Food Research International 54 (2013) 17381745

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Food Research International

journal homepage: www.elsevier.com/locate/foodres

Stability of whippable oil-in-water emulsions: Effect of monoglycerides

on crystallization of palm kernel oil
Merete B. Munk a,, Alejandro G. Marangoni b, Hanne K. Ludvigsen a, Viggo Norn a, Jes C. Knudsen c, Jens Risbo c,
Richard Ipsen c, Mogens L. Andersen c
a
b
c

Palsgaard A/S, Palsgaardvej 10, DK-7130 Juelsminde, Denmark

Department of Food Science, University of Guelph, 50 Stone Road East, Guelph N1G 2W1, Ontario, Canada
Department of Food Science, University of Copenhagen, Rolighedsvej 30, DK-1958 Frederiksberg C, Denmark

a r t i c l e

i n f o

Article history:
Received 12 June 2013
Accepted 5 September 2013
Keywords:
Palm kernel oil
Emulsion stability
Lipid crystallization
Monoglycerides
LACTEM
Polymorphism

a b s t r a c t
The effect of selected monoglycerides on droplet morphology and rheology of palm kernel oil emulsions was
studied. Combination of lactic acid ester of monoglycerides (LACTEM) and unsaturated monoglycerides (GMU)
yielded highly viscous emulsions caused by partial coalescence. LACTEM combined with saturated monoglycerides (GMS) or diacetyl tartaric acid ester of monoglycerides (DATEM) gave low-viscous emulsions. The rheological behavior was found not to be related to polymorphic transformations of fat in the emulsions.
A possible relationship between structural changes of emulsions and the effect of monoglyceride crystallization
behavior of PKO was examined by studying the monoglycerides' inuence on crystallization of bulk PKO. Polymorphic transformation, crystallization kinetics and melting/crystallization prole of bulk PKO were analyzed.
The monoglycerides had minor effect on crystallization behavior of bulk PKO compared to the major impact on
emulsion stability. This indicates that emulsiers act differently in bulk and dispersed fat. For PKO the structural
changes of the emulsions were independent of the effects of monoglycerides on the crystallization of bulk fat.
2013 Elsevier Ltd. All rights reserved.

1. Introduction
Making a successful whippable fat-based emulsion requires a delicate balance between conicting physical properties. On the one hand,
to achieve voluminous and stable foam of a whipped aerated emulsion,
the fat droplets must interact strongly with each other and form a stable
semi-rigid network, which stabilizes foam structure. Partial coalescence
plays a crucial role for foam structure formation by creating a network
of fat droplets that stabilize air bubbles in foams (Barfod & Krog, 1987;
Goff, 1997). On the other hand, prior to whipping, the emulsion must
be pourable and thus have a perpetual low viscosity, which requires
that attractive forces between droplets are weak in order to avoid occulation, droplet coalescence and consequent creaming of larger aggregates. In addition, the emulsion should preferably have long-term
stability, implying resistance to agitation and thermal uctuations occurring during transportation and storage. Ideally, formation of a strong
network of partly coalesced droplets should not happen until whipping
is initiated, and should not give rise to undesirable solidication,
clumping or creaming of the emulsion.
To ensure stable foam with long shelf life, various types of monoglycerides are often used as emulsifying agents in whippable emulsions.
The high surface activity of monoglycerides is assumed to displace
proteins at the fat droplet interface. Consequently, steric repulsion is
Corresponding author. Tel.: +45 41125539.
E-mail address: bogelund@life.ku.dk (M.B. Munk).
0963-9969/$ see front matter 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodres.2013.09.001

reduced and a new thinner monolayer of monoglyceride surrounds

the interface, which is more susceptible for crystal penetration from
one fat droplet into another. As a result, partial coalesced fat aggregates,
which make up the foam network, are more likely to be formed. However, some monoglycerides seem to cause unintended properties for
emulsions before the whipping. Unsaturated monoglyceride (GMU)
and lactic acid ester of monoglyceride (LACTEM) have been reported
to cause increased viscosity of emulsions both under shear and quiescent conditions (Allen, Murray, & Dickinson, 2008; Barfod, Krog,
Larsen, & Buchheim, 1991; Davies, Dickinson, & Bee, 2000; Davies,
Dickinson, & Bee, 2001).
Emulsiers will affect the adsorbed interfacial layer around emulsied droplets, which is decisive for the nature of droplet interactions
among themselves and with other components in the emulsion.
Additionally, emulsiers can play a secondary role due to their inuence
on crystallization of the emulsied fat. Emulsiers can also affect
crystallization behavior including nucleation, crystal growth, recrystallization of polymorphic forms and melting proles of bulk fat
(Smith, Bhaggan, Talbot, & van Malssen, 2011). Crystallization of fat in
the dispersed droplets can lead to changes of physical properties of
emulsions, such as increased viscosity. The objective of the present
work was to investigate how physical stability of whippable palm
kernel oil (PKO) emulsions prior to whipping was affected by different
combinations of monoglycerides. Physical stability in this context refers
to a perpetual pourability of the non-whipped emulsions, which requires that the dispersed droplets remain nite and spherical. Droplet

M.B. Munk et al. / Food Research International 54 (2013) 17381745

size determinations, confocal microscopy, and rheological measurements were used to study physical stability of emulsions. Furthermore,
the effect of the monoglycerides on crystallization of bulk PKO was
examined with the aim of elucidating a possible relationship between
undesirable textural changes of non-whipped emulsions and the monoglycerides' effect on crystallization behavior of PKO. The latter was
examined by DSC, pNMR and powder X-ray diffraction.
2. Materials & methods
2.1. Materials
Emulsiers were of commercial food grade and all were provided by
Palsgaard A/S (Juelsminde, Denmark): Lactic acid ester of monoglyceride (LACTEM) made from fully hydrogenated palm oil and rape-seed
oil (C16:0 and C18:0 N97% of fatty acids); unsaturated monoglycerides
(GMU) made from sunower oil (C18:1 N81%); >saturated monoglycerides (GMS) made from fully hydrogenated palm oil and rape-seed
oil (C16:0 and C18:0 N 97%); and diacetyl tartaric acid ester of monoglyceride (DATEM) made from palm oil and sunower oil (C16:0,
C18:1, and C18:2 comprised 93% of fatty acids). The stabilizer mixture
(Palsgaard A/S, Juelsminde, Denmark) contained microcrystalline cellulose (MCC), sodium carboxymethylcellulose (CMC) and disodium phosphate. Hydrogenated palm kernel oil (PKO) was obtained from AAK
(Karlshamn, Sweden) and sodium caseinate from DMV International
(Veghel, The Netherlands). Sugar was purchased from Nordic Sugar
(Nakskov, Denmark) and sorbitol from Roquette (Cedex, France).
2.2. Emulsion and bulk PKO blend preparation
2.2.1. Emulsions
Sodium caseinate (0.6 wt.%), stabilizer mixture (0.6 wt.%) and sugar
(10 wt.%) were dispersed in water under continuous stirring and
put aside for 4 h to hydrate proteins. Melted PKO (25 wt.%), LACTEM
(0.55 wt.%) and GMU (0.15 wt.%), GMS (0.15 wt.%) or DATEM
(0.10 wt.%) were mixed with the heated water phase (~70 C) and the
complete mix was re-heated to 82 C. A pre-emulsion was conducted
by mixing with a high-shear blender (Ultra-Turrax, IKA, NC, USA) for
approximately 20 s. Homogenization was subsequently carried out on a
two-stage high-pressure valve homogenizer (APV Rannie LAB-12.50,
SPX, Silkeborg, Denmark) at 150/50 bar. Emulsions were rapidly cooled
to 56 C by a plate heat exchanger (APV U2, SPX, Silkeborg, Denmark)
connected to the homogenizer. The emulsions were immediately poured
into small beakers and stored at 5 C. Control emulsions with sodium
caseinate as the only emulsier were prepared in a similar way.
2.2.2. Bulk PKO blends
GMU (5.0 wt.%), GMS (5.0 wt.%) or DATEM (5.0 wt.%), either singly
or in combination with LACTEM (5.0 wt.%), was added to melted PKO
and the blend was stirred to ensure homogenous distribution and heated to 80 C to remove crystal history. The blend was applied to an
appropriate container (DSC-vial, NMR-tube or X-ray plate) depending
on analysis method and immediately analyzed by DSC, NMR or X-ray.

1739

Heidelberg, Germany). Total concentration of 1 ppm BODIPY 493/503

(4,4-diuoro-1,3,5,7,8-pentamethyl-4-bora-3a,4a-diaza-s-indacene)
(Invitrogen, Carlsbad, USA) dissolved in dimethyl sulfoxide was added
to emulsions to stain lipid. Excitation wavelength was 488 nm and
emission bandwidth was 500570 nm. A water immersion objective
(HCX PL APO lambda blue 63.0 1.20 water UV) was used and image
resolution was set to 1024 1024 pixels. A thin layer of stained emulsion was applied on a large standard cover glass and covered by another
cover glass to avoid evaporation/drying.
2.3.3. Rheological measurements
An ARG2 rheometer (TA-Instruments, West Sussex, England, UK)
tted with a Peltier temperature control connected to a water bath
was used for all rheological measurements. Steady state ow experiments with increasing shear rate from 0.01 to 100 s1 were performed
at 5 C and 20 C to determine viscosity. A cone (diameter 40 mm, inclination 1.59) and plate geometry was applied. Some emulsions
exhibited a very rm texture and were not owable. These were
molded into discs of diameter 20 mm and characterized by a stress
sweep with increasing oscillation stress from 0.1 to 1000 Pa at a constant shear frequency of 1 Hz. Plate (diameter 20 mm) and a plate geometry, both with roughened surfaces, to avoid slip was used.
Viscosity measurements were performed at least in duplicate whereas
small amplitude oscillatory shear was repeated 5 times.
2.4. Characterization of fat crystallization
2.4.1. Differential scanning calorimetry
Melting and crystallization temperatures were determined for both
emulsions and PKO blends by the use of DSC1 Stare System (Mettler
Toledo, Schwerzenbach, Switzerland). Samples were initially held at
5 C for 10 min, then heated to 80 C at 2 C/min and subsequently
re-cooled to 5 C at 2 C/min. Empty pans were used as reference and
each sample was analyzed in duplicate.
2.4.2. Crystallization kinetics
Solid fat content (SFC) of PKO blends containing emulsiers was
determined as a function of time at isothermal conditions by the use
of pulsed NMR spectroscopy (Bruker Minispec, Bruker, Germany).
Tubes of samples were placed in water baths at 10 C and 20 C and
SFC was measured at appropriate intervals till crystallization appeared
to be completed as the crystalline fraction reached a constant level.
25 replicates of each sample were measured.
The results were tted to the Avrami equation by non-linear regression of Origin 7 (OriginLab, Northampton, USA):
SFC t
kt n
1e
:
SFC max

Where SFCt is the crystalline content (%) at time t and SFCmax the
maximum crystalline content (%) achieved at the particular temperature. k is the crystallization rate constant, and n the Avrami exponent
which is dependent on crystal growth mechanism with respect to
nucleation and growth dimensions.

2.3. Characterization of emulsions

2.3.1. Determination of droplet size by light scattering
Droplet size distribution was measured 2, 8 and 15 days after emulsion preparation (Mastersizer Microplus, Malvern Instruments, Malvern,
U.K.). Measurements were conducted at room temperature on emulsions diluted approximately 1:100 with de-ionized water. At least 3 replicates were measured for each emulsion.
2.3.2. Confocal laser scanning microscopy
Structure and morphology of fat globules in emulsions were examined by an inverse confocal laser scanning microscope (Leica TCS SP5,

2.4.3. Polymorphic transformation

The polymorphism of crystals was examined for both emulsions and
PKO blends by powder X-ray diffractometer (Rigaku, Tokyo, Japan). Radiation at = 1.54 was generated from a copper anode and operated
at 40 kV and 44 mA. The scanning velocity was 0.05/min for emulsions
and 1/min for PKO blends in the area of 1.235 2. The diffractometer
was protected by a Multiex chamber, which was equipped with Peltier
temperature control connected to a water bath. Emulsions were examined at 20 C without any preceding treatment and also after exposing
the emulsion to stirring with a spatula. At least two diffraction spectra
were recorded for each emulsion. Fully melted PKO blends were cooled

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M.B. Munk et al. / Food Research International 54 (2013) 17381745

to either 5 C or 20 C and after 10 or 15 min respectively the diffraction

spectrum was recorded. The samples of PKO blends were kept and
recorded the following day, day 7 and day 21.

2.5. Statistical analysis

The results of the analyses are reported as the mean and standard
deviation calculated from the replicates. The level of signicance for
melting and crystallization temperature is calculated by one way
ANOVA and is given by the P-value. Intervals of condences for n and
k of the Avrami model is calculated from the Jacobian matrix which is
an output of the non-linear tting procedure.
3. Results
3.1. Rheological properties of emulsions
Four different palm kernel oil emulsions were made using LACTEM
as one of the emulsiers. The emulsions also contained caseinate, stabilizer and sugar in order to mimic the composition of a commercial
whippable emulsion. LACTEM was included because it promotes fat
droplet aggregation (Westerbeek, 1989) and therefore is widely used
in whipped toppings and dessert products (Si, 1991). In addition,
three of the emulsions contained DATEM, unsaturated (GMU) or saturated monoglycerides (GMS), since LACTEM is often used in combination with other monoglycerides in commercial products. Furthermore,
the specic combinations of LACTEM and the other monoglycerides
were selected in order to create large variations in emulsion stability
in terms of changes in viscosity and texture. An emulsion without
added monoglycerides was prepared as a control. The differences
among the emulsions were immediately discernible after preparation as
observed by the products ranging from low-viscous homogeneous emulsions to thick pastes and clumping emulsions. Emulsions with LACTEM
alone were found to yield semi-solid emulsions. The properties were
initially qualitatively assessed by stirring the emulsions with a spatula at
room temperature or at 5 C. Stirring the LACTEM emulsion at 5 C drastically reduced the viscosity, whereas stirring the emulsion at room
temperature gave a thick paste. In addition, the process causing the instability of this emulsion appeared to be reversible, as a pourable emulsion
could be obtained again by cooling the thick paste to 5 C followed by stirring. Combining LACTEM with GMU gave very rm emulsions that
appeared not to be sensitive to any combination of stirring and temperature. In fact, clotting appeared already within 1015 min after the preparation of the emulsion and after 1-2 h at 5 C complete solidication
occurred. On the other hand, combinations of GMS or DATEM together
with LACTEM yielded perpetual pourable emulsions. The control emulsion made without any added monoglyceride resulted in a low-viscous
emulsion that was not affected by stirring and temperature.
The observed qualitative changes in viscosity and texture were characterized by rheological measurements (Fig. 1). At 5 C and at low shear
rates, the apparent viscosity of the emulsion containing LACTEM was
~50 Pas (Fig. 1a). The apparent viscosity decreased with increasing
shear rate, demonstrating shear thinning. However, the apparent viscosities were 5 to 10 times higher than for the control emulsion without
monoglycerides. The two emulsions with either added GMS or DATEM
also demonstrated shear thinning behavior and had viscosities between
the control emulsion and the LACTEM-only emulsion at all shear rates.
At 20 C only the control emulsion demonstrated a true shear thinning effect comparable to measurements at 5 C (Fig. 1b). The viscosity
of the emulsion with LACTEM could not be determined at 20 C since
the texture of the emulsion was too rm. A lumpy texture in emulsions
caused by LACTEM has previously been reported (Allen et al., 2008), and
is probably due to partial coalescence as a consequence of protein displacement on droplet interface (Thivilliers-Arvis, Laurichesse, Schmitt,
& Leal-Calderon, 2010). Emulsions with LACTEM + GMS exhibited a
sudden shear thickening behavior at 20 C at shear rates around

Fig. 1. Monoglycerides' inuence on rheological behavior of emulsions. : Control, :

LACTEM + DATEM, : LACTEM + GMS, : LACTEM, : LACTEM + GMU. a: Viscosity
at 5 C, LACTEM + GMU was too viscous to be measured. b: Viscosity at 20 C, emulsions
with LACTEM and LACTEM + GMU were too viscous to be measured. c: The elastic
modulus at 20 C of high viscous emulsions, LACTEM and LACTEM + GMU.

0.03 s1 (Fig. 1b). Yet, the viscosity was signicantly lower compared
to emulsions with LACTEM. Addition of DATEM prevented shear
induced modications until a certain degree, and at sufcient high
shear rates a slight increase of viscosity occurred indicating subtle
shear thickening effects.
The instability of emulsions containing LACTEM + GMU was very
evident since they exhibited complete solidication at all temperatures.
The rm emulsions remained resistant to shear torque at 2 105 mN/m
at 5 C and 20 C, and thus viscosity was not a measurable parameter.

M.B. Munk et al. / Food Research International 54 (2013) 17381745

Fig. 2. The effect of monoglycerides on droplet size distribution in emulsions, (solid

black) control(solid black),
LACTEM (long dash),
The line style of "LACTEM +
GMS" should be grey LACTEM + GMS (solid grey), LACTEM + DATEM (short dash),
LACTEM + GMU (dot).

Instead, rheological properties were characterized by small amplitude oscillatory shear. An elastic modulus, G 3.2 0.4 106 Pa
(Fig. 1c) was found, and similar for the rm LACTEM emulsion at
20 C with G 4.5 3 106 Pa. Although no statistical signicant
differences in G were obtained between emulsions containing
LACTEM and LACTEM + GMU, it should be noted that emulsions
containing LACTEM + GMU were three times rmer than emulsions
made only with LACTEM (measured by back extrusion; data not
shown). The higher stress and/or longer duration required to induce
structural changes in emulsions with LACTEM + GMS as compared
to LACTEM + GMU is in agreement with Barfod et al. (1991) who
observed a high stability of emulsions containing GMS in contrast
to emulsions with GMU.
3.2. Droplet size
The droplet size distributions were clearly related to the viscosity of
the emulsions. A monomial and fairly narrow distribution of small

1741

droplets was characteristic for the three low-viscosity emulsions made

without added monoglycerides, a combination of LACTEM + GMS, or
a combination of LACTEM + DATEM (Fig. 2). The mean diameters,
d32, were respectively 0.98 0.03 m, 0.84 0.03 m and 0.65
0.01 m for the three emulsions after storage at 5 C for 2 days. The
two highly viscous and rm emulsions made either with LACTEM or
LACTEM + GMU were characterized by very broad droplet distributions. The broad distributions of mainly large droplets suggest the
presence of aggregates in emulsions. Prior to size distribution measurements, emulsions were diluted with water. Despite the fact that the
continuous phases of all the emulsions were water, dilutions of the
rm emulsions made with LACTEM and LACTEM + GMU were practically impossible despite vigorous shaking using a vortex mixer. This
demonstrated extreme sensitivity towards agitation and thermal uctuation since both types of emulsions, but particularly LACTEM + GMU,
appeared as large non-dispersible ocks in water as a consequence of
the preparatory steps of measurements even though the samples
were held at 5 C. Droplet size distributions were also measured after
8 and 15 days of storage at 5 C but no signicant changes occurred
during this period.
The differences among the emulsions in terms of microstructure
were clearly visualized by confocal laser scanning microscopy (Fig. 3).
The three low-viscous emulsions, without added monoglycerides,
with LACTEM + GMS or LACTEM + DATEM, all exhibited a structure
of small spherical fat globules (Fig. 3a, e and f), supporting the results
from droplet size distributions. Conversely, the fat globules in the
high viscous emulsion containing LACTEM + GMU exhibited large
irregular aggregates (Fig. 3d). The emulsion with only LACTEM
also contained aggregated droplets, and the sizes of the aggregates
increased drastically when the emulsion was subjected to shear at
room temperature (Fig. 3b and c). The physical mechanism of the
latter aggregate enlargement was most likely occulation, since
reduced viscosity could be regained by cooling and shearing. On
the other hand, the large coherent aggregates of the emulsions
with LACTEM + GMU could not similarly be changed to give a
low-viscous emulsion and thus probably resulted from partial
coalescence.

Fig. 3. Micro-structure investigated by CLSM of emulsions containing various monoglycerides. Before microscopic analysis the emulsions were stored at 5 C. The fat fraction is stained and
the scale bar equals 10 m. a: Control, b: LACTEM, low viscous, c: LACTEM, high viscous due to shear at room temperature, d: LACTEM + GMU, e: LACTEM + GMS, f: LACTEM + DATEM.

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M.B. Munk et al. / Food Research International 54 (2013) 17381745

Fig. 4. Melting and crystallization thermograms of bulk PKO. Arrows indicate dened
melting and crystallization temperatures.

3.3. DSC analysis of emulsions and bulk PKO

Partial coalescence of lipid droplets in emulsions requires that the
lipids are partially crystallized. Since the emulsiers potentially could
modify the crystallization behavior of the palm kernel oil, the effect of
the different monoglyceride combinations on the melting and crystallization behaviors of both bulk and emulsied PKO was studied by DSC.
For the study of bulk PKO, the monoglycerides were added both
separately in a concentration of 5 wt.% or in combination with 5 wt.%
LACTEM. The concentration of monoglycerides in the bulk fat was
higher than in the emulsions for two reasons. First, it was assumed
that an excessive concentration would make the crystallization effect
of monoglycerides more distinct if any. Second, the local concentration
of monoglyceride was much higher at the droplet interface compared to
the total concentration in the emulsions and therefore it was presumed
that a higher monoglyceride concentration in bulk PKO would reect
the crystallization behavior more realistic.
The endothermic DSC thermogram of pure PKO showed that PKO
melted over a broad temperature range between 5 C and 43 C in
accordance with the observations by Siew (2001) (Fig. 4). The onset
melting temperatures could not be determined reliably due to a very
gradual onset of melting. Melting temperatures were therefore dened
as the peak temperature of the lowest endothermic peak as indicated in
Fig. 4. In agreement with previous work, the presence of monoglycerides did not signicantly affect the melting behavior of neither bulk
nor emulsied PKO (P N 0.05), Table 1 (Barfod, Schrader, & Buchheim,
2000).
In the cooling experiments, most of the emulsions and bulk PKO
samples showed two exothermic peaks caused by crystallization; a
minor one at 2223 C and a major one at 1918 C (Fig. 4
and Table 1). Apart from GMS, the monoglycerides did not have

signicant effect on the crystallization temperature of bulk PKO samples

(P N 0.05). Only addition of GMS increased crystallization temperature
signicantly of bulk PKO (P b 0.001), probably due to GMS being a
high melting component (mp. N 60 C). Formation of crystals early in
the cooling experiment can have acted as seeds for nucleation of PKO.
In contrast, the crystallization temperature of emulsied PKO without
added monoglycerides was signicantly reduced compared to bulk
PKO most likely as a consequence of supercooling. The entire fat fraction
in the control emulsion crystallized around 8 C, which indicated
supercooling due to the absence of nucleus monoglycerides and the
presence of a thick protecting layer of sodium caseinate blocking intrusion of nucleating crystals (Palanuwech & Coupland, 2003). Similar to
the bulk PKO-blends, emulsions containing LACTEM, GMU and GMS
displayed a small exothermic peak around 2228 C, and a second
exothermic peak at temperature N 8 C (Table 1). Destabilized emulsion droplets are expected to yield a thermogram comprising the crystallization peaks of both the bulk PKO and the dispersed PKO, that is
23 C, 19 C and 8 C respectively (Palanuwech & Coupland, 2003;
Vanapalli, Palanuwech, & Coupland, 2002). Since none of the emulsions
containing monoglycerides were crystallizing at 8 C, droplet destabilization was apparently not the cause for the increased crystallization
temperature as compared to the control emulsion. Emulsion destabilization implies that the positions of the peaks corresponding to bulk
and emulsied fat are steady, and therefore the single crystallization
peak at 11 C for emulsions containing LACTEM + DATEM can neither
be explained by droplet destabilization. The different crystallization
behavior between emulsions with monoglycerides and the control
emulsion is probably caused by monoglycerides that catalyze the nucleation event by intervening as heteronuclei and thereby increase the
crystallization temperature. Davies, Dickinson, and Bee reported similar
effects on crystallization of both bulk and emulsied fat with addition of
emulsiers (Davies et al., 2001).

3.4. Crystallization kinetics of bulk PKO

Crystallization of bulk PKO at 5 C occurred too fast to follow experimentally and therefore the monoglycerides' inuence on crystallization kinetics of bulk PKO was examined at 10 C and 20 C by pulsed
nuclear magnetic resonance (pNMR). By tting the isothermal SFC as
function of time to the Avrami model (Eq. (1)), estimates could be
obtained for the crystallization rate constant (Avrami constant k), and
the index of crystallization related to the type of nucleation and dimensionality of growth (Avrami exponent n) (Table 2). The maximum
crystalline content was within 6070% at 10 C and 8289% at 20 C
for all emulsions. Each monoglyceride was added in concentrations of
5 wt.% as no effect could be detected at lower concentrations. GMU
and DATEM did not affect crystallization of PKO at either temperature
and values of k and n similar to those of pure PKO were obtained
(Fig. 5 and Table 2). Both LACTEM and GMS increased the crystallization
rate of PKO. The increase in crystallization rate was also observed when

Table 1
Melting and crystallization temperatures of bulk PKO and emulsions containing various monoglycerides. Melting temperature was dened as the peak of the rst endothermic peak. Most
samples displayed two exothermic crystallization peaks, a minor (~22 C) and a large one (~19 C).
Emulsier

No monoglycerides
LACTEM
GMU
GMU + LACTEM
GMS
GMS + LACTEM
DATEM
DATEM + LACTEM

Bulk PKO

Emulsion

Tpeak melting (C)

Tpeak crystallization (C)

Tpeak melting (C)

Tpeak crystallization (C)

23.2
23.8
24.0
23.3
24.4
23.4
22.9
23.4

22.8
23.5
22.1
22.7
25.7
26.3
23.3
23.3

24.0 0.4
23.7 0.6

7.8 0.1
21.9 0.1/17.4 0.2

23.8 0.5

21.8 0.8/17.7 0.6

23.5 0.2

27.9 0.4/15 0.2

24.0 0.6

11.4 0.2

0.8
0.6
0.0
1.1
0.1
0.0
0.5
0.3

0.3/19.1
0.3/19.1
0.2/18.3
0.5/18.1
0.2/19.7
0.5/20.9
0.2/19.7
0.1/18.9

0.2
0.2
0.3
0.1
1.0
0.2
0.1
0.4

M.B. Munk et al. / Food Research International 54 (2013) 17381745

1743

Table 2
Avrami exponent n and Avrami constant k for crystallization of PKO containing various monoglycerides at 10 C and 20 C.
PKO

10 C

20 C

n
No monoglycerides
LACTEM
GMU
GMS
DATEM
LACTEM + GMU
LACTEM + GMS
LACTEM + DATEM

1.8
1.5
1.9
1.6
1.9
1.6
1.3
1.5

0.1
0.1
0.1
0.1
0.1
0.2
0.1
0.1

6.8
6.4
5.7
2.9
4.9
3.4
1.6
4.2

10
104
105
104
105
104
103
104

LACTEM was mixed with other emulsiers, although the effect was
moderated with GMU and DATEM. In contrast, combination of GMS
and LACTEM resulted in the highest crystallization rate and the lowest
value of n at 10 C.
For all PKO blends the Avrami exponent, n, assumed a value
between 1.3 and 1.9. This indicate a similar crystal growth mechanism, which according to polarized light microscopy (PLM) (data
not shown) most likely was a rod-like growth from sporadic nuclei
(n = 2) (Marangoni, 2005). Thus, the tested monoglycerides
seemed not to have an effect on nucleation and growth dimensions.
However, the possibility that GMS induced rod-like growth from
instantaneous nuclei (n = 1) (Marangoni, 2005) cannot be rejected
since initial crystallization for this blend had occurred prior to the
PLM examination. Image analysis (data not shown), however,
suggested similar nucleation and morphology among the blends.

Fig. 5. Effect of monoglycerides on the initial crystallization kinetics of PKO at 10 C and

20 C. : No monoglycerides,: LACTEM, : GMS, : DATEM, : GMU. Insets: The
complete crystallization at 10 C and 20 C followed over 1 h and 2 h, respectively.

3.5
3.5
2.9
1.2
3.0
3.5
4.1
2.3

10
104
105
104
105
104
104
104

1.9
1.4
1.9
1.6
1.9
1.6
1.3
1.7

0.2
0.2
0.2
0.1
0.2
0.2
0.1
0.2

2.3
4.4
1.5
1.2
1.6
9.2
5.0
5.0

105
104
105
104
105
105
104
105

2.0
3.6
1.5
8.6
2.2
9.9
2.7
6.4

105
104
105
105
105
105
104
105

3.5. Polymorphic transformations of emulsions and bulk PKO

The polymorphic forms were identied by reections in powder Xray diffraction experiments corresponding to characteristic d-spacing.
The -form is characterized by d = 4.15 , is characterized by
d = 3.8 and 4.2 and characterized by d = 4.6 (AOCS, 1995).
The transformation of emulsions from pourable to thick pastes upon
shear made it plausible to believe that the transformation came along
with recrystallization of the fat. Therefore X-ray diffraction was carried
out twice on emulsions. First emulsions were not exposed to any
preceding treatments and second emulsions were stirred with a spatula
until viscosity had increased. The prevalence of polymorphic forms was
indifferent although the textural properties of the emulsions had
changed, Fig. 6. All emulsions regardless the type of monoglycerides
and pourability consisted of -polymorphs with traces of .
For the PKO blends the X-ray diffractograms differed a bit more,
although all blends contained a mixture of - and -polymorphs.
The ratio of / depended on added monoglyceride, cooling temperature and time after temperature quench. The diffractograms were
superimposed, which complicated a straightforward quantication of
the relative amount of each polymeric forms using information from
the short spacing part of the diffractograms. In such case long spacing
d-values in the SAXS regime have previously been used for quantication of the polymorphs (de Graef, van Puyvelde, Goderis, &
Dewettinck, 2009; Fredrick, Foubert, Van De Sype, & Dewettinck,
2008; Kalnin, Schafer, Amenitsch, & Ollivon, 2004). In the present case
a clear correlation was found between the reection at 4.15 and a
reection at 43 , and the latter was thus also attributed to the polymorph. Likewise, a correlation was established between reections
at 3.8/4.2 and 35 , which again were attributed to the -polymorph.

Fig. 6. Crystal polymorphism of emulsion containing LACTEM + GMS. The upper

diffractogram displays a low-viscous pourable emulsion with no pre-treatment and the
lower diffractogram displays the emulsion stirred into a thick paste. The peaks
corresponding to (d = 4.2 and 3.8 ) and (d = 4.15 ) are pointed out.

1744

M.B. Munk et al. / Food Research International 54 (2013) 17381745

Table 3
The relative ratio of -crystals compared to -crystals in bulk PKO with addition of
monoglycerides as a function of storage time at 5 C and 20 C. After day 7 the peak
corresponding to was not detectable (n.d.) for most samples and therefore results
from day 21 are not included in the table.
Emulsier

No monoglycerides
LACTEM
GMU
GMU + LACTEM
GMS
GMS + LACTEM
DATEM
DATEM + LACTEM

5 C

20 C

Day 0

Day 1

Day 7

Day 0

Day 1

Day 7

0.9
3.0
1.0
1.0
1.6
6.4
1.2
1.0

1.3
3.2
2.0
1.8
2.2
9.5
1.9
2.9

1.4
3.0
4.0
3.1
2.1
8.0
n.d.
4.1

1.5
3.4
2.0
1.5
2.0
2.3
3.4
1.5

1.9
4.5
6.5
2.1
2.3
4.2
n.d.
2.0

2.1
4.3
5.9
n.d.
2.8
8.8
n.d.
n.d.

The relative rate of polymorphic transformation from to at temperatures of 5 C and 20 C was followed for a duration of 3 weeks, and
the results are shown in Table 3. The time development of the polymorphic transformation from to is studied by examining the ratio:


0 Apeakd35

;
Apeakd43

which expresses the ratio between the amount the two polymorphic
forms.
Initially all PKO blends comprised a mix of and , but during three
weeks was transformed into , the most stable polymorph of PKO
(Rossell, 1975). The preference of over is most likely due to the
wide range of fatty acid chain lengths in triglycerides of PKO (Sato,
2001; Timms, 1984). The initial fraction of , as well as polymorphic
transformation rate, was clearly dependent on crystallization temperature, storage time and addition of monoglycerides. Using a storage temperature of 5 C caused a higher fraction of -polymorphs as compared
to 20 C, due to low thermodynamic stability and low melting point of
(Sato & Ueno, 2011). Subsequent recrystallization into -polymorphs
occurred, and after 7 days the reection corresponding to was not
distinguishable for most samples. Several of the tested monoglycerides
favored initial formation of directly from the melt. This applied especially to LACTEM, LACTEM + GMS and DATEM, although for the latter
this was only observed at 20 C, Table 3. In addition, all monoglycerides,
to varying degrees, accelerated the polymorphic transformation in
agreement with previous studies (Aronhime, Sarig, & Garti, 1988;
Fredrick et al., 2008; Smith et al., 2011). Overall, all tested monoglycerides promoted faster formation of the most stable polymorph. This
might be related to the fact that except anionic DATEM, all applied
monoglycerides were non-ionic which are suggested to induce transformation of crystals into more stable polymorphs (Bunjes, Koch, &
Westesen, 2003). However, the monoglycerides appeared to be divided
into two groups: GMU and DATEM (used in pure form or in combination
with LACTEM) accelerated gradually the transformation from to ,
whereas GMS and LACTEM readily caused a high level of directly
from the melt which remained fairly constant during the remaining experimental period (Fig. 7).
4. Discussion
The stabilities of the investigated emulsions were strongly dependent on the type of monoglycerides added. Both droplet size distributions and microscopic inspection revealed formation of large fat
aggregates in the two high-viscosity emulsions made with LACTEM or
LACTEM + GMU. Furthermore, increased viscosity as a consequence
of shear and increased temperature was linked to enlargement of the
aggregates. The clusters of droplets remained non-dispersable in a
water solution despite vigorous shaking, and therefore the aggregates

Fig. 7. Polymorphic transformations of to -crystals during storage for 20 days at 5 C

for PKO without monoglycerides (), with LACTEM + GMU () and with
LACTEM + GMS ().

were unlikely to be held together by occulation. Furthermore, the

partial crystalline state of the dispersed fat, the irregular shapes of the
aggregates and the increase in emulsion viscosity indicated that
coalescence was not the responsible instability mechanism, since coalesced droplets remain spherical and do not affect viscosity (Fredrick,
Walstra, & Dewettinck, 2010). Partial coalescence is therefore a likely
mechanism of aggregation of fat droplets and the change of the
emulsion textures. While GMU increased the instability induced by
LACTEM, partial coalescence was somehow prevented by using a combination of LACTEM together with GMS or DATEM. The mechanism behind the enhanced stability remains unknown, but GMS and DATEM
might increase the thickness of the adsorbed layer around the fat droplets, as a thicker layer counteracts partial coalescence (McClements,
Dickinson, Ngan, Sella, et al., 1993; Palanuwech & Coupland, 2003). Susceptibility to partial coalescence was not related to polymorphic transformations of lipid crystals since the polymorphism was identical for
all emulsions and remained unaltered despite large rheological changes.
The impact of the monoglycerides on emulsions was only assessed at a
given set of concentrations highly inspired by the compounding of
commercial emulsier mixtures. Further studies are required to establish if the reported effects will be valid for other concentrations of
monoglycerides as well.
A prerequisite for partial coalescence is fat crystallization in the dispersed droplets. Besides being present at the interface, crystals should
be large and oriented perpendicular to the interfacial plane. The probability of a crystal piercing through the interface of another droplet is
higher when the orientation is perpendicular to the interfacial plane
(Rousseau, 2000). For occurrence of partial coalescence, large crystal
sizes are preferable since penetration into neighboring droplets would
be more efcient. Vanboekel and Walstra (1981) found that large droplet sizes increased the rate of partial coalescence due to the fact that
crystals can achieve larger sizes in larger droplets. Fast cooling or efcient undercooling could therefore contribute to suppression of partial
coalescence as these methods inhibit crystal growth and generate
small crystals (Ghosh & Rousseau, 2010). This might explain the stability of the emulsion containing LACTEM + DATEM, which displayed a
considerably higher undercooling prior to crystallization compared to
the other emulsions containing monoglycerides (Table 1). Furthermore,
partial coalescence increases with increasing solid fat content as a consequence of high prevalence of crystals able to protrude and pierce
through adjacent droplets (Boode, Walstra, & Degrootmostert, 1993).
However, the polarity of crystal faces is also of importance as protruding
crystals should be wetted by oil instead of water to initiate partial coalescence (Vanboekel & Walstra, 1981). Partial coalescence also implies
that fat droplets are able to approach one another, thus implying that
electrostatic repulsion provided by interfacial sodium caseinate must
be reduced. It is generally assumed that monoglycerides displace

M.B. Munk et al. / Food Research International 54 (2013) 17381745

proteins on the droplet interface and consequently the fat droplets tend
to agglomerate. If protein displacement is the cause of the observed
morphology and rheological changes, GMU can be expected to displace
caseinate from the droplet interface more efciently than GMS and
DATEM. However, the present study provides no information about
competitive displacement and further analyses are required to clarify
this aspect.
In contrast to the large effects on emulsion stability, the monoglycerides tested only affected the overall crystallization rate of bulk PKO to a
minor extent. However, polymorphic transformations to stable PKO
crystals were accelerated by addition of all monoglycerides in the
bulk. Furthermore, both LACTEM and GMS increased the rate of crystallization, while GMS also initiated the nucleation stage causing crystallization to proceed at a higher temperature. In conclusion, the impact of
monoglycerides on bulk PKO crystallization is far less noticeable compared to the pronounced effect observed in the dispersed state (emulsions). In particular, GMU in combination with LACTEM had a major
impact on the stability of the emulsions. However, only a minor effect
was observed when LACTEM + GMU were added to bulk PKO.
5. Conclusion
This study shows that physical stability of emulsions is strongly dependent on the choice of emulsiers. However, the results also point to
the fact that the effects of emulsiers in bulk fat and emulsied fat are different. Furthermore it emphasizes that emulsiers either exert a different
effect when oriented at the wateroil interface compared to orientation
in bulk fat or affect interactions between droplets rather than affecting individual droplets (McClements & Dungan, 1997). In conclusion, the different processes leading to instability of emulsions, as illustrated by
changes in droplet size, droplet morphology and rheological properties,
cannot be explained by investigating a simplied bulk fat model system.
Acknowledgment
The authors gratefully acknowledge the Danish Agency for Science,
Technology and Innovation for nancial support.
Appendix A. Supplementary data
Supplementary data to this article can be found online at http://dx.
doi.org/10.1016/j.foodres.2013.09.001.
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