographs of native potato starch (A, 1000), carboxymethyl potato starches with a DS of 0.32 (B, 1000) prepared with microwave-assisted
method, and carboxymethyl potato starches with a DS of 0.30 (C, 1000) prepared with conventional method.
difficult. From the above analysis, the optimal temperature was
45 °C.
3.2. Fourier transform infrared (FT-IR) spectroscopy
The infrared spectra of NPS (a), MAT-CMS (b, with a DS of 0.32)
and CT-CMS (c, with a DS of 0.30) are presented in Fig.1. FT-IR spec-
troscopy shows similarity in MAT-CMS and CT-CMS. The band
stretch around 3400 cm1 (c, 3413.47 cm1; b, 3414.70 cm1) is
attributed to hydrogen-bonded hydroxyls on the starch molecules.
The band at m = 2924 cm1 is attributed to –CH2 symmetrical
stretching vibrations. The bond at m = 1643 cm1 in the native
starch is assigned to scissoring of two O–H bonds of absorbed
water molecules. The spectral group from 1020 to 1159 cm1, typ-
ical starch, is preserved in native starch and CMS. In the CMS, the
intense bands at m = 1614 cm1 (c), m = 1622 cm1 (b), m =
1429 cm1 (c), and m = 1419 cm1 (b) are assigned to carbonyl
functional group. These new bands confirms that carboxymethyla-
tion of starch did take place. Similar observations are reported for
kudzu starch (Wang et al., 2010) and yam starch after carboxy-
methylation (Wang, Gao, & Li, 2009).
3.3. Starch granule morphology
The scanning electron micrographs (SEM) of NPS (A), MAT-CMS
(B and C, with a DS of 0.32) and CT-CMS (D and E, with a DS of 0.30)
are shown in Fig. 2. SEM showed the carboxymethylation to change
the structure of starch granules, compared with native starch. Na-
tive starch granules were smooth, round in shape with sizes rang-
ing from 15 to 100 lm. Although the treatment brought noticeable
changes to the granular structure of CMS, the integrity of the starch
granules was still preserved, i.e. each granule still maintained its
individual entity. After carboxymethylation, both in MAT-CMS
 J. Liu et al. / Food Chemistry 133 (2012) 1196–1205
and CT-CMS, some granules got together to form big groups and
the granular surface become rough and scaly (Fig.2). As compared
to CT-CMS, the surface of granules of MAT-CMS did show a greater
collapse in the structure. Most granules of modified starch seemed
to be heavily distorted and appeared as folded structures with their
outer sides drawn inwards, assuming the shape of a doughnut. It is
possible that the changes observed on the modified starch granules
were due to the loss of crystalline structure and the breakage of
chemical bonds in starch molecules affected by strong alkaline
environment together with heat treatment. This means that the
carboxymethyl reaction not only took place on the surface of starch
granules but also within. Similar observations have been reported
for carboxymethyl starches in the literature (Kittipongpatana,
Sirithunyalug, & Laenger, 2006; Wang et al., 2009). Meanwhile,
Lee et al. (2010) found carboxymethylation of starch to result in
reduction in the molecular weight due to molecular degradation
by the alkaline treatment.
3.4. X-ray diffractometry (XRD)
The wide-angle X-ray diffractograms obtained for NPS (a), MAT-
CMS (b, with a DS of 0.32) and CT-CMS (c, with a DS of 0.30) are
presented in Fig.3. XRD patterns of NPS exhibited strong diffraction
peaks at 5.1, 15.6, 17.3, 22.9, and 24.3 2h angles, which was a char-
acteristic B-type crystallinity pattern in Fig.3(a). Hydrogen bonds
are responsible for forming crystal structure of starch molecules
and maintain the stability of crystal structure. After carboxymeth-
ylation, in both MAT-CMS and CT-CMS, the carboxymethyl groups
on the anhydroglucose unit remarkably reduced the crystallinity as
seen in Fig.3(b and c). This observation also corroborates the mor-
phological studies. It is reasonable that the strong alkaline condi-
tion with heat treatment caused granular disintegration during
the modification (Lawal et al., 2007).
3.5. Differential scanning calorimetric (DSC)
The DSC thermograms of NPS (a), MAT-CMS (b, with a DS of
0.32) and CT-CMS (c, with a DS of 0.30) are presented in Fig. 4.
The T0, Tp, Tc for the NPS are 62.9, 72.7, 87.1 °C, repectively. The
gelatinization enthalpy for the NPS (DH) was 10.366 J/g. As ob-
served, gelatinization enthalpy of both MAT-CMS and CT-CMS
could not be determined. It is indicative that carboxymathylation
tremendously affected the starch crystallinty and eventually in-
Fig. 3. Wide angle X-ray diffraction pattern of native potato starch (a) and
carboxymethyl potato starch prepared with microwave-assisted method (b, with a
DS of 0.32) and conventional method (c, with a DS of 0.30).
1201
Fig. 4. The DSC curves of native potato starch (a) and carboxymethyl potato starch
prepared with microwave-assisted method (b, with a DS of 0.32) and conventional
method (c, with a DS of 0.30).
creased the starch amorphous region. The loss of crystallinty cor-
roborates the previous X-ray diffraction studies. Similar
observations have been also reported for water yam starch (Lawal
et al., 2008a) and pigeon pea starch (Lawal et al., 2008b). Starch
gelatinization is a process that involves the uncoiling of double
helices and melting of the starch crystallites in the starch crystal-
line region. However, the processes involved in carboxymethyla-
tion tremendously affected the starch crystallinity thus making
the granules largely amorphous. The results of XRD and DSC
showed that the loss of crystalline structures inside the granules
may result in rough surface granules observed on the SEM images
and finally cause the starch granules to become ‘‘pregelatinized’’. A
study on crystalline properties of starch also reported that pregel-
atinized maize starches, which are water-soluble, lost the crystal-
line structures in the starch granules (Zhang, Yang, Li, & Gao,
2011). Chen and Jane (1994) explained that the strong alkaline
conditions in such treatment transformed hydroxyl groups of the
starch molecules (St–OH) into alkoxide groups (St–O). The repul-
sion between negative charges resulted in swelling of the starch
granules and exerted a tension on neighbouring crystallites of
starch molecules. This led to dissociation of double-helical regions
and the breakup of the crystalline structure. The starch granules
thus became weaker and soluble in cold water.
3.6. In vitro digestibility
As shown in the starch hydrolysis curve (Fig. 5), NPS, MAT-CMS
as well as CT-CMS were hydrolysed quickly during the early stage
and then reached a plateau. This may due to the swelling of starch
after gelatinization through boiling subsequent to which pancre-
atic a-amylase can easily get into the inner structure of starch
(Chung et al., 2008). However, the rate of starch digestion was re-
duced by carboxymethylation during the early stage (about
30 min). After 120 min, the degree of hydrolysis became substan-
tially different among the starches tested. In the late stage of
hydrolysis (180 min), the NPS showed higher level of hydrolysis
than the CMS (DS P 0.3). Similar observation was reported for
starch after etherification and oxidation (Wolf, Bauer, & Fahey,
1999). This was probably because the substituted groups were in-
ert to the enzymatic action, and thus provided CMS the resistance
to enzymatic hydrolysis (Han & BeMiller, 2007; Hoover, Hannouz,
& Sosulski, 1988; Tester, Karkalas, & Qi, 2004).
1202 J. Liu et al. / Food Chemistry 133 (2012) 1196–1205
Fig. 5. Starch hydrolysis patterns of the native and carboxymethyl potato starch in
the gelatinized state after heating in boiling water bath for 20 min prior to
treatment with porcine pancreatic a-amylase for 3 h at 37 °C. NPS, native potato
starch; MAT-CMS, carboxymethyl potato starch prepared with microwave-assisted
method; CT-CMS, carboxymethyl starch prepared with conventional method.
Starch can be classified into three nutritional fractions after en-
zyme hydrolysis: rapidly digestible starch (RDS), which causes a
rapid increase in blood glucose; slowly digestible starch (SDS),
which is digested slowly but completely in the human small intes-
tine; resistant starch (RS), which cannot be digested in the small
intestine but can be in the large intestine (Englyst et al., 1992).
The starch nutritional fractions which were determined by the
method of Englyst are presented in Table 2. The NPS had the high-
est amount of RDS (83.2%) among the tested starches. The MAT-
CMS and CT-CMS had significantly lower RDS content (63.7% and
64.6%) than NPS. The observations in the present study suggested
that carboxymethylation of starch increased the amount of SDS
and RS as compared to the NPS. As the DS of the CMS increased
from 0.05 to 0.32, no significant difference in the amount of SDS
and RS was observed. The different nutritional fractions between
the NPS and CMS may be caused by alteration in the structure.
The relationship between crystallinity and enzymatic resistance
is different for native starch and chemically modified starch. Crys-
tallinity in native starch is mainly responsible for the resistance to
enzyme hydrolysis. When crystalline regions decreased, the hydro-
lysis of native starch increased. However, modified starch shows
opposite situation. Although decreased crystallinity enhanced the
accessibility of the starch chains to the enzyme, the hydroxyl group
of glucose substituted by carboxymethyl group could possibly hin-
Table 2
Amount of rapidly (RDS) and slowly (SDS) digestible starches, and resistant starch
(RS) in gelatinized native and carboxymethyl potato starch.A,B
StarchC RDS (%) SDS (%) RS (%)
NPS 83.2 ± 1.1b 6.0 ± 1.2a 10.8 ± 0.8a
MAT-CMS (DS = 0.32) 63.8 ± 2.9a 16.2 ± 3.2b 20.0 ± 2.0b
MAT-CMS (DS = 0.05) 64.6 ± 3.6a 20.8 ± 3.8b 14.6 ± 2.5b
CT-CMS (DS = 0.30) 63.7 ± 4.2a 15.9 ± 3.8b 20.4 ± 2.7b
CT-CMS (DS = 0.08) 64.5 ± 3.2a 20.1 ± 1.6b 15.4 ± 1.6b
A
Values are expressed as mean ± standard deviation of triplicate analysis.
B
Values bearing different superscript lowercase letters in same column are sig-
nificantly different (P < 0.05) by Tukey’s HSD test.
C
NPS, native potato starch; MAT-CMS, carboxymethyl starch prepared with
microwave-assisted method; CT-CMS, carboxymethyl starch prepared with con-
ventional method; DS, degree of substitution.
der the proper positioning of the substrate into the active site of
a-amylase and also effectively restrict enzyme attack on adjacent
glycosidic bonds of unsubstituted glucose residues (Chung, Lim,
& Lim, 2006; Hoover & Sosulski, 1985). Woo and seib (2002) and
Han and BeMiller (2007) observed that the digestion rate of
cross-linked, hydroxypropylated and acetylated starch was
reduced as the melting enthalpy decreased.
3.7. Hydrolysis kinetics and estimated glycemic index
The in vitro digestion results, including equilibrium concentra-
tion (C1) at 180 min, kinetic constant (k), hydrolysis index (HI),
and estimated glycemic index (GI), are listed in Table 3. The equi-
librium concentration (C1) represented the proportion of the
amount of hydrolysis of starch in 180 min to total starch. The
C1 was founded to be negatively related the DS of the modified
starches, and a higher DS (P0.3) of carboxymethylation brought
about a significantly decreased C1. However, modification meth-
ods (conventional or microwave-assistance) did not have signifi-
cant influence on C1 of modified starches with similar DS. The
kinetic constant (k), which reflected the rate of hydrolysis, was sig-
nificantly higher in the NPS (0.126) than both MAT-CMS and CT-
CMS (0.047–0.062, respectively). The differences in equilibrium
concentration (C1) and kinetic constant (k) indicated that carboxy-
methylation of starch hindered the hydrolysis of starch by porcine
pancreatic a-amylase. However, MAT-CMS and CT-CMS did not
show significant difference in equilibrium concentration (C1) and
kinetic constant (k) at similar DS. Hydrolysis index (HI) is a useful
tool from a nutritional point of view for comparison of starch
digestibility (Sandhu & Lim, 2008). This parameter expresses the
digestibility of the starch in foods in relation to the digestibility
of starch in white bread and is used for the estimation of the gly-
cemic index (GI). The HI of MAT-CMS ranged from 110 to 117
and from 107 to 115 for CT-CMS (Table 3). The GI value of NPS
(109.7) was significant higher than that of MAT-CMS and CT-CMS
(98.7–104.3). Most modified starches such as acetylated starch,
oxidised starch and hydroxypropylated starch have been reported
to have lower HI and GI than native starches (Chung et al., 2008). It
was founded in Table 3 that CT-CMS had lower GI than MAT-CMS
with similar DS, which implied that CT-CMS is rather more resis-
tant to hydrolysis than MAT-CMS. This result may be due to the
differences in substitution position of carboxymethyl groups in
glucose residues between MAT-CMS and CT-CMS. Östergård et al.
(1988) observed that substitution with chemical group occurs at
carbon atom 2 (C2) in the glucose residues is more resistant to
hydrolysis by a-amylase in vitro, than substitution occurs at carbon
atom 6 (C6). The different behave of starch on resistant to hydroly-
sis between CT-CMS and MAT-CMS may also due to the degrada-
tion of starch in MAT-CMS caused by microwave.
3.8. Freeze–thaw stability
The freeze–thaw stability of NPS, MAT-CMS and CT-CMS pastes
are shown in Fig.6. The syneresis values of CMS pastes remained
unchanged during the first two cycles and only slightly increased
in subsequent four cycles. Similar results were reported for car-
boxymethyl starches of mungbean, corn and waxy amaranth
(Bhattacharyya et al., 1995b; Kittipongpatana et al., 2006) In con-
trast, NPS paste showed lack of freeze–thaw stability as its syner-
esis increased sharply from 10.5% to 30.9% and a phase separation
was observed throughout the observed period. The improved
freeze–thaw stability of CMS gels could be ascribed to starch chain
alignment retarded by the incorporation of hydroxyl groups during
storage, resulting in decreased retrogradation.
 J. Liu et al. / Food Chemistry 133 (2012) 1196–1205 1203

Table 3
Equilibrium concentration (C1) of enzymatic hydrolysis at 180 min, kinetic constant (k), hydrolysis index (HI) and estimated glycemic index (GI) for gelatinized native and
carboxymethyl potato starch.A,B

StarchC C1 (%) kD HIE GIF

b b e
NPS 89.8 ± 0.9 0.126 ± 0.012 127.5 ± 0.1 109.7 ± 0.1e
MAT-CMS (DS = 0.32) 81.4 ± 2.9a 0.062 ± 0.011a 110.0 ± 0.2b 100.1 ± 0.1b
MAT-CMS (DS = 0.05) 89.8 ± 2.4b 0.047 ± 0.006a 117.6 ± 0.2d 104.3 ± 0.1d
CT-CMS (DS = 0.30) 80.3 ± 2.9a 0.056 ± 0.009a 107.4 ± 0.2a 98.7 ± 0.1a
CT-CMS (DS = 0.08) 87.1 ± 3.7b 0.051 ± 0.006a 115.3 ± 0.3c 103.0 ± 0.1c
A
Values are expressed as mean ± standard deviation of triplicate analysis.
B
Values bearing different superscript lowercase letters in same column are significantly different (P < 0.05) by Tukey’s HSD test.
C
NPS, native potato starch; MAT-CMS, carboxymethyl starch prepared with microwave-assisted method; CT-CMS, carboxymethyl starch prepared with conventional
method; DS, degree of substitution.
D
k was determined by the equation of C ¼ C 1 ð1  ekt Þ.
E
HI was calculated as the percentage of total glucose released from the sample, to that released from white bread.
F
GI was calculated from equation proposed by Goni et al. (1997).

Fig. 7. Clarity of the native and carboxymethyl potato starch pastes stored at 4 °C.
NPS, native potato starch; MAT-CMS, carboxymethyl potato starch prepared with
microwave-assisted method; CT-CMS, carboxymethyl starch prepared with con-
ventional method.

Fig. 6. Freeze–thaw stability of the native and carboxymethyl potato starch pastes.
NPS, native potato starch; MAT-CMS, carboxymethyl potato starch prepared with
microwave-assisted method; CT-CMS, carboxymethyl starch prepared with con-
ventional method.
3.9. Paste clarity
Paste clarity of NPS, MAT-CMS and CT-CMS pastes as a function
of storage time is shown in Fig.7. The transmittance values
(P85.1%) of CMS pastes were much higher than that of NPS
(42.4–67.2%) and also increased with an increase in DS, indicating
that the higher paste clarity of the carboxymethyl starches was
caused by higher hydroxyl content. The higher paste clarity of
CMS can be explained by the greater swelling caused by the incor-
poration of hydrophilic hydroxyl groups, which dissociate the
starch molecules and ultimately inhibit the association of starch
chains after pasting, resulting in more transmittance of light. The
paste clarity of CMS was stable over storage time at 4 °C while a
significant decrease was observed for NPS.
3.10. Paste viscosity
Viscosity changes as function of shear and temperature of na-
tive and modified starch pastes indicate on complex rheological
behaviour (Fig.8). The NPS paste was highly viscous as compared
to that of CMS indicated that carboxymethylation caused a de-
crease in viscosity of pastes. It is due to degradation of polymer
(partial depolymerization, partial cleavage of glycosic bonds, low-
ered molecular mass) during modification as mentioned previ-
ously. In this context, the lower paste viscosity of MAT-CMS
compared to that of CT-CMS reflected the fact that microwave
caused the degradation of starch. However, the paste viscosity of
CMS increased with an increase in DS could be interpreted by
the presence of negative charges in their molecules. Due to the
repulsion of negatively charged carboxymethyl groups, starch mol-
ecules tended to exist in a more expanded state; thus occupying
more space for gyration and exhibiting higher viscosity
(Sangseethong, Ketsilp, & Klanarong, 2005). Also in cases of acety-
lated and oxidised starch lowered viscosity was observed (Berski
et al., 2011; Lawal, 2004). Viscosity changes as function of shear
of NPS and CMS behave like shear-thickening in which the viscos-
ity decreased with increasing shear rate (Fig. 8A). As seen in
Fig. 8(B), the paste viscosity of NPS decreased dramatically as paste
temperature elevated from 25 to 85 °C, while reasonably stable
paste viscosity was observed for CMS.
It is well known that the paste clarity, freeze–thaw stability and
viscosity of starch and its derivates are very important parameters
for food applications. Improved paste clarity of CMS is a useful
property in the manufacture of some foods like jellies, sausages
and fruit pastes, which require transparency. CMS with high DS
(P0.3), which shows excellent paste clarity and viscosity over tem-
perature varied form 25 to 85 °C, is suitable for use as thickening
agent in milk products, fruit juice, as well as hot beverage. Also,
CMS with high DS can be a better choice for the preparation of
1204 J. Liu et al. / Food Chemistry 133 (2012) 1196–1205

Fig. 8. Paste viscosity of the native and carboxymethyl potato starch as a function of shear rate (A) and temperature (B). NPS, native potato starch; MAT-CMS, carboxymethyl
potato starch prepared with microwave-assisted method; CT-CMS, carboxymethyl starch prepared with conventional method.

low GI food for diabetes patients and nutritional food, because of
its resistance to enzymatic hydrolysis. CMS with low DS (60.08)
can be used as coating agent for fruits and vagetables preservation,
which is relatively non-viscous. In addition, the loss of crystallinity
opens up a potential utilisation of CMS as pregelatinized starch in
instant food. This superior freeze–thaw stability makes CMS suit-
able for use in custards, puddings and pie-fillings which are frozen
stored.
4. Conclusions
This synthesis of microwave assisted carboxymethylation of po-
tato starch and its characterisation showed temperature, ratio of
water to solvent in the reaction medium, amount of MCA as well
as amount of NaOH to influence the DS of the CMS. A reaction time
of 1–25 min in a microwave reactor was sufficient to obtain CMS
with a DS range of 0.06–0.32. For similar DS, microwave assisted
carboxymethylation of potato starch could be carried out in
25 min as compared to 150 min for conventional carboxymethyla-
tion. The in vitro digestibility and estimated glycemic index of
MAT-CMS and CT-CMS were significantly lower. Higher DS gave
a higher content of RS and lowered value of GI by decreasing SDS
content. All CMS samples exhibited better freeze–thaw stability.
The viscosity of CMS increased with DS. The NPS paste was more
viscous as compared to that of CMS, while CMS paste gave a stable
viscosity over the temperature range of 25–85 °C.
Acknowledgements
This research was conducted by financial supported from the
National Spark Program (2010GA811002), and National High-tech
R&D Program (863 Program) of China (2011AA100805). We also
thank the JingTian Agriculture Development Co. Ltd. (Chongqing,
China) for providing the potato starch.
References
Bernfeld, P. (1955). Amylases, a and b. In S. P. Colowick & N. O. Kaplan (Eds.),
Methods in enzymology (pp. 149–158). New York: Academic Press.
Berski, W., Ptaszeka, A., Ptaszeka, P., Ziobroa, R., Kowalskia, G., Grzesika, M., et al.
(2011). Pasting and rheological properties of oat starch and its derivatives.
Carbohydrate Polymers, 2, 665–671.
Bhattacharyya, D., Singhal, R. S., & Kulkarni, P. R. (1995a). A comparative account of
synthesis of sodium carboxymethyl starch prepared from corn and waxy
amaranth starch. Carbohydrate Polymers, 27, 247–253.
 J. Liu et al. / Food Chemistry 133 (2012) 1196–1205
Bhattacharyya, D., Singhal, R. S., & Kulkarni, P. R. (1995b). Physicochemical
properties of carboxymethyl starch prepared from corn and waxy amaranth
starch. Carbohydrate Polymers, 27, 167–169.
Björck, I., Gunnarsson, A., & Östergård, K. (1988). A study of native and chemically
modified potato starch. Part II: Digestibility in the rat intestinal tract. Starch/
Stärke, 41, 128–134.
Chen, J., & Jane, J. (1994). Properties of granular cold-water-soluble starches
prepared by alcoholic-alkaline treatments. Cereal Chemistry, 71, 623–626.
Chung, H. J., Lim, H. S., & Lim, S. T. (2006). Effect of partial gelatinization and
retrogradation on the enzymatic digestion of waxy rice starch. Journal of Cereal
Science, 43, 353–359.
Chung, H. J., Shin, D. H., & Lim, S. T. (2008). In vitro starch digestibility and estimated
glycemic index of chemically modified corn starches. Food Research
International, 41, 579–585.
Craig, S. S., Maningat, C. C., Seib, P. A., & Hoseney, R. C. (1989). Starch paste clarity.
Cereal Chemistry, 66, 173–182.
Englyst, H. N., Kingman, S. M., & Cummings, J. H. (1992). Classification and
measurement of nutritionally important starch fractions. European Journal of
Clinical Nutrition, 46, 30–50.
Goni, I., Garcia-Alonso, A., & Saura-Calixto, F. (1997). A starch hydrolysis procedure
to estimate glycemic index. Nutrition Research, 17, 427–437.
Granfeldt, Y., Björck, I., Drews, A., & Tovar, J. (1992). An in vitro procedure based on
chewing to predict metabolic responses to starch in cereal and legume
products. European Journal of Clinical Nutrition, 46, 649–660.
Han, J. A., & BeMiller, J. N. (2007). Preparation and physical characteristics of slowly
digesting modified food starches. Carbohydrate Polymers, 67, 366–374.
Hebeish, A., & Khalil, M. I. (1988). Chemical factors affecting preparation of
carboxymethyl starch. Starch/Stärke, 50, 147–150.
Hoover, R., Hannouz, D., & Sosulski, F. W. (1988). Effects of hydroxypropylation on
thermal properties, starch digestibility and freeze–thaw stability of field pea
(Pisum sativum cv Trapper). Starch/Stärke, 40, 383–387.
Hoover, R., & Sosulski, F. (1985). A comparative study of the effect of acetylation on
starches of Phaseolus vulgaris biotypes. Starch/Stärke, 37, 397–404.
Jyothi, A. N., Rajasekharan, K. N., Moorthy, S. N., & Sreekumar, J. (2005). Microwave-
assisted synthesis and characterization of succinate derivatives of cassava
(Manihot esculenta Crantz) starch. Starch/Stärke, 57, 556–563.
Khalil, M. I., Hashem, A., & Hebeish, A. (2006). Carboxymethylation of maize starch.
Starch/Stärke, 42, 60–63.
Kittipongpatana, O. S., Sirithunyalug, J., & Laenger, R. (2006). Preparation and
physicochemical properties of sodium carboxymethyl mungbean starches.
Carbohydrate Polymers, 63, 105–112.
Lawal, O. S. (2004). Succinyl and acetyl starch derivatives of a hybrid maize:
Physicochemical characteristics and retrogradation properties monitored by
differential scanning calorimetry. Carbohydrate Research, 16, 2673–2682.
Lawal, O. S., Lechner, M. D., Hartmann, B., & Kulicke, W. M. (2007). Carboxymethyl
cocoyam starch: Synthesis, characterization and influence of reaction
parameters. Starch/Stärke, 59, 224–233.
Lawal, O. S., Lechner, M. D., & Kulicke, W. M. (2008a). Single and multi-step
carboxymethylation of water yam (Dioscorea alata) starch: Synthesis and
characterization. International Journal of Biological Macromolecules, 42, 429–435.
Lawal, O. S., Lechner, M. D., & Kulicke, W. M. (2008b). The synthesis conditions,
characterizations and thermal degradation studies of an etherified starch from
an unconventional source. Polymer Degradation and Stability, 93, 1520–1528.
Lee, S., Kim, S. T., Pant, B. R., Kwen, H. D., Song, H. H., Lee, S. K., et al. (2010).
Carboxymethylation of corn starch and characterization using asymmetrical
flow field-flow fractionation coupled with multiangle light scattering. Journal of
Chromatography A, 1217, 4623–4628.
Lewandowicz, G., Fornal, J., Walkowski, A., MaÎczynÂski, M., Urbaniak, G., &
SzymanÂska, G. (2000). Starch esters obtained by microwave radiation-
structure and functionality. Industrial Crops and Products, 11, 249–257.
1205
Li, K. L., Xia, L. X., Li, J., Pang, J., Cao, G. Y., & Xi, Z. W. (2001). Salt-assisted acid
hydrolysis of starch to D-glucose under microwave irradiation. Carbohydrate
Research, 331, 9–12.
Miao, M., Zhang, T., Mu, W. M., & Jiang, B. (2010). Effect of controlled gelatinization
in excess water on digestibility of waxy maize starch. Food Chemistry, 119,
41–48.
Noda, T., Takigawa, S., Matsuura-Endo, C., Suzuki, T., Hashimoto, N., Kottearachchi,
N. S., et al. (2008). Factors affecting the digestibility of raw and gelatinized
potato starches. Food Chemistry, 110, 465–470.
Ornanong, S. K., Jakkapan, S., & Reinhard, L. (2006). Preparation and
physicochemical properties of sodium carboxymethyl mungbean starches.
Carbohydrate Polymers, 63, 105–112.
Östergård, K., Björck, I., & Gunnarsson, A. (1988). A study of native and chemically
modified potato starch. Part I: Analysis and enzymic availability in vitro. Starch/
Stärke, 40, 58–66.
Qiu, H. Y., & He, L. M. (1999). Synthesis and properties study of carboxymethyl
cassava starch. Polymers for Advanced Technologies, 10, 468–472.
Sandhu, K. S., & Lim, S. T. (2008). Structural characteristics and in vitro digestibility
of mango kernel starches (Mangifera indica L.). Food Chemistry, 107, 92–97.
Sangseethong, K., Ketsilp, S., & Klanarong, S. (2005). The role of reaction parameters
on the preparation and properties of carboxymethyl cassava starch. Starch/
Stärke, 57, 84–93.
Shogren, R. L., & Biswas, A. (2006). Preparation of water-soluble and water-
swellable starch acetates using microwave heating. Carbohydrate Polymers, 64,
16–21.
Singh, V., & Tiwari, A. (2008). Microwave-accelerated methylation of starch.
Carbohydrate Research, 343, 151–154.
Stojanovic, Z., Jeremic, K., & Jovanovic, S. (2000). Synthesis of carboxymethyl starch.
Starch/Stärke, 52, 413–419.
Stojanovic, Z., Jeremic, K., Jovanovic, S., & Lechner, M. D. (2005). A Comparison of
some methods for the determination of the degree of substitution of
carboxymethyl starch. Starch/Stärke, 57, 79–83.
Tester, R. F., Karkalas, J., & Qi, X. (2004). Starch structure and digestibility enzyme–
substrate relationship. World’s Poultry Science Journal, 60, 186–195.
Tijsen, C. J., Kolk, H. J., Stamhuis, E. J., & Beenackers, A. A. C. M. (1999). Optimisation
of the process conditions for the modification of starch. Chemical Engineering
Science, 54, 2765–2772.
Tijsen, C. J., Kolk, H. J., Stamhuis, E. J., & Beenackers, A. A. C. M. (2001a). An
experimental study on the carboxymethylation of granular potato starch in
non-aqueous media. Carbohydrate Polymers, 45, 219–226.
Tijsen, C. J., Kolk, H. J., Stamhuis, E. J., & Beenackers, A. A. C. M. (2001b). Design of a
continuous process for the production of highly substituted granular
carboxymethyl starch. Chemical Engineering Science, 56, 411–418.
Wang, Y. L., Gao, W. Y., & Li, X. (2009). Carboxymethyl Chinese yam starch:
Synthesis, characterization, and influence of reaction parameters. Carbohydrate
Research, 344, 1764–1769.
Wang, L. F., Pan, S. Y., Hu, H., Miao, W. H., & Xu, X. Y. (2010). Synthesis and
properties of carboxymethyl kudzu root starch. Carbohydrate Polymers, 80,
174–179.
Wolf, B. W., Bauer, L. L., & Fahey, G. C. (1999). Effects of chemical modification on
in vitro rate and extent of food starch digestion: An attempt to discover a slowly
digested starch. Journal of Agriculture and Food Chemistry, 47, 4178–4183.
Woo, K. S., & Seib, P. A. (2002). Cross-linked resistant starch: Preparation and
properties. Cereal Chemistry, 79, 819–825.
Wootton, M., & Chaudhry, M. A. (1981). In vitro digestion of hydroxypropyl
derivatives of wheat starch. Starch/Stärke, 33, 135–137.
Zhang, L. M., Yang, X., Li, S., & Gao, W. Y. (2011). Preparation, physicochemical
characterization and in vitro digestibility on solid complex of maize starches
with quercetin. LWT – Food Science and Technology, 44, 787–792.