292 DOI 10.1002/star.

200500468 Starch/Stärke 58 (2006) 292–299

Alummoottil N. Jyothi Effect of Cross-linking with Epichlorohydrin on the

Subramoney N. Moorthy
Kallikat N. Rajasekharan
Division of Crop Utilization,
Central Tuber Crops
Research Institute,
Sreekariyam,
Thiruvananthapuram,
Kerala, India
Properties of Cassava (Manihot esculenta Crantz)
Starch
Cassava starch was cross-linked with epichlorohydrin (EPI) at 457C for 2 h in three
different media which include water, water in the presence of a phase transfer catalyst
(PTC) and N,N-dimethylformamide (DMF). The products were characterized by deter-
mining their physicochemical, thermal and retrogradation properties. In aqueous me-
dium, the use of a PTC, tetrabutylammonium bromide (TBAB) produced derivatives
with higher degree of cross-linking than those prepared without the use of the catalyst.
The degree of cross-linking was found to be higher using the same concentration of
EPI when the reaction was carried out in DMF. At low levels of cross-linking, the peak
viscosity of the cross-linked starches increased in comparison to that of the native
starch. With increasing degree of cross-linking, the peak viscosity showed a significant
reduction. The swelling volume, solubility and light transmittance of the starch pastes
were lower for the modified starches. The cross-linked starches showed slightly
reduced values for the gelatinization temperatures, Tonset, Tpeak and Tend. The enthalpy
of gelatinization of the modified starches increased with increase in the degree of
cross-linking. The modified starches exhibited higher water-binding capacities (WBC)
than the native starch; but with increase in the degree of cross-linking, there was a
gradual decrease in WBC. The in vitro alpha amylase digestibility of the modified
starches decreased gradually with increase in the level of cross-linking.

Keywords: Cassava starch; Cross-linking; Epichlorohydrin; Physicochemical proper-

ties; Thermal properties; Enzyme digestibility; Retrogradation

1 Introduction bonds in the granules with new covalent bonds. As a result,

Cassava starch has many remarkable properties such as
high paste viscosity, high paste clarity and freeze-thaw
stability which are advantageous to many industries. The
native starch with high purity can readily be modified by
physical, chemical or enzyme processes to afford prod-
ucts with improved starch functionality and consequently
Research Paper
cross-linked starch is more resistant to acid, heat and
shearing than native starch and is thus suitable for applica-
tions such as in canned food [3]. The starch-related literature
contains several patents covering the use of a variety of bi-
or polyfunctional chemicals for starch cross-linking. The
most widely used cross-linking reagents include adipic and
acetic mixed anhydrides, phosphorus oxychloride, sodium

these products find diverse applications [1]. trimetaphosphate and epichlorohydrin (EPI). Cross-linking
of starch with EPI is the most common method used in
Chemical modification of starch is being used to introduce
polysaccharide chemistry [4]. Reaction of starch with EPI
desirable properties to starch for specific applications.
gives distarch glycerols [5, 19]. The rate and extent of reac-
Cross-linking and substitution of starch are two such meth-
tion with EPI was studied by Hamerstrand et al. by deter-
ods utilized in food industry. Cross-linked starches con-
mining the quantity of unreacted EPI [5]. The theoretical
stitute a major class of modified starches. Cross-linking of
number of anhydroglucose units per cross-link was found to
starch by reaction with bi- or polyfunctional reagents is
vary from 50 to 2360. Rutledge et al. cross-linked rice with
widely used in numerous industrial applications such as
preparation of wet-rub-resistant starch paper coatings, EPI and developed a rice suitable for canning [6]. Kuniak and
Marchessault reported that depending upon conditions of
permanent textile sizes and water-resistant adhesives [2].
time, temperature and mole ratio of the reactants, 5 – 25% of
Cross-linking reinforces the already present hydrogen
the applied EPI is bound in the starch as a glycerol mono-
ether substituent [4]. Distarch glycerols are highly resistant
Correspondence: Alummoottil N. Jyothi, Division of Crop Utili-
to pH and mechanical shear due to the formation of ether
zation, Central Tuber Crops Research Institute, Sreekariyam,
Thiruvananthapuram – 695 017, Kerala, India. E-mail: linkages between the cross-links and the hydroxyl groups in
sreejyothi_in@yahoo.com. the starch. The granules of root and tuber starches become

© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.starch-journal.com

Starch/Stärke 58 (2006) 292–299 Effect of Epichlorohydrin Cross-linking on Properties of Cassava Starch
more highly hydrated upon heating in hot water and develop
higher peak viscosities than that of cereal starches. How-
ever, on prolonged heating, their viscosities drop sharply
and cause marked viscosity breakdown. It is also possible to
minimize or prevent granule rupture and loss in viscosity by
cross-linking [2]. Literature available on the cross-linking
reaction of cassava starch with EPI is sparse and hence an
attempt has been made to study systematically the mod-
ification of cassava starch with different concentrations of
EPI in aqueous as well as non-aqueous media and charac-
terization of the products. Another objective was to investi-
gate the effectiveness of a PTC in the cross-linking reaction
of cassava starch under heterogeneous conditions.
2 Materials and Methods
Cassava starch was obtained from Sagoserve (Salem,
Tamilnadu, India). Epichlorohydrin was purchased from
Sisco Research Laboratories (Mumbai, India). All the
other chemicals used were of analytical grade.
2.1 Synthesis of cross-linked starches
The cross-linking reaction was carried out in water, water
in the presence of the phase transfer catalyst, tetra-
butylammonium bromide (TBAB) and DMF. Cassava
starch (20 g) was slurried by adding distilled water or DMF
(40 mL) so as to obtain a solid concentration of 50% (w/v)
and the paste was stirred mechanically. The pH of the
slurry was adjusted to 10.5 using dilute sodium hydroxide
solution (0.1 M) and EPI was then added to give 0.25,
0.50, 0.75 and 1.0% (0.042, 0.084, 0.126 and 0.168 mL,
respectively) final concentrations based on the starch dry
weight. The flasks were stoppered well and the contents
were stirred on a magnetic stirrer at 457C for 2 h. Upon
completion of the reaction, the pH of the slurry was
adjusted to 6.0 using 0.1 M HCl, filtered and the product
was washed several times (56100 mL) with distilled
water. The slurry was then filtered by suction and the
product was dried overnight in an air oven at 457C. In one
set of experiments, TBAB (10% based on starch dry
weight) was also added before the addition of EPI fol-
lowed by the usual steps. The products obtained in these
experiments have been labeled S1 to S12.
2.2 Analysis of the products
2.2.1 FTIR spectroscopy
The FTIR spectra of the starch samples were taken in an
ABB FTIR instrument (Model FTLA 2000, ABB Ltd, Zürich,
Switzerland) using KBr pellets.
© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
293
2.2.2 Degree of cross-linking
The degree of cross-linking of the modified starches was
determined from the viscosity values, according to the
method of Chatakanonda et al. [7]. The peak viscosity of
the starch samples were recorded using a Rapid Visco
Analyser (RVA-4, Newport Scientific, Warriewood, NSW,
Australia). The starch slurry (10% by weight) was heated
from 50 to 957C at 127C/min at 160 rpm, and was then
held at 957C for 2 min. Afterwards the paste was cooled to
507C at 127C/min and finally kept at 507C for 2 min. The
degree of cross-linking was calculated as follows [7]:
½A
B
Degree of cross-linking =  100
A
where, A is the peak viscosity in RV units of the native
starch and B is that of the cross-linked starch.
2.2.3 Determination of physicochemical
properties
The pasting properties of the native and modified starches
were determined using RVA as described in Section 2.2.1.
The peak, breakdown, final and set back viscosities and
pasting temperature were recorded. The breakdown ratio
was calculated as the ratio of breakdown to peak viscosity.
The swelling volume and solubility in water at 907C and
paste clarity of the native and cross-linked starches were
determined according to standard procedures [8, 9, 10].
The water-binding capacity was determined by the
method described by Yamazaki [11] and modified by
Medcalf and Gilles [12] as follows: a suspension of starch
(5 g) in distilled water (75 mL) was agitated for 1 h and
centrifuged (30006g) for 10 min. The water layer was
decanted from the wet starch, which was drained for 10
min and the wet starch was weighed. The water-binding
capacity (WBC) was calculated as percentage, from the
weight of the water bound by the starch sample.
2.2.4 Thermal properties
The gelatinization properties were determined using a Met-
tler Toledo DSC 822e instrument (Mettler-Toledo, Grei-
fensee, Switzerland). The starch sample (5 mg) was
weighed out into a preweighed aluminum pan and distilled
water (10 mL) was added. The pan was sealed and trans-
fered to the heating pan of the DSC. An empty pan was used
as reference and the sample was heated from 20 to 1007C at
a rate 107C/min and cooled back to 207C at the same rate.
Gelatinization properties viz., onset gelatinization tempera-
ture (To), the temperature at peak maximum (Tp), offset or
end temperature (Te) and enthalpy of gelatinization (DH)
were recorded by means of the built in software.
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294 A. N. Jyothi et al. Starch/Stärke 58 (2006) 292–299

2.2.5 In vitro enzyme digestibility Tab. 1. Degree of cross-linking of cassava starch mod-
ified with epichlorohydrin.
The in vitro a-amylase digestibility of the native and mod-
ified starches was determined using porcine pancreatic Sample Conc. of Medium Degree of
amylase with a digestibility unit of 500,000 (13.4 units EPI [%] cross-linking
amylase/ mg solids at pH 6–9, Sigma, St. Louis, USA)
S1 0.25 water ND#
according to the method of Singh et al., [13], modified by S2 0.50 water ND
Padmaja (personnel communication). The detailed pro- S3 0.75 water 35.960.81e
cedure has been described in a previous paper [14]. S4 1.0 water 82.460.89c
S5 0.25 water 1TBAB* ND
S6 0.50 water 1TBAB ND
2.2.6 Retrogradation (% syneresis) S7 0.75 water 1TBAB 82.460.06c
S8 1.0 water 1TBAB 95.360.08b
Starch suspension (2%, w/v) was heated at 85–907C for S9 0.25 DMF{ 9.361.45f
30 min in a water bath followed by rapid cooling in an ice S10 0.50 DMF 42.560.33d
bath to room temperature. The starch paste was stored for S11 0.75 DMF 95.660.20b
24, 48, 72, 96 and 102 days at 47C. Syneresis of the starch S12 1.0 DMF 99.660.02a
paste was measured as the percentage amount of water
released after centrifugation at 27006g for 15 min [15]. * Tetrabutylammonium bromide. { Dimethylformamide.
#
Not detectable.

2.2.7 Statistical analysis

All the determinations were done in triplicate and the data
were analyzed statistically using the package of SAS 8.01
to perform ANOVA and Duncan Multiple Range Test
(DMRT). The mean values are presented in tables with the
ranking. Simple correlation was done to study the corre-
lation between the degree of cross-linking and the physi-
cochemical properties of the modified starches.
3 Results and Discussion
of cross-linking of theses starches was not great enough
to restrict the swelling of the granules and these exhibited
higher peak viscosity values than that of the native starch.
However, an increase in the amount of EPI used to 0.75
and 1% reagent resulted in higher degree of cross-linking.
Reactions carried out in DMF even with low levels of EPI,
resulted in products with reduced swelling and viscosity
due to higher levels of cross-links. There was a gradual
increase in the degree of cross-linking with the increase in
EPI concentration used. For example, at a 1% reagent
level, nearly 99% cross-linking was observed in DMF.

In our studies, since we have used a heterogeneous condi-

3.1 FTIR spectroscopy
The IR spectra of the modified starches showed the typical
peaks for the starch backbone. However, the O-H stretching
band in the region 3600–3000 cm21 was found to be
broadened and became less sharp in the spectra of the
modified starches, in comparison to that of the native starch.
In addition, in the IR spectra of the cross-linked starches, the
characteristic absorption bands of starch at 1020 cm21,
1080 cm21 and 1150 cm21, which are due to C-O stretching
vibrations, were highly diminished in intensity.
tion and the ratio of starch to EPI was high, the expected
product was a distarch glycerol. According to earlier litera-
ture, the reaction of EPI with starch in presence of alkali led
to the formation of glycerol mono and diethers [5, 16, 17].
Roberts [16] suggested a three-stage mechanism for the
reaction. Based on kinetic evidences, Kartha and Srivastava
[16] also proposed a three-step mechanism as follows:

3.2 Degree of cross-linking

The degree of cross-linking of the modified starches was

determined from the RVA viscosity values and the mean
values are presented in Tab. 1. It has been observed that
at lower levels of EPI (0.25 and 0.5%), the degree of
cross-linking was not detectable by the viscosity method
used, for the modified starches prepared in aqueous me-
dium either alone or in the presence of TBAB. The degree
In the case of reactions under homogeneous conditions,
it was observed that the intermediate (i) can undergo side
reaction via opening the epoxide ring by OH2 to form the
monostarch glycerol (iii).

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Starch/Stärke 58 (2006) 292–299 Effect of Epichlorohydrin Cross-linking on Properties of Cassava Starch 295

Tab. 2. Viscosity (RVA) characteristics of native and

cross-linked cassava starches.

Sample Peak Break- Final Setback Pasting

viscosity down viscosity viscosity temp.
[mPa s] ratio [mPa s] [mPa s] [7C]
However, in heterogeneous reactions, where a higher
S1 4302.0c 0.227bcde 6059.5a 2735.0ab 70.1f
ratio of starch to EPI was employed, the distarch glycerol
S2 4264.5c 0.366b 4955.0c 2244.0c 70.6ef
(ii) is the major product [17]. S3 1864.5f 0.149de 3105.5e 1531.0d 72.3de
S4 508.0h 0.098de 958.5g 500.5f 74.7bc
Most of the chemical modifications of starch have been
S5 4608.5a 0.344bc 6190.5a 2915.5a 71.0ef
carried out in heterogeneous systems and often the yields
S6 4461.5b 0.238bcde 5677.0b 2278.5c 73.0cd
are poor. By using PTC such as quaternary ammonium or S7 507.0h 0.091e 917.5g 456.5fg 75.9b
phosphonium salts, it should be possible not only to S8 137.0i 0.124de 294.0h 174.0fgh 80.0a
improve the yield of the desired products of reactions of S9 2612.0e 0.275bcd 4312.0d 2414.5bc 71.1ef
starch with reagents like acetyl chloride, benzoyl chloride, S10 1657.0g 0.203bcde 2977.0e 1656.5d 70.3ef
stearoyl chloride, benzyl chloride, methyl chloride and S11 126.5i 0.155de 181.0h 74.5gh NR{
propylene oxide, but also it enables the reaction of starch S12 11.5j 0.177cde 1.0h 1.5h NR{
with hydrophobic reagents [18–20]. In the present study, Native 2880.5d 0.668a 1938.5f 981.0e 69.1f
starch
the presence of TBAB in the aqueous medium, favored
the cross-linking reaction of cassava starch with EPI and
* Means with the same superscripts are not significantly
the products thus obtained were seen to possess a higher
different.
level of cross-linking than those prepared in the absence {
Not recorded.
of the catalyst. According to the mechanism proposed by
Kartha and Srivastava [17], the reaction of starch with
epichlorohydrin in the alkaline heterogeneous medium was observed even at much lower concentrations of the
occurs through the formation of the anion StO2. Thus the reagent, which had led to a further lowered viscosity.
key step in the cross-linking reaction is the formation of Strengthening the bonding between starch chains
the anion on the starch granules in the slurry. We suggest through cross-links might have increased the resistance
that this step was facilitated by the presence of the phase of the granules to swelling leading to lower peak viscosity.
transfer catalyst in its hydroxide form at the interface be- Fig. 1 shows the RVA pasting profiles of the native and
tween the starch granules and the bulk reaction medium. cross-linked starches. The samples S4, S8 and S11
Daly et al. [21] have suggested that in the alkylation of showed no peaks in the RVA viscosity patterns, due to a
cellulose, tetramethylammonium chloride acts as a cata- higher level of cross-linking and consequent inhibition of
lyst by virtue of the formation of a salt saturated additional the swelling of the granules. Introducing low degrees of
‘microphase’ at the biopolymer surface which provides a cross-linking in starch has been reported to increase the
favorable environment for the reaction. In the reaction of peak and cold paste viscosities of starch [17]. According
starch with epichlorohydrin in presence of TBAB, a similar to Wurzburg [2] corn starch containing lower levels of
effect could be expected, which increases the ease of cross-links exhibited a somewhat higher viscosity com-
cross-linking reaction. pared to that of the native starch and the viscosity con-
tinued to increase on prolonged heating. However, at
higher levels of cross-links (at 1 cross-link per 440 anhy-
3.3 Viscosity characteristics droglucose units), there were sufficient cross-links to in-
hibit the swelling of the starch and there was a decrease in
The RVA viscosity characteristics of native and cross- viscosity. A decrease in the peak viscosity of cross-linked
linked starches are shown in Tab. 2. As discussed earlier, potato starches with increase in the concentration of epi-
the starches cross-linked with 0.25 and 0.5% EPI in the chlorohydrin was reported [22].
aqueous medium showed an increase in the peak vis-
cosity. However, at higher reagent concentrations of EPI The fall in viscosity breakdown of the modified starches
(0.75 and 1%), a drastic fall in peak viscosity was could be attributed to the formation of cross-links be-
observed. Further, such a decrease in peak viscosity was tween starch molecules which strengthens the swollen
more evident in samples prepared in presence of TBAB. granules against breakage under conditions of high tem-
The cross-linked starches prepared in DMF showed a perature and shear. With cross-linked starches, the gran-
gradual decrease in viscosity with increase in the con- ule may swell as the hydrogen bonds are weakened when
centration of EPI. Here, a higher degree of cross-linking continuously heated in water. However, after they rupture,

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296 A. N. Jyothi et al.
the covalent cross-links may provide sufficient granule
integrity to keep the swollen granules intact and minimize or
prevent loss in viscosity [2]. At lower levels of cross-linking
the setback viscosity was found to be higher. This could be
attributed to the observed low peak viscosity values. How-
ever, with an increase in the degree of cross-linking the set-
back viscosity also decreased. The final viscosities of most
of the cross-linked starches were observed to be signifi-
cantly higher than that of the native starch. However, sam-
ples S4, S7, S8, S11 and S12 showed lower final viscosities,
which could again be correlated to the observed peak vis-
cosity values for these samples. The pasting temperatures
of the cross-linked starches were significantly increased in
comparison to that of the native starch. Cross-linking of
molecular chains makes the starch granules more ordered
and consequently more energy would be required for swel-
ling [17]. With increase in the level of cross-linking the gela-
tinization temperature also increased. In the case of samples
S11 and S12 no pasting temperature was recorded using RVA.
3.4 Swelling volume and solubility
The swelling volumes of the native and cross-linked
starches are shown in Tab. 3. The cross-linked starches
showed lower swelling volumes in comparison to the
© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Starch/Stärke 58 (2006) 292–299
Fig 1. RVA pasting profiles of native
and cross-linked cassava starches.
native starch. A significant positive correlation between the
degree of cross-linking and swelling volume was seen. The
use of a higher concentration of the EPI, which resulted in
more cross-links, thereby imposed constraints on the
swelling behavior of the modified starches. The results are
also consistent with earlier reports [23, 24]. Liu et al. had
reported a considerable decrease in the swelling power of
cross-linked waxy rice starch [24]. According to Kartha and
Srivastava [17] with increase in the degree of cross-linking,
the granular structure of starch becomes more compact
and swelling and solubility are reduced.
All the cross-linked starches exhibited lower solubilities in
water at 907C than the native starch (Tab. 3). The solubility
decreased with an increase in reagent concentration,
which could be attributed to the increase in the degree of
cross-linking and is also consistent with the swelling
properties observed. The solubility of the native starch
was 20.2% and those of the modified starches in the
three sets of experiments were 3.2–14.4, 5.7–12.4 and
7.7–13.7%, respectively. Due to the strengthening of the
starch granules through cross-links, less disintegration
takes place during gelatinization and there is lowering of
solubles. Cross-linking reduces the solubles which leach
out of the swollen granules on cooking and the extent of
solubilization is affected by the degree of cross-linking;
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Starch/Stärke 58 (2006) 292–299 Effect of Epichlorohydrin Cross-linking on Properties of Cassava Starch 297

Tab. 3. The physicochemical properties and in vitro en- transmittance compared to the native starch. The starch
zyme digestibility of native and cross-linked cas- pastes treated with 1% EPI showed the lowest paste
sava starches. clarity. This could be explained on the basis of differences
in the swelling behavior. Granules held together by cross-
Sample Swelling Solubility Light Water Digesti-
links showed reduced swelling and the more dense gran-
vol. [%] transmit- binding bility [%]
[mL/g] tance [%] capacity [%] ules reflect light rather than transmit it, resulting in a lower
clarity [27–29]. The native starch granules had completely
S1 18.5b* 14.4a 8.9b 90.4c 74.7c disintegrated after gelatinization, whereas cross-linked
S2 15.5cd 10.9ef 6.1c 88.8c 68.8cd starches remained almost intact, thus reducing clarity [30].
S3 12.5efg 6.6gh 3.9ef 88.5c 58.3ef
S4 10.0gh 3.2i 3.6f 84.0d 54.7fg
S5 15.0cde 12.4cd 5.8c 98.7b 101.7a 3.7 In vitro enzyme digestibility
S6 14.0de 10.1f 4.4d 94.4b 97.7ab
S7 9.5h 6.0h 2.4hi 83.3d 71.4cd The modified starches with low degree of substitution exhib-
S8 9.0 h 5.7h 2.0i 75.8e 64.9de ited slightly higher digestibilities when compared to native
S9 17.0bc 13.7bc 4.3de 103.4a 90.3b
starch. However, there was a gradual decrease in the enzyme
S10 13.5def 13.7bc 4.1de 94.1b 73.1cd
susceptibility of cross-linked starches with increase in
S11 11.0fgh 11.9de 3.2g 84.3d 47.4g
S12 8.8h 7.7g 2.5h 84.5d 33.0h degree of cross-linking (Tab. 3). The samples S11 and S12
Native 27.0a 20.2a 22.6a 68.3f 72.6cd showed very low enzyme digestibilities (47.4 and 33%,
starch respectively). The decrease in enzyme susceptibility of
cross-linked starches could be attributed to the stabilization
* Means with the same superscripts are not significantly of starch granules through cross-links which inhibits the
different. action of a-amylase. Cross-linking within the level approved
for modified food starches is reported to have no significant
effect on the in vitro digestibility of the starch [2]. Walker and
the fewer the cross-links the higher would be the solubles
ElHarith have shown that cross-linking has no significant
[2]. According to earlier reports also, the cross-linked
effect on the digestibility of distarch phosphates [31].
starches exhibited a lower solubility than the native starch
According to Huber and BeMiller, the cross-linking of starch
[22, 24]. According to Liu et al., who observed lowering of
restricts the entrance of a-amylase through the channels that
solubility of cross-linked waxy rice starches, cross-linking
lead to the interior of cereal starches and therefore there was
restricts swelling of granules and will also lower solubility
a decrease in the enzyme digestibility [32]. The cross-linking
by increasing chain binding [24].
interferes with the formation of the a-amylase and starch
complex [33]. Because cross-linking of starch stabilizes the
3.5 Water-binding capacity granule structure and restricts swelling, highly cross-linked
starch is very difficult to hydrolyze by a-amylase [34].
The water-binding capacities of the native and cross-
linked starches are presented in Tab. 3. The modified
starches with higher degree of cross-linking exhibited a 3.8 Thermal properties
decrease in the WBC. However, in comparison to native
The DSC gelatinization parameters of the native and
starch (68.3%), the cross-linked starches exhibited higher
cross-linked cassava starches are shown in Tab. 4. All the
WBC values. On the other hand, Yook et al. has reported a
samples exhibited a decrease in gelatinization tempera-
decrease in the water uptake of rice starch upon cross-
tures, To, Tp and Te, except the sample S12, with the highest
linking with EPI [25]. Kaur observed a considerable low-
degree of cross-linking, for which the gelatinization tem-
ering of WBC of cross-linked potato starches in compar-
peratures were almost same as those of the native starch
ison to the native counterparts [22]. Choi and Kerr
(Tab. 4). However, in each set of the experiment, the
reported that cross-linking at low concentration of phos-
sample with higher degree of cross-linking exhibited
phorus oxychloride has little effect on the dynamic water
higher To and Tp values than those with lower degree of
properties of wheat starch [26].
cross-linking. Increased cross-linking caused a decrease
in Te for the modified starches except that the sample S12
3.6 Light transmittance (% T) had somewhat higher value than S11. The heat of gelati-
nization was increased with increase in the degree of
The light transmittance of the cross-linked starch disper- cross-linking. In comparison to native starch, all the
sion was lower than that of the native starch (Tab. 3). The modified starches, except S1 showed higher DH values.
higher the concentration of EPI used, the lower was the Heat of gelatinization reflects the energy required for dis-

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298 A. N. Jyothi et al. Starch/Stärke 58 (2006) 292–299

Tab. 4. Thermal properties (DSC) of native and cross-

linked cassava starches.

Sample To [7C] Tp [7C] Te [7C] Gelatinization

enthalpy [J/g]

S1 64.960.25c * 70.760.33c 79.060.04b 9.460.83c

S4 65.960.35b 71.760.38b 78.660.47c 10.460.34ab
S5 65.960.11b 71.460.28b 79.160.13ab 11.060.17ab
S8 66.560.42b 71.860.47b 78.360.88c 11.160.01a
S11 65.460.09b 71.560.04b 78.960.03b 10.260.08bc
S12 67.360.11a 72.760.08a 79.260.07ab 11.160.01a
Native 66.860.01a 72.460.02a 79.660.21a 10.260.22ab
starch
* Means with the same superscripts did not differ sig-
nificantly.
rupting the starch granule structure and since cross-linking
reinforces the starch granules, more heat will be required
for gelatinization [24, 34]. Liu et al. reported that there was
a slight decrease in the gelatinization temperatures and the
heat of gelatinization in the case of cross-linked normal rice
starch [24]. However, they have observed that the effect of
cross-linking on the thermal parameters differ among waxy
and non-waxy rice starches [24]. Cross-linking increased
the onset gelatinization temperature and DH of waxy rice
starch, but Tp and Tc (conclusion temperature) were slightly
lowered. Yeh and Yeh also reported that cross-linking
increased the heat of gelatinization for rice starch [34].
However, according to Yook et al. the cross-linked rices
exhibited a lower value of heat of gelatinization than native
rice because cross-linking reduced the portion of starch
granules that could be gelatinized [25]. The cross-linked
wheat starches (distarch phosphate and aluminum octenyl
succinate) were reported to have virtually unchanged To
and Te values compared to unmodified starch but Tp was
slightly increased [35]. In contrast, these increases in Tp
were observed to be small, which may be due to an
increase in the rigidity of the cross-linked starches [35].
Chatakanonda et al. observed no effect of cross-linking on
the glass transition temperature of starch gels [7]. On the
basis of DSC studies, Woo and Seib reported that the
cross-linked starches showed higher gelatinization tem-
peratures and lower enthalpies than their parent starches
[33]. Therefore, according to earlier reports, the effect of
cross-linking on gelatinization properties has been found
to depend on the nature of the starch source.
3.9 Retrogradation properties
The retrogradation of the native and cross-linked starch
gels increased with increase in storage duration at 47C
(Fig. 2). The cross-linked starches S4 and S8 show more
Fig 2. Effect of storage duration (at 47C) on the syneresis
(%) of native and cross-linked cassava starch pastes.
pronounced syneresis than the native starch. Cross-link-
ing results in ordered structure in the starch pastes, thus
resulting in a higher degree of retrogradation. The retro-
gradation properties of starches are indirectly influenced
by the structural arrangement of starch chains, which in
turn, influence the extent of granule breakdown during
gelatinization and the interaction that occur between
starch chains during gel storage [36]. The restricted mo-
bility of the cross-linked amylopectin branches could
have retarded the re-association of the starch chains and
inhibited the recrystallization [37]. Liu et al. observed a
decrease in the retrogradation of non-waxy cross-linked
rice starch, but an increase in waxy starch [24].
4 Conclusions
Reaction of cassava starch with epichlorohydrin in different
reaction media produced a range of derivatives with dif-
ferent levels of cross-linking. Cross-linking in presence of
DMF resulted in products with higher degree of cross-link-
ing than either in water alone or in water in the presence of
TBAB. However, in aqueous medium, the use of TBAB, the
phase transfer catalyst, was found to produce derivatives
with higher degree of cross-linking than those prepared
without the catalyst. Cross-linking has been found to be a
technique for modifying the properties of cassava starch
which offers a means for reinforcing the granules to the
extent where intact granules can be used as such under
conditions which would swell granules of the native starch.
Such applications include usages as surgical dusting
powders, carriers, absorbents and ion exchange resins.
References
[1] K. Sriroth, K. Piyachomwan, K. Sangseethong, C. Oates:
Modification of Cassava starch. Paper presented at X Inter-
national Starch Convention, Cracow, Poland, 11–14 June
2002.
[2] O. B. Wurzburg: Cross-linked starches, in Modified Starches:
Properties and Uses (Ed O.B. Wurzburg) CRC Press,, Florida
1986, 42–51.

© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.starch-journal.com

Starch/Stärke 58 (2006) 292–299 Effect of Epichlorohydrin Cross-linking on Properties of Cassava Starch
[3] R. C. Hoseney: Starch, in Principles of Cereal Science and
Technology, 2nd ed. Am. Assoc. Cereal Chem. Inc., St. Paul,
Minnesota, 1994, 54–57.
[4] L. Kuniak, R. H. Marchessault: Study of the cross-linking
reaction between epichlorohydrin and starch. Stärke 1972,
24, 110–116.
[5] G. E. Hamerstrand, B. T. Hofreiter, C. L. Mehltretter: Deter-
mination of extent of reaction between epichlorohydrin and
starch. Cereal Chem. 1960, 37, 519.
[6] J. E. Rutledge, M. N. Islam, W. H. James: Improved canning
stability of rice by chemical modification. Cereal Chem.
1972, 49, 430–436.
[7] P. Chatakanonda, S. Varavinit, P. Chinachoti: Relationship of
gelatinization and recrystallization of cross-linked rice to
glass transition temperature. Cereal. Chem. 2000, 77, 315–
319.
[8] G. B. Crosbie: The relationship between starch swelling
properties, paste viscosity and boiled noodle quality in
wheat flours. J. Cereal Sci. 1991, 13, 145–150.
[9] T. J. Schoch: Swelling power and solubility of granular
starches, in Methods in Carbohydrate Chemistry (Eds. R. L.
Whistler, J. R. Smith, M. L. Wolform) Academic Press, New
York, 1964.
[10] L. A. Bello-Perez, O. Paredes-Lopez: Starch and amylo-
pectin- Effects of solutes on clarity of pastes. Starch/Stärke
1996, 48(6), 205–207.
[11] W. T. Yamazaki: An alkaline water retention capacity test for
the evaluation of cookie baking potentialities of soft winter
wheat flour. Cereal Chem. 1953, 30, 242–246.
[12] D. G. Medcalf, K. A. Gilles: Wheat starches. 1. Comparison
of physicochemical properties. Cereal Chem. 1965, 42,
558–568.
[13] U. Singh, M. S. Kherdekar, R. Jambunathan: Studies on Desi
and Kabuli chickpea (Cicer arietinum L) cultivars: The levels
of amylase inhibitors, levels of oligosaccharides and in- vitro
starch digestibility. J. Food. Sci. 1982, 47, 510–512.
[14] A. N. Jyothi, K. N. Rajasekharan, S. N. Moorthy, J. Sreeku-
mar: Synthesis and characterization of low DS succinate
derivatives of cassava (Manihot esculenta Crantz) starch.
Starch/Stärke 2005, 57(7), 319–324.
[15] L. Kaur: Physicochemical properties of potatoes in relation
to thermal and functional properties of their starches. PhD
Thesis, Guru Nanak Dev University, Punjab, India, 2004, 59.
[16] H. T. Roberts, in Starch Chemistry and Technology (Ed. R. L.
Whistler) Academic Press, New York, 1965, 482.
[17] K. P. R. Kartha, H. C. Srivastava: Reaction of epichlorohydrin
with carbohydrate polymers. Starch/Stärke 1985, 37, 297–
306.
[18] C. M. Starks, C. L. Liotta, in Phase Transfer Catalysis: Prin-
ciples and Techniques. Academic Press, New York, 1978.
[19] J. Berry, G. Gram, G. Decodts, A. Loupy, P. Pigeon, J. San-
soulet: Tetrahedron Lett. 1982, 23(51), 5407.
[20] H. C. Srivasthava: Starch research: The Indian perspective,
in New Developments in Carbohydrates and Related Natural
Products (Eds. M. J. Mulky, A. Pandey) Oxford and IBH
Publishing Co. Pvt. Ltd, New Delhi, 1995, 34.
[21] W. H. Daly et al.,Polym. Sci. Technol. 1984, 24, 45, Cited in: E.
V. Dehmlow: Tetramethylammonium Salts as Phase Transfer
© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
299
Catalysts, Phases – The Sachem Phase Transfer
Catalysis review, issue (1). www.sacheminc.com/lifesciences/
ptc/phases/Phaseso1.pdf()
[22] L. Kaur: Physicochemical properties of potatoes in relation
to thermal and functional properties of their starches. PhD
Thesis, Guru Nanak Dev University, Punjab, India, 2004,
140–146.
[23] J. B. Hirsch, J. L. Kokini: Understanding the mechanism of
cross-linking agents (POCl3, STMP and EPI) through swel-
ling behavior and pasting properties of cross-linked waxy
maize starches. Cereal. Chem. 2002, 79, 102–107.
[24] H. Liu, L. Ramsden, H. Corke: Physical properties of cross-
linked and acetylated normal and waxy rice starch. Starch/
Stärke 1999, 51(7), 249–252.
[25] C. Yook, U. H. Pek, K. H. Park: Gelatinization and retro-
gradation characteristics of hydroxypropylated and cross-
linked rices. J. Food. Sci. 1993, 58, 405–407.
[26] S. G. M. Choi, W. L. Kerr: Effects of chemical modification of
wheat starch on molecular mobility as studied by pulsed 1H-
NMR. Lebensm.-Wiss. Technol. 2003, 51, 1–8.
[27] I. Reddy, P. A. Seib: Modified waxy wheat starch compared
to modified waxy corn starch. J. Cereal. Sci. 2000, 31, 25–
39.
[28] H. Lim, P. A. Seib: Location of phosphate esters in a wheat
starch phosphate by P-nuclear magnetic resonance spec-
troscopy. Cereal Chem. 1993, 70, 145–152.
[29] Y. Wu, P. A. Seib: Acetylated and hydroxypropylated dis-
tarch phosphates from waxy barley: paste properties and
freeze thaw stability. Cereal. Chem. 1990, 67, 202–208.
[30] K. Morikawa, K. Nishinari: Effect of concentration depend-
ence of retrogradation behavior of dispersions for native and
chemically modified potato starch. Food Hydrocoll. 2000,
14, 395–401.
[31] R. Walker, E. A. ElHarith: Nutritional and toxicological prop-
erties of some raw and modified starches. Ann. Nutr. Ali-
ment. 1978, 32, 671.
[32] K. C. Huber, J. N. BeMiller: Channels of maize and sorghum
starch granules. Carbohydr. Polym. 2000, 41, 269–276.
[33] K. S. Woo, P. A. Seib: Cross-linked resistant starch: Prepa-
ration and properties. Cereal Chem. 2002, 79, 819–825.
[34] A.-I. Yeh, S.-L. Yeh: Some characteristics of hydroxy-
propylated and cross-linked rice starch. Cereal Chem. 1993,
70(5), 596–601.
[35] M. Wootton, A. Bamunuarachchi: Application of differential
scanning calorimetry to starch gelatinization I. Commercial
native and modified starches. Starch Stärke 1979, 31, 201–
204.
[36] C. Perera, R. Hoover, A. M. Martin: The effect of hydroxy-
propylation on the structure and physicochemical properties
of native, defatted and heat moisture treated potato star-
ches. Food Res. Int. 1997, 30, 235–247.
[37] P. Chatakanonda, S. Varavinit, P. Chinachoti: Effect of cross-
linking on thermal and microscopic transitions of rice starch.
Lebens.-Wiss. Technol. 2000, 33, 276–284.
(Received: October 28, 2005)
(Revised: March 1, 2006)
(Accepted: March 3, 2006)
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