The Potential Application of Rice Bran Wax

Oleogel to Replace Solid Fat and Enhance

Unsaturated Fat Content in Ice Cream
Daniele C. Zulim Botega, Alejandro G. Marangoni, Alexandra K. Smith, and H. Douglas Goff
C: Food Chemistry

Abstract: The development of structure in ice cream, characterized by its smooth texture and resistance to collapse
during melting, depends, in part, on the presence of solid fat during the whipping and freezing steps. The objective
of this study was to investigate the potential application of 10% rice bran wax (RBW) oleogel, comprised 90% high-
oleic sunflower oil and 10% RBW, to replace solid fat in ice cream. A commercial blend of 80% saturated mono-
and diglycerides and 20% polysorbate 80 was used as the emulsifier. Standard ice cream measurements, cryo-scanning
electron microscopy (cryo-SEM), differential scanning calorimetry (DSC), and transmission electron microscopy (TEM)
were used to evaluate the formation of structure in ice cream. RBW oleogel produced higher levels of overrun when
compared to a liquid oil ice cream sample, creating a lighter sample with good texture and appearance. However, those
results were not associated with higher meltdown resistance. Microscopy revealed larger aggregation of RBW oleogel fat
droplets at the air cell interface and distortion of the shape of air cells and fat droplets. Although the RBW oleogel did
not develop sufficient structure in ice cream to maintain shape during meltdown when a mono- and diglycerides and
polysorbate 80 blend was used as the emulsifier, micro- and ultrastructure investigations suggested that RBW oleogel did
induce formation of a fat globule network in ice cream, suggesting that further optimization could lead to an alternative
to saturated fat sources for ice cream applications.

Keywords: fat structure, fat substitute, ice cream, oleogel, rice bran wax

Practical Application: The objective of this study was to investigate the potential application of 10% rice bran wax
(RBW) oleogel, comprised 90% high-oleic sunflower oil (HOSO) and 10% RBW, to replace solid fat in ice cream. This
would provide ice cream manufacturers with an alternative to solid fat sources, which could expand the potential for fat
source selection beyond either natural saturated fats or hydrogenated oils, and reduce saturated fat intake for consumers. It
was found that RBW oleogels created better structure than HOSO alone but not as good as milk fat. Further optimization
of the oleogel manufacture and mix composition may improve product structure.

Introduction to utilize domestic oils in ice cream applications would be of great

A considerable amount of the ice cream in the global market is
formulated with nondairy fat. Natural saturated fat sources such
as palm kernel oil, palm oil or coconut oil, or hydrogenated oils,
are mostly used (Marshall and others 2003). There are 2 issues
that emerge from this. The health risk associated with the con-
sumption of trans fat and saturated fat has been intensely discussed
during the last few decades. Current governmental dietary recom-
mendations suggest a reduction of saturated fat and elimination of
trans fat to reduce the risk of cardiovascular diseases and other
diet-related disorders (USDA and US HHS 2011; Health Canada
2011). However, current knowledge suggests there is no direct
relation between saturated fat intake and increased risk of car-
diovascular disease (Siri-Tarino and others 2010). Nevertheless,
increased consumption of (poly)unsaturated oil to replace satu-
rated fat provides numerous benefits to health (National Institute
of Health 2005). In addition to health concerns, a reduction in
reliance on imported natural sources of saturated fat and the ability
MS 20130205 Submitted 2/12/2013, Accepted 5/2/2013. All authors are with
Dept. of Food Science, Univ. of Guelph, Guelph, Ontario, Canada N1G 2W1.
Direct inquiries to author Goff (E-mail: dgoff@uoguelph.ca).
interest to manufacturers.
Ice cream is a complex food composed of several structural el-
ements that confer the desirable texture. The whipping/freezing
process of ice cream is responsible for incorporating air bubbles
and for applying intense shear that promotes the partial coales-
cence of fat droplets, thus creating a fat globule network (Marshall
and others 2003; Goff 2006). The presence of a minimum amount
of crystalline fat is necessary to preserve the round shape of the
fat droplets even after destabilization, which allows for the de-
velopment of structure-forming aggregates by partial coalescence,
as opposed to coalescence of fat, which is not structure-forming
(Goff 1997a,b; Rousseau 2000; Coupland 2002; Marshall and oth-
ers 2003; Fredrick and others 2010). Therefore, solid fat has an
important role in the formation and stability of the structure of ice
cream and the reduction of solid fat content in ice cream cannot
be accomplished by simply replacing the solid fat by liquid oil.
Oleogelation, a novel structuring method, has been of great in-
terest recently because of its ability to gel a large volume of edible
liquid oils (approximately 98%) with relatively low concentration
of oleogelators, thus conferring features of a solid fat without
the need of large amounts of saturated fat. A 3-dimensional net-
work, formed by an oleogelator, immobilizes the liquid oil to

R
C 2013 Institute of Food Technologists

C1334 Journal of Food Science r Vol. 78, Nr. 9, 2013 doi: 10.1111/1750-3841.12175
Further reproduction without permission is prohibited
Rice bran wax oleogel in ice cream . . .

create a hard material (Penetti and others 2007; Rogers and others
2009; Marangoni and Garti 2011; Co and Marangoni 2012). Re-
cently, some food grade oleogelators have been suggested (Bot
and Agterof 2006; Wright and Marangoni 2006, 2007; Toro
Vazquez and others 2007; Rogers and others 2009; Zetzl and
others 2012). However, the application of innovative oleogelators
and the progress in the application of oleogels in food is still very
oleogel was determined to be 1.474 (J357 Automatic Refractome-
ter, Rudolph Research Analytical, Hackettstown, N.J., U.S.A.).
Protein adsorption analysis—dumas method. The fat
phase was separated from the ice cream emulsions by centrifu-
gation (Optima LE-80K Ultracentrifuge, Beckman Coulter, Inc.,
U.S.A.) at 20 ◦ C, 12000 rpm for 40 min. The supernatant layer
was removed and deposited on a filter paper (Qualitative P5,

C: Food Chemistry
limited. Fisher Scientific Company, Ottawa, ON, Canada). A FP-528 Pro-
The purpose of this research was to investigate the potential tein/Nitrogen Analyzer (LECO Corp., St. Joseph, Mich., U.S.A.)
application of rice bran wax (RBW) oleogels in ice cream. The was used to determine the nitrogen content of the sample. The
study examined the fat structure in ice creams formulated with adsorbed protein was calculated by dividing the protein concen-
RBW oleogel compared to a traditional milk fat ice cream and tration of the supernatant layer (mg protein/g fat) by the specific
a liquid oil ice cream using standard methods. The micro- and surface area (m2 /g fat) determined by light scattering.
ultrastructure of the ice creams were evaluated using cryo-scanning Meltdown stability. The frozen samples, in their original
electron microscopy and transmission electron microscopy. shape (150 mL), were placed on a stainless steel mesh grid placed
over a tripod stand at room temperature and allowed to melt. The
Materials and Methods melted serum that passed through the mesh was collected and
weighed every 10 min for a total of 90 min. The meltdown rate
RBW oleogel preparation was calculated from the percentage of ice cream melted per min.
RBW (10%; Koster Keunen LLC, Watertown, Conn., U.S.A.)
was combined with high-oleic sunflower oil (HOSO; 90%; Differential scanning calorimetry analysis
Nealanders International Inc., Mississauga, ON, Canada). The Bulk RBW, HOSO, and RBW oleogel were analyzed, as
temperature of the blend was increased to 80 ◦ C and the blend well as ice cream mixes and frozen ice creams. Calibration was
was vigorously stirred. carried out using an indium metal standard (m.p. 156.6 ◦ C,
Hf = 28.45 J/g). An empty hermetically sealed aluminum pan
Ice cream preparation was used as a reference. The pans containing the frozen samples
◦
The ice cream mixes were prepared with 10% fat, 10% milk were prepared in a walk-in freezer (–18 C). Frozen ice cream
3
solids not fat (from skim milk powder, Parmalat Canada Inc., pieces of 2 to 3 mm were placed and sealed into aluminum
Toronto, ON, Canada), 12% sucrose (Lantic Inc., Montreal QC, pans and rapidly transferred to a differential scanning calorimeter
Canada), 4% corn syrup solids (42DE, Casco Inc., Etobicoke, ON, (DSC Q1000, TA Instruments, U.S.A.) with the sample chamber
◦
Canada), 0.1% guar gum, 0.015% carrageenan, 0 to 0.2% (in 0.05% previously cooled to –25 C. Samples were heated from –25 to
◦
increments) Polmo emulsifier (all from Danisco Inc., Scarbor- 80 C, equilibrated at that temperature for 5 min and cooled to
R

◦ ◦
ough, ON, Canada), and the balance with water. Polmo is a com- –50 C at the rate of 5 C/min. After analysis, the pan contain-
mercial blend consisting of 80% saturated mono- and diglycerides ing the sample was removed from the equipment and weighed.
and 20% polysorbate 80. As fat sources, butter (80% fat, unsalted, Ice cream mixes and bulk RBW, HOSO, and RBW oleogel were
Gay Lea Foods Co-operative Ltd, Guelph, ON, Canada), HOSO, weighed and sealed into aluminum pans at room temperature. The
◦
or RBW oleogel were used. Pasteurization was performed at samples were heated to 100 C, equilibrated at that temperature
◦ ◦ ◦
85 C for 30 s. Mixes were homogenized using a 2-stage ho- for 5 min, cooled to –60 C at the rate of 5 C/min, equilibrated
◦ ◦
mogenizer (31MR Laboratory Homogenizer, APV Gaulin Inc., at –60 C, and heated to 100 C at a rate of 5 ◦ C/min. Ice cream
◦ ◦
Wilmington, Mass., U.S.A.) at 20.4/6.9 MPa. The mixes were mixes were heated to 80 C instead of 100 C to avoid evaporation
cooled and aged overnight at 5 ◦ C. of water. Thermograms were analyzed with TA Universal Analysis
◦
Ice cream mix was frozen in a Taylor batch freezer (Model Software to obtain the crystallization and melting peak (Tm , C)
104–27 Batch Ice Cream Freezer, Taylor Company, Rockton, Ill., temperatures.
U.S.A.). The mix was frozen for 5.5 min (to approximately –5 ◦ C)
followed by further whipping resulting in a total process time of
10.5 min. Ice cream was placed into a hardening room at –30 ◦ C.
All mix formulations were prepared in triplicate.

Standard ice cream measurements

Overrun. A container of fixed volume (227 mL) was weighed
and the overrun percentage was determined by the following
formula:

OR% =
(weight of ice cream mix−weight of frozen ice cream)×100%
.
weight of frozen ice cream

Particle size distribution. Ice cream mixes and melted frozen

ice cream (allowed to melt at 5 ◦ C for 24 h) were analyzed by
diluting a single drop of mix in the sample chamber of the Mas-
tersizer 2000 (Malvern Instruments, Malvern, Worcs, U.K.) in the
proportion of 1:1000 (mix/water). The refractive index of RBW
Figure 1–The fat globule size distributions of 10% rice bran wax (RBW)
oleogel (10% RBW and 90% high-oleic sunflower oil, HOSO) ice cream mix
with 0% (), 0.05% (), 0.10% (o), 0.15% (△), and 0.20% (•) Polmo
emulsifier. Size distributions for 10% milk fat ice cream (dashed line—no
symbol) and 10% HOSO ice cream (solid line—no symbol) are also shown.

Vol. 78, Nr. 9, 2013 r Journal of Food Science C1335

 Rice bran wax oleogel in ice cream . . .
Table 1–Effect of increasing PolmoR emulsifier (80% saturated mono- and diglycerides, and 20% polysorbate 80) concentration in
rice bran wax (RBW) oleogel ice cream on protein adsorption at the fat droplet interface, overrun and fat globule size of frozen
ice creams. Parameters are compared to a typical milk fat ice cream and a liquid oil (high-oleic sunflower oil, HOSO) ice cream.
Fat source Polmo concentration (%)
Milk fat 0.20
HOSO 0.20
RBW 0.20
C: Food Chemistry
RBW 0.15
RBW 0.10
RBW 0.05
RBW 0 4.9a
Means followed by the same letter are not significantly different (P > 0.05).
Cryo-scanning electron microscopy
A small sample of frozen ice cream immersed in liquid nitrogen
(–196 ◦ C) was placed into a copper double screw specimen holder
(Caldwell and others 1992). The holder containing the speci-
mens was rapidly transferred into the cryogenic preparation unit
(Emitech K1250X, Quorum Technologies Ltd., Ashford, Kent,
U.K.) under vacuum, where samples at –196 ◦ C were fractured.
After fracturing, sublimation was carried out at –80 ◦ C for 5 min.
The fractured samples were sputter-coated with gold (30 nm) at
–160 ◦ C and then transferred under vacuum into the cold stage
(≤–140 ◦ C) of the scanning electron microscope (S-570, Hitachi
Ltd., Tokyo, Japan). Samples were visualized at 10 kV accelerat-
ing voltage and pictures were captured using Quartz PCI (Ver-
sion 7.0, Quartz Imaging Co., Rexdale, ON, Canada) at different
magnifications.
Air cell size distribution and shape
Air cell distribution was studied using cryo-SEM as described
by Chang and Hartel (2002). Samples were observed at 300×
magnification. For this test, sublimation was not performed, which
enhanced differentiation between ice crystals and air cells. Pictures
were analyzed using Adobe Photoshop image analysis software
(Photoshop CS5 Extended, Adobe Systems Inc., Sao Jose, Calif.,
U.S.A.).
Figure 2–Meltdown behavior of frozen ice creams formulated with 10%
rice bran wax (RBW) oleogel (10% RBW and 90% high-oleic sunflower oil,
HOSO) with 0% (), 0.05% (∗), 0.1% (♦), 0.15% (o), and 0.2% (•) Polmo
emulsifier, compared to 10% HOSO () and 10% milk fat () controls
formulated with 0.2% Polmo.
C1336 Journal of Food Science r Vol. 78, Nr. 9, 2013
Adsorbed protein (mg/m2 ) Overrun (%) d4,3 (µm) frozen ice cream
3.6e 73.1c 11.30c
4.0b,c,e 32.0a 10.32c
3.0d 50.8b 11.27c
3.7c,e 49.7b 6.32b
4.1b,c,e 55.8b 3.68a,b
4.5a,b 58.3b 1.49a
50.2b 1.05a
Transmission electron microscopy
Analyses were performed according to the method described
by Goff and others (1987), with some modifications. Ice cream
mixes were combined with a 2% Ultra-low Gelling Tempera-
ture Agarose, previously prepared and cooled to 25 ◦ C, (Type
IX–A, Sigma-Aldrich Co., St. Louis, Mo., U.S.A.) in the propor-
tion of 1:3 (v:v), respectively. The mix was equilibrated at 4 ◦ C
overnight. Small pieces of the hardened agarose gel containing the
ice cream mix were fixed with 4% glutaraldehyde solution in 0.1
M phosphate buffer (pH 7.0) for 24 h, at room temperature. The
samples were rinsed with phosphate buffer several times. After
rinsing, the samples were postfixed in 1% osmium solution in 0.1
M phosphate/imidazole buffer (1:1 v/v; pH 7.0) for 48 h at room
temperature. The samples were rinsed with phosphate/imidazole
buffer and dehydrated by aqueous dilutions of ethanol up to 100%
ethanol and finally embedded in Spurr’s resin. Samples were in-
serted in molds and polymerized at 70 ◦ C overnight. The blocks
were cut in thin sections (90-nm thickness) and transferred to a
grid (Formvar/Carbon Coated Copper). The samples in the grid
were double stained with uranyl acetate and lead citrate. Samples
were viewed using a transmission electron microscope (Philips
EM300, Eindhoven, the Netherlands) operating at 80 kV.
Results and Discussion
Ice cream mix characterization
The properties of ice cream mix and frozen ice cream contain-
ing RBW oleogel (10% RBW) were evaluated and compared to 2
control ice creams containing the typical milk fat or an unsaturated
oil (HOSO) as the fat source. To confirm whether the homoge-
nized RBW oleogel droplets were gelled, mixes were centrifuged
with conditions optimized to separate the droplets without induc-
ing coalescence (40 ◦ C, low speed, added salt to increase aqueous-
phase density). The lipid phase was removed from the centrifuge
tubes and characterized visually. The lipid phase from the HOSO
sample was a liquid whereas the lipid phases from both milk fat
and the RBW oleogel were solid. This provided evidence that the
RBW was sufficiently dispersed within the oil droplets to form an
oleogel. Oil droplet size analysis revealed a bimodal distribution
for the sample formulated with RBW oleogel with particle size
lower than 10 µm, compared to monomodal normal distribu-
tions presented by the controls with particle size lower than 2 µm
(Figure 1). The 2nd peak of larger sizes decreased when concen-
tration of emulsifier was increased.
Protein adsorbed to the oil interface is reported in Table 1.
As expected, the protein per surface area of oil in the RBW
oleogel emulsions decreased as the concentration of emulsifier
increased, from 4.9 mg/m2 with no emulsifier to 3.0 mg/m2 at
Rice bran wax oleogel in ice cream . . .

0.2% emulsifier. This value was lower than either the milk fat
or HOSO controls. This could lead to a higher instability of the
emulsion during the whipping/freezing stage, which would pro-
mote enhanced aggregation of this new system. Feijter and others
(1987) stated that the degree of protein displacement is related to
surfactant concentration and type. However, according to them,
the type of fat does not have a large effect on protein displacement.
studies and reviews (Bolliger and others 2000b; Barfod 2001;
Koxholt and others 2001; Muse and Hartel 2004) was not ob-
served in RBW oleogel samples formulated with Polmo emul-
sifier. This suggests that the destabilization phenomena observed
were not sufficient to establish a 3-dimensional network structure,
perhaps due to a lack of interaction with air interfaces or a lack of
interaction amongst droplets.

C: Food Chemistry
The characteristics described above for the RBW oleogel ice
cream emulsion are acceptable parameters desired in a well- Microstructure characterization
homogenized and stable milk fat ice cream emulsion (Koxholt Differential scanning calorimetry was used to determine if
and others 2001), which should lead to satisfactory destabilization RBW was dispersed in HOSO instead of being separated from
during the freezing process. it after the oleogel was dispersed in the ice cream emulsion. The
RBW oleogel melting curve revealed 2 peaks that corresponded
Frozen ice cream characterization to the melting point of RBW (70.2 ◦ C) and HOSO (–73 ◦ C),
Frozen ice cream was characterized according to the amount while the pure RBW melting curve showed the presence of only
of air incorporated, the destabilization of fat during the freez- 1 peak (81.3 ◦ C), which corresponded to the melting point of the
ing/whipping process and its meltdown stability. The overrun wax. When RBW is dissolved in HOSO, the melting temperature
of the oleogel ice creams was higher than the HOSO control, of RBW is depressed significantly (11.1 ◦ C) due to colligative
thus creating a lighter sample with better texture and appearance effects, that is, melting point depression, while the crystallization
(Table 1). Oil spreading at the air interface results in air bubble temperature is depressed due to inhibition of nucleation in dis-
collapse, and these results suggest that the olegel samples improved persed systems (Hartel 2001; Coupland 2002). When the RBW
the foam structure of ice cream compared to HOSO samples.
However, overrun was not as high as that found for the milk
fat control, suggesting that aeration and air stability was not fully
promoted by the gelled oil compared to crystalline fat.
A significant increase in the mean particle diameter (d4,3 ) was
seen when the concentration of Polmo emulsifier was increased
from 0 to 0.2% in the RBW oleogel samples, which supports
the results for surface protein displacement (Table 1). However, at
0.2% Polmo emulsifier, the fat system does not seem to influence
the degree of destabilization since no difference in mean parti-
cle diameter was found between samples with different types of
fat (P > 0.05). The size distributions were all similar in appear-
ance, showing a bimodal peak representing individual fat droplets
and fat aggregates. The presence of polysorbate 80 as an emul-
sifier has been associated with high levels of fat destabilization
during freezing (Goff and others 1987; Goff and Jordan 1989;
Bolliger and others 2000a). However, both coalescence and par-
tial coalescence can lead to enhanced fat aggregation but differ-
ent levels of ice cream colloidal stability (Mendez-Velasco and
Goff 2012a,b).
No improvement was seen in meltdown resistance for samples
formulated with RBW oleogel (10% RBW) compared to HOSO
(Figure 2). No significant (P > 0.05) difference in meltdown
stability was observed between RBW oleogel ice cream samples
formulated with different concentrations of emulsifier. All samples Figure 4–Cryo-scanning electron micrographs of rice bran wax (RBW)
oleogel (10% RBW and 90% high-oleic sunflower oil, HOSO) ice creams
melted considerably faster than the milk fat control. formulated with 0.05 (A, C) and 0.2% (B, D) emulsifier concentration at 2
The expected relationship between protein adsorption, fat different magnifications each (scale bar: A and B = 3 µm; C and D = 100
destabilization, and meltdown stability presented in numerous µm). a, air cell; i, ice crystal; s, serum phase; f, fat; i∗ , air cell interface.

Figure 3–Cryo-scanning electron micrographs of

ice cream formulated with different types of fat:
milk fat (A), HOSO (B), and rice bran wax (RBW)
oleogel (10% RBW and 90% high-oleic
sunflower oil, HOSO) (C), all with the same
emulsifier concentration (0.2%). The
micrographs depict the air cell interface covered
by fat. a, air cell; c, crystallized or platelet fat; f,
fat; s, serum phase. Scale bar = 6 µm.

Vol. 78, Nr. 9, 2013 r Journal of Food Science C1337

 Rice bran wax oleogel in ice cream . . .

Table 2–Parameters for air cell distribution (mean equivalent diameter) and shape (circularity of air cells) of ice cream formulated
with 0.2% emulsifier and 10% milk fat or 10% rice bran wax (RBW) oleogel (comprised of 90% high-oleic sunflower oil and 10%
RBW) measured from cryo-scanning electron microscopy images.

10% Milk fat ice cream 10% RBW oleogel ice cream
Equivalent diameter Circularity of air cells Equivalent diameter Circularity of air cells
Sample size 353 353 314 276
C: Food Chemistry

Median (µm) 4.68 0.85 9.04 0.81

Mean (µm) 8.95 0.84 10.61 0.78
Std. dev. (µm) 14.72 0.07 11.96 0.11
Min. (µm) 1.39 0.34 1.51 0.30
Max. (µm) 98.35 0.95 103.0 0.93
Std. error 0.78 0.00 0.67 0.01
Lower 95% CI of mean 7.40 0.83 9.28 0.76
Upper 95% CI of mean 10.49 0.84 11.93 0.80
CI, confidence interval.

oleogel ice cream emulsion was analyzed, the melting tempera-
ture of RBW in the ice cream emulsion was similar (approximately
70 ◦ C) to the melting temperature of RBW in the oleogel. The
effect of melting point depression was evident for both systems,
which suggested that in both bulk oleogel and emulsions, RBW
is in solution in HOSO.
Aggregation of fat at the air cell interface in RBW oleogel
ice cream was evident by cryo-SEM for different sources of fat
(Figure 3). Milk fat ice cream (Figure 3A) shows the presence
of a large number of spherical droplets that protrude out of the
air bubble interface. Ice cream formulated with HOSO was also
investigated as a control sample (Figure 3B). The smooth mem-
brane visible around the air cell was indicative of oil spreading at
the interface. A fat network was not formed in that case, thereby
compromising incorporation of air and stabilization of the air cell.
RBW oleogel seemed to aggregate at the interface forming a rough
membrane but with no definition of droplet shape (Figure 3C).
RBW oleogel samples also showed platelets of fat that protruded
from the fat layer around the air interface. The aggregation of fat
at the air cell interface in RBW oleogel ice cream at different
concentrations of emulsifier was also evident (Figure 4). Samples
formulated with lower concentration of emulsifier (0.05%) had al-
most no aggregation of fat at the air cell interface (Figure 4A). The
serum of the same sample, however, displayed a large number of
separate spherical fat droplets instead of clusters. When emulsifier
was added at higher concentration (0.2%), more fat aggregation at
the air interface was observed and the presence of single fat droplets
at the serum phase was reduced (Figure 4B). An increase in fat
concentration at the air cell interface was observed at lower magni-
fication, which increased the distortion of the air cell (Figure 4D)
when compared to an ice cream with lower fat aggregation at the
air cell (Figure 4C).
Air bubble sizes in an RBW oleogel ice cream were analyzed
and compared to the milk fat sample. Quantitative analyses of the
air cell distribution revealed that both samples had large popu-
lations of air cells with small diameters (Table 2). Although the
results for mean equivalent diameter were statistically different (P
< 0.05) between the 2 samples, the difference was minor com-
pared to HOSO ice cream samples, which showed very large air
bubbles that were difficult to quantify (results not shown). Bol-
liger and others (2000a) and Eisner and others (2005) stated that
the formation of air cells with small diameters created stable foam
with better resistance against melting. Circularity of air cells in
wax oleogel ice cream was also quantified and compared to milk
fat ice cream (Table 2). More surface distortion (lower circularity)
was seen in the RBW oleogel ice creams.

Figure 5–Transmission electron micrographs of

rice bran wax (RBW) oleogel (10% RBW and
90% high-oleic sunflower oil) ice cream mix
formulated without emulsifier (A and B) and
with Polmo emulsifier (C and D) showing the
internal structure of the oleogel droplets with
RBW crystals. Scale bar = 500 nm (A) and 1 µm
(B, C, and D).

C1338 Journal of Food Science r Vol. 78, Nr. 9, 2013

Rice bran wax oleogel in ice cream . . .
The ultrastructure of RBW oleogel droplets in ice cream was
investigated by TEM, which illustrated the presence of wax crys-
tals within the fat droplets (Figure 5), which were sufficient to
gel the fat phase, as shown earlier. The crystallization of RBW is
represented by the lighter thread inside the droplet, since pene-
tration of osmium is hindered by the solid state of the wax. The
existence of crystals in small droplets is a good indication that
the RBW was efficiently distributed. Casein micelles can also be
clearly identified around the membrane of the fat droplet.
Figure 5A and B represent an RBW oleogel ice cream mix
formulated with 0% of emulsifier, and Figure 5C and D were taken
from samples formulated with 0.2% of emulsifier. In the absence
of emulsifier, fat droplets remained spherical while the presence
of emulsifier led to increased deformation of the droplets.
Conclusion
RBW oleogel was effectively emulsified into ice cream mix and
led to the formation of small fat droplet sizes and gelled droplets.
Proteins adsorbed to the surface of the droplets and emulsifiers
were effective at displacing the proteins from the droplet surfaces.
The frozen ice cream with RBW oleogel produced higher levels
of overrun compared to HOSO. Air bubbles were slightly larger
than the milk fat controls, but much smaller than air bubbles found
in the HOSO samples. Similar levels of fat destabilization were
observed between RBW oleogel ice creams and milk fat controls.
An increase in fat aggregation at the air cell interface was also
observed, which was similar to milk fat but contrasted sharply from
the oil spreading seen in the HOSO sample. Fat aggregation at the
air interface was enhanced by increasing emulsifier concentration.
These results all suggested that the oleogel droplets were behaving
in ice cream more like crystallized fat droplets than like liquid oil.
Fat droplet destabilization and aggregation at the air cell surface
(Muse and Hartel 2004; Goff 2006) and small air bubbles should
lead to a reduction in the rate of structural collapse during melting.
However, the fat structure that was formed by the oleogel droplets
was not sufficient to cause the expected delay in structural collapse.
Further understanding of the factors that contribute to oleogel
structure could lead to the successful application of RBW oleogels
in ice cream.
Acknowledgments
Appreciation is expressed to Nestlé Product Technology Cen-
tre, Beauvais, France, for financial support and to the research
associates of Nestlé, Max Puaud, and Virginie DeBoishebert, for
their valuable assistance, which was important to the preparation
and completion of this work.
References
Barfod NM. 2001. The emulsifier effect. Dairy Ind Int 66:32–4.
Bolliger S, Kornbrust B, Goff HD, Tharp BW, Windhab EJ. 2000a. Influence of emulsifiers on
ice cream produced by conventional freezing and low-temperature extrusion processing. Int
Dairy J 10:497–504.
Bolliger S, Goff HD, Tharp BW. 2000b. Correlation between colloidal properties of ice cream
mix and ice cream. Int Dairy J 10:303–9.
Bot A, Agterof WGM. 2006. Structuring of edible oils by mixtures of γ -oryzanol with β-
sitosterol or related phytosterols. J Am Oil Chem Soc 83:513–21.
Caldwell KB, Goff HD, Stanley DW. 1992. A low-temperature scanning electron microscopy
study of ice cream. I. Techniques and general microstructure. Food Struct 11:1–9.
Chang Y, Hartel RW. 2002. Measurement of air cell distributions in dairy foams. Int Dairy J
12:463–72.
Co E, Marangoni AG. 2012. Organogels: an alternative edible oil-structuring method. J Amer
Oil Chem Soc 89:749–80.
C: Food Chemistry
Coupland JN. 2002. Crystallization in emulsions. Curr Opin Colloid Interface Sci 7:445–50.
Eisner MD, Wildmoser H, Windhab EJ. 2005. Air cell microstructuring in a high viscous ice
cream matrix. Colloids Surf A 263:390–9.
Feijter JA, Benjamins J, Tamboer M. 1987. Adsorption displacement of protein by surfactants in
oil-in-water emulsions. Colloids Surfaces 27:243–66.
Fredrick E, Walstra P, Dewettinck K. 2010. Factors governing partial coalescence in oil-in-water
emulsions. Adv Colloid Interfac 153:30–42.
Goff HD. 1997a. Colloidal aspects of ice cream-a review. Int Dairy J 7:363–73.
Goff HD. 1997b. Instability and partial coalescence in whippable dairy emulsions. J Dairy Sci
80:2620–30.
Goff HD. 2006. Ice cream. In: Fox PF, McSweeney PLH, editors. Advanced dairy chemistry,
Vol 2: Lipids. 3rd ed. New York: Springer. p 441–50.
Goff HD, Jordan WK. 1989. Action of emulsifiers in promoting fat destabilization during the
manufacture of ice cream. J Dairy Sci 72:18–29.
Goff HD, Liboff M, Jordan WK, Kinsella JE. 1987. The effects of polysorbate 80 on the fat
emulsion in ice cream mix: evidence from transmission electron microscopy studies. Food
Microstruct 6:193–8.
Hartel RW. 2001. Crystallization in foods. Gaithersburg: Aspen Publishing. p 325.
Health Canada: Saturated and Trans Fats. Available from: http://www.hc-sc.gc.ca/
fn-an/label-etiquet/nutrition/cons/fats-sat-trans-graisses-eng.php. Accessed 2011
December 1.
Koxholt MMR, Eisenmann B, Hinrichs J. 2001. Effect of fat globule sizes on the meltdown of
ice cream. J Dairy Sci 84:31–7.
Marangoni AG, Garti N. 2011. An overview of the past, present and future of organogels. In:
Marangoni AG, Garti N, editors. Edible oleogels: structure and health implications. Urbana,
IL: AOCS Press. p 1–18.
Marshall RT, Goff HD, Hartel RW. 2003. Ice cream. 6th ed. New York: Kluwer Aca-
demic/Plenum Publishers. p. 371.
Mendez-Velasco C, Goff HD. 2012a. Fat aggregation in ice cream: a study on the types of fat
interactions. Food Hydrocoll 29:152–9.
Mendez-Velasco C, Goff HD. 2012b. Fat structures as affected by unsaturated or saturated
monoglyceride and their effect on ice cream structure, texture and stability. Internat Dairy
J 24:33–9.
Muse M R, Hartel RW. 2004. Ice cream structural elements that affect melting rate and hardness.
J Dairy Sci 87:1–10.
National Institute of Health. 2005. Omega-3 fatty acids, fish oil, alpha-linolenic acid.
Available from: http://www.nlm.nih.gov/medlineplus/druginfo/natural/patient-fishoil.html.
Accessed 2005 August 1.
Pernetti M, van Malssen KF, Floter E, Bot A. 2007. Structuring edible oils by alternatives to
crystalline fat. Curr Opin Colloid Interface Sci 84:989–1000.
Rogers MA, Wright AJ, Marangoni AG. 2009. Nanostructuring fiber morphology and solvent
inclusions in 12-hydroxystearic acid/ canola oil oleogels. Curr Opin Colloid Interface Sci
14:33–42.
Rousseau D. 2000. Fat crystals and emulsion stability-a review. Food Res Int 33:3–14.
Siri-Tarino PW, Sun Q, Hu FB, Krauss RM. 2010. Meta-analysis of prospective cohort studies
evaluating the association of saturated fat with cardiovascular disease. Am J Clin Nutr 91:535–
46.
Toro-Vazquez JF, Morales-Rueda JA, Dibildox-Alvarado E, Charó-Alonso M, Alonzo-Macias
M, González-Chávez MM. 2007. Thermal and textural properties of oleogels developed by
candelilla wax in safflower oil. J Am Oil Chem Soc 84:989–1000.
[USDA and US HHS] U.S. Department of Agriculture and U.S. Department of Health and Hu-
man Services. 2011. Dietary guidelines for americans, 2010. [Internet]. Washington, DC: U.S.
Government Printing Office. Available from: http://health.gov/dietaryguidelines/2010.asp.
Accessed 2011 January 31.
Wright AJ, Marangoni AG. 2006. Formation, structure, and rheological properties of ricinelaidic
acid–vegetable oil oleogels. J Am Oil Chem Soc 83:497–503.
Wright AJ, Marangoni AG. 2007. Time, temperature, and concentration dependence of
ricinelaidic acid–canola oil organogelation. J Am Oil Chem Soc 84:3–9.
Zetzl AK, Marangoni AG, Barbut S. 2012. Mechanical properties of ethylcellulose oleogels
and their potential for saturated fat reduction in comminuted meat products. Food Funct
3:327–37.
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