Food Hydrocolloids 24 (2010) 619e625

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Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Effect of cross-linking on the physicochemical and physiological properties of

corn starch
Seung Hyun Koo, Kwang Yeon Lee, Hyeon Gyu Lee*
Department of Food and Nutrition, Hanyang University, 17 Haengdang-dong, Seongdong-gu, Seoul 133-791, Republic of Korea

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

Article history: Corn starch was chemically modied by cross-linking with STMP/STPP (99:1. w/w) and the physico-
Received 28 September 2009 chemical and physiological properties (in vitro and in vivo) of the cross-linked corn starch were inves-
Accepted 25 February 2010 tigated as a function of the degree of cross-linking. Cross-linking decreased the solubility, swelling factor,
and paste clarity of corn starch. While the swelling factor was highly correlated with the degree of cross-
Keywords: linking (R2 0.878), the X-ray diffraction patterns did not show any signicant alteration in the crys-
Cross-linked corn starch
tallinity of corn starch. It was shown by SEM measurement that a black zone was observed on the surface
Resistant starch
of crossed-linked starch granules, which did not occur with native starch. When mice were fed the diets
Physicochemical
Physiological
containing the corn starch with low (CLCS-5) and high (CLCS-12) degree of cross-linking (51.3 and 99.1%,
respectively), signicant effects on the nal body weight, weight gain as well as perirenal weight of the
mice (p < 0.05) were observed. Also, signicant decreases in total lipid, triglyceride, and total cholesterol
concentrations in serum were detected in CLCS-5 and CLCS-12 groups (p < 0.05). While total lipid level in
the liver decreased with increasing degree of cross-linking, the triglyceride level was not affected by the
supplementation with both of CLCS-5 and CLCS-12 corn starch samples.
2010 Elsevier Ltd. All rights reserved.

1. Introduction 2002). Among these methods, cross-linking has been commonly

Starch, as a major reserve substance of plant sources, contrib-
utes to the appearance, structure and quality of food products
(Aparicio-Saguilan et al., 2005; Lawala & Adebowaleb, 2005). From
the nutritional point of view, it is classied into three major frac-
tions according to the timeline in the gastrointestinal tract which
are rapidly digestible starch (RDS), slowly digestible starch (SDS),
and resistant starch (RS) (Englyst, Kingman, & Cummings, 1992).
Out of them, resistant starch has been subdivided into four types:
physically inaccessible starch (RS1), granular starch (RS2), retro-
graded or high amylose starch (RS3) and chemically modied
starch (RS4) (Xie, Liu, & Cui, 2006). In particular, it has been known
that the chemically modied starch inhibits the in vitro digestibility
of starch (Hood & Anderson, 1976; Leegwater & Luten, 1971) and
also provides markedly changed physicochemical properties in
a number of foods (Song, He, Ruan, & Chen, 2006).
The chemical modication of starch can be achieved by a variety
of different chemical reactions such as acid hydrolysis, oxidation,
etherication, esterication and cross-linking (Jayakody & Hoover,
* Corresponding author. Tel: 82 2 2220 1202; fax: 82 2 2292 1226.
E-mail address: hyeonlee@hanyang.ac.kr (H.G. Lee).
used to modify native starch where various cross-linking agents
such as sodium trimetaphosphate (STMP), sodium tripolyphos-
phate (STPP), epichlorohydrin (EPI) and phosphoryl chloride (POCl3)
have been used (Ratnayake & Jackson, 2008), being capable of
forming either ether or ester inter-molecular linkages between
hydroxyl groups on starch molecules (Rutenberg & Solarek, 1984).
Also, the cross-linking of starch was reported to be affected by
various factors which include starch source, cross-linking reagent
concentration and composition, the extent of substitution, pH,
reaction time, and temperature (Chung, Woo, & Lim, 2004; Lim &
Seib, 1993). In the food industry, starch modication with cross-
linking is commonly used to provide stabilized granular structure
and restricted swelling (Ratnayake & Jackson, 2008) as well as
nutritionally benecial effects (Woo, 1999; Wurzburg, 1986). Thus,
most of the preceding studies have focused on the physicochemical
and nutritional properties of cross-linked starches (Woo, 1999;
Wurzburg, 1986). However, little information exists regarding the
biological response (in vitro as well as in vivo) of cross-linked starch
as a function of the degree of cross-linking.
In the present study, the objectives were to prepare for corn
starch with different degree of cross-linking, to investigate the
effects of the degree of cross-linking on the physicochemical
properties and nutritional fractions (RDS, SDS and RS) of the corn

0268-005X/$ e see front matter 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodhyd.2010.02.009
620 S.H. Koo et al. / Food Hydrocolloids 24 (2010) 619e625
starch, and nally to determine the comparative effects of cross-
linked corn starches with low and high cross-linking on the weight
gain and lipid prole in mice.
2. Materials and methods
2.1. Materials
Corn starch, sodium trimetaphosphate (STMP), sodium tripoly-
phosphate (STPP), a-amylase (A-3176, Type VI: From porcine
pancreas) and porcine pancreatin (P-1750) were obtained from
SigmaeAldrich Chemical Co. (St. Louis, MO, USA). Amyloglucosi-
dase (AMG 300 L, activity 300 AGU/mL) was obtained from Novo-
zymes (Bagsvaerd, Denmark). All other chemicals were analytical
grade reagents.
2.2. Preparation of cross-linked corn starch
Cross-linked starches were prepared according to the method of
Woo and Seib (2002). Corn starch (50 g) was mixed with different
amounts (5, 10, and 12% (w/w), based on dry weight of starch) of
a mixture of STMP/STPP (99/1% w/w) and dissolved in 70 mL water,
which were designated as CLCS-5, CLCS-10, and CLCS-12, respec-
tively. After the pH was adjusted to 11.0 with 0.1 N NaOH, the slurry
was kept at 45  C for 3 h in a shaking water bath. The suspensions
were neutralized to pH 6.0 with 0.1 N HCl, washed with distilled
water four times, and dried at 40  C for 24 h in an oven. The dried
samples were then grounded in a mortar and sieved (100 mesh).
The starch subjected to the cross-linking condition without
a mixture of STMP/STPP was used as a control in each experiment.
2.3. Determination of the degree of cross-linking
The degree of cross-linking of modied starches was deter-
mined from the viscosity values, according to the procedure of
Kaur, Singh, and Singh (2006). The peak viscosity of modied starch
slurries (25% by weight) was measured from a controlled-stress
rheometer equipped with a starch pasting cell. (AR1000, TA
Instruments, New castle, DE, USA). A programmed heating and
cooling cycle to obtain the peak viscosity was employed as follows:
starch slurries were heated from 50 to 95  C at 11  C/min, and then
held at 95  C for 2 min. Afterwards the paste was cooled down to
50  C at 11  C/min and nally kept at 50  C for 2 min. The degree of
cross-linking was calculated by using the equation below:
Degree of cross  linking% A  B=A  100
where A is the peak viscosity of the control sample (without STMP/
STPP), and B is the peak viscosity of the cross-linked starch.
2.4. Paste clarity
The paste clarity of native and modied starches was determined
according to the method of Reddy and Seib (1999). Starch (0.05 g,
db) was suspended in distilled water (5 mL) in a glass-stoppered
tube and heated at 95  C for 30 min with shaking every 5 min. After
cooling, the starch clarity was measured on a spectrophotometer
(Shimadzu model UV-160A) at 650 nm against water blank.
2.5. Swelling factor
The swelling factor was measured by the method of Tester and
Morrison (1990). A starch sample (100 mg) was suspended in
5 mL of distilled water and kept in a shaking water bath at 70  C for
30 min. The tubes were then cooled rapidly to 20  C, 0.5 mL of blue
dextran solution (5 mg/mL) was added, and the contents were
mixed well. After centrifugation at 3000  g for 10 min, the absor-
bance of the supernatant was read at 620 nm.
2.6. Solubility
Solubility was measured according to the method of Schoch
(1964). Starch sample (0.5 g, db) was heated with 40 mL of water
at desired temperatures (50e90  C) for 30 min. After centrifugation
at 3000  g for 15 min, the supernatant was dried at 120  C for 2 h
and the remnant was weighed. The solubility (%) was determined
as the weight ratio of the dried supernatant to the dry starch.
2.7. In vitro digestibility
The in vitro digestibility of cross-linked corn starch was deter-
mined according to the method of Englyst et al. (1992). Enzyme
solution was prepared as follows: Pancreatin (1 g) was added to
12 mL of distilled water, mixed for 10 min, and then centrifuged at
1500  g for 10 min. The supernatant (10 mL) and amyloglucosi-
dase (0.2 mL) were transferred to a volumetric ask and the nal
volume was adjusted to 12 mL with distilled water. This enzyme
solution was freshly made for each digestibility test. Starch samples
(30 mg) and 0.75 mL of sodium acetate buffer (pH 5.2) was added
into each micro tube (2 mL) which was placed in a shaking incu-
bator at 37  C for 10 min. The prepared enzyme solution (0.75 mL)
was added to each tube which was incubated in the shaking
incubator of 250 rpm for 20 min and then boiled for 10 min to stop
the reaction. Then, the glucose concentration in the supernatant
was measured using dinitrosalicylic acid (DNS) method to calculate
the content of RDS (rapidly digestible starch). Also, the glucose
concentration after 120 min incubation was determined in the
same manner to obtain the content of SDS. Then, the content of RS
was calculated by differences (RS 100  RDS  SDS).
2.8. X-ray diffraction (XRD) measurement
X-ray diffraction analysis was performed with an X-ray diffraction
image processor (D/Max-1200, Rigaku, Japan) (analysis parameters:
50 kV and 90 mA, CuK radiation l 0.154 nm, nickel lter). The
sample was scanned through the 2q range from 5 to 40 .
2.9. Scanning electron microscopy (SEM)
The granular structure was observed using a scanning electron
microscope (SEM). Starch samples were placed in an oven at 40  C
for a few days for dehydration. The dehydrated samples were then
evenly distributed on SEM specimen stubs with double adhesive
tape and coated with a 10 nm gold layer. SEM analysis was con-
ducted using a JEOL-6330F scanning electron microscope (JEOL,
Ltd., Tokyo, Japan). Representative micrographs were taken for each
sample at magnications of 200 (left) and 500 (right).
2.10. Animal experiments
Four-week old male mice (C57BL/6J) were purchased from
Central Lab. Animal Inc. (Seoul, Korea). After a weeks acclimatiza-
tion period on standard rodent chow (Samyang, Seoul, Korea), the
animals were stratied by weight and assigned to one of the four
diets; a normal diet (ND, AIN-76A puried rodent diet 65% corn
starch #111753 (Dyets Inc., Bethlehem, PA, USA), a high-fat diet
(HFD, 40% beef tallow modied AIN-76A puried rodent diet
#101556 (Dyets Inc., Bethlehem, PA, USA))), and two experimental
diets (high-fat diet contained with corn starched crossed-linked
with 5% (CLCS-5) and 12% (CLCS-12) of STMP/STPP having the
 S.H. Koo et al. / Food Hydrocolloids 24 (2010) 619e625 621

composition of 44% starch. 13% protein, and 3.97 kcal/g energy) Table 2
(Table 1). While the mice were housed in cages with wire-mesh Degree of cross-linking, paste clarity and swelling factor of native and cross-linked
corn starch prepared with different level of STMP/STPP.
bottoms in a conditioned room (23  C; relative humidity 55%), they
were provided with food and water ad libitum and maintained on Degree of cross-linking (%) Paste clarity (% T650) Swelling factor
each diet for a 6-week period. Food intakes were measured twice Control 0 30.15  1.34a 38.92
weekly and their body weights were recorded once a week. The CLCS-5 51.3 1.01  0.20b 33.51
CLCS-10 98.1 0.86  0.13b 20.25
care and treatment of mice were approved by the Hanyang
CLCS-12 99.1 0.86  0.13b 13.17
University Lab Animal Care Committee, which were in accordance
All values are means of triplicate determinations  standard deviation.
with the Korean Guide for the Care and Use of Laboratory Animals.

2.11. Serum collection and organ preparations
After fasted for 24 h prior to being sacriced, the mice were
anesthetized with dry ice and dissected. Blood collected from the
heart, was placed at room temperature for 2 h and then centrifuged
at 1500  g for 30 min at 4  C to obtain serum. The adipose tissue
(perirenal and epididymal), liver, kidney and epididymis were
excised from each mouse and the wet weights were measured. The
separated serum and organs were then snap frozen in liquid
nitrogen and stored at 70  C for further analysis.
2.12. Lipid prole
Serum lipid proles (total lipid, triglyceride, total cholesterol)
were measured by blood chemistry analyzer (Olympus AU 400,
Japan). Total lipids from the livers were extracted with chloroform:
methanol (2:1, v/v) by a modied method of Folch, Lees, and
Stanley (1956). Liver total cholesterol and triglyceride contents
were assayed by enzymatic colorimetric procedures using
a commercial kit (Asan Pharmaceutical Co., Seoul, Korea).
2.13. Statistical analysis
All experiments were performed in triplicate and Statistical
Package for the Social Science (SPSS, Version 12.0, 2004, SPSS Inc.,
Chicago, IL, USA) was used to statistically analyze the results. The
results were expressed as the mean  SD and one-way analysis of
variance (ANOVA) followed by Duncans multiple comparison test
was performed.
3. Results and discussion
3.1. Degree of cross-linking, paste clarity and swelling factor
The degree of cross-linking of corn starches which were modi-
ed by different amounts of STMP/STPP (0, 5, 10, and 12%) was
determined to be 0, 51.3, 98.1, 99.1% (Table 2). There seemed to be
a proportionally increase in the degree of cross-linking with
increasing concentration of STMP/STPP up to 10%, which then
leveled off from 10 to 12%.
The paste clarity at T650 of the control starch exhibited 30.15%
(Table 2). However, the use of STMP/STPP cross-linking reagent led
to a substantial decrease in the paste clarity. Corn starch crossed-
linked with 5% STMP/STPP showed 1.01% past clarity and the other
samples exhibited 0.86% paste clarity. There was not however
signicant differences among the cross-linked samples (p > 0.05).
These results were consistent with the reports by Lim et al. (1993)
and Kaur et al. (2006) also found that cross-linked starches showed
lower pastes clarity than their counterpart native starches. The
signicant decrease in the paste clarity was possibly attributed to
a change in the starch granular structure by cross-linking
(Morikawa & Nishinari, 2000a). That is, compared to native starch,
the structure of cross-linked starches seemed to be almost intact
after heating at 95  C, resulting in the decreased paste clarity (Kaur
et al., 2006; Morikawa & Nishinari, 2000b). Furthermore, it has
been suggested that the reduced swelling of cross-linked starches
might be partly responsible for their reduced paste clarity (Kaur
et al., 2006; Reddy & Seib, 2000).
The swelling factor measured at 70  C decreased signicantly
with increasing level of the cross-linking reagent (Table 2). This
result was in agreement with the nding of Mirmoghtadaie,
Kadivar, and Shahedi (2009) who reported the reduced swelling
factor of cross-linked oat starch with increasing degree of cross-
linking. Regression analysis exhibited that the swelling factor was
linearly related to the concentration of cross-linking reagent used
(R2 0.878). It is well known that cross-linking strengths the
bonding between starch chains, thus allowing them to resist
against swelling. Therefore, the reduced swelling factor would be
related to the formation of inter-molecular bridges by phosphorous
residual after cross-linking reaction (Chung et al., 2004; Janzen,
1969; Woo, 1999; Wurzburg, 1986).

Table 1 50
Composition of experimental diets (g/kg diets).

Ingredient ND Experimental groupsa

40
HFD CLCS-5 CLCS-12
Casein 200 200 200 200
0%
Solubility (%)

DL-methionine 3 3 3 3 30 5%
Corn starch 650 150 e e
Sucrose 0 150 150 150 10%
CLCS-5 e e 150 e 20 12%
CLCS-12 e e e 150
Corn oil 50 e e e
Beef tallow e 400 400 400 10
Cellulose 50 50 50 50
Mineral Mix S10022G 35 35 35 35
Vitamin Mix V10037 10 10 10 10 0
Choline bitartrate 2 2 2 2
50C 60C 70C 80C 90C
Total calories (kcal/kg) 3592 5542 5542 5542
Total (g/kg) 1000 1000 1000 1000
Temperature (C)
a
ND, Normal diet; HFD, High-fat diet; CLCS, High fat diet containing with cross- Fig. 1. Solubility of native and cross-linked corn starch prepared with different levels
linked corn starch with different degree of cross-linking. of cross-linking with the mixture STMP/STPP.
622 S.H. Koo et al. / Food Hydrocolloids 24 (2010) 619e625

Table 3 3.3. Fraction of RDS, SDS and RS

RDS, SDS, and RS contents derived from native and cross-linked corn starch
prepared with different level of STMP/STPP.
RDS content (58.05% / 28.63%) and SDS content (36.46% /
Starch fraction (%) 12.69%) signicantly decreased, but the RS content (5.50% /
RDS SDS RS 58.68%) increased with increasing concentration of cross-linking
Control 58.05  3.6a 36.46  7.07a 5.50  4.55c reagent (Table 3). It is well known that the contents of starch
CLCS-5 49.05  5.46a 33.96  2.37a 16.98  3.10b fractions (RDS, SDS and RS) can be affected by various factors such
CLCS-10 33.51  8.03b 16.17  4.77b 50.31  4.86a as amylose content, swelling power, granule surface area and starch
CLCS-12 28.63  8.2b 12.69  2.37b 58.68  6.87a
crystalline structure (Shin, Kim, Ha, Lee, & Moon, 2005, Spence &
Jane, 1999). Specially, when they were plotted against the
swelling factor, a highly positive correlation was observed for RDS
(R2 0.983) and SDS (R2 0.972). Since the swelling of starch
3.2. Water solubility
granule is likely to enhance the access of digestive enzymes to its
inner structure, it appeared that the cross-linked starch with lower
When the solubility of native and cross-linked corn starches was
swelling factor (Fig. 2) was less degraded by the attack of the
measured as a function of temperature (50e90  C) (Fig. 1), there was
digestive enzymes, giving rise to a decrease in RDS and SDS
an overall tendency that the solubility decreased with increasing
contents. However, RS content was inversely related to the swelling
degree of cross-linking at all temperatures tested. A similar pattern
factor (R2 0.920) since the restricted swelling by cross-links can
was reported by Kaur et al. (2006) for potato starches modied
reduce enzymatic hydrolysis. These results were conrmed by
with EPI and POCl3 at different concentrations. It suggested that
previous studies (Chung, Shin, & Lim, 2008; Chung et al., 2004; Han
decreased solubility by cross-linking would be possibly due to
& Bemiller, 2007; Shin et al., 2005; Woo, 1999).
increased density of cross-links in the starch structure which
seemed to cause less disintegration of starch granules during gela-
tinization (Jyothi, Moorthy, & Rajasekharan, 2006). 3.4. XRD

The X-ray diffraction patterns of native and cross-linked

starches are shown in Fig. 3. There was no pronounced difference
between the native and modied starches. That is, the native and
70
modied starches showed similar diffraction patterns with peaks
60 y = 1.142x + 12.07
R = 0.983 at 15.02, 17.18 and 23.73 (2q), which are typical characteristics of
50 A-type starch (Zobel, 1988). This result suggested that cross-link-
RDS (%)

40 ing with STMP/STPP up to 12% did not dramatically alter the

30 crystalline pattern of corn starch. This observation was in agree-
ment with the nding of Hoover and Sosulski (1986), who reported
20
that cross-linking mainly took place in the amorphous regions of
10 starch granule and did not change the crystalline patterns of
0 starches.
10 20 30 40
Swlling factor (SF)
3.5. SEM
50
y = 1.009x - 1.904
R = 0.972
The scanning electron micrographs (SEM) of native and cross-
40 linked corn starches are shown in Fig. 4. SEM investigation showed
SDS (%)

30

20

10

0
10 20 30 40
Swlling factor (SF)

70
60 y = -1.856x + 76.99
R = 0.920
50
40
RS (%)

30
20
10
0
10 20 30 40
Swlling factor (SF)

Fig. 2. Linear regression of swelling factor and starch fraction (RDS, SDS and RS) of Fig. 3. X-ray diffraction patterns (XRD) of native and cross-linked corn starch prepared
cross-linked corn starch prepared with different levels of cross-linking with the with different levels of cross-linking with the mixture STMP/STPP. (A) Native, (B) CLCS-
mixture STMP/STPP. 5, (C) CLCS-10, (D) CLCS-12.
 S.H. Koo et al. / Food Hydrocolloids 24 (2010) 619e625 623

Fig. 4. Scanning electron microphotograph (SEM) of native and cross-linked corn starch prepared with different levels of cross-linking with the mixture STMP/STPP. (A) Native, (B)
CLCS-5, (C) CLCS-10, (D) CLCS-12. (Right: 200; Left: 500, bar 10 mm)

that the cross-linking caused slight changes in the structure of
starch granules compared with native starch. Native starch gran-
ules (Fig. 4A) were polygonal in shape with well-dened edges,
whereas cross-linked starch granules (Fig. 4B-D) exhibited
a slightly rough surface and black zone on the surface. These results
were consistent with the reports by Carmona-Garcia, Sanchez-
Rivera, Mndez-Montealvo, Garza-Montoya, and Bello-Prez
(2009) who found that cross-linked banana starch with STMP/
STPP showed black zones on the surface along the longitudinal axis.
Singh, Kaur, and McCarthy (2007) suggested that the black zones
seemed to indicate a slight fragmentation and the formation of
a deep groove in the starch granules.
3.6. Animal studies
3.6.1. Body weight, food intake, tissue and fat pads weight
The body weight, food intake, tissue and fat pads weight in
mice fed different diets are shown in Table 4. At the beginning of
the study, the mice fed normal diet (ND) had less initial weight
than the other groups which were fed the high-fat diet while there
was no signicant difference in body weight among the three
groups. However, the addition of the CLCS-5 and CLCS-12 to the
high-fat diet (HFD) had signicant (p < 0.05) effects on the mice
nal body weight which was signicantly (p < 0.05) reduced
compared to the HFD group. However, no signicant difference
624 S.H. Koo et al. / Food Hydrocolloids 24 (2010) 619e625

Table 4
Body weight, food intake, tissue and fat pads weight of C57BL/6J mice fed high-fat diet after 6 weeks of treatment with experimental diets.

ND HFD CLCS-5 CLCS-12

Initial weight, g 24.00  0.69b 32.03  1.73a 30.00  1.94a 29.50  3.78a
Final weight, g 26.52  0.41c 38.64  1.79a 34.08  1.02b 33.61  1.99b
Weight gain, % 7.11  2.82b 20.67  0.93a 7.42  3.11b 9.47  1.96b

Food intake, g/day 109.10  7.49a 94.40  0.28ab 78.90  1.27b 98.70  21.35ab
Food efciency, g gain/g food 0.016  0.001c 0.094  0.000a 0.084  0.001ab 0.071  0.015b

Tissue weight
Liver, g 0.97  0.04c 1.58  0.27a 1.24  0.18b 1.32  0.34b
Kidney, g 0.32  0.03b 0.34  0.02ab 0.34  0.02ab 0.34  0.03a
Epididymis, g 0.24  0.09ns 0.21  0.01ns 0.19  0.06ns 0.20  0.04ns

Fat pad weight

Perirenal fat, g 0.11  0.06c 0.29  0.08a 0.26  0.08a 0.19  0.07b
Epididymis, g 0.48  0.12b 2.06  0.35a 1.82  0.40a 1.71  0.53a

Initial weight: mean body weight at beginning the experimental diets.

Weight gain (%) [(Final weight (g) - initial weight (g))/initial weight (g)]  100.
Energy intake (kcal) mean food consumption (g)  dietary metabolize energy (kcal).
Food efciency mean body weight gain (g)/mean food intake (g).
a-c
Values with different subscripts within the same column are signicantly different among samples at a 0.05 level by Duncans multiple range test.

was detectable between CLCS-5 and -12 groups. Gain weights
were 20.67  0.93%, 7.42  3.11% and 9.47  1.96% for mice fed
HFD, CLCS-5 and -12 groups, respectively. In addition, a 64.10% and
54.18% reduction in the body weight was observed in the CLCS-5
and -12 groups, respectively, compared with the HFD group
(p < 0.05) at the end of the trial. This result was favorably
compared with the study of Jeong, Kim, Kang, and Kim (2002)
where the diets containing resistant starch caused a reduced
weight gain by 7%. It would be explained by the fact that resistant
starch decrease energy absorption, thus giving rise to a decrease in
epididymal fat pads and serum triacylglycerol concentration (de
Deckere, Kloots, & van Amelsvroot, 1995). However, Bjorck,
Gunnarsson, and Ostergard (1989) reported no signicant effect
of the diets containing cross-linked and acetylated corn and rice
starches on body weight loss in normal and diabetic rats. Food
intake was lower from the mice on the CLCS-5 diet, compared
with the other groups, even though there were no signicant
differences. Greater food efciency was observed greater from the
mice on the HFD (0.094  0.000), CLCS-5 (0.084  0.001) and
CLCS-12 (0.071  0.015) diets, as compared to mice fed the normal
diet (0.016  0.001). All the groups fed the high-fat diet exhibited
higher liver weight than ND. Specically, the liver weight of the
HFD group was greater by 63% than that of the ND group. Less liver
weight was observed in the CLCS groups than HFD where no
signicant differences in kidney and epididymis weights were
detected among the groups (p < 0.05). Previous studies (Cheng &
Lai, 2000; Ebihara, Shiraishi, & Okuma, 1998; Ranhotra, Gelroth,
& Glaser, 1996) showed that the diet containing resistant starch
reduced the organ weight such as liver. Hence, these results would
result possibly from the inclusion of CLCS in the diets which
belongs to resistant starch,
3.6.2. Lipidic parameters in serum and liver
The changes of plasma and liver lipids concentration are shown
Table 5. The serum total lipid value was signicantly lower in the
mice fed CLCS-5 (303.50  22.20 mg/dL) and CLCS-12
(294.45  17.65 mg/dL) diets than those fed the HFD diet
(337.81  8.19 mg/dL) (p < 0.05). A signicant decrease in serum
triglyceride was observed in CLCS-12 groups compared to other
groups (p < 0.05). Serum total lipid and triglyceride level appeared
to decrease with increasing degree of cross-linking. However, the
differences between CLCS-5 and CLCS-1v2 were not signicant. The
diets containing resistant starch are shown to reduce weight gain as
well as level of blood lipid in previous studies (Kim, Chung, Kang,
Kim, & Park, 2003; Martinez-Flores, Chang, Martinez-Bustos,
& Sgarbieri, 2004; Ranhotra et al. 1996), which are thus favorably
compared with our results. Serum total cholesterol levels were
lower by 8.3% and 5.7% in the mice fed CLCS-5 and CLCS-12, as
compared to the group fed HFD. Moreover, the mice fed the diets
containing cross-linked corn starch exhibited lower level of LDL-
cholesterol than the HFD group while no signicant difference in
the HDL-cholesterol concentration was observed. In addition, the
level of HDL-cholesterol was higher in the groups fed the HFD diets
while the ND group exhibited the lowest value (p < 0.05). Ranhotra
et al. (1996) reported that the reduction of total cholesterol by RS
was derived from a decrease in both levels of HDL- and LDL-
cholesterol. Thus, it might suggest that the increased level of total
serum cholesterol in the groups fed the high fat diet appeared to
raise both levels of both LDL and HDL.
Total liver lipid values were signicantly lower in mice fed
the CLCS-5 (79.47  19.72 mg/g) and CLCS-12 (80.60  8.81 mg/g)
than those fed HFD (241.50  15.65 mg/g) (p < 0.05). The supple-
mentation with cross-linked corn starch seemed to cause more

Table 5
Lipidic parameter in serum and liver of C57BL/6J mice fed high-fat diet after 6 weeks of treatment with experimental diets.

ND HFD CLCS-5 CLCS-12

Serum
Total lipid, mg/dL 333.01  12.78a 337.81  8.19a 303.50  22.20b 294.45  17.65b
Triglyceride, mg/dL 149.23  9.66a 98.89  4.63b 86.76  11.44bc 80.13  8.20c
Tota cholesterol, mg/dL 121.94  5.53c 155.38  4.43a 142.56  10.06b 146.55  9.26ab
HDL-cholesterol, mg/dL 76.30  4.21b 97.20  2.25a 89.50  4.28a 91.90  5.88a
LDL-cholesterol, mg/dL 45.64  1.64c 58.18  2.25a 53.06  6.12b 54.65  4.63b

Liver
Total lipid, mg/g liver 114.13  7.29b 241.50  15.65a 79.47  19.72b 80.60  8.81b
Triglyceride, mg/g liver 19.22  2.44b 28.14  2.37a 32.28  3.41ab 30.51  3.54a
Tota cholesterol, mg/g liver 1.16  0.36a 1.07  0.43a 1.08  0.41ab 0.73  0.31b
aec
Values with different subscripts within the same raw are signicantly different among samples at a 0.05 level by Duncans multiple range test.
 S.H. Koo et al. / Food Hydrocolloids 24 (2010) 619e625
excretion of triglyceride in the liver even though a statistical
difference was not observed. Also, a signicant decrease (p < 0.05)
in liver total cholesterol occurred in the mice fed CLCS-12 diet.
Acknowledgement
This work was supported by the Academic Research & Devel-
opment Program funded by the Youlchon Foundation.
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