LWT - Food Science and Technology 138 (2021) 110593

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LWT
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Palm-based fat crystallized at different temperatures with and without

high-intensity ultrasound in batch and in a scraped surface heat exchanger
Thais Lomonaco Teodoro da Silva a, b, *, Sabine Danthine b, Silvana Martini a, **
a
Department of Nutrition, Dietetics, and Food Sciences, Utah State University, 8700 Old Main Hill, Logan, UT, 84322-8700, USA
b
Science des Aliments et Formulation, Gembloux Agro-Bio Tech, ULiège, Gembloux, Belgium

A R T I C L E I N F O A B S T R A C T

Keywords: The objective of this study was to compare the sonocrystallization process of a palm-based fat in batch to a
Sonocrystallization continuous set-up using a scraped surface heat exchanger (SSHE). The sample was crystallized at 32, 30, 28, and
Batch 26 ◦ C (Tc). High-intensity ultrasound (HIU, 20 kHz, 12.7 mm horn, 50% amplitude, 57W) was applied using 10s
Scraped surface heat exchanger
pulses. After crystallization, samples were stored (48 h/25 ◦ C) and analyzed for crystal microstructure, melting
Low saturated fat
behavior, oil binding capacity (OBC), hardness, and viscoelastic properties. HIU improved all physical properties
of the material when crystallized in batch at all Tc; however, SSHE at 26 ◦ C was the only condition improved by
HIU. G’ and hardness were the highest in sonicated SSHE samples crystallized at 26 ◦ C (141 kPa, 4.5N) and these
values were higher than the ones obtained in batch. OBC was also improved in sonicated SSHE samples (77%)
but in lower magnitude compared to the OBC obtained in batch (82%). Crystal size was not affected by HIU in
the SSHE, but a reduction in crystal size was observed due to HIU in batch for all Tc. Similar behavior was
observed for Tp, however, Ton and ΔH were not affected by HIU.

1. Introduction sonocrystallization using high-intensity ultrasound (HIU) (Ueno, Ristic,

Lipids are commonly used in foods, pharmaceutical, and cosmetic
industries since they provide desired physical properties to final prod­
ucts. Palm oil and its fractions are among the most studied and used oils
worldwide because of their availability and the unique functional
properties inherent to these products, allowing their use in a wide range
of products including margarines, shortenings, baking, and confection­
ary fats (DeMan, 1998). Previous studies on the effect of external factors
such as shear on the crystallization of palm oil demonstrated that shear
enhanced the primary crystallization and changed its microstructure (de
Graef, Puyvelde, Goderis, & Dewettinck, 2009).
Nutrition concerns regarding consumption of high levels of saturated
fats as found in the palm oil and its high melting point fractions, have
resulted in a growing demand to reduce the saturated fatty acid (SFA)
content in fat-based products. This reduction requires increased crys­
tallization rates and strengthening the crystal network of trans free and
low saturated fat products by applying external factors to the crystalli­
zation processes (Sato & Ueno, 2011).
One alternative explored by food scientists was the use of
Higaki, & Sato, 2003). Studies have shown that the vibrational energy
input from cavitation bubbles promotes nucleation over greater heat and
mass transfer. In addition, HIU promotes heterogeneous or secondary
nucleation, resulting in a change of micro- and macroscopic properties
of the sonicated fat (Suzuki, Lee, Padilla, & Martini, 2010). Most of the
studies on lipid sonocrystallization were performed in a batch system
(Chen, Zhang, Sun, Wang, & Xu, 2013; Suzuki et al., 2010; Ueno et al.,
2003). However, few studies have evaluated the effect of HIU in
continuous systems including studies in a flow cell for palm (Ye &
Martini, 2015) and milk fat blend (Gregersen et al., 2019), and in a
scraped surface heat exchanger (SSHE) for a soybean and palm-based fat
(da Silva, Danthine & Martini, 2020b; da Silva & Martini, 2019, 2020b,
2020a).
Overall, sonocrystallization studies have shown that the effect of
sonication is dependent not only on sonication conditions but also on the
type of sample used. Kadamne, Ifeduba, Akoh, and Martini (2017b,
2017a) showed that sonication of stearic based samples resulted in
greater changes in physical properties compared to sonication of
palm-based ones. Some of our previous studies evaluated the effects of

* Corresponding author. Department of Nutrition, Dietetics, and Food Sciences, Utah State University, 8700 Old Main Hill, Logan, UT, 84322-8700, USA.
** Corresponding author.
E-mail addresses: thais_lts@yahoo.com.br (T.L.T. da Silva), silvana.martini@usu.edu (S. Martini).

https://doi.org/10.1016/j.lwt.2020.110593
Received 12 August 2020; Received in revised form 23 October 2020; Accepted 14 November 2020
Available online 21 November 2020
0023-6438/© 2020 Elsevier Ltd. All rights reserved.
T.L.T. da Silva et al. LWT 138 (2021) 110593

Table 1 2. Material and methods

Triacylglycerol composition.
TAG species Percentage (%) 2.1. Material
LnLnL 0.80 ± 0.01
LnLL 3.78 ± 0.05 Interesterified palm olein (IPO, melting point 44.8 ◦ C), and soybean
LLL 10.41 ± 0.16 oil (SBO, melting point − 12.8 ◦ C) were donated by ADM. They were
LnLO 1.74 ± 0.04 mixed in the proportion 60IPO:40SBO to reduce the level of SFA to
LLO 8.08 ± 0.09 approximately 32%. The major fatty acid composition of the blend based
PLL 7.96 ± 0.04
LnLSt 0.42 ± 0.01
on the specification sheet provided by the supplier was 27.9% palmitic,
OOL 6.26 ± 0.12 4.2% stearic, 35.0% oleic, 27.4% linoleic and 3.1% linolenic. The solid
POL 11.97 ± 0.21 fat content of the blend is 29.4% (10 ◦ C), 15.0% (20 ◦ C), 7.3% (30 ◦ C)
PLP 4.88 ± 0.11 and 2.1% (40 ◦ C).
MPP 0.28 ± 0.01
OOO 4.75 ± 0.05
POO 14.66 ± 0.20 2.2. Crystallization
POP 13.59 ± 0.22
PPP 3.89 ± 0.11 Batch crystallization: Samples (100g) were melted and kept in the
StOO 1.69 ± 0.21 oven (75 ◦ C/30min) to erase crystalline memory. Samples were then
POSt 3.11 ± 0.36
PPSt 1.71 ± 0.61
placed in a double-walled crystallization cell connected to a
StOSt 0.36 ± 0.22 temperature-controlled circulating water bath and stirred with a mag­
Total TAGs by group netic stirrer (100 rpm) for the first 10 min. Samples were crystallized at
UUU 35.82 ± 0.51 26, 28, 30 and 32 ◦ C (Tc) for 90min. The time point when the first
SUU 36.49 ± 0.05
crystals appeared (onset of crystallization) was observed with the naked
SUS 21.94 ± 0.18
SSS 5.74 ± 0.64 eye as the presence of slight turbidity, and an induction time was fixed
for this point (Suzuki et al., 2010). HIU (20 kHz, 10s, 50% amplitude,
Ln: linolenic acid, L: linoleic acid, O: oleic acid, P:
57W, 12.7 mm horn, Q500 system, Qsonica, Newtown, CT, USA) was
palmitic acid, M: myristic acid, St: stearic acid. U: un­
applied at the onset of crystallization to maximize the induction of
saturated, S: saturated.
secondary nucleation which occurred at 12min for Tc = 26 ◦ C, 15min for
Tc = 28 ◦ C, 22 min for Tc = 30 ◦ C, and 32min for Tc = 32 ◦ C. In all cases
HIU was applied for 10s (Ye & Martini, 2015). After 90 min of crystal­
lization in the cell at constant Tc (26, 28, 30, and 32 ◦ C), samples were
stored in an incubator at 25 ◦ C for 48h. Samples processed in the same
way as described above but without the use of sonication were used as
controls. All crystallization conditions were replicated three times.
SSHE crystallization: The same temperatures as those described for
the batch system (Tc = 26, 28, 30, 32 ◦ C) were used in the SSHE
experiment. The sample was melted in a pot placed on top of a con­
vection stove at 60 ◦ C/30min. An overhead stirrer was used to ensure
appropriate heat transfer. The melted sample was pumped through the
SSHE system at 11L/h. The SSHE (Armfield Limited, England, model
FT25BBPA–IF–C) was composed of a progressive cavity pump (20L/h
capacity, 01–71L/4-TF), followed by two barrels with shear agitation
(344 rpm, Tc), and one pin worker (208 rpm, no temperature control) as
illustrated in da Silva and Martini (2019). HIU was applied by placing a
65 mL water-jacketed flow cell (flow cell #4650; Qsonica, Newtown, CT,
USA) attached to the HIU horn at the end of SSHE (after the pin worker)
Fig. 1. Melting point curve of the palm-based blend. to assure no shear was applied after sonication, similar to what was
performed in the batch system. HIU was applied with a 12.7 mm horn
using 50% amplitude and 10s pulses (10sON/10sOFF) (da Silva &
HIU processing parameters and position in the SSHE in a soybean
Martini, 2020b). All crystallization conditions were performed in trip­
based-fat (da Silva & Martini, 2019, 2020a; da Silva, Danthine et al.,
licate. For each crystallization condition tested a sonicated and a
2020b). These studies showed that when used in a SSHE a
non-sonicated sample (control) were collected. Although at least 1500g
soybean-based fat crystallized at a lower supercooling resulted in the
of each condition was produced to ensure that the SSHE was full of the
greatest improvement on the physical properties due to HIU (da Silva,
treated fat, only 100g of sample (3 pulses of 10s) was collected and
Danthine et al., 2020b). For a palm-based fat, although HIU seems to
stored it in an incubator at 25 ◦ C for 48h.
have an effect on physical properties when used in the SSHE (da Silva &
It is important to notice that the same HIU conditions such as fre­
Martini, 2020b), the magnitude of this improvement was lower
quency, horn size, sonication duration and amplitude were used in both
compared to the findings for a soybean based. This issue is normally not
systems. The only difference between both systems is that in the SSHE
found in batch sonocrystallized samples, and due to the importance of
sonication was applied using 10 s pulses to ensure that the sample
palm oil in foods and the need to reduce SFA in fats, the objective of this
moving through the cell is subjected to only 10 s of acoustic waves
study was to compare and evaluate the effects of HIU in an palm-based
(residence time in the cell ~20 s).
fat with low content of SFA (32%) in a batch and in a SSHE system using
different supercooling levels to evaluate the difference in physical
2.3. Methods
properties based on different crystallization systems.

2.3.1. Triacylglycerol composition (TAG)

The TAG composition was analyzed by reversed-phase HPLC based
on the official AOCS method Ce 5b-89 (AOCS, 2009). Minor practical

2
T.L.T. da Silva et al.
Fig. 2. Images of the crystalline network obtained from polarized light microscopy at 25 ◦ C after crystallization in batch or in a SSHE at different temperatures with
and without HIU and stored at 25 ◦ C for 48h. Magnification 10x.
adjustments to the flow rate and mobile phase composition were made
as described below. The analysis was performed on a Waters HPLC
system (Zellik, Belgium), equipped with two stainless steel Nova-Pak
C18 columns (4 μm, 3.9 × 150mm) (Waters, Belgium). The mobile
phase was an isocratic solvent mixture of acetone and acetonitrile
(62.5:37.5) with a flow rate of 1.2 mL/min; the injection volume was 20
μl. The samples (0.04g) were dissolved in methanol/chloroform (1.6 mL;
1:1 v:v) and a differential refractometer was utilized for detection. The
equivalent carbon number (ECN) was used to predict the elution order.
ECN = CN-2(DB), where CN is the total carbon number and DB is the
total number of double bonds in the fatty acids. Peaks were integrated by
using the program Empower Pro with a generic Apex Track method and
areas were correlated with the quantities of TAG in the sample. Peak
areas below 4000 area counts were not considered.
2.3.2. Polarized light microscopy
After storage at 25 ◦ C for 48h a small portion of the crystallized
samples were placed on a slide and covered with a cover-slide. The slides
were placed on a polarized light microscope (Olympus BX41 micro­
scope, Olympus, Tokyo, Japan) equipped with a digital camera (Infinity
3
LWT 138 (2021) 110593
2, Lumenera Scientific, Ottawa, ON, Canada). Images were obtained and
used to characterize crystal morphology. The diameters of the crystals
were measured using the software Image-Pro® Premier E 9.2 (Media
Cybernetics, USA). For each crystallization condition four images were
obtained.
2.3.3. Oil binding capacity
After storage at 25 ◦ C for 48h, 1g of the crystallized sample was
placed in a pre-weighed microcentrifuge tube and centrifuged at 10,000
rpm for 15 min at room temperature using a microcentrifuge (Ther­
moscientific, Sorvall micro 17 centrifuge, Germany). After centrifuga­
tion, the microcentrifuge tube was inverted upside down and left to
drain for 3min to release the liquid oil not entrapped in the crystalline
network. The oil binding capacity (OBC) was performed in quadrupli­
cate and calculated according to equation (1).
( )
wbefore − wafter
OBC(%) = 100 − * 100 (1)
Wsample
where Wbefore is the initial weight of the tube and the sample, Wafter is the
T.L.T. da Silva et al. LWT 138 (2021) 110593

Table 2 removed and cut into 1 cm height cylinders. Hardness was measured
Crystal size obtained from polarized light microscopy images reported as using a Texture Profile Analyzer (Model TA, XT Plus, Texture Technol­
diameter (μm, mean ± standard deviation) at 25 ◦ C after crystallization in batch ogies Corp., Hamilton, MA, United States) with a two-step 25%
or in a SSHE at different temperatures with and without sonication and stored at compression test and a 5 cm cylinder probe. Five tubes were prepared
25 ◦ C for 48h. for hardness measurement for each crystallization run.
Temperature of crystallization (◦ C) Process condition Diameter (μm)

26 Batch control 7.5 ± 1.83d 2.3.7. Statistical analysis

Batch HIU 4.4 ± 0.08f Crystallization runs were performed in triplicate and analytical
SSHE control 5.3 ± 0.25 ef measurements were done in replicates as described above. Data reported
SSHE HIU 5.2 0.22 ef
±
are mean values and standard deviation of the mean. Statistical differ­
28 Batch control 10.0 ± 1.25c ences between treatments were evaluated using two-way ANOVA (α =
Batch HIU 5.2 ± 0.32 ef 0.05) using Tukey as a post-hoc test (GraphPad Prism, version 8.0, La
SSHE control 5.1 ± 0.09 ef
SSHE HIU 5.3 ± 0.16 ef
Jolla, CA). For all analyses, crystallization temperature and process
conditions (batch vs. SSHE) were used as the two factors.
30 Batch control 18.2 ± 3.40b
Batch HIU 5.9 ± 0.20 def
SSHE control 5.3 ± 0.09 ef 3. Results and discussion
SSHE HIU 5.6 ± 0.08 def

32 Batch control 22.0 ± 2.52a

Batch HIU 6.5 ± 0.66 de
SSHE control 5.9 ± 0.32 def
SSHE HIU 5.5 ± 0.25 def
*Means followed by a common letter are not significantly different by the Tukey-
test at the 5% level of significance.
final weight of the tube and the sample (after centrifugation and
draining of the oil), Wsample is the weight of the sample.
2.3.4. Melting behavior
The melting profile and melting point were measured in a Differen­
tial Scanning Calorimetry (DSC). DSC was calibrated at 5 ◦ C/min using
Indium as a standard. The melting point of the sample was measured by
placing the sample (10–15 mg) in a Tzero DSC pan, covered with a Tzero
lid, sealed hermetically, and placed in the DSC oven at 25 ◦ C. The sample
was heated at 5 ◦ C/min to 80 ◦ C, kept at 80 ◦ C for 30 min to erase
crystalline memory, and cooled to − 20 ◦ C at 5 ◦ C/min and kept at − 20
◦
C for 90 min to allow for complete crystallization. Samples were then
melted to 80 ◦ C at 5 ◦ C/min. The peak temperature (Tp) obtained from
the last melting step was used to quantify the melting point.
To evaluate the melting behavior of the samples after storage at 25 ◦ C
for 48h the sample was stabilized in the DSC at 25 ◦ C for 1 min and then
heated from 25 to 80 ◦ C using a 5 ◦ C/min rate. The parameters used to
quantify the melting behavior of the samples were onset temperature
(Ton), peak temperature (Tp), and change in enthalpy associated with the
melting process (ΔH). This measurement was done in triplicate for each
crystallization run.
2.3.5. Viscoelasticity
The viscoelasticity of samples crystallized with and without HIU
after storage at 25 ◦ C for 48h was measured using an AR-G2 Rheometer
(TA Instruments, New Castle, Delaware) operating with air purge
(30psi). Measurements were performed using a 40 mm standard steel
parallel plate and a 2,000 μm gap. The instrument was operated by the
Rheology Advantage Instrument Control software, where viscoelastic
properties such as elastic (G′ ) and viscous (G′′ ) moduli were measured
using a strain sweep oscillation from 0.0008 to 10% strain at 25 ◦ C. The
frequency was kept constant at 1Hz and G’ and G” reported are means of
the values obtained in the linear viscoelastic region (LVR). The Rheology
Advantage software (TA Instruments, New Castle, Delaware) was used
for data analysis. All experiments were performed in quadruplicate for
each crystallization run.
2.3.6. Texture profile analyzer (TPA)
Hardness was measured after storing the samples in 1 cm diameter
plastic tubes at 5 ◦ C for 48h. Samples were not stored at 25 ◦ C since they
were too soft to perform compression tests. After storage, samples were
3.1. Triacylglycerol composition
The main TAGs found were LLL, POL, POO and POP (L: linoleic acid,
O: oleic acid, P: palmitic acid, Table 1). In terms of TAG types, tri-
unsaturated (UUU) and di-unsaturated (SUU) TAGs (S: saturated, U:
unsaturated fatty acids) were found in higher amounts (~35% each)
followed by disaturated (SUS,~20%) and trisaturated (SSS, ~6%) group
(Table 1).
The melting point curve shows two main peaks sub-divided in two
sub-peaks each (Fig. 1). The first peak had one shoulder with Tp around
− 4.5 ◦ C (black filled arrow) representing some UUU TAGs. The second
shoulder of the first peak had a Tp at 6.9 ◦ C (black non-filled arrow)
representing the melting of high melting point UUU and low melting
point SUU. The second peak contains high melting point TAGs in the first
shoulder (Tp of 19.5 ◦ C, grey filled arrow) and might be composed by the
SUU with high melting points and SUS TAGs. The last shoulder (grey
non-filled arrow) at higher temperatures (Tp of 41.5 ◦ C, melting point)
might be due to the smaller concentration of SSS TAGs.
3.2. Microscopy and macroscopy images
All non-sonicated samples from the batch crystallization showed big
round crystals (Fig. 2) that changed in size depending on the Tc used
(Table 2). This type of crystal morphology and size are similar to those
previously observed by Ye and Martini (2015) and Chen et al. (2013) for
palm oil. Sonication applied in batch changed crystal morphology
(Fig. 2) and reduced crystal sizes (Table 2) as previously reported in the
literature (Chen et al., 2013; Suzuki et al., 2010). Sonicated samples
showed small needle-like crystals that were more organized and evenly
distributed throughout the fat crystal network, and as lower the Tc
smaller the crystal. Bubbles generated during acoustic cavitation can
either oscillate or collapse forming a new population of bubbles that
induce primary nucleation, or release shock waves and create high shear
forces breaking down big crystals already formed and inducing sec­
ondary nucleation (Suzuki et al., 2010). In our study, HIU was applied at
the onset of crystallization where primary nucleation has already been
formed and therefore HIU induced secondary nucleation inhibiting or
delaying crystal growth and/or breaking growing crystals into many
smaller ones. Similar changes in crystalline organization have been re­
ported in the literature using sonocrystallization at the onset of crys­
tallization when HIU was applied for a short period of time (Chen et al.,
2013; da Silva, Cooper, Lee, Gibon, & Martini, 2020a).
No big round crystals were obtained in the non-sonicated samples
crystallized in the SSHE and small needles like the ones obtained in
sonicated samples in batch were observed. De Graef et al. (2009)
observed in a batch that smaller and more crystals were formed under
continuous isothermal shear (1 or 5s− 1), while big crystals were
observed under static conditions. Acevedo & Marangoni (2014) ob­
tained opposite results when using a soybean-based sample in a lab-scale

4
T.L.T. da Silva et al.
Fig. 3. Macroscopic view of crystalline networks obtained at 25 ◦ C after crystallization in batch or in a SSHE at different temperatures with and without HIU and
stored at 25 ◦ C for 48h.
SSHE system with wall temperature of 0 ◦ C and 40 rpm agitation, sug­
gesting that the time, amount, duration, and crystallization system used
for shear influences crystal size. The reduction in crystal size in the SSHE
was due to the shaved agitation that promoted faster cooling, induced
nucleation, and reduced crystal growth. No differences in crystal size or
morphology were observed due to sonication at all Tc in the SSHE, as
observed by (da Silva, Danthine et al., 2020b) this result does not
necessarily indicate that HIU was not effective, but that SSHE has a
bigger effect on tailoring crystal size.
Samples crystallized in batch at all Tc without sonication had a soft
texture that flowed easily and did not maintain its shape when placed in
a watch-glass (Fig. 3). Big crystals usually hinder the formation of a well-
structured crystalline network due to the small contact area and the
weak interactions among them (Kanagaratnam et al., 2013). After son­
ication, all samples crystallized in batch were visibly harder than the
5
LWT 138 (2021) 110593
non-sonicated ones, and sonicated samples crystallized in batch at 26,
28, and 30 ◦ C looked harder than the ones crystallized at 32 ◦ C. These
differences in visual hardness are attributed to the crystal sizes discussed
above. Samples crystallized in the SSHE without sonication were visibly
harder than samples crystallized without HIU in batch and they looked
as hard as the batch sonicated samples. After sonication, only SSHE
samples crystallized at 26 and 28 ◦ C kept this visual appearance while
samples crystallized with HIU in the SSHE at 30 and 32 ◦ C were visibly
softer, even though no differences were observed in the crystals
morphology and size.
3.3. Oil binding capacity (OBC)
The non-sonicated samples crystallized in batch, except for the one
crystallized at 32 ◦ C, showed lower OBC (p < 0.05; Fig. 4) than samples
T.L.T. da Silva et al. LWT 138 (2021) 110593

Fig. 4. Oil binding capacity (OBC) obtained at 25 ◦ C after crystallization in batch or in a SSHE at different temperatures with and without HIU and stored at 25 ◦ C
for 48h.

Fig. 5. Viscoelastic parameters G’ (A) and G” (B) obtained at 25 ◦ C after crystallization in batch or in a SSHE at different temperatures with and without HIU and
stored at 25 ◦ C for 48h.

crystallized in the SSHE. These results suggest that SSHE induced
nucleation and created smaller crystals that can entrap more liquid oil
when samples are crystallized between 26 and 30 ◦ C. In a previous study
in a palm blend with high levels of SFA, samples with bigger crystals had
a poorer OBC due to the lower surface area of the bigger crystals and
weaker interactions of the crystalline network (Kanagaratnam et al.,
2013).
The lack of effect on OBC for the samples crystallized in the SSHE at
32 ◦ C could be due to the slow crystallization process of the sample (De
Graef et al., 2009). When the sample was crystallized at 32 ◦ C, the higher

6
T.L.T. da Silva et al. LWT 138 (2021) 110593

Fig. 6. Hardness (N) measured at 5 ◦ C after crystallization in batch or in a SSHE at different temperatures with and without HIU and stored at 5 ◦ C for 48h.

OBC for all Tc (p < 0.05). The highest values were found for samples
Table 3 crystallized at 26, 28, and 30 ◦ C and the lowest for samples crystallized
Melting parameters obtained from DSC curves at 25 ◦ C after crystallization in at 32 ◦ C. An increase in OBC was observed only in sonicated samples in
batch or in a SSHE at different temperatures with and without HIU and stored at
the SSHE at 26 ◦ C and this improvement was not as big as the one
25 ◦ C for 48h.
observed for samples crystallized in batch. When samples were soni­
Temperature of Process Parameters cated in the SSHE at 28 ◦ C no change in the OBC was observed. However,
crystallization(◦ C) condition
Ton (◦ C) Tp (◦ C) ΔH (J/g) sonication at higher temperatures, 30 and 32 ◦ C, in the SSHE resulted in
26 Batch control 27.26 ± 39.17 ± 15.54 ±
a lower OBC compared to the non-sonicated samples. A previous study
0.35a 0.26 bc 2.53a in our group found that HIU improved OBC in low saturated fats because
Batch HIU 28.12 ± 37.72 ± 13.72 ± cavitation and bubbles created during sonication induced secondary
1.30a 0.48 cd 1.36a nucleation and crystallization of low-melting-point TAGs that would not
SSHE control 27.58 ± 37.43 ± 13.83 ±
crystallize without sonication (da Silva, Cooper et al., 2020a). The TAGs
0.36a 0.42d 1.67a
SSHE HIU 27.98 ± 37.85 ± 13.10 ± presented in high amounts due to sonication in (da Silva, Cooper et al.,
0.45a 0.22 cd 0.69a 2020a) were LLL, OLLn, LLO, PLL and POL, similar to the TAGs present
28 Batch control 27.76 ± 40.61 ± 14.82 ±
in this study (Table 1), suggesting that either they were better entrapped
0.14a 0.53 ab 1.26a or induced to crystallize. Shear that induced nucleation in the SSHE and
Batch HIU 27.54 ± 37.77 ± 14.32 ± assisted in the formation of the crystalline network can also break it, if
0.25a 0.34 cd 0.74a applied under inappropriate conditions. This could explain the small
SSHE control 27.87 ± 37.53 ± 13.57 ±
effect of SSHE on OBC values, where the shear experienced in the sample
0.18a 0.30d 0.93a
SSHE HIU 28.23 ± 37.27 ± 13.10 ± after crystallization improved the plasticity of the sample and might
0.53a 0.71d 0.85a have interfered with oil entrapment.
30 Batch control 28.03 ± 41.93 ± 9.68 ±
0.68a 1.65a 1.78b 3.4. Viscoelasticity
Batch HIU 28.23 ± 38.40 ± 15.24 ±
1.17a 1.44 cd 0.70a
SSHE control 27.78 ± 37.19 ± 14.08 ± All G′ values were higher than G” indicating an elastic behavior
0.43a 0.45d 0.44a (Fig. 5A–B). The highest G′ and G” were found for samples crystallized
SSHE HIU 27.94 ± 41.81 ± 14.45 ± with HIU at 26 ◦ C in the SSHE. For G”, no other condition besides 26 ◦ C
0.44a 0.92a 1.59a
with HIU in the SSHE showed the same high G” value. Nevertheless,
32 Batch control 27.73 ± 41.62 ± 13.38 ± sonication in batch or in the SSHE at 26 ◦ C showed statically similar G′
0.71a 0.94a 1.05a values (p > 0.05). At 26 ◦ C sonicated samples in batch or in the SSHE
Batch HIU 28.17 ± 37.34 ± 14.36 ±
0.16a 0.25d 1.11a
presented a higher G’ (p < 0.05), however the improvement in G′ due to
SSHE control 28.31 ± 42.13 ± 14.06 ± HIU in batch was not significantly different from the sample without
0.53a 0.82a 0.25a HIU in the SSHE. This was also true for G′ and G” at 28 ◦ C and G′ at 30 ◦ C
SSHE HIU 28.20 ± 41.15 ± 13.59 ± (p > 0.05). At 32 ◦ C, the sample crystallized in the SSHE without HIU
0.38a 0.30a 1.91a
had a higher G′ compared to the samples crystallized at the same Tc with
*Means followed by a common letter are not significantly different by the Tukey- HIU in batch. The high shear and efficient heat transfer in the SSHE
test at the 5% level of significance. explain the improvement in physical properties such as G’, G”, and OBC,
observed in samples crystallized using a SSHE compared to a batch
temperature, low driving force, and low residence time in the SSHE was system.
not enough to create a significant amount of nuclei before collecting the Similar to the tendency observed for OBC, G′ and G”, samples crys­
sample from the SSHE. This low number of nuclei hampered the for­ tallized in batch were always improved with sonication, similar to re­
mation of a strong network during storage such as the one formed at sults previously reported for other fat sources (da Silva, Cooper et al.,
lower temperatures. 2020a; Ye & Martini, 2015). When samples were crystallized at 30 and
Sonicated samples in batch showed a significant improvement in 32 ◦ C in the SSHE, sonication reduced sample viscoelasticity (p < 0.05).
Unlike tendencies observed in OBC, the significant reduction in G′ due to

7
T.L.T. da Silva et al.
Fig. 7. Melting behavior obtained after crystallization in batch or in a SSHE at different temperatures with and without HIU and stored at 25 ◦ C for 48h.
sonication in the SSHE was already observed at 28 ◦ C. This behavior
could be attributed to excessive sonication for this crystallization con­
dition, which also prove the better effect of HIU on inducing secondary
nucleation, since effective improvement of G’ was found only at 26 ◦ C,
where primary nucleation was already established (da Silva and Martini,
2020b).
3.5. Hardness
Similar to the results discussed for G’, the hardest samples were
obtained in the SSHE with HIU at 26 ◦ C, followed by the sonicated
samples crystallized in batch (26–30 ◦ C) and the non-sonicated samples
crystallized in the SSHE (Fig. 6). The softer samples were non-sonicated
and crystallized in batch.
For samples crystallized in batch, sonicated samples were always
harder than non-sonicated ones at all Tc (p < 0.05), as previously found
for different fat sources (da Silva, Cooper et al., 2020a; Suzuki et al.,
2010). For samples crystallized in the SSHE, harder samples were ob­
tained with sonication at 26 ◦ C while softer samples were obtained at 28
◦
C and no difference in hardness was obtained at 30 and 32 ◦ C. The lack
of effect of sonication in hardness values in a continuous system was also
found for a milk fat (Gregersen et al., 2019). This might have happened
for two reasons:(1) a slight increase in temperature due to sonication
melted the few crystals formed in the SSHE at high temperatures (28–32
◦
C). It is possible that when the sample was crystallized at lower tem­
peratures, such as 26 ◦ C, more crystals were present and the slight in­
crease in temperature generated during sonication was not sufficient to
melt the crystals; (2) insufficient nuclei were present in the sample at
high temperatures and HIU was not able to induce secondary crystalli­
zation. In addition, storage at a lower temperature (5 ◦ C) resulted in a
higher supercooling that can hinder the effects generated during
sonication.
3.6. Melting behavior
Melting enthalpies and Ton were not significantly different (Table 3,
8
LWT 138 (2021) 110593
p > 0.05) but both sonication and SSHE crystallization affected the
melting profiles (Fig. 7) and Tp (Table 3). All control samples in batch
showed one main melting peak and a small shoulder at lower temper­
atures (grey filled arrow).
When HIU was applied in batch, two behaviors were observed: (1) at
lower temperatures (26 and 28 ◦ C) the shoulder became insignificant
and (2) a reduction in Tp to values between 37 and 38 ◦ C was observed.
At 30 and 32 ◦ C a reduction in Tp due to sonication was also observed
and Tp was like the ones obtained at 26 and 28 ◦ C. However, the melting
profile obtained at 30 ◦ C showed an additional shoulder (non-filled grey
arrow) which was more evident at 32 ◦ C. It is possible that sonication
melted some of the growing crystals at 30 and 32 ◦ C and that nucleation
occurred in these samples when stored at 25 ◦ C and not during pro­
cessing. Nuclei formed during storage generated a crystalline network
different from the one generated during processing which is evidenced
as shoulders in the melting profile.
The non-sonicated samples crystallized in the SSHE at 26 ◦ C showed
a melting profile similar to the non-sonicated in batch with a shoulder
around 32 ◦ C and a main peak at the same Tp as the sonicated sample in a
batch, confirming that SSHE promoted similar changes in the sample in
a continuous system than HIU in batch system. No changes were
observed in the melting profile or in Tp after sonication in the SSHE
suggesting that even though cavitation might have increased sample
temperature as discussed above, nucleation formed in the SSHE at 26 ◦ C
was sufficient to allow HIU bubbles to induce secondary nucleation and
improve other physical properties without changing the amount of
crystalline material (same ΔH and SFC) or melting behavior. Similar
results were found using different HIU conditions in a SSHE for a
soybean-based fat (da Silva & Martini, 2019).
At 28 ◦ C the non-sonicated SSHE sample had an additional shoulder
at higher temperatures 45 ◦ C (black arrow), even though this sample had
the same melting parameters (Ton, Tp and ΔH) as the sonicated ones in
batch. After sonication, this shoulder was less pronounced. Similar re­
sults were observed by Kadamne, Ifeduba, Akoh, and Martini (2017a)
where sonication promoted incorporation of high-melting point TAGs
into the crystalline matrix forming a more organized crystalline network
T.L.T. da Silva et al.
of a stearic acid-based interesterified sample. When the sample was
crystallized at 28 ◦ C few nucleation sites were generated in the SSHE and
these shoulders represent nuclei and crystals formed during storage.
During sonication, the presence of these peaks were not as pronounced,
suggesting that sonication helped form a more uniform crystalline
network compared to the non-sonicated samples.
Although samples showed the same Ton at 30 and 32 ◦ C, a broader
melting profile with various shoulders was observed and Tp was similar
to non-sonicated samples in batch, which confirms our theory of static
crystallization under storage. Due to the similarity in the melting profile
observed at 30 and 32 ◦ C we believe that almost all crystallization for
these temperatures occurred during storage and that the bubbles
generated during sonication were not able to induce crystallization
during storage.
4. Conclusion
Crystallization in batch and in a continuous sheared system (SSHE)
generated different crystalline materials. HIU can act as a crystallization
modifier in both crystallization systems. In general, sonication induced
secondary nucleation by inducing more nucleation or formation of
smaller nuclei. This effect forms a more uniform crystal network that is
more elastic, harder, and capable of entrapping more liquid oil. When
samples are processed in batch a significant effect of HIU is observed in
all supercoolings tested, which can be attributed to the lower efficiency
of the batch process on inducing nucleation, and on the fact that HIU,
was always applied at the onset of crystallization, which is not always
the case in the SSHE. Nevertheless, when samples were crystallized in a
SSHE, heat transfer is more efficient, and an improved crystallization is
observed even in the absence of sonication. However, if the crystalli­
zation settings are sufficient to induce a significant number of nuclei in
the SSHE before sonication, as observed at 26 ◦ C, HIU is efficient at
improving physical properties in the SSHE even further. Results from
this study show that the best values of OBC, G’, G”, and hardness were
obtained when samples were crystallized and sonicated at 26 ◦ C in the
SSHE. The SSHE used is a good representation of what is used nowadays
in industry for fat crystallization and the results found in the study
suggest that it is possible to combine sonication with a SSHE for a low-
saturated palm-based fat and obtain better physical properties than in a
batch lab-scale.
CRediT authorship contribution statement
Thais Lomonaco Teodoro da Silva: Conceptualization, Methodol­
ogy, Formal analysis, Investigation, Validation, Writing - review &
editing, was responsible for develop the conceptualization and meth­
odology. Made the formal analysis, investigation and validation and also
wrote, revised & edited the manuscript. Sabine Danthine: Investiga­
tion, Writing - review & editing, was responsible for investigation,
revision and edition of the manuscript. Silvana Martini: Supervision,
was the Supervisor, Project administration, Funding acquisition,
Conceptualization, Validation, Writing - review & editing, responsible
for project administration and funding acquisition, also participated on
conceptualization, validation and manuscript writing and revising steps.
LWT 138 (2021) 110593
Declaration of competing interest
The authors declare no conflict of interest, and that this paper was
not submitted to any other journal besides LWT- Food Science and
Technology.
Acknowledgements
This project was supported and approved by the Utah Agricultural
Experiment Station (USA) as project number #9293. Financial support
for this research was provided by University Technology Acceleration
Grant (UTAG) program (Utah Science Technology and Research
(USTAR) initiative, USA) as grant number 18065UTAG001.
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