Starch/Strke 61 (2009) 291299 DOI 10.1002/star.

200800103 291

Vicente Espinosa-Solisa Physicochemical Characteristics of Starches from

Jay-lin Janeb
Luis A. Bello-Pereza
a
Centro de Desarrollo
de Productos Biticos del IPN,
Yautepec, Morelos, Mxico
b
Department of Food Science
and Human Nutrition,
Iowa State University,
Ames, IA, USA
Unripe Fruits of Mango and Banana
Mango and banana starches were isolated from unripe fruits and their morphology;
thermal and pasting properties; molar mass and chain length distribution were deter-
mined. Mango starch granules were spherical or dome-shaped and split, while banana
starch had elongated granules with a lenticular shape. Amylopectin of both fruit starches
had a lower molar mass than maize starch amylopectin; however, mango amylopectin
had the highest gyration radius. Banana amylopectin showed the lowest percentage of
short chains [degree of polymerization (DP) 612] and the highest level of long chains
(DP  37); mango amylopectin presented the highest fraction of short chains, but the
level of longest chains was intermediate between those of banana and maize amylo-
pectins. Banana starch presented the highest average gelatinization temperature fol-
lowed by mango starch and maize starch had the lowest value; a similar pattern was
found for the gelatinization enthalpy. The two fruit starches had a lower pasting temper-
ature than maize starch, but the former samples showed higher peak and final viscos-
ities than maize starch. Structural differences identified in the fruit starches explain their
physicochemical characteristics such as thermal and pasting behavior.

Keywords: Mango; Banana; Structure; Amylopectin

1 Introduction they reported the physicochemical and morphological

Most of the structural and physicochemical characteriza-
tions of starch have been performed in wheat, maize,
potato and rice starches because of their commercial
importance. In contrast, little is known about structural
and molecular characteristics of starch from unconven-
tional sources, such as green banana and mango fruits.
However, diverse studies, where their physicochemical
and functional properties were tested, have demon-
strated potential application [17].
Banana (Musa paradisiaca L.) and mango (Mangifera
indica L.) are climacteric fruits. In Mxico and many other
properties. Recently, the structural characteristics and in
vitro digestibility of mango kernel starch from the varieties
chausa and kuppi were reported [9]. Limited studies on
starch structural characteristics of banana variety macho
[10] and mango Tommy Atkins variety were reported [10].
However, the fine structure of amylopectin of these star-
ches has not been studied in detail. This information is
important to relate the structures to physicochemical and
functional characteristics such as pasting behavior, gelati-
nization and retrogradation phenomena. The present work
analyzed the structural and physicochemical properties of
two starches from unconventional sources: banana variety
macho and mango variety Tommy Atkins, using scan-

countries they are consumed in the ripe state. For this
reason, large quantities of fruits are lost during shipping
resulting from an improper postharvest handling. The
large starch contents of these fruits at unripened state
(7080%) make the fruits a potential source of starch for
various applications [1].
Research Paper
ning electron microscopy (SEM), iodine titration, high-per-
formance size-exclusion chromatography equipped with
multi-angle light-laser scattering and refractive index
detectors (HPSEC-MALLS-RI), fluorophore-assisted cap-
illary electrophoresis (FACE), differential scanning calorim-
etry (DSC) and pasting properties.

Banana and mango starches have not been extensively

studied but there have been investigations of different
varieties of these fruits. Kuar et al. [8] studied starch iso- 2 Materials and Methods
lated from mango kernels (seeds) from five varieties and
2.1 Starch isolation

Unripened bananas (Musa paradisiaca L.) from the variety

Correspondence: Luis A. Bello-Perez, Centro de Desarrollo de
Productos Biticos del IPN, Apartado postal 24 C.P., 62731, Yau-
tepec, Morelos, Mxico. Phone: 152-735-3942020, Fax: 152-
735-3941896, e-mail: labellop@ipn.mx.
macho were purchased in the local market in Cuautla
(Mexico) immediately after harvesting without any post-
harvest treatment. The skin color and size were the pa-

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292 V. Espinosa-Solis et al.
rameters used for cutting of the fruit. Unripened mangos
(Mangifera indica L.) from the cv. Tommy Atkins were
also purchased in the local market. Both starches were
isolated using a pilot-scale procedure proposed by Kim et
al. [11]. The fruits were peeled, cut into 56 cm cubes (100
kg total weight), immediately submerged in aqueous citric
acid solution (0.5 g/L) and then macerated at low speed in
a Waring blender (10 kg of fruit to 10 L of solution) for 2 min.
The homogenate was consecutively sieved through
screens (20, 40, 100 and 200 US mesh) until the wash
water (distilled) was clear, and then it was centrifuged in a
semicontinuous centrifuge (Veronesi model BSGAR 1500,
Verona, Italy) at 10,7506g. The sediments from the 100
and 200 US mesh screens were washed and then cen-
trifuged. The processes of sieving and centrifuging for
starch isolation and purification were repeated three times.
The starch sediments were dried in a spray dryer (Niro
Atomizer model P-6.3. Copenhagen, Denmark), with a
feeding temperature of 1301507C, a solids concentration
in the feeding line of 300400 g/kg and an air outlet tem-
perature of 70807C. The powder was ground to pass a US
No 100 sieve and stored at room temperature (257C) in a
glass container. Normal maize starch was obtained from
Arancia, productos de maz (Toluca, Estado de Mexico).
2.2 Apparent amylose content
Starch samples were defatted using a Soxhlet apparatus
and 85% methanol solution for 24 h. The samples were
washed with ethanol and recovered by filtration. Iodine
affinities of defatted starch were measured using an auto-
matic potentiometric titrator (702 SM Tirino, Metrohm,
Herisau, Switzerland) following the method previously
reported [12]. Apparent amylose contents were calculated
by dividing the iodine affinity of defatted starch by 20% [13].
2.3 Starch granule morphology
Starch granules, spread on silver tape and mounted on a
brass disk, were coated with gold/palladium (60/40).
Sample images were observed at 15006magnification
under a scanning electron microscope (JOEL, model
JSM-5800LV, Tokyo, Japan) at Bessey Microscopy Facil-
ity, Iowa State University.
2.4 Molar mass and gyration radius of
amylopectin
Weight-average molar mass and z-average gyration
radius of amylopectin were determined using HPSEC-
MALLS-RI. Starch samples were prepared as described
by Yoo and Jane [14]. The HPSEC system consisted of a
2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Starch/Strke 61 (2009) 291299
HP 1050 series isocratic pump (Hewlett Packard, Valley
Forge, PA, USA), a multi-angle laser-light scattering
detector (Dawn DSP-F, Wyatt Tech. Co., Santa Barbara,
CA, USA) and a HP 1047A refractive index detector
(Hewlett Packard, Valley Forge, PA, USA). To separate
amylopectin from amylose, a Shodex OH pak KB-guard
column and KB-806 and KB-804 analytical columns
(Showa Denko K.K., JM Science, Grand Island, NY, USA)
were used. Operating conditions and data analysis were
the same as described by Yoo and Jane [15], except that
the flow rate used was 0.5 mL/min and sample con-
centration was 0.3 mg/mL.
2.5 Amylopectin branch chain-length
distribution
Starch was fractionated using GPC, following the method of
Song and Jane [16]. The amylopectin fractions were col-
lected, evaporated, and precipitated with ethanol. Isolated
amylopectin was analyzed for chain-length distribution
using FACE as essentially described by [17, 18]. Amylo-
pectin was dispersed in 90% dimethyl sulfoxide (DMSO)
solution, precipitated by ethanol, and centrifuged at
67506g for 15 min. Amylopectin was suspended in water
to give 2 mg amylopectin per milliliter water. The mixture
was heated in a boiling water bath for 30 min and then
cooled down to room temperature. Eighty microliters of the
mixture was added with 19 mL acetate buffer solution (50
mM, pH 3.5) containing 0.02% sodium azide, and digested
with 1 unit isoamylase from Pseudomonas sp. (Megazyme
International, Wicklow, Ireland) at 407C for 2 h. The digested
sample was heated in a boiling water bath for 10 min to
inactivate the isoamylase. Fifty microliters of the sample
were evaporated to dryness in a centrifugal vacuum evap-
orator. The reductive amination reaction of the reducing end
was performed by adding 2 mL of 0.2 M aqueous 8-amino-
1,3,6-pyrenetrisulfonic acid (APTS: Sigma. Aldrich Co., St.
Louis, MO, USA) and 2 mL of 1 M aqueous sodium cyano-
borohydride to the dry sample. The mixture was incubated
at 407C for 18 h, and then 46 mL of deionized water were
added. An aliquot of 10 mL was diluted to 200 mL with
deionized water. Analysis of chain-length distribution of
amylopectin was conduced using a Beckman P/ACE cap-
illary electrophoresis instrument with the 50 cm length
N-CHO coated capillary (50 cm diameter) and a separation
Gel Buffer-N (ProteomeLab Carbohydrate labeling and
analysis kit: Beckman Coulter, Inc., CA, USA) sample was
introduced by pressure injection 5 s at 3447 Pa (0.5 psi).
2.6 Thermal properties of starch
Thermal properties of starch were determined using a
differential scanning calorimeter (DSC-7, Perkin-Elmer,
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Starch/Strke 61 (2009) 291299 Characteristics of Starches from Unripe Fruits of Mango and Banana 293

Norwalk, CT, USA) [19]. Starch (2 mg, dsb) was accurately
weighed in an aluminum pan, mixed with 6 mg of deion-
ized water and sealed. The sample was allowed to equili-
brate for 1 h and scanned at a rate of 107C/min over a
temperature range of 101107C. An empty pan was used
as the reference. The rate of starch retrogradation was
determined using the same gelatinized samples, stored at
47C for seven days, and analyzed using the same process
described for gelatinization. All thermal analyses were
conducted in triplicate for each sample.
2.7 Pasting properties of starch
Starch pasting properties were analyzed using a Rapid Visco
Analyser (RVA-4, Foss North America, Eden Prairie, MN,
USA) [19]. A starch suspension (8%, w/w, dsb), in triplicate
for each sample, was prepared by weighing starch (2.24 g,
dsb) into a RVA canister and making up the total weight to 28
g with deionized water. The starch suspension was equili-
brated at 307C for 1 min, heated at a rate of 6.07C/min to
957C, maintained at 957C for 5.5 min, and then cooled to
507C at a rate of 6.07C/min. Constant paddle rotating speed
(160 rpm) was used throughout the entire analysis.
banana starch was in agreement with that reported for
banana starch isolated from the same variety (37%) [23]
and from another variety (40%) [24], using the same
method. The amylose content of the mango fruit starch
was similar to that used for resistant starch preparation
from the same variety Tommy Atkins (28.7%) [7]. How-
ever, two other varieties, criollo (12.9%) and manila
(13.3%) presented lower apparent amylose content [6].
3.2 Starch granule morphology
Scanning electron micrographs of normal maize (A),
mango fruit (B) and banana (C) starch granules are shown
in Fig. 1. Mango starch granules were spherical or dome-
shaped and split. Mango starch granules present some
indentations that could be due to non-uniform growth
within starch granule or collapse during drying. This kind

3 Results and Discussion

3.1 Apparent amylose content

Iodine affinities and corresponding apparent amylose

contents of normal maize, mango fruit and banana star-
ches are shown in Tab. 1. Apparent amylose was different
for the starches analyzed. Banana starch had the largest
apparent amylose content (36.2%), followed by mango
(31.1%) and normal maize starch (29.7%). The difference
in amylose content affect the physicochemical and func-
tional properties, because starches with higher amylose
contents produce firmer and more opaque gels with

Tab. 1. Iodine affinities and apparent amylose content for

defatted starches.

Source Iodine affinitya Apparent amylose

contentb [%]

Normal maize 5.93 6 0.06 29.7

Mango 6.21 6 0.19 31.1
Banana 7.24 6 0.24 36.2

a) Averaged from three replicates 6 standard deviation.

b) Calculated as C= IAS/0.20 where C is the percentage of Fig. 1. Scanning electron micrographs of normal maize
apparent amylose content and IAS is the iodine affinity (A), mango (B) and banana (C) starches (15006) (scale
of the whole defatted starch. bar = 20 mm).

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294 V. Espinosa-Solis et al.
of morphology has been found for other fruits such as
winter squash [25], apple [26] and in cassava [27]. The
granule sizes of mango starch were similar to those of the
small granules population of maize starch, between 510
mm. This range was in agreement with those of starches
isolated from two mango varieties [6]. Banana starch had
elongated granules, with a lenticular shape, where the
average longitudinal dimension was 40 mm and the short
radius around 20 mm, which is in agreement with other
reports of granule size and shape of banana starches [2,
28]. The granule size of normal maize starch ranged be-
tween 1015 mm, with a higher heterogeneity showing
round, oval and polygonal shapes. Sizes and shapes of
starch granules influence some physicochemical, func-
tional and nutritional characteristics, because larger
granules develop a high paste viscosity and small gran-
ules are more digestible.
3.3 Molar mass and gyration radius of
amylopectin
Weight-average molar mass (Mw) and gyration radius (Rz)
of normal maize, mango and banana amylopectins are
shown in Tab. 2. Normal maize and mango amylopectins
presented similar Mw values, and these were higher than
those of banana amylopectin. This pattern is in agree-
ment with the Rz value, mango amylopectin displayed the
largest gyration radius (298 nm) and banana amylopectin
had the smallest (267 nm). The Mw for normal maize
obtained in this study agreed with data in the literature
[15]. The molar mass and gyration radius of banana amy-
lopectin were slightly higher than that in the literature [15],
which could be attributed to different varieties of banana
starches. The Mw of amylopectin of the three starches
decreases with increasing amylose content. A similar
inverse correlation has being found in other studies [9, 10,
15]. It was reported that the molar mass has influence on
the pasting viscosity of the starch [29]. Studies in amylo-
pectin of diverse botanical origins gave Mw ranging be-
tween 5.46107 and 1.16108 g/mol and Rz between 171
and 242 nm [30]. Maize amylopectin solubilized using a
Tab. 2. Average amylopectin molecular weight and gyra-
tion radius of starches.
Source Mw6108 [g/mol]a,b Rz [nm]a,c
Normal maize 5.154 6 0.065 281.10 6 1.980
Mango 5.013 6 0.177 297.85 6 1.202
Banana 3.371 6 0.179 267.10 6 5.515
a) Averaged from two replicates.
b) Weight-average molar mass.
c) z-average radius of gyration.
2009 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Starch/Strke 61 (2009) 291299
microwave heating for 35 s had Mw of 2.26108 g/mol and
Rz of 229 nm [31]. Amylopectin of diverse botanical sour-
ces presented Mw between 2.96107 and 3.56108 g/mol,
and Rz between 95-340 nm [32], and Mw between 1.3 and
56.86108 g/mol, and Rz between 201-782 nm [15]. Amy-
lopectin of Tef starch isolated from diverse cultivars pre-
sented Mw 1.0 and 1.46108 g/mol, and Rz between 156
and 205 nm [29]. The studies above mentioned used a
HPSEC-MALLS-RI system similar to that utilized in this
work.
3.4 Amylopectin branch chain-length
distribution
Amylopectin branch chain-length distribution for the
starches of normal maize, mango and banana are shown
in Fig. 2 and summarized in Tab. 3. Normal maize has few
short chains of DP 6 and content in chains of DP 7 and 8
gradually increases. This is characteristic of cereal star-
ches. Banana starch also showed an increase in chains of
DP 6-8, but it more pronounced than in case of normal
maize. Mango starch displays a higher population of short
chains of DP 7 than DP 6 and 8. This branch chain dis-
tribution has been found for some tubers, root, and
legume starches [19].
In this study, mango and normal maize starches, both had
A-type polymorphism, consisted of more short chains (DP
612), mango (25.7%) and normal maize (24.0%), and
fewer long chain (DP  37), mango (16.6%) and normal
maize (13.4%), in comparison with banana starch (C-
type) [10] that displayed a lower proportion of short
chains (20.5%) and a higher proportion of long chain
(18.6%). This is in agreement with the results of Jane et al.
[19], who reported that A-type starches had larger pro-
portions of short chains (DP 612) and smaller proportion
of long chains (DP  37) than B type starches.
In general, the three starch samples had the largest pro-
portion of chains with degree of polymerization (DP) be-
tween 1324, followed by short chains with DP between
612 and chains with DP higher than 37. Similar pattern
was found for cereal starches, in which the amylopectin
had larger proportion of short chains. Slight differences
were observed in the average chain lengths, with DP 21.5,
22.3 and 23.9 for normal maize, mango and banana
starch, respectively. The average chain length for banana
amylopectin found was comparable with that of ginkgo
amylopectin, both are C-type starches [33]. Some mo-
lecular and physicochemical characteristics, such as ret-
rogradation tendency, temperature and enthalpy of gela-
tinization, crystallinity percentage, etc., are related to the
proportion of short chains of amylopectin [31].
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Starch/Strke 61 (2009) 291299 Characteristics of Starches from Unripe Fruits of Mango and Banana 295

Fig. 2. Amylopectin chain

length distribution of normal
maize (A), mango (B) and
banana (C) starches, meas-
ured by fluorophore-assisted
capillary electrophoresis
(FACE).

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296 V. Espinosa-Solis et al. Starch/Strke 61 (2009) 291299

Tab. 3. Branch chain-length [CL] distribution of amylopectins.

Source Average Distribution [%] Highest

CL DP 6-12 DP 13-24 DP 25-36 DP  37 detectable DP

Normal maize 21.5 24.0 50.2 12.3 13.4 80

Mango 22.3 25.7 46.1 11.6 16.6 84
Banana 23.9 20.5 48.1 12.9 18.6 89

Averaged from two replicates.

3.5 Thermal properties
Gelatinization characteristics of the three starches stud-
ied showed differences; normal maize starch had the
lowest gelatinization temperature whereas banana starch
had the highest. A similar trend was found for the enthalpy
changes of the starches (Tab. 4). Gelatinization of starch is
the loss of the crystalline structure of the starch granule,
which mainly results from crystalline amylopectin. The
highest gelatinization temperature of banana starch can
be attributed to its smaller proportion of short branch
chains and larger proportion of long branch chain found in
the amylopectin molecule (Tab. 3). The different the
branch chain-length pattern of banana amylopectin
agreed with the X-ray diffraction pattern (C-type). It has
been reported that the type of the X-ray diffraction pattern
relates to the branch chain-length distribution of the
amylopectin [34]. Starches of the A-type crystalline
structure consist of more short branch chains than those
of the C-type crystalline structure.
Retrogradation properties (Tab. 4) showed that the crys-
tals formed during storage of gelatinized banana starch
had the largest thermal stability because they dissociated
at higher temperature and with a larger enthalpy change;
this result indicated that a higher degree of crystallinity
was produced in banana starch than in the other starches
analyzed. It is also noticed that the dissociation tempera-
tures of the retrogradation starch samples were lower
than the gelatinization temperatures, because during the
retrogradation small and/or less perfect crystals were
formed [35, 36]. Mango starch displayed the least per-
centage of retrogradation and banana starch the highest.
The lower degree of retrogradation of mango starch could
be attributed to the larger proportion of short branch
chain of amylopectin [37, 38] and its lower lipid content
than normal maize starch. Fast retrogradation of banana
starch was reported; after 14 h of storage at low moisture
content some peaks of crystallinity were found and the
polymorphism of the retrograded banana starch was A-
type [36].
3.6 Pasting properties
The pasting properties of starch are influenced by other
components present in the starch suspension during
heating as well as the interactions between the starch
molecules in the granule [39-41]. When a starch suspen-
sion is heated at a constant rate, the viscosity increases
gradually until a maximum value is reached (Fig. 3). The
pasting temperatures for the starch samples were in the
order: normal maize . banana . mango (Tab. 5). Banana
starch showed the largest peak viscosity and normal
maize the lowest. The difference between the DSC gela-
tinization onset temperature (T0) and the pasting temper-
ature of a particular starch was greater for normal maize
starch than for mango and banana starches. This diffe-

Tab. 4. Thermal properties of native and retrograded starch from normal maize, mango and banana.

Source Starch gelatinization Starch retrogradation

T0 [7C]* TP [7C]* TC [7C]* H [J/g]* T0 [7C]* TP [7C]* TC [7C]* H [J/g]* % Retro-
gradation

Normal maize 63.9 6 0.2 68.7 6 0.3 73.2 6 0.4 13.4 6 0.3 37.3 6 0.5 50.2 6 0.2 61.5 6 0.8 7.2 6 0.6 53.6
Mango 66.5 6 0.2 71.3 6 0.3 76.1 6 0.3 14.0 6 0.7 39.9 6 2.3 52.7 6 1.4 62.3 6 0.1 5.4 6 0.0 38.4
Banana 70.9 6 0.6 76.5 6 0.9 83.3 6 1.0 16.5 6 0.7 42.1 6 0.6 56.5 6 2.4 67.7 6 0.4 9.4 6 1.9 56.9

* = Onset temperature [T0], peak temperature [Tp], completion temperature [Tc] and enthalpy change [H]. Values are
averages of three replicates of each sample.

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Starch/Strke 61 (2009) 291299 Characteristics of Starches from Unripe Fruits of Mango and Banana 297

Fig. 3. Rapid Visco Analyser profiles of normal maize (o), mango (n) and banana (^) starches (8% dsb).

Tab. 5. Pasting properties of starches measured by Rapid Visco Analyser.c

Sourcea Pasting temperature [7C] Viscosity [RVU]b

Peak Breakdown Final Setback

Normal maize 84.0 6 1.5 149.8 6 0.5 68.3 6 0.8 145.1 6 0.7 63.6 6 1.0
Mango 71.3 6 0.2 194.1 6 3.2 50.2 6 3.4 239.1 6 2.7 95.2 6 2.0
Banana 79.3 6 0.3 215.8 6 2.7 33.5 6 2.2 323.8 6 6.3 141.7 6 5.4

a) Mixture consisted of 8% (w/w, dsb) starch in water.

b) Measured in Rapid Visco Analyser Units.
c) Averaged from three replicates.

rence could be attributed to the restricting complexes
present in normal maize starch. The peak viscosity of
mango starch was obtained at a lower temperature than
banana and normal maize starches, and the viscosity
maintained a plateau until 957C. The maximum of the
peak viscosity reflects the ability of starch granules to
swell freely before their physical breakdown [42]. When
starch is heated in the presence of water, the granules are
swollen while some components, including amylose and
small amylopectin, diffuse out, resulting in swollen and
dispersed particles present in a continuous phase [43].
The starch dispersion properties were more affected by
the branch chain-length distribution of amylopectin mol-
ecule than by the Mw [44]. During holding at 957C under
shear the viscosity of starch pastes decreased for all
three starches, resulting from the breakdown of some
swollen starch granules. Stevenson et al. [26] found a
correlation between breakdown viscosity and starch
structures. Their results show that starches, which display
less breakdown viscosity, consist of amylopectin with
more branch chains of DP  37 and less branch chains of
DP (13-24) and larger apparent amylose content. The
breakdown viscosity values of the three starches deter-
mined is this study were in the following order: normal
maize . mango . banana, which agreed with the results
of Stevenson et al. [26]. When the temperature dropped
the viscosity increased, which resulted from network for-
mation between amylose and amylopectin while retaining
a certain amount of water [45, 46] and resulted in a gel-
like characteristic.

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298 V. Espinosa-Solis et al.
4 Conclusions
The investigated fruit starches had a lower molar mass
than maize starch but mango starch showed the highest
gyration radius. Banana starch presented the lowest
amount of short chains and the highest level of long
chains. Mango starch had the highest level of short
chains. The average temperature and enthalpy of gelati-
nization were higher in the two fruit starches than in maize
starch. Banana starch showed the highest retrogradation
and mango the lowest. The fruit starches presented lower
pasting temperature but higher peak and final viscosity
than maize starch. This pattern is related to the higher
amylose content of the fruit starches. The structural dif-
ferences determined in the unconventional starches
explain their physicochemical characteristics.
Acknowledgements
We appreciate the financial support from SIP-IPN,
COFAA-IPN and EDI-IPN, and the technical assistance of
Dr. Sathaporn Srichuwong and Hongxin Jiang. One of the
authors (VES) also acknowledges a scholarship from
CONACYT-Mxico.
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(Received: November 4, 2008)
(Revised: December 3, 2008)
(Accepted: December 4, 2008)
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