Documenti di Didattica
Documenti di Professioni
Documenti di Cultura
www.elsevier.com/locate/carbpol
Abstract
The effect of sucrose on gelation of oxidised starch (partially depolymerised amylopectin with ,1 carboxyl group per 30 residues) has
been studied by small-deformation oscillatory measurements (1 rad s 21; 0.5 strain) of storage and loss moduli (G 0 and G 00 ). Solutions were
prepared at 958C, cooled to 58C at 18C/min, and held at 58C for 500 min. In the first series of experiments, the combined concentration of
sucrose and starch was held fixed at 65 wt%. Sharp increases in moduli, attributable to double-helix formation, were observed during cooling,
and moved to progressively higher temperature as the concentration of starch was increased (from 10 to 40 wt%). At all concentrations,
however, G 00 on reaching 58C was higher than G 0 , indicating that intermolecular interactions at this stage are largely topological. Formation
of true gel networks G 0 q G 00 was observed during holding at 58C, and is attributed to intermolecular helixhelix aggregation. The
holding time at the onset of gelation decreased as the concentration (c) of starch was increased, and at concentrations in the range ,20 to
,30 wt% showed the c 22-dependence expected for a simple dimerisation process. At higher concentrations the slope became steeper,
consistent with some limited aggregation during cooling. In a second series of experiments, where c was held fixed at 40 wt% and sucrose
concentration was varied (between 0 and 25 wt%), the time-course of gelation at 58C showed little change with increasing sucrose content,
but there were large increases in the moduli attained on completion of cooling. Measurement of the same samples by differential scanning
calorimetry showed that the extent of ordering during cooling varied from ,2 to ,15% of the helix content in the ungelatinised granules as
the sucrose concentration was increase from 0 to 25 wt%. Replacement of sucrose by glucose or fructose decreased the rate of conformational
ordering during cooling, in the sequence: fructose , glucose , sucrose; but on holding at 58C the order was reversed, with fructose causing
rapid gelation and sucrose having least effect. A similar inverse correlation between ordering and aggregation has been observed for other
biopolymer systems, and is tentatively ascribed to stability of individual helices inhibiting slight changes in conformation required for
efficient packing. q 2000 Elsevier Science Ltd. All rights reserved.
Keywords: Gelation; Oxidised starch; Conformational ordering; Polysaccharide aggregation
1. Introduction
Conformational ordering and intermolecular association
of biopolymers can often be promoted by introduction of
sugars and related polyols as cosolute. This phenomenon
can be explained, in part, by the associated reduction in
water content. Water is a particularly effective solvent for
biopolymers, because of its ability to form hydrogen bonds
with hydroxyl groups and other polar substituents of the
polymer chains. Partial replacement by a material with
less capacity for hydrogen bonding would therefore be
expected to promote polymerpolymer association by reducing competition from polymersolvent interactions.
Different cosolutes, however, can differ substantially in
the magnitude of the changes they produce, although their
order of effectiveness with different biopolymers is often the
* Corresponding author. Tel.: 1353-21-903625; fax: 1353-21-270001.
E-mail address: ed.morris@ucc.ie (E.R. Morris).
0144-8617/00/$ - see front matter q 2000 Elsevier Science Ltd. All rights reserved.
PII: S0144-861 7(99)00158-7
262
Fig. 1. Changes in (a) G 0 and (b) G 00 (1 rad s 21; 0.5% strain) during cooling
from 95 to 58C at l8C/min for mixtures of oxidised starch with sucrose at a
combined concentration of 65 wt%, illustrated for starch concentrations
(wt%) of 10 (S), 15 (V), 20 (W), 22.5 (X), 25 (K), 30 (O), 35 (A) and
40 (B).
3. Results
3.1. Rheology at 65 wt% total solids
Fig. 1 shows the changes in G 0 (Fig. 1a) and G 00 (Fig. 1b)
observed during cooling from 95 to 58C for mixtures of
oxidised starch with sucrose at a combined concentration
of 65 wt% (i.e. with water content held constant at 35 wt%).
The starch concentrations used range from 10 to 40 wt%
(with 5525 wt% sucrose). The effect of starch concentration on the values of G 0 and G 00 attained on completion of
cooling is shown in Fig. 2a. Both figures have been
263
presented previously in an accompanying paper on co-gelation with pectin (Evageliou et al., 2000b), where they were
used for direct comparison with the corresponding mixed
systems; Fig. 1, however, includes traces for two additional
starch concentrations (22.5 and 35 wt%) which were not
shown in the previous paper. The changes in moduli for
the same preparations during holding for 500 min at 58C
are illustrated in Fig. 3, and the effect of starch concentration on the final values on G 0 and G 00 recorded at the end of
the holding period are shown in Fig. 2b.
As shown in Fig. 1a, the samples all have significant
elastic character (G 0 ) at the loading temperature of 958C,
which, as will be demonstrated later by results from DSC, is
well above the melting temperature for amylopectin double
helices, even in the presence of high concentrations of
sucrose. The elastic response must therefore come from
physical entaglement (interpenetration) of neighbouring
molecules. In the initial stages of cooling, most of the
samples show a slight reduction in G 0 , which can be
explained by reduction in hydrodynamic volume as the
temperature is decreased (Yamakawa, 1971), with consequent reduction in the degree of entanglement. At the highest starch concentrations, however, this initial reduction is
swamped by a large increase in G 0 , attributable to the onset
of double-helix formation. The increase in G 0 is accompanied by a corresponding increase in G 00 (Fig. 1b), and both
are displaced to progressively lower temperature as the
concentration of oxidised starch is decreased.
On completion of cooling to 58C (at 18/min), the viscoelastic response (Fig. 2a) at the lower end of the range of
starch concentrations studied is predominantly solution-like
G 0 q G 00 : At higher concentrations the moduli converge,
but G 00 remains slightly higher than G 0 up to the highest
concentration of oxidised starch used (40 wt%, in combination with 25 wt% sucrose). It seems likely, therefore, that
the intermolecular interactions on completion of cooling are
still largely topological, and that the increases in moduli
(Fig. 1) result from association of flexible, disordered
strands into stiff, extended helices, making it more difficult
for individual molecules to move through the surrounding
matrix of neighbouring chains. After holding for 500 min at
58C (Fig. 2b), the samples prepared at starch concentrations
below ,20 wt% log c < 1:3 remain predominantly
viscous G 00 . G 0 ; but at higher concentrations G 0 rises
steeply above G 00 , demonstrating formation of an extensively crosslinked network during the holding period at 58C.
As found during cooling (Fig. 1a), the changes in G 0 on
holding at 58C (Fig. 3a) occur in two stages: an initial slight
reduction followed by a sharp increase, which is displaced
to progressively longer times with decreasing concentration
of oxidised starch and becomes undetectable at starch
concentrations below ,15 wt%. As discussed above, the
increase in elastic response can be attributed to intermolecular association by helixhelix aggregation. The
initial reduction may also be due to aggregation of helical
sequences, but within the same molecule, thus reducing the
264
Fig. 3. Changes in (a) G 0 and (b) G 00 (1 rad s 21; 0.5% strain) during holding
at 58C after cooling from 958C at 18C/min for mixtures of oxidised starch
with sucrose at a combined concentration of 65 wt%, illustrated for starch
concentrations (wt%) of 20 (W), 22.5 (X), 25 (K), 30 (O), 35 (A) and 40 (B).
Fig. 4. Changes in tan d (1 rad s 21; 0.5% strain) during holding at 58C after
cooling from 958C at 18C/min for mixtures of oxidised starch with sucrose
at a combined concentration of 65 wt%, illustrated for starch concentrations
(wt%) of 20 (W), 22.5 (X), 25 (K), 30 (O), 35 (A) and 40 (B).
265
27.5 wt% starch (with 42.5 and 37.5 wt% sucrose, respectively), but are omitted for clarity. The intercepts with the
axis at dlog G 0 =dt 0; which define the transition from
decreasing to increasing values of G 0 , could be determined
with good precision (to within about ^5%). The time at 58C
required to reach this point will be denoted as t0 (onset time
for gel formation).
The variation of t0 with starch concentration (c) is shown
in Fig. 7a. There is a smooth, increasing progression to
higher values of t0 with decreasing concentration of starch.
Fig. 7b shows the same data plotted on logarithmic axes.
The decrease in log t0 with increasing log c is virtually
linear across the concentration range from 20 to ,30 wt%
starch, with a slope close to 22. At higher concentrations of
starch (35 and 40 wt%), the decrease in log t0 becomes
progressively steeper, which may reflect the increasing
content of ordered structure already present in these samples
at the start of the holding period (i.e. formed during the
cooling process shown in Fig. 1).
3.2. Effect of sucrose on rheology of 40 wt% oxidised starch
266
Fig. 6. Rate of change in log G 0 during holding at 58C (Fig. 3a) for mixtures
of oxidised starch with sucrose at a combined concentration of 65 wt%,
illustrated for starch concentrations (wt%) of 20 (W), 25 (K), 30 (O), 35 (A)
and 40 (B). The points of intersection with the horizontal axis at
dlog G 0 =dt 0 define the transition from decreasing to increasing values
of G 0 .
Fig. 8. Changes in (a) G 0 and (b) G 00 (1 rad s 21; 0.5% strain) during cooling
from 95 to 58C at 18C/min for 40 wt% oxidised starch in the presence of
sucrose at concentrations (wt%) of 0 (W), 5 (X), 10 (K), 15 (O), 20 (A) and
25 (B).
temperature over a period of ,25 min, weighed for balancing with the reference pan, loaded into the calorimeter,
cooled to 58C at 18C/min, and held at 58C for ,25 min to
allow the instrument to equilibrate. These conditions were
chosen to give roughly the same extent of structure formation as in the early stages of the holding period in the rheological experiments (Fig. 10). The samples were then
scanned to 988C at a heating rate of 0.58C/min, which represents a compromise between the conflicting aims of avoiding excessive thermal overshoot while minimising further
structure formation in the early stages of heating (where the
sample is at low temperature). The resulting traces are
shown in Fig. 11. All samples show a single, well-defined
endotherm, which increases in magnitude and extends to
progressively higher temperature as the concentration of
sucrose is raised. The variation of transition enthalpy
(DH) per gram of starch with increasing concentration of
267
268
Fig. 10. Changes in (a) G 0 and (b) G 00 (1 rad s 21; 0.5% strain) during
holding at 58C after cooling from 95 at 18C/min for 40 wt% oxidised starch
in the presence of sucrose at concentrations (wt%) of: 0 (W), 5 (X), 10 (K),
15 (O), 20 (A) and 25 (B).
269
4. Discussion
The comparisons shown in Figs. 2 and 9 of the values of
G 0 and G 00 recorded on completion of cooling to 58C, and
after holding at 58C for a further 500 min, strongly suggest
that the increases in moduli observed during cooling (Figs. 1
and 8) arise predominantly from expansion and stiffening of
individual molecules (with consequent enhancement of
topological interactions), and that formation of a true
crosslinked gel network by intermolecular helixhelix
aggregation occurs largely during the holding period at 58C.
For the samples prepared at 65 wt% total solids, the
holding time (to) at the onset of gelation for starch concentrations between ,20 and ,30 wt% follows (Fig. 7b)
the c 22-dependence expected for a simple dimerisation
process. At higher concentrations the slope becomes steeper,
but, as suggested previously, this probably reflects an
increase in the extent of intermolecular association that
occurs during cooling, with consequent reduction in gel
times at 58C.
A much steeper concentration-dependence of gel time
(,c 25.2) was observed in an investigation by Clark (1995),
using partially depolymerised amylopectin prepared by acid
hydrolysis and dissolved in water. The small content of
carboxyl groups in the oxidised starch sample used in the
present work is unlikely to be a significant factor. Introduction of increasing concentrations of sucrose with decreasing
270
Fig. 13. Gelatinisation endotherm (0.58C/min) for oxidised starch (30 wt%
in water).
Fig. 14. DSC endotherms (0.58C/min) for 30 wt% oxidised starch in combination with 35 wt% sucrose after holding at 58C for 30 min (W), 1.5 h (X),
4 h (A) and 6 h (B).
Fig. 15. Effect of holding time at 58C on the enthalpy (DH) of the endothermic transitions obtained (Fig. 14) for 30 wt% oxidised starch in combination with 35 wt% sucrose.
271
Fig. 16. Changes in (a) G 0 and (b) G 00 (1 rad s 21; 0.5% strain) during
cooling from 95 to 58C at 18C/min for 30 wt% oxidised starch in mixtures
with sucrose (K), glucose (B) or fructose (W) at a concentration of
36.84 wt%.
Fig. 17. Changes in (a) G 0 and (b) G 00 (1 rad s 21; 0.5% strain) during holding at 58C after cooling from 958C at 18C/min for 30 wt% oxidised starch in
mixtures with sucrose (K), glucose (B) or fructose (W) at a concentration of
36.84 wt%.
272