International Dairy Journal 9 (1999) 537}545

Rheological properties of acid milk gels as a!ected by the nature

of the fat globule surface material and heat treatment of milk
Y.H. Cho, J.A. Lucey, H. Singh*
Institute of Food, Nutrition and Human Health, Massey University, Private Bag 11-222, Palmerston North, New Zealand
Received 16 February 1999; accepted 25 June 1999

Abstract

The e!ects of di!erent fat globule surface materials on the rheological properties of acid milk gels, formed by acidi"cation with
glucono-d-lactone at 303C, were investigated. Acid gels were prepared from recombined milks (&3.4% fat) containing fat globules
stabilized with low-heat, medium-heat or high-heat skim milk powder (SMP), sodium caseinate, whey protein concentrate (WPC),
heated WPC or Tween 60. Unheated reconstituted skim milk (URSM) or milk heated at 803C for 30 min (HRSM) was used in the
preparation of recombined milks. In URSM or HRSM systems, gels containing fat globules stabilized by non-interacting materials
(&structure breaker') (Tween and unheated WPC) had low storage moduli (G) compared with interacting materials (&structure
promoter') (SMP, sodium caseinate and heated WPC). HRSM systems had higher G values than URSM. Gels containing fat globules
stabilized by sodium caseinate or heated WPC had the higher G values than those containing SMP-stabilized fat globules in both
URSM and HRSM systems.  1999 Elsevier Science Ltd. All rights reserved.

Keywords: Fat globule membrane; Rheology; Acid milk gelation

1. Introduction bed at the fat globule surface in#uence the properties of

Fermented milk products are produced throughout
the world with yoghurt being one of the most popular.
The formation and properties of acid milk gels have been
recently reviewed (Lucey & Singh, 1997). Milk heat treat-
ment and homogenization are important processing vari-
ables in the production of fermented milk products, such
as yoghurt (Tamime & Robinson, 1985; Puhan, 1988;
Tamime & Marshall, 1997). In many parts of the world,
recombined milks are used to make fermented milk prod-
ucts. Recombined milk is produced when anhydrous
milk fat is emulsi"ed with an aqueous solution of skim
milk powder (SMP). During homogenization, the fat
surface area increases markedly and a new adsorbed
layer consisting of milk proteins (casein micelles, casein
sub-units and whey proteins) is formed around the fat
globules. The concentration and type of proteins adsor-
* Corresponding author. Tel.: #64-6-3504401; fax: #64-6-3504645.
E-mail address: hsingh@massey.ac.nz (H. Singh)
 Current address: Department of Food Science, University of Wis-
consin-Madison, 1605 Lindon Drive, Madison, WI53706-1565, USA.
the recombined milk.
Fundamental rheological studies on the e!ects of fat
globule surface material on acid milk gels have been
reported (van Vliet & Dentener-Kikkert, 1982; van Vliet,
1988). The nature of the surface material on fat globules
determines the types of interaction that can occur be-
tween fat globules and the protein matrix. Fat globules
may act as an inert "ller (&structure breaker') when the
native fat globule membrane is intact since it would not
interact with casein particles. However, the &arti"cial' fat
globule membranes (consisting of mainly caseins and
some serum proteins) in homogenized fresh milk would
interact positively (&structure promoter') with the protein
network (van Vliet & Dentener-Kikkert, 1982; van Vliet,
1988). However, little work has been reported on the
rheological properties of acid gels made from recombined
milk in which the fat globules were stabilized by a wide
range of materials, such as whey protein concentrates
(WPC), di!erent types of SMP and non-protein mate-
rials.
The e!ects of heat treatment on the rheological
and microstructural properties of acid skim milk gels
have been reported previously (van Vliet & Keetels,
1995; Lucey, Teo, Munro & Singh, 1997; Lucey, Teo,

0958-6946/99/$ - see front matter  1999 Elsevier Science Ltd. All rights reserved.
PII: S 0 9 5 8 - 6 9 4 6 ( 9 9 ) 0 0 1 2 3 - 5
538 Y.H. Cho et al. / International Dairy Journal 9 (1999) 537}545

Munro & Singh, 1998b; Lucey, Tamehana, Singh &
Munro, 1998c). Heat treatment of recombined milk had
a greater e!ect on the rheological properties of acid milk
gels than fat content (Lucey, Munro & Singh, 1998a).
Our objectives were to determine the e!ects of a combi-
nation of di!erent fat globule membrane materials and
heat treatment of the reconstituted skim milk on
rheological properties of acid milk gels.
2. Materials and methods
2.1. Materials
Low-, medium- and high-heat SMP were obtained
from the New Zealand Dairy Research Institute,
Palmerston North. The whey protein nitrogen indices of
these powders were 8.0, 2.7 and 1.5 mg g\ powder, re-
spectively. Spray-dried whey protein concentrate (WPC,
ALACEN 392), and sodium caseinate (ALANATE 180)
powders were obtained from the New Zealand Dairy
Board, (Wellington, New Zealand). Fresh frozen milk fat
for recombination (FFMR), which contained 99.95%
milk fat and 0.05% moisture, was also obtained from the
New Zealand Dairy Board. Tween威 60 (polyoxyethylene
sorbitanmonostearate) was purchased from BDH Chem-
icals (BDH Ltd., Poole, England). All the chemicals used
were of analytical grade obtained from either BDH
Chemicals (BDH Ltd., Poole, England) or Sigma Chem-
ical Co. (St. Louis, MO, USA) unless speci"ed otherwise.
Reconstituted low-heat SMP (RSM) was prepared by
dissolving &16 g of SMP in 100 g of distilled water. In
some cases RSM was heated (HRSM) to 803C in a pilot
scale UHT plant (Spiral #ow, indirect UHT plant, Alfa-
Laval, Australia) and held for 30 min by transferring the
samples to beakers, which were placed in a thermostati-
cally controlled water bath. The beakers were covered
with aluminium foil to prevent evaporation and stirred
occasionally. After heat treatment, milks were rapidly
cooled to 303C with agitation by immersion in ice water.
Recombined milks were prepared by mixing 364 g of
unheated RSM (URSM) or HRSM with 196 g of emul-
sion and stirring for 5 h at room temperature. The experi-
mental protocol used is shown in Fig. 1.
2.4. Characterization of emulsions
Samples of emulsions were diluted with demineralized
water or 2% (w/w) SDS bu!er (2% SDS, 0.05 M EDTA in
water) in the ratio of 1 : 3 sample to bu!er. After 30 min
mixing, the fat globule size distributions of the emulsions
were determined using a Malvern MasterSizer E (Mal-
vern Instruments Ltd, Malvern, Worcester, UK) as de-
scribed by Srinivasan, Singh and Munro (1996).

2.2. Preparation of protein solutions

Appropriate quantities of low-, medium- or high-heat

SMP, sodium caseinate or WPC were dissolved in de-
mineralized water to give a protein concentration of
&2.0% (w/w). Solutions were stirred for 5 h at &253C,
and 0.02% NaN was added. In some instances, WPC

solutions were heated at 803C for 30 min to denature the
whey proteins using a temperature-controlled water
bath.

2.3. Preparation of recombined milks

The protein solution was mixed with appropriate

quantities of FFMR, the mixture heated to 553C, and
then passed through a Rannie two-stage valve homogen-
izer (Model Lab, type 12.50H, DK-2620, Albertslund,
Denmark) without applying any pressure. This produced
a temporary oil-in-water emulsion. The mixture was then
homogenized at 20.7 and 3.5 MPa for the "rst and sec-
ond stages, respectively. The resulting emulsion was
cooled to room temperature and then used for the prep-
aration of recombined milk. Typically, these emulsions
contained&2.0% protein and 10% (w/w) milk fat. In one Fig. 1. Experimental protocol for the preparation of recombined milk
case, an emulsion was prepared using 0.5% Tween 60. systems.
 Y.H. Cho et al. / International Dairy Journal 9 (1999) 537}545
The amount of protein adsorbed onto the fat surface
was determined by measuring the protein content of the
washed cream layer after separation of the dispersed and
aqueous phases by centrifugation (Sharma, Singh
& Taylor, 1996; Srinivasan et al., 1996). The emulsion
samples were centrifuged at 20,000 g for 20 min at 203C
(Sorvall RC5C centrifuge, Dupont Company, Newton,
CT). The cream layer was carefully removed, dispersed in
distilled water for 1 h and re-centrifuged under the same
conditions. The protein load, as mg protein/m fat sur-
face area, was calculated from the protein present in the
washed cream layers and the fat surface area, calculated
from the average volume}surface particle diameter (d )

determined by the Mastersizer results (Sharma et al.,
1996).
2.5. Compositional analysis
Total protein was measured by determining total ni-
trogen by the macro-Kjeldahl method (AOAC, 1996a)
and multiplying by a factor of 6.38. The samples were
digested and distilled using a Kjeltec system (Tecator,
Sweden). Total fat was determined using the Rose-Got-
tlieb gravimetric method for milk (International Dairy
Federation, IDF 1C, 1987) and cream (IDF 16C, 1987).
Total solids content was obtained using an air-oven
method (AOAC, 1996b). The temperature of the oven
was kept at approximately 1053C for overnight drying of
samples.
2.6. Preparation of acid gels
Glucono-d-lactone (GDL) was used as an acid precur-
sor. Recombined milks were acidi"ed at the rate of 1.4%
(wt/vol) GDL at 303C as reported (Lucey et al., 1997).
The pH of the gels was &4.6 after incubation for 16 h.
2.7. Rheological measurements
Gel formation was monitored by dynamic measure-
ments using a constant-strain-controlled rheometer
(Bohlin VOR, Bohlin Reologi AB, Lund, Sweden) in an
oscillatory mode as described by Lucey et al. (1997). The
measuring system and test conditions were frequency
0.1 Hz, maximum strain (0.01 and coaxial cylinders (dia
25 and 27.5 mm). For unheated samples, the
0.46 mN m\ torsion bar was used while for the heated
samples the 4.29 mN m\ bar was used. On addition of
GDL to the milk the mixture was stirred for 2 min and
then 13 ml of the mixture was transferred to the
rheometer. Vegetable oil was added on the surface to
prevent evaporation. Gelation was de"ned as the point
when gels had a G of 51 Pa (Lucey et al., 1997). The loss
tangent (tan d) is the tangent of the loss angle (d) and is
the ratio of the loss modulus (G) to the storage modulus
(G), i.e. tan d"G/G. The e!ect of timescale on the
539
rheological properties was determined by varying the
frequency from 0.001 to 1 Hz. All experiments were done
in duplicate.
3. Results
3.1. Composition of emulsions
The fat globule size distributions of emulsions were
similar, i.e. d varied from 0.66 to 0.48 lm, under the

homogenization conditions used in the present study
(results not shown). Xiong, Aguilera and Kinsella (1991)
reported that emulsions containing small-sized fat glob-
ules reinforced gels more than emulsions comprized of
large globules at an equal fat concentration, re#ecting the
importance of the size of globules for the preparation of
gels. However, the relatively small range of fat globule
size distributions for all samples in our study minimized
any e!ects of a variation in the sizes of fat globules on gel
properties.
The protein loads (mg protein/m fat surface) of vari-
ous emulsions varied from 1 to 7 mg m\ (Table 1).
Emulsions containing SMP had higher protein loads
than other types of emulsions. Emulsions made with
low-heat SMP had the highest protein load while those
made with unheated WPC had the lowest load. The
protein load values obtained for unheated WPC and
sodium caseinate stabilized emulsions are in agreement
with earlier studies (Oortwijn & Walstra, 1979; Mulvihill
& Murphy, 1991; Sharma & Singh, 1998) and suggest
that these proteins are adsorbed in the form of a mono-
layer with some unfolding. The protein load of emulsions
made with heated WPC was higher than those made with
unheated WPC. This is probably due to the presence of
denatured and aggregated whey proteins in the heated
WPC, which subsequently adsorb on the fat globule
surface during homogenization. This is consistent with
the results of Oortwijn and Walstra (1979) who reported
Table 1
Protein load (mg m\) of emulsions containing 10% milk fat and
di!erent types of protein material
Type of emulsifying agent Protein load
(mg m\)
Low-heat SMP
7.05
Medium-heat SMP
4.64
High-heat SMP
5.09
Sodium caseinate 1.26
WPC 1.13
Heated WPC 1.99
Means of duplicates.

Skim milk powder.
Whey protein concentrate.
540 Y.H. Cho et al. / International Dairy Journal 9 (1999) 537}545

that heat treatment of whey prior to homogenization Table 2

with milk fat resulted in an increased protein load. Protein, fat and total solids contents of recombined milks prepared by
The large di!erences in protein loads observed be- mixing unheated (URSM) or heated (803C for 30 min) reconstituted
skim milk (HRSM) and emulsions stabilized by various materials
tween di!erent emulsions are probably due to the size
and type of particles adsorbed on the fat globules during Type of Type of Protein Fat Total solids
homogenization. In emulsions made with SMP there was emulsion used reconstituted (%) (%) (%)
adsorption of large casein micelles particles on to fat skim milk
globules as well as some serum protein (Walstra & Oor-
Low-heat URSM 5.33 3.39 16.00
twijn, 1982; Sharma et al., 1996). In contrast, sodium SMP
HRSM 5.20 3.45 15.89
caseinate and WPC solutions contain smaller, &soluble'
proteins that form more compact surface layers than Medium-heat URSM 5.22 3.32 16.42
casein micelles. This would result in lower protein loads. SMP
HRSM 5.17 3.43 16.46
If the surface of fat globules was covered with a layer of
High-heat URSM 5.27 3.47 15.69
completely spread protein molecules, the protein load SMP
HRSM 5.06 3.31 15.78
would be little more than 1 mg m\ (Oortwijn
& Walstra, 1979). The interfacial layer in caseinate-stabil- Sodium URSM 5.23 3.52 14.47
ized emulsions appears to be a monolayer so that even if caseinate HRSM 5.30 3.44 14.63
the protein was originally present as an aggregate it
WPC URSM 5.23 3.54 13.78
seems that dissociation of these aggregates occurs as the HRSM 5.12 3.47 13.91
protein adsorbs to the oil/water interface (Dalgleish,
1997). Surface coverage by caseinate molecules depends Heated WPC URSM 5.28 3.43 14.55
on the protein (and oil) content and on the homogeniz- HRSM 5.54 3.44 14.89
ation conditions. Protein loads for caseinate-stabilized
Tween 60 URSM 4.16 3.51 13.34
emulsion droplets can vary from 1 to 3 mg m\ (Dal- HRSM 4.48 3.55 13.45
gleish, 1997; Sharma & Singh, 1998).
Means of duplicates.
3.2. Composition of recombined milks
Skim milk powder.
Whey protein concentrate.
The protein, fat and total solids contents of recom-
bined milks containing fat globules stabilized by di!erent
types of materials are shown in Table 2. The protein The pH at gelation showed similar trends, i.e. higher pH
contents of various recombined milks were similar except at gelation in heated systems compared to unheated ones.
for the recombined milks containing fat globules stabil- Acid milk gels containing fat globules stabilized by
ized by Tween 60 in which the protein content was lower heated WPC had very short gelation times and high
(since only a small concentration of Tween 60 was needed gelation pH values in both URSM and HRSM systems
as an emulsi"er in this system). The fat content was in the con"rming that denatured whey proteins are responsible
range 3.3}3.5%. The total solids contents varied from for the increased pH of gelation of acidi"ed milk (Lucey
13.3 to 16.5% between di!erent samples as a result of et al., 1998c).
constant protein and fat contents. The samples made The G values as a function of time for various gels that
using SMP-based emulsions had higher total solids be- were made with URSM containing di!erent types of fat
cause of the extra lactose and minerals present. globules are shown in Fig. 2. Recombined milks contain-
ing fat globules stabilized by Tween 60 and WPC had the
3.3. Rheological properties of recombined milks lowest G values. The G versus time pro"les for the
various types of SMPs were similar but sodium caseinate
The e!ects of the type of fat globule membrane mate- gave higher values while heated WPC had the highest G
rial and heat treatment of reconstituted skim milk on the values. The G values, at 16 h, of acid milk gels were in the
gelation time and pH at gelation are shown in Table 3. order: WPC(Tween 60(high-heat SMP"medium
Both the pH of gelation and gelation time were greatly heat SMP"low-heat SMP(sodium caseinate(heated
in#uenced by heat treatment of reconstituted skim milk WPC (Table 3). Xiong and Kinsella (1991) investigated
which resulted in an increase in the pH at gelation and the e!ect of di!erent fat globules membranes on the G
a reduction in the gelation time. These "ndings are in of acid milk gels formed by heating milk that had been
agreement with those reported by other authors (Heertje, acidi"ed to pH 4.6 at 23C. They found that the G of these
Visser & Smits, 1985; Horne & Davidson, 1993; Lucey gels were in the order: Tween 80(low-heat SMP(so-
et al., 1997). No signi"cant di!erences were observed in dium caseinate(whey protein isolate.
the gelation of acid milk gels containing fat globules In HRSM systems, gel formation was faster and the
stabilized by low-heat, medium-heat or high-heat SMP. values for G were higher compared with gels made from
 Y.H. Cho et al. / International Dairy Journal 9 (1999) 537}545 541

Table 3
Rheological properties of gels made from recombined milks prepared
by mixing unheated (URSM) or heated (803C for 30 min) reconstituted
skim milk (HRSM) and emulsions stabilized by various materials. Gels
made at 303C with glucono-d-lactone (GDL)

Type of Type of Gelation
Storage

emulsifying reconstituted modulus
,
agent used skim milk Time (ks) pH G (Pa)

Low-heat URSM 23.5 4.87 30

SMP HRSM 10.0 5.07 184

Medium- URSM 22.5 4.85 28

heat
SMP HRSM 9.6 5.09 319

High-heat URSM 22.0 4.84 26

SMP HRSM 9.0 5.09 330
Fig. 3. Storage modulus (G) as a function of time for acid milk gels
Sodium URSM 18.5 4.84 49 containing fat globules stabilized by low-heat (䊏), medium-heat (䊐),
caseinate HRSM 10.5 5.12 478 high-heat SMP (䉱), sodium caseinate (;), WPC (*), heated WPC (䢇)
or Tween 60 (䉭) in heated (803C for 30 min) RSM. Gels were made by
WPC URSM 27.0 4.75 13 acidi"cation with glucono-d-lactone at 303C.
HRSM 7.8 5.12 196

Heated URSM 7.2 5.17 85

WPC
HRSM 6.0 5.30 523 URSM (Fig. 3). The G of all gels increased rapidly
initially after gel formation (especially for HRSM sys-
Tween 60 URSM 20.0 4.98 13 tems) and tended to #atten only after long aging (Figs. 2
HRSM 7.8 5.18 196 and 3). Heating RSM, at 803C, resulted in a large
increase in G, con"rming the results of Lucey et al. (1997,
Point when gels had a storage modulus 51 Pa.

Means of duplicates. 1998c). In recombined milks made from HRSM, G
Measured 16 h after addition of GDL. values (at 16 h) of acid milk gels were in the order:
Skim milk powder. low-heat SMP"Tween 60"WPC(medium-heat
Whey protein concentrate. SMP"high-heat SMP(sodium caseinate(heated
WPC (Table 3). The G values of HRSM systems, 16 h
after addition of GDL, were in the range 180}540 Pa,
whereas URSM systems produced gels with G values in
the range &20}90 Pa (Table 3). Lucey et al. (1997) also
found that heating milks at temperatures 5803C in-
creased the G value compared to unheated milk
(&15 Pa) and produced gels with G values in the range
350}450 Pa (lower G values than present study probably
due to lower total solids content).
The rheological properties of acid gels depend on the
time scale of the applied deformation (Roefs & van Vliet,
1990; Lucey et al., 1997). Plotting log G versus log fre-
quency gave straight lines with slopes between 0.14 and
0.17 (results not shown); similar values were reported for
skim milk gels by Roefs and van Vliet (1990) and Lucey
et al. (1997). The acid gels made in HRSM tended to give
slightly lower values for the slope; a similar trend was
noticed by Lucey et al. (1998a). Tan d decreased slightly
with increasing frequency for acid gels made in URSM
(Fig. 4) or HRSM systems (Fig. 5), which is in agreement
Fig. 2. Storage modulus (G) as a function of time for acid milk gels with the results obtained for acid gels made from recom-
containing fat globules stabilized by low-heat (䊏), medium-heat (䊐),
high-heat SMP (䉱), sodium caseinate (;), WPC (*), heated WPC (䢇) bined milk by Lucey et al. (1998a). In both URSM and
or Tween 60 (䉭) in URSM. Gels were made by acidi"cation with HRSM systems gels containing fat globules stabilized
glucono-d-lactone at 303C. with heated WPC as an emulsi"er had the lowest tan d
542 Y.H. Cho et al. / International Dairy Journal 9 (1999) 537}545
Fig. 4. Loss tangent (tan d) as a function of frequency for acid milk gels
containing fat globules stabilized by low-heat (䊏), medium-heat (䊐),
high-heat SMP (䉱), sodium caseinate (;), WPC (*), heated WPC (䢇)
or Tween 60 (䉭) in URSM. Gels were made by acidi"cation with
glucono-d-lactone at 303C.
Fig. 5. Loss tangent (tan d) as a function of frequency for acid milk gels
containing fat globules stabilized by low-heat (䊏), medium-heat (䊐),
high-heat SMP (䉱), sodium caseinate (;), WPC (*), heated WPC (䢇)
or Tween 60 (䉭) in heated (803C for 30 min) RSM. Gels were made by
acidi"cation with glucono-d-lactone at 303C.
value at all frequencies while gels containing fat globules
stabilized with low-heat SMP or Tween 60 had the high-
est tan d at all frequencies.
4. Discussion
The main factor responsible for the formation of acid-
induced gels from unheated milk is the reduction in the
net negative charge on casein particles as the isoelectric
point is approached; in these systems, the gel matrix
consists exclusively of casein particles (Lucey & Singh,
1997). When milk is heated above 703C, denatured whey
proteins associate with casein micelles via the formation
of intermolecular disulphide bonds (Singh, 1995) and
denatured whey proteins also participate in the gel
matrix (Lucey et al., 1998c). Cross-linking or bridging, by
denatured whey protein associated with the casein
micelles, is mainly responsible for the greatly increased G
of acid gels made from HRSM (Lucey et al., 1998c). In the
present study acid gels were made using both URSM and
HRSM systems, thus we also have two di!erent types of
gel matrices.
Our results clearly show that the type of adsorbed
layer around the fat globules strongly in#uences the
rheological properties of acid milk gels made from both
HRSM and URSM systems. The material adsorbed at
the fat globule surface can interact positively (&structure
promoter') with the gel matrix, which would be re#ected
in an increase in G value of the gel. Alternatively, the
adsorbed material may not interact with the matrix
(&structure breaker') which could be re#ected in a reduc-
tion in the G value of the gel. In recombined milk
systems made from URSM, it is clear that the fat globules
stabilized by various SMPs, sodium caseinate and heated
WPC interacted positively with the casein matrix, as
indicated by an increase in the G values (Fig. 2).
There was little di!erence in the G values of acid gels
made from recombined milk containing fat globules sta-
bilized by di!erent types of SMP but the gels containing
sodium caseinate stabilized fat globules had higher G
values than gels containing SMP-stabilized fat globules.
Surprisingly, acid gels made from URSM containing fat
globules stabilized by heated WPC had the highest G
values.
Fat globules stabilized by SMP solutions have been
shown to consist mainly of casein micelles, casein sub-
units and some whey proteins (Sharma et al., 1996) while
those stabilized by sodium caseinate contain all caseins
(Srinivasan et al., 1996). Sodium caseinate stabilized
globules would provide a greater total surface (contact)
area between particles in the gel compared with SMP-
stabilized globules as part of these globule surfaces are
covered with &non-interacting' whey proteins. The in-
creased number of interactions between sodium caseinate
stabilized fat globules and the casein-based gel network
would lead to increase in G values. It has been reported
(Modler, Larmond, Lin, Froehlich & Emmons, 1983;
Guinee, Mullins, Reville & Cotter, 1995) that at similar
protein levels, yoghurts enriched with sodium caseinate
show higher viscosities or "rmness compared with yo-
ghurts enriched with SMP. Roefs (1986) reported that, at
similar casein levels, acid GDL-induced milk gels made
from sodium caseinate had higher G values than those
made from low-heat SMP.
 Y.H. Cho et al. / International Dairy Journal 9 (1999) 537}545 543

Fig. 6. A schematic presentation of acid milk gels made from URSM with fat globules stabilized by di!erent materials. (A) Gel matrix is formed by
association of casein particles. Fat globules stabilized by interacting materials (sodium caseinate or SMPs) are incorporated into the casein-based gel
network. (B) Fat globules stabilized by heated WPC interact with each other as the isoelectric point of whey protein is approached. (C) Fat globules
stabilized by non-interacting materials (Tween 60 or WPC) "ll space between strands and clusters within the gel.

When the fat globules stabilized by denatured whey
proteins (i.e. by heated WPC) were incorporated into
URSM systems, it is likely that these globules would
become susceptible to aggregation as the isoelectric point
of whey proteins (pH&5.2) is approached. This may
result in the formation of a separate gel network consist-
ing of fat globules cross-linked by whey proteins. Alter-
natively, there may be a formation of fat globule clusters
cross-linked by denatured whey proteins, which become
associated with the casein-based network by electrostatic
interactions as the pH decreases further. In either case,
there would be an increase in the extent of cross-linking
and contact area, resulting in an increase in G. A sche-
matic representation of acid gels made from URSM with
fat globules stabilized by di!erent materials is shown in
Fig. 6.
In recombined milk systems made from URSM, WPC
and Tween 60 did not interact with the casein matrix
resulting in gels with low G values (Fig. 2). It has been
reported (Aguilera, Kinsella & Libo!, 1993; McClements,
Monahan & Kinsella, 1993) that small molecule surfac-
tants, i.e. Tween, do not cross-link with milk protein gel
networks; instead they may even weaken it. Xiong and
Kinsella (1991) reported that milk fat emulsi"ed with
Tween 80 had only a small e!ect on the G of acid milk
gels in contrast to protein-stabilized emulsions. They
suggested that fat globules stabilized with Tween 80 were
loosely entrapped in the gel matrix and acted as an inert
"ller. It has recently been reported that native whey
proteins do not interact with casein particles during
acidi"cation of unheated milk and act as a destructive
"ller or as a &structure breaker' in acid milk gels (Lucey,
Teo, Munro & Singh, 1999).
In recombined milk systems made from HRSM, the
formation of a gel during acidi"cation is largely deter-
mined by interactions of denatured whey proteins with
casein particles (Lucey et al., 1998c). Higher G of acid
gels made from HRSM containing fat globules stabilized
by high-heat SMP compared to those stabilized by low-
heat SMP can be explained by di!erences in the com-
position of fat globule surfaces. Fat globules stabilized by
high- and medium-heat SMP had a higher concentration
of denatured whey proteins on their surfaces than those
stabilized by low-heat SMP, and consequently provided
additional cross-links in the gel in HRSM systems as the
denatured whey proteins are an important cross-linking
agents (Lucey et al., 1998c). It would be expected
that heated WPC stabilized fat globules act as large
544 Y.H. Cho et al. / International Dairy Journal 9 (1999) 537}545

Fig. 7. A schematic presentation of acid milk gels made from HRSM with fat globules stabilized by di!erent materials. (A) Gel matrix is formed by
association of denatured whey proteins attached to casein micelles. Fat globules stabilized by sodium caseinate interact with each other by
casein}casein interactions. (B) Fat globules stabilized by interacting materials (heated WPC, medium- or high-heat SMP) are incorporated into the
denatured whey-protein-based gel network. (C) Fat globules stabilized by non-interacting materials (Tween 60 or WPC) "ll space between strands and
clusters within the gel.

denatured whey protein particles and interact strongly
within the gel network, resulting in a large increase in G.
The increase in G observed by the incorporation of
sodium caseinate stabilized fat globules in HRSM is
rather surprising. It is possible that sodium caseinate
stabilized fat globules could aggregate with each other
close to the isoelectric point of casein and these particles
could become incorporated into the gel network. A sche-
matic representation of acid gels made from HRSM with
fat globules stabilized by di!erent materials is shown in
Fig. 7.
Varying the timescale of the applied deformation pro-
vided information on the nature of the bonds in the gel
network (van Vliet, van Dijk, Zoon & Walstra, 1991).
The slope of the log G versus log frequency curves was
not greatly in#uenced by the nature of the fat globule
surface suggesting that the overall nature of the bonds in
the gels did not vary greatly. Tan d decreased with in-
creasing frequency for all types of gels (Figs. 4 and 5)
suggesting that gels become more elastic in character at
higher frequencies (shorter timescales). In both URSM
and HRSM systems, gels containing fat globules stabil-
ized with heated WPC as an emulsi"er had the lowest
tan d at all frequencies suggesting that this type of gel had
the most elastic character.
In conclusion, the e!ects of emulsi"ed fat globules on
the properties of acid milk gels depend on their surface
composition; non-interacting materials such as Tween
and WPC act as a "ller while interacting materials such
as SMP, sodium caseinate and heated WPC can reinforce
the gel matrix resulting in an increase in the G of acid
milk gels.
Acknowledgements
This work was supported by the Foundation for Re-
search Science and Technology, New Zealand. The
authors thank P. A. Munro for useful comments on this
subject.
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