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Rheological Properties of Starch and Whey Protein Isolate Gels

C.W.P. Carvalho, C.I. Onwulata and P.M. Tomasula
Food Science and Technology International 2007; 13; 207
DOI: 10.1177/1082013207079897

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Rheological Properties of Starch and Whey Protein Isolate Gels

C.W.P. Carvalho2, C.I. Onwulata1* and P.M. Tomasula1

1Dairy Processing and Products Research Unit, Eastern Regional Research Center, Agricultural Research
Service, U.S. Department of Agriculture, 600 E. Mermaid Lane, Wyndmoor, PA 19038, USA
2Embrapa Food Technology, Av. das Américas, 29501, Guaratiba, 23020-470 Rio de Janeiro, Brazil

The paste viscosity of starches Amioca (~0.5% amylose), native corn (~27% amylose), Hylon VII (~70%
amylose), tapioca (~19% amylose), and their whey protein isolate (WPI; 50/50) mixtures at varying
solid concentration of 2.5, 5, 10, 15 and 20% were determined. At higher gel concentration (20%) during
the cooling cycle, WPI paste measured by rapid visco-analyser (RVA) showed a tremendous increase
in viscosity over that of the micro visco-amylograph (MVA), 5784 and 184 cP, respectively. RVA gels had
a more uniform network of coarse particles (~250–500 nm) than the fine structures (~50 nm) in those
of the MVA. Inclusion of WPI reduced the paste viscosity of all starches by at least 50%, except for
Hylon VII at 20% solid concentration. The strength of Hylon VII and corn starch gels was reduced by
WPI. In contrast, gels of WPI/tapioca (G⬘ ⫽ 45.4 Pa) and WPI/amioca (G⬘ ⫽ 18.3 Pa) had similar rheolog-
ical properties as their pure starch control (G⬘ ⫽ 47.4 Pa and G⬘ ⫽ 15.3 Pa for Tapioca and Amioca,
respectively).

Key Words: whey protein, starch, viscosity, gel, rheology

INTRODUCTION the two polysaccharides vary according to the botanical

The growing research interest in polysaccharide-
protein systems is due to their numerous applications,
as gelling agents, thickeners, emulsifiers, texture modi-
fiers, stabilizers in the food, pharmaceutical, cosmetics,
textiles and in biomedical industries (Nishinari et al.,
2000). One polysaccharide-protein system of major
interest is the starch-whey system, featured in foods as
yoghurts, puddings, snacks, breads, pasta and breakfast
cereals (Marshall and Chrastil, 1992). They are the main
components of many industrially processed foods that
improve their texture and taste properties. Whey pro-
teins are widely used as food ingredients, because they
are highly nutritious and have useful functional proper-
ties; their gelling property is considered one of the most
important (Vardhanabhuti et al., 2001).
Starch granules are composed of two types of ␣-
glucans, amylose and amylopectin, which represent
approximately 98–99% of the dry weight. The ratios of
*To whom correspondence should be sent
(email: Charles.Onwulata@ars.usda.gov),
Received 17 May 2006; revised 21 September 2006.
Food Sci Tech Int 2007; 13(3):207–216
© SAGE Publications 2007
Los Angeles, London, New Delhi and Singapore
ISSN: 1082-0132
DOI: 10.1177/10820132013207079897
origin of the starch (Tester et al., 2004). ‘Waxy’ starches
contain less than 15% amylose, an example is Amioca™.
‘Normal’ starch contain between 17–25%, examples are
tapioca starch (19%) and corn starch (27%) (Taggart,
2004) and ‘high’ (amylo-)amylose corn starch has greater
than about 40% amylose, an example is Hylon® VII,
which contains about 70% amylose. In a recent work, it
was indicated that amylose plays an important role in the
formation of complexes with protein and lipids by
increasing the viscosity of the mixture at the cooling
paste viscosity stage (Zhang and Hamaker, 2003).
Paste viscosity is one of the fundamental functional
characteristic of starch during heating of starch–water
suspensions and it is considered the basis for food appli-
cation (Meares et al., 2004), it also reveals the extent of
interaction among a mixture of food components (Zhang
and Hamaker, 2003). It is used, for instance, in identifying
starchy crop varieties (Meares et al., 2004), for examining
starch functionality changes upon processing (Zhang and
Hamaker, 2003) and monitoring physico-chemical modi-
fications of starches (Sriburi and Hill, 2000).
There are a few studies of mixture of whey protein
and starch sources such as corn starch (Shim and
Mulvaney, 2001), cassava starch (Aguilera and Rojas,
1996) and cross-liked waxy maize starch (Ravindra et al.,
2004). The use of simple techniques such as with micro
visco-amylograph (MVA) and rapid visco-analyser
(RVA) has contributed greatly to enhance food quality

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208 C.W.P. CARVALHO ET AL.
(Suh and Jane, 2003). MVA and RVA paste viscosity
analysers are excellent low cost tools used to follow
important physical changes during thermal events, which
is common in food preparation. Paste viscosity values are
used to determine quality consistency in raw materials or
processed food systems. Additionally, the effects of addi-
tives/ingredients on protein-starch interactions can also
be well monitored using paste viscosity profiles generated
from instruments like MVA and RVA.
In the past, RVA has been used to produce gels for
determining fundamental rheology of starch/whey sys-
tems (Shim and Mulvaney, 2001). The aim of this work
was to evaluate the paste viscosity of several starch
sources containing different levels of amylose and their
mixture with whey protein isolate using MVA and
RVA, and to characterize the rheology and microstruc-
ture of gels formed at the end of the thermal analysis
conducted using these instruments.
MATERIAL AND METHODS
Whey protein isolate (WPI) (Provon 190, ~90% pro-
tein) was purchased from Glanbia Foods (Richfield,
ID). Native corn starch (Pure-dent B700, ~27% amy-
lose) was donated by Grain Processing Corporation
(Muscatine, IA). Native tapioca starch (~19% amylose),
amioca™ (~98% amylopectin) and Hylon® VII (~70%
amylose) were donated by National Starch Food
Innovation (Bridgewater, NJ).
Gel Preparation
Gels of different solids concentration, 2.5, 5, 10, 15
and 20% of starch and WPI (50:50) were obtained by
collecting the material at the end of the heat-cool cycle
of the visco-analyzers, MVA and RVA. Moisture content
of each sample was used to calculate the weight of the
powder needed for analyses using the Micro
ViscoAmyloGraph (MVA) (C.W. Brabender Instru-
ments, South Hackensack, NJ, USA) or the Rapid
ViscoAnalyzer (RVA-4) (Newport Scientific,
Warriewood, Australia). To ensure variable solid-to-
liquid ratio for sample ranging from 2.5 to 20%, based
on the moisture content of the solids (fixed at 14%, wet
basis), the total weight of the samples were kept con-
stant, 100 g and 28 g for MVA and RVA, respectively.
Starch, WPI powders and deionized water (pH ~5.8)
weighed to ⫾0.001 g were mixed. Readings of pH,
before and after gel preparation ranged from 4.4 to 6.3
and it was not controlled during gel preparation. For
WPI/Starch (50:50) mixtures, a low speed mixer
(~100 rpm, maximum velocity) fitted with a plastic
paddle was used to create a homogenous paste prior to
gel preparation. It was found that mixing in the visco-
analysers was not adequate to obtain uniform paste, par-
ticularly, when whey protein was present. If the paddle
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mixer device was not used, whey protein tended to
phase separate on the top of the final gel.
Methods
MVA Measurement
Depending on the concentration and moisture,
2.7–22.3 g aliquots of pure starch or mixed starch and
WPI samples (14% moisture, wet basis) were trans-
ferred into a stainless steel container and then stirred at
300 rpm rotational speed. The temperature profile used
was: hold at 50°C for 3.5 min, ramp to 95°C at
7.5°C/min, hold at 95°C for 3 min, cool back to 50°C at
7.5°C/min, hold at 50°C for 1 min. Each analysis took
approximately 21 min and was done in duplicate. RVA
analysis required lower heating time ratio (7.5°C/min)
than MVA (11.3°C/min) because of the design (MVA
larger sample container made of stainless steel, approx-
imately three times larger than the aluminum canister
used in the RVA). As soon as the software indicated
that the test ended, the samples were transferred into
air tight plastic containers and stored at ~5°C for
approximately 18 h before the dynamic oscillatory tests.
Previously, the results of MVA and RVA could not
be compared because of differences in the unit of meas-
urement (Suh and Jane, 2003). In this study, viscosity
measurements were compared because MVA viscosity
reading (mPas) was converted to units of cP.
RVA Measurement
The paste viscosity in cP was measured and recorded
with the software Thermoclyne for Windows version 2.2.
The temperature profile chosen is the approved method
ICC Standard No. 162 (Sriburi and Hill, 2000). The pro-
file was: hold at 50°C for 1 min, ramp to 95°C at
~12°C/min, hold at 95°C for 2 min and 40 s, cool back to
50°C at ~12°C/min, hold at 50°C for 2 min at rotation
speed of 160 rpm. Each analysis took 13 min and was
done in duplicate. Immediately after analysis, the RVA
paddle was carefully removed from the sample con-
tainer and covered with Parafilm (American National
Can, Chicago, IL) and stored at ~5°C for approximately
18 h before the dynamic oscillatory tests.
The paste viscosity responses of both MVA and RVA
curves were: onset paste temperature (initial increase of
viscosity), peak (maximum viscosity during heating),
breakdown (difference between the peak viscosity and
the lowest viscosity after heating ramp) and setback
(difference between the maximum viscosity during
cooling and the lowest viscosity after the heating ramp).
Dynamic Oscillatory Measurement
The samples, stored in the refrigerator overnight at
~5°C, were kept in their original containers and left at
 Rheology of Starch and Whey Protein Isolate Gels
room temperature (~20°C) to equilibrate for 2 h prior
to analysis. The rheological studies were carried out at
room temperature (20 ⫾ 1.5°C) using a controlled
stress AR-2000 rheometer (TA Instruments, New
Castle, DE, USA) with parallel plates. According to
their consistency, the gels were either pipetted (liquid-
like) or cut slice into a disk measuring 25 mm in diame-
ter and 2.5–5 mm thick. Strain sweep tests were per-
formed to determine the linear viscoelastic range of the
sample and 0.4% strain was deemed appropriate in
order to assure preservation of the gel structure.
Frequency sweeps were then run from 0.1 to 100 rad/s.
Values of elastic or storage modulus (G⬘) and viscous or
loss modulus (G⬙) were obtained at 10 rad/s.
Scanning Electron Microscopy
Depending on the gel consistency, the samples were
either transferred into dialysis tubing (weak gel) using a
syringe or cut into pieces with a stainless steel razor blade.
Both types of samples were immersed in a solution of
2.5% glutaraldehyde-0.1 M imidazole buffer (pH 7.0)
overnight and further processed by freeze-fracture
(Brooks et al., 1975). The samples were dehydrated in a
graded ethanol series (50, 70, 90 and 100%) and frozen in
liquid nitrogen.After 5 min of freezing, samples were frac-
tured manually with the edge of a cooled scalpel blade.
Sample fragments were thawed in absolute ethanol and
critical-point dried from liquid carbon dioxide. Dry frag-
ments were mounted on aluminum specimen stubs with
colloidal silver adhesive (Electron Microscopy Sciences,
Ft. Washington, PA, USA) and coated with a thin layer of
gold by dc sputtering. Images of topographical features in
fracture faces were made using a model Quanta F200
(FEI Company, Hillsboro, OR, USA) scanning electron
microscope at an accelerating voltage of 10 kV.
209
RESULTS AND DISCUSSION
Paste Viscosity of Starches and WPI/Starch Mixes
The responses of the WPI and starches and
WPI/starch mixtures (10% total solid concentration) to
heating are presented in Table 1. At this concentration
the paste viscosity profiles of the starches were smooth
curves without sharp peaks. Above 15% total solid con-
centration the profiles were uneven curves (data not
shown). This was the case, except for Hylon VII that
showed very low viscosity throughout. Corn starch, dur-
ing the cooling stage, showed inconsistent viscosity
response as it tended to form stronger and more brittle
gel structure that was broken and re-formed many times
during the continuous mixing imposed by the visco-
analysers. The paste viscosities of all RVA readings were
higher than MVA except setback values for tapioca and
WPI/Tapioca setback. Also, onset pasting temperatures
of RVA were generally 0–9°C higher except for that of
WPI/amioca and WPI/Tapioca which were similar. Suh
and Jane (2003) comparing MVA and RVA paste profile
of several starches also found that RVA values were
higher than MVA for pasting temperature of corn and
Amioca. These general findings were attributed to the
differences in the mixing mechanism of the visco-analy-
sers (Suh and Jane, 2003). Although the two instruments
are designed differently, they measure similar responses.
Trends determined generally hold true for both instru-
ments, only the magnitude of the values differ. As MVA
and RVA showed a similar trend in paste viscosity read-
ings, only the RVA data are discussed in depth.
Tapioca and amioca starches showed the highest value
of peak viscosity (gelatinization), 4891 and 4151 cP,
respectively, followed relatively close by corn, 3206 cP
(Table 1). Starch gelatinization is a complex phenomenon

Table 1. Paste behaviours of starches and WPI/starch mixtures in MVA and RVA at 10% concentration.
Pasting Property

MVA Viscosity (cP) RVA Viscosity (cP)

Peak2 Break Down Setback Onset Peak2 Break Down Setback Onset
Pasting Pasting
Product1 T (°C) T (°C)

WPI 27.0 ⫾ 0 — — 72.8 ⫾ 0.1 86.5 ⫾ 36 — — 77.5 ⫾ 0.1

39.0 ⫾ 0 — — — 162.0 ⫾ 43 — —
Tapioca 1333.0 ⫾ 15.6 605.5 ⫾ 10.6 894.5 ⫾ 21.9 66.2 ⫾ 0.4 4891.0 ⫾ 99.0 2983.5 ⫾ 31.8 811.5 ⫾ 13.4 68.6 ⫾ 0.6
WPI/tapioca 475.0 ⫾ 39.6 182.0 ⫾ 25.5 356.0 ⫾ 24.0 68.1 ⫾ 0.4 1188.5 ⫾ 34.6 351.0 ⫾ 22.6 359.5 ⫾ 19.1 68.3 ⫾ 0.6
Amioca 1100.5 ⫾ 3.5 502.0 ⫾ 4.2 211.0 ⫾ 1.4 67.6 ⫾ 0.2 4151.0 ⫾ 103.2 2406.5 ⫾ 187.4 604.0 ⫾ 91.9 70.2 ⫾ 0.0
WPI/amioca 535.0 ⫾ 7.1 208.5 ⫾ 4.9 94.0 ⫾ 5.7 69.3 ⫾ 0.4 1976.0 ⫾ 31.1 1006.0 ⫾ 36.8 333.5 ⫾ 20.5 69.0 ⫾ 0.6
Corn 965.5 ⫾ 2.1 247.0 ⫾ 5.7 335.5 ⫾ 0.7 69.9 ⫾ 0.4 3206.5 ⫾ 40.3 994.0 ⫾ 65.1 1511.5 ⫾ 31.8 72.3 ⫾ 0.6
WPI/corn 259.0 ⫾ 3.6 3.0 ⫾ 4.4 47.3 ⫾ 6.4 70.9 ⫾ 0.1 797.7 ⫾ 14.5 12.0 ⫾ 3.6 261.3 ⫾ 10.1 76.5 ⫾ 0.9
Hylon VII 28.0 ⫾ 1.4 3.0 ⫾ 3.0 4.0 ⫾ 0.0 85.9 ⫾ 0.8 46.0 ⫾ 8.5 20.5 ⫾ 9.2 20.5 ⫾ 2.1 94.8 ⫾ 0.4
WPI/Hylon 45.5 ⫾ 2.1 — 32.0 ⫾ 0.0 74.7 ⫾ 0.1 134.5 ⫾ 0.7 15.0 ⫾ 7.1 127.5 ⫾ 13.4 77.5 ⫾ 0.1
1The amylose contents of tapioca, amioca, corn and Hylon VII is approximately 19%, 0.5%, 27% and 70%, respectively.
2WPI paste viscosity showed two peaks, one at the heating stage and other at the final constant temperature cycle of 95°C.

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210 C.W.P. CARVALHO ET AL.
that occurs when the internal crystalline structure of the
starch granule is lost by heating in the presence of an
excess of water (Batey and Curtin, 2000). The visco-
analyser peak viscosity is also known as gelatinisation
peak (Becker et al., 2001); as higher peak viscosity indi-
cates high amylopectin content (crystalline structure).
Mixing WPI with starches, the highest decrease of peak
viscosity was in tapioca, followed by corn and amioca; in
contrast, it increased in Hylon VII, showing more of an
additive effect. Breakdown, an indication of how easily
the granular structure of starch breaks (Varavinit et al.,
2003), was higher in tapioca, followed by amioca and corn,
2983, 2406 and 994 cP, respectively. Mixtures of starch and
WPI showed lower breakdown than pure starch; amioca
and tapioca starches were the most affected. The effect of
whey protein could be attributed to a plasticizing
response that prevented the starch granule from swelling,
maintaining gelation even during the cooling cycle
(Vardhanabhuti et al., 2001). As expected, setback viscos-
ity, which is associated with amylose content and the
degree of retrogradation of starch pastes was found
higher for corn starch. Setback reading, which is also asso-
ciated with the magnitude of peak viscosity, was highest
for Hylon VII, followed by corn. In mixtures with WPI,
the setback values were also reduced, except for Hylon
VII. The relative decrease of setback viscosity was in this
order: Hylon VII, corn, tapioca and amioca, this suggestes
a synergetic behaviour between amylose retrogradation
and protein aggregation in the cooling cycle. These results
are in concordance with the amylose content of the
starches Hylon VII (~70%), corn (~27%), tapioca
(~21%) and amioca (~0.5%).
The paste viscosity of Hylon VII at 10% concentration
showed the highest onset temperature and the lowest
peak viscosity of the starches (Table 1). This also was
observed at 15 and 20% concentrations (Figure 1). The
mixing of Hylon VII with WPI contributed to increased
paste viscosity which could be explained by the lower vis-
cosity of Hylon VII at 10% concentration.As the concen-
tration increased to 20% (B), WPI decreased the viscos-
ity of the system; at 15% concentration, the viscosity of
mixture seemed very much controlled by WPI gelation,
particularly during the cooling stage. The onset pasting
temperature of WPI/Hylon VII (74.7°C, MVA) was initi-
ated by the denaturation of WPI (72.8°C, MVA), since
the onset temperature of Hylon VII is much higher
(85.9°C). Richardson et al. (2000) reported that amylose
may act as a restraint to swelling. The low gelatinisation
peak and high onset temperature of Hylon VII could be
explained not only by the high amylose content, but also
by the overall reduced level of amylopectin, the starch
component mainly responsible for its granular structure.
Paste Viscosity of WPI
The paste viscosity curve of WPI analysed with MVA
was similar to a very dilute starch system (data not
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Figure 1. Effect of WPI and concentration on RVA
profile. (A) 20% Hylon VII, (B) 20% WPI/Hylon VII, (C)
15% WPI/Hylon VII, (D) 15% Hylon VII.
shown). For example, the maximum paste viscosity of
low tapioca starch concentration of 2.5% (82.5 cP) dur-
ing cooling stage would match the viscosity profile of
WPI at 10% (84.0 cP). Although neither the RVA or
MVA viscosity profile of WPI has been found in the lit-
erature, the dynamic scanning calorimetry (DSC)
curves of tapioca starch and WPI reported by Aguilera
and Rojas (1996) reveal that the denaturation peak of
pure WPI in excess water at pH 6.0, occurs few degrees
higher (74.5°C) than the gelatinisation peak of tapioca
starch (66.0°C). Also reasonable correlation was found
between DSC curves and RVA viscosity curves (Qian
and Kuhn, 1999). In this work, the pasting onset temper-
ature of WPI was 72.8°C (MVA) and 77.5°C (RVA).
These values are very close to the onset temperatures
reported by Aguilera and Rojas (1996), believed to be
the initial denaturation steps. As for tapioca starch, the
MVA onset temperature was 66.2°C (Table 1).
A dramatic increase in paste viscosity during cooling
to 50°C and a slight decrease while holding at 50°C was
observed for WPI pastes (Figure 2). The formation of a
secondary peak/bump after initial increase of viscosity
which coincides with the end of holding cycle at 95°C,
just before the cooling cycle, can be seen in Figure 2.
Similar behaviour was observed earlier with the viscos-
ity and gelation curves of heat-induced WPI paste stud-
ied using a rheometer fitted with a cup and bob attach-
ment (Vardhanabhuti et al., 2001; Havea et al., 2004).
Both RVA and MVA paste viscosity curves of whey
protein suggest a two-stage mechanism of gelation with
primary and secondary thermally induced peaks
(Figure 2). Similar finding was reported for thermally
inducing unfolding of native protein (Clark et al., 2001).
As the concentration of WPI increased, a tremen-
dous increase in viscosity was observed particularly at
the final cooling stage (note the magnitude of the
 Rheology of Starch and Whey Protein Isolate Gels 211

(a) 180 100 (b) 6000 100

90 5500 90
160 20%
5000
80 80
140 4500
70 70

Temperature (°C)

Temperature (°C)
120 Secondary peak 4000

Viscosity (cP)
Viscosity (cP)

60 3500 60
100 20%
50 3000 50
80 15% 2500 15%
40 40
60 2000 Secondary peak
10% 30 30
1500
40 20 20
5% 1000 10%
20 10 500 10
2.5%
0 0 0 0
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14
Time (min) Time (min)

Figure 2. MVA and RVA paste viscosity of varied concentrations of WPI alone. (a) MVA paste profile. (b) RVA
paste profile (2.5 and 5% concentrations were suppressed in this plot due to their small magnitude).

viscosity of the RVA profile in Figure 2b). Even at 20% Apart from Hylon VII, RVA produced gels were
concentration this was not observed in the MVA paste more rigid than MVA as shown by the higher values of
curves. This can be attributed to differences in design G⬘ and complex viscosity (␩*) (Table 2). The G⬘ of pure
(Lai et al., 2000). Using the Brabender Visco- WPI gels produced in the RVA was 10-fold greater
Amylograph, which uses a similar principle as the than MVA gels at 10% of concentration. As seen in
MVA, Thiewes and Steeneken (1997) reported a lower Figure 3, as the solids concentration increases, RVA
breakdown viscosity for native starch samples than that gels show a larger increase in G⬘ above 10% and a
of RVA. The microstructure of the WPI gels resulting plateau at 20%. In contrast, MVA gels showed a grad-
from the two visco-analysers (MVA and RVA) show ual and progressive increase in G⬘. Large G⬘ values for
distinct features that illustrate the differences in their RVA gels indicate that its mixing mechanism may has-
design and mode of analysis. ten the process of network formation. High gel strength
values observed for Hylon VII and native corn may be
Dynamic Rheological Data related to the retrogradation of amylose. Lionetto et al.
(2005) reported similar increases in gel strength for ret-
The results of dynamic rheological studies of 10% rograded starch. Although, the amylose content of tapi-
gels measured at 10 rad/s are presented in Table 2. As oca is slightly less than corn, its elastic modulus (G⬘)
recorded, G⬘ is the elastic modulus or storage modulus, values were low similar to that of Amioca, which has
G⬙ is the loss modulus in simple shear deformation, ␩* much lower amylose content. Adding WPI to tapioca
is the complex viscosity and tan ␦ is the ratio between starch did not substantially reduce G⬘, as happened
G⬙ and G⬘. with corn and Hylon VII starches, but addition of WPI

Table 2. Responses of dynamic rheological measurement of 10% gels of WPI, starches and WPI/starches from
MVA and RVA at frequency of 10 rad/s.
MVA RVA

Product1 G⬘ G⬙ ␩* Tan ␦ G⬘ G⬙ ␩* Tan ␦

WPI 133.1 ⫾ 50.6 37.9 ⫾ 17.4 13.8 ⫾ 5.3 0.3 ⫾ 0.0 1468.0 ⫾ 258.6 243.1 ⫾ 37.1 148.8 ⫾ 26.1 0.2 ⫾ 0.0
Tapioca 47.4 ⫾ 1.2 21.9 ⫾ 0.7 5.2 ⫾ 0.1 0.5 ⫾ 0.0 95.2 ⫾ 10.8 32.6 ⫾ 7.9 10.1 ⫾ 1.2 0.3 ⫾ 0.0
WPI/tapioca 45.4 ⫾ 6.8 15.0 ⫾ 0.7 4.8 ⫾ 0.7 0.3 ⫾ 0.0 68.7 ⫾ 13.4 18.9 ⫾ 2.3 7.1 ⫾ 1.4 0.3 ⫾ 0.0
Amioca 15.3 ⫾ 0.3 10.1 ⫾ 0.4 1.8 ⫾ 0.0 0.6 ⫾ 0.0 18.2 ⫾ 0.1 10.1 ⫾ 0.3 2.1 ⫾ 0.0 0.6 ⫾ 0.0
WPI/amioca 18.3 ⫾ 2.8 10.1 ⫾ 1.1 2.9 ⫾ 0.3 0.5 ⫾ 0.0 33.4 ⫾ 1.8 14.0 ⫾ 0.6 3.6 ⫾ 0.2 0.4 ⫾ 0.0
Corn 1539.2 ⫾ 237.1 59.2 ⫾ 13.9 154.0 ⫾ 23.7 0.0 ⫾ 0.0 1894.3 ⫾ 194.5 3.6 ⫾ 8.4 189.6 ⫾ 19.5 0.0 ⫾ 0.0
WPI/corn 200.7 ⫾ 12.2 34.8 ⫾ 2.2 20.4 ⫾ 1.2 0.2 ⫾ 0.0 340.8 ⫾ 38.8 37.8 ⫾ 4.3 34.3 ⫾ 3.9 0.3 ⫾ 0.0
Hylon VII 2257.7 ⫾ 112.5 129.9 ⫾ 8.2 226.1 ⫾ 11.3 0.0 ⫾ 0.0 1537.0 ⫾ 403.8 74.8 ⫾ 18.7 153.8 ⫾ 10.4 0.3 ⫾ 0.1
WPI/Hylon VII 319.8 ⫾ 48.8 49.5 ⫾ 4.0 32.4 ⫾ 4.9 0.2 ⫾ 00 528.5 ⫾ 90.7 75.4 ⫾ 9.2 53.4 ⫾ 9.1 0.1 ⫾ 0.0
1The amylose contents of tapioca, amioca, corn and Hylon VII is approximately 19%, 0.5%, 27% and 70%, respectively.

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212 C.W.P. CARVALHO ET AL.
100000 (a) 100000
10000
10000
1000

Log G′ (Pa)
Log G′ (Pa)

1000 100

10
100
1

10 0.1
0 5 10 15 20 25

1 Concentration (%)
0 5 10 15 20 25
Concentration (%) (b)
100000

Figure 3. Effect of concentration and visco-analyser 10000

type on the rheological property of heat-induced WPI
1000

Log G′ (Pa)
gel at 10 rad/s. (䉱) WPI from MV. (䊏) WPI from RVA.
100

approximately doubled the elastic component of
Amioca, which is very rich in amylopectin.
In the particular case of high amylose starch, Hylon
VII, the rigid component, G⬘ was the highest amongst
the gels, although its paste viscosity was the lowest. This
is due to amylose rearrangement also called retrograda-
tion (Lionetto et al., 2005); our gels were stored
overnight at 5°C allowing further interaction which may
have formed reinforced entanglements of adjacent
amylose molecules. Similar mechanism for the forma-
tion of strong gels has been reported by McGrane et al.
(2004). Substantial increases in the rigid component
(G⬘) of the gels (at frequency of 10 rad/s) as concentra-
tion increased can be seen in Figure 4. Corn starch and
Hylon VII (above 5%) had the highest values whereas
amioca and tapioca had the lowest, one order of magni-
tude lower. Lai and Liao (2002) reported a lower G⬘ for
tapioca gel compared to corn and wheat, and this was
attributed to their low amylose content. Low G⬘ values
for amioca can be explained by its low amylose content,
but tapioca has substantially high amylose however it
behaves similar to amioca.
The effect of frequency on the elastic modulus G⬘ of
WPI and WPI/Starch mixtures at 10% concentration is
shown in Figure 5. For corn and Hylon VII, which
showed the highest rigid component value G⬘, there is no
frequency dependence up to a frequency of 10 rad/s, a
typical behaviour for true gels (McGrane et al., 2004).
This is also in agreement with the work of Yang et al.
(2004). However, at higher frequencies, particularly for
amioca and tapioca gels, the frequency dependence
affected their rheological property to a higher degree.
Shim and Mulvaney (2001) showed independence of
complex modulus (G⬘) particularly at high frequencies
for starch and water systems resulting in stronger gels
characterised as an entangled polymer system in the
plateau region. But,Aguilera and Rojas (1996) suggested
that WPI functions as inactive filler in starch matrices
Downloaded from http://fst.sagepub.com at RMIT UNIVERSITY on March 30, 2010
10
1
0.1
0 5 10 15 20 25
Concentration (%)
Figure 4. Effect of concentration on the MVA elastic
component (G⬘) at 10 rad/s (a) and RVA (b) of gels.
(䉭) Corn; (䉱) WPI/corn; (䊐) Hylon VII; (䊏) WPI/Hylon
VII; (䊊) tapioca; (䊉) WPI/tapioca; (䉫) amioca; (䉬)
WPI/amioca.
producing weaker gels. Weak gel structures of starch-
protein mixture can be seen in Figure 5 for Hylon VII
and corn starch mixed with WPI. Gels of tapioca and
amioca showed large dependence on frequency, but the
presence of WPI, in contrast, increased the elastic mod-
ules of amioca and MVA tapioca suggesting that WPI
acted as active filler. So, the effect of adding WPI
depends on the starch type.
Microstructure
The micrographs of pure WPI gels produced by
MVA and RVA are presented in Figure 6. WPI gels pro-
duced in the MVA formed a multiphasic structure with
two distinctive components, one smooth and the other
rough (Figure 6A). The interface between these two
structures is shown in Figure 6B. The smooth surface is
constituted of very fine structure distinguished only at
very high magnification (100,000 ⫻ ) (Figure 6C) as a
network of small ‘strings of beads’ or nanospherical
particles (~50 nm). These nanospherical units linked
to each other as fine strands are well described in the
work of Ikeda and Morris (2002). In Figure 6D, RVA
produced WPI gel showed a dominant structure of
larger clusters of protein aggregates of varying sizes,
 Rheology of Starch and Whey Protein Isolate Gels
(a) 10000
1000
100
G′ (Pa)
10
1
0.1
0.1 1 10
w (rad/s)
(b) 10000
1000
G′ (Pa)
100
10
1 gels. Tapioca starch developed a less opaque and weak
0.1 1 10
w (rad/s)
Figure 5. Effect of frequency (␻) on G⬘ for MVA (a)
and RVA (b) gels. (䉭) Corn; (䉱) WPI/corn; (䊐) Hylon
VII; (䊏) WPI/hylon VII; (䊊) tapioca; (䊉) WPI/tapioca;
(䉫) amioca; (䉬) WPI/amioca.
each unit ranging from ~250 to ~500 nm, and at least
five-fold larger than a unit of the fine strand protein net-
work in Figure 6C. These large clusters are the only
dominant structure of WPI gel produced in the RVA.
The large structures can be associated with the rheolog-
ical properties as correlating with higher elastic modu-
lus (G⬘) for WPI gels produced by RVA. Therefore, the
agglomerated protein structure of WPI gel produced in
the RVA (Figure 6D, E, F) is presumably mainly
responsible for the gel strength. The formation of this
more structured gel may be attributed to the mixing
mechanism of the RVA as well as the heating regime
used to produce the gels. The RVA mixing system is
characterized by spinning a pitched paddle impeller pri-
marily designed to keep starch slurries in suspension
(Lai et al., 2000) at a higher heating/cooling ratio of
11.3°C/min, whereas for MVA, the sample holder
rotates while a fixed square bar with a hollow structure
mixes the sample (Suh and Jane, 2003) at a lower heat-
ing/cooling ratio. Running WPI in the RVA at the same
temperature profile used in the MVA produced lower
gelation viscosity (data not shown). However, RVA
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213
WPI pastes were still much higher in viscosity during
the cooling cycle. It is possible that the gelation mecha-
nism is very much related to the mode of spinning the
sample and also the rate of heating/cooling.
SEM of pure tapioca and corn starches show very
distinct structures (Figure 7). Pure tapioca gel has a
smooth structure, well defined by the holes left where
air and water were entrapped in the original gel (Figure
7A). Absent are remnants of granular structure as seen
in the corn starch gels (Figure 7C), where micropores
predominantly appear inside fractured granular starch
100 structures. The presence of granular starch remnants
indicates that corn starch was more resistant to the ther-
mal-mechanical damage in the visco-analyser. In con-
trast, tapioca gels showed a more regular network with
no remnants of granular starch structure under the
tested conditions. It is possible that tapioca starch gela-
tinised at lower temperature (66.2°C) than corn
(69.9°C) and the onset temperatures can be linked to
early degradation in normal thermal shearing processes
(Carvalho and Mitchell, 2000).
The differences in the microstructure of tapioca and
corn were reflected in the viscoelastic properties of the
100
gel with low G⬘ and for corn starch a more opaque and
stronger gel with higher elastic component G⬘ (Table 2).
The presence of starch remnants in corn gels (Figure
7C) reinforced the gel structure. Acting as filler between
the retrograded amylose entanglements, a more com-
pact structure was formed which was much larger than
the WPI network (Figure 7B, D). Shim and Mulvaney
(2001) reported similar findings showing that increasing
temperature of WPI/corn gels resulted in higher disso-
lution of starch granules and formation of more exten-
sive amylose network with aggregated WPI sandwiched
between the starch granules. Aguilera and Rojas (1996)
reported that WPI reinforced tapioca gels supported
the highly fragile or disintegrated granules of tapioca.
They also hypothesised that swollen tapioca starch
granules would entrap more water after gelatinisation
which could also explain the large number of open
structures in dehydrated pure tapioca gels (Figure 7A).
As the concentration of swollen granules increases they
no longer occupy discrete spaces but become a predom-
inant phase interfering with the formation of a continu-
ous WPI network (Figure 7B).
The differences between the starches in relation to
their amylose content played an important role in
determining the rheological properties of the gels.
Addition of WPI influenced paste viscosity, gel forma-
tion and the final texture of gels. High amylose starch
gels, Hylon VII and corn, were affected more by the
presence of WPI which seemed to work as a plasticiser
by preventing molecular rearrangement of amylose
leading to reduced rigidity. In contrast, in very low amy-
lose starches, particularly amioca, WPI seemed to work
as filler increasing rigidity (G⬘).
214 C.W.P. CARVALHO ET AL.

Figure 6. Scanning electron microscopy of dehydrated gels of WPI at 15% concentration. (a) MVA gel at 2500⫻.
(b) MVA gel at 50,000⫻. (c) MVA gel at 100,000⫻. (d) RVA gel at 2,500⫻. (e) RVA gel at 25,000⫻. (f) RVA gel at
50,000⫻.

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 Rheology of Starch and Whey Protein Isolate Gels 215

Figure 7. Scanning electronic microscopy dehydrated gels. (a) Pure tapioca (20%). (b) WPI/tapioca (15%).
(c) Pure corn (20%). (d) WPI/corn (15%).

ACKNOWLEDGEMENTS Brooks S.E.H., Reynolds P., Audretsch J.J. and Haggis G.

The assistance of Dr Peter Cooke and Guoping Bao
with the microscopy is gratefully acknowledged.
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