LWT - Food Science and Technology 202 (2024) 116303

Contents lists available at ScienceDirect

LWT
journal homepage: www.elsevier.com/locate/lwt

Impact of ultrasound on nutrition recovery and physicochemical properties

of oat beverage
Esther Kwok a, Jingqi Yang b, Parinaz Taheri c, Lingyun Chen a, *
a
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, T6G 2P5, Canada
b
Food Processing Development Centre, Government of Alberta, Leduc, T9E 7C5, Canada
c
Earth’s Own Food Group, 700-4445 Lougheed Highway, Burnaby, BC, V5C 0E4, Canada

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

Keywords: Oat beverage has become increasingly popular, yet its processing removes valuable nutritional components and
Ultrasound results in waste stream. This study evaluated the use of ultrasound as a pre-treatment technology to improve
Oat beverage nutrient recovery in oat beverage preparation. Various ultrasound power levels (60, 240, and 960 W) and
Oat protein
treatment times (1, 5, and 9 min) were investigated for their effects on physical and chemical changes during oat
Oat β-glucan
Physicochemical properties
beverage preparation. Ultrasound effectively reduced the size of larger oat grain fibrous particles while largely
leaving smaller size compounds unaffected. Partial disintegration of cell wall structures was also observed under
confocal laser scanning microscopy. This resulted in a significant increase (up to 39%) in β-glucan recovery with
minor alteration to its molecular weight in oat beverage, preserving its inherent health benefits. Protein recovery
was not improved, but ultrasound led to partial unfolding of tertiary structure, improving protein solubility and
foaming properties of oat beverage, which could be beneficial in developing coffee-based beverages and desserts.
The shear-thinning behavior of oat beverage was maintained after ultrasound treatment, benefiting processing
efficiency and mouthfeel. Overall, these findings highlight the potential of ultrasound as a sustainable pre-
treatment technology to enhance the nutritional quality and physicochemical properties of oat beverage.

1. Introduction including wet or dry grinding, enzymatic hydrolysis with α-amylase and
separation (Yu et al., 2023). According to Triantafyllou (2014), wet
The global growth of the plant-based beverage sector is propelled by grinding yielded a 24% higher protein content than dry grinding. This
environmental sustainability consideration, their potential health ben­ method is preferable for industry as it is more effective in releasing
efits (including cholesterol lowering effect, and antioxidative and anti- nutritional components, particularly proteins and β-glucan, from the oat
inflammatory properties) and dairy allergies (Siddiqui et al., 2023). endosperm. Subsequently, α-amylase is introduced to hydrolyze oat
Besides soy and tree nuts-based beverages, oat beverage has emerged as starch, limiting starch gelatinization that tends to induce high viscosity
one of the most popular plant-based beverages in North American and enhances the fluidity of oat beverage (Yu et al., 2023). Afterward,
markets. Oats are an excellent source of β-glucan, a water soluble dietary the solid residues are separated from the oat beverage by centrifugation
fiber, which is well known for its health beneficial effects including or descanting. However, during this step, a significant amount of valu­
blood-glucose and blood-cholesterol lowering ability, and satiety effect able compounds, such as dietary fiber and protein, is lost in the residues
(Cui, Jia, Zhao, Hou, & Zhou, 2023). The Food and Drug Administration due to inefficient grinding and extraction. Similar inefficiencies are
(FDA) and Health Canada both have claimed that consume a 3 g per day observed in other cereal grain processing, such as sorghum (Amoura,
of oat β-glucan would reduce the risk of coronary heart dieases by Mokrane, & Nadjemi, 2020) and corn (Ozturk, Kaasgaard, Palmén,
lowering blood-cholesterol (Hughes & Grafenauer, 2021). Additionally, Vidal, & Hamaker, 2021). Therefore, developing a more efficient pro­
oats contain good amount of protein with a higher lysine content cessing technology to enhance nutrients recovery in oat beverage is
compared to other cereals, along with a well-balanced amino acid important.
essential profile (Cui et al., 2023). Ultrasound, an environmentally friendly technology, has garnered
The production of oat beverage involves several processing steps, increased attention in food processing for its ability to enhance

* Corresponding author.
E-mail addresses: [email protected] (E. Kwok), [email protected] (L. Chen).

https://doi.org/10.1016/j.lwt.2024.116303
Received 21 February 2024; Received in revised form 2 June 2024; Accepted 4 June 2024
Available online 5 June 2024
0023-6438/© 2024 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).
E. Kwok et al. LWT 202 (2024) 116303

extraction yields and modify physicochemical properties of plant-based 2.3. Enzymatic treatment of the oat slurry
materials. The cavitation effect of ultrasound ruptures plant cell walls,
enhancing the extractability of intercellular materials through high The oat slurries (100 mL) were adjusted to pH 7 using 1 mol/L NaOH,
shear rates and the collapse of microbubbles in a liquid medium (Yusoff, then hydrolyzed with 1 mL α-amylase at 60 ◦ C for 30 min, followed by
Mat Taher, Rahmat, & Chua, 2022). Several studies had revealed that inactivation in a 90 ◦ C water bath for 10 min. After cooling, the oat
ultrasound improved the extractability of intracellular materials from slurries were tested for particle size analysis and observed by an optic
oat including β-glucan (Chen et al., 2018) and protein (Perez-Vega et al., microscopy with a AxioCam ERc 5s digital camera (Zeiss Pimostar 1,
2023) compared to conventional methods. Hematian Sourki, Koocheki Jena, Germany) with a 10 × magnification. Afterward, the slurries were
& Elahi (2017) observed ultrasound disintegrated barley β-glucan centrifugated at 2000 rpm for 2 min to collect the oat beverage (su­
structure, and Falsafi et al. (2019) found similar effects on oat starch, pernatant) were stored were stored at 4 ◦ C or lyophilized for subsequent
improving paste fluidity. Li and Xiong (2021) utilized ultrasound to analysis. The oat beverage samples had a final pH range of 6.26–6.40.
modify oat protein isolates, observing a reduction in particle size and an The residues were also collected, which were then stored at 4 ◦ C for
increase in protein solubility and functional properties. Other studies CLSM analysis. Sodium azide (0.02 mg/mL) was added in all oat
have demonstrated ultrasound as a post-preparation process improved beverage samples and residues as an antimicrobial reagent. The oat
the physical stability of plant-based beverages, preventing creaming and beverage sample without ultrasound treatment was also prepared as
component phase-separation (Lu et al., 2019; Vallath & Shanmugam, control.
2022). However, the use of ultrasound as a pre-treatment technology to
improve β-glucan and protein content, and its impact of physicochem­ 2.4. Particle size distribution
ical properties of oat beverage, remains unclear.
This work aimed to evaluate ultrasound as a pre-treatment technique The particle size distributions of oat slurries and oat beverages were
for oat beverage processing. The impact of ultrasound power levels (60, determined by a Malvern Mastersizer 3000 (Malvern Instruments Ltd.,
240, and 960 W) and treatment durations (1, 5, and 9 min) on a series of Worchester, UK) with Hydro LV extension at 25 ◦ C. The samples were
component changes during oat beverage preparation was studied added dropwise to the instrument with distilled water as a dispersion
including the extraction content of protein and β-glucan. Additionally, medium until the obscuration rate of 10–12% was obtained. The
the oat grain size reduction and cell wall disintegration were studied by refractive index of particle and water set as was 1.47 and 1.33, respec­
particle size measurement and confocal laser scanning microscopy tively. The volume surface (D43) was recorded.
(CLSM) observation. Moreover, the protein and β-glucan structure
changes in the ultrasound treated oat beverage were characterized by 2.5. Rheological analysis
sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-
PAGE), size exclusive-high-performance liquid chromatography (SE- The flow behavior of the oat beverage samples was studied using a
HPLC), Fourier-transform infrared spectroscopy (FT-IR) and intrinsic DHR-3 rheometer (TA Instruments, New Castle, DE, USA) equipped with
fluorescence analysis. The generated knowledge will provide scientific a 40 mm diameter parallel plate and the gap size was set at 1000 μm. The
insight into the oat protein and β-glucan extraction and modification by viscosity was measured over the shear rate range of 0.1–1000 1/s at
ultrasound as a pre-treatment technology during oat beverage prepa­ 25 ◦ C.
ration. Increasing nutrients recoveries through ultrasound will provide a Flow behavior index (n) and consistency coefficient (k) values were
more sustainable method to help oat beverage industry in improving the calculated by fitting the power-law model (Eq. (1)) (Hematian Sourki,
product nutritive quality and enhancing competitiveness in the global Koocheki, & Elahi, 2017):
market.
τ = kγ̇n (1)
2. Materials and methods where τ is the shear stress (Pa), k is the consistency coefficient (Pa.sn), γ̇
is the shear rate (s− 1), and n is the flow behavior index (dimensionless).
2.1. Materials
2.6. Protein characterization
Dehulled oat grains (Avena sativa) were purchased from a Wedge
Farms Ltd., Manitoba, Canada. The α-amylase from Bacillus spp. (EC 2.6.1. Protein content and solubility
3.2.1.1., 500 U/mL), acid fusion and calcofluor white used for confocal The protein content in the oat beverages was determined by using a
laser microscopic analysis were purchased from Sigma-Aldrich Co. combustion nitrogen analyzer (CN-628, Leco Corporation, St Joseph MI,
(Oakville, Ontario, Canada). The β-glucan assay kit used for β-glucan USA) with a factor of 5.83 used for protein conversion. The total solid
content determination was purchased from Megazyme International Ltd content of oat beverage samples was determined by oven-drying
(Bray, Co. Wicklow, Ireland). Distilled water was used for the oat method. The protein content (mg/mL) in oat beverages was calculated
beverage preparation, and Milli-Q water (Millipore, Billerica, Ma, USA) from the following equation (Eq. (2)):
was used for analysis. All chemicals were reagent graded.
(mg ) Protein content of oat beverage (% db)
Protein content =
2.2. Preparation of oat slurry and ultrasound treatment mL 100
(mg )
× total solid content of oat beverage (2)
The 0.2 g/mL oat slurry was prepared with oat grains (stored at 4 ◦ C) mL
mixed with distilled water and grounded with a laboratory blender The protein solubility of the oat beverages was measured by Eckert
(Blender 7012 S, Warning Commercial Blender, USA) for 2 min. The oat et al. (2019) with modifications. Aqueous oat beverage was mixed in
slurry then was treated by an ultrasonic processor (Model JY98 – IIIDN, equal volume of Milli-Q water for 30 min, and then centrifuged at 3500
Ningbo Scientz Biotechnology Co., Ltd., China) equipped with a tita­ rpm for 30 min to collect the precipitate for lyophilization and
nium probe (diameter 3/4 inch). The samples were treated at different weighting. The protein solubility (%) was calculated from the following
power output levels (60, 240, and 960 W) for 1, 5, and 9 min (pulse equation (Eq. (3)):
duration: on-time, 5 s; off-time, 5 s). The temperature was controlled by ( )
mass of insoluble protein
using an ice bath as not exceeding 45 ◦ C. Protein solubility (%) = 1 − × 100%
mass of protein in oat beverage
(3)

2
E. Kwok et al. LWT 202 (2024) 116303

Fig. 1. Effect of ultrasound (60, 240, and 960 W) for 1, 5, and 9 min on the (A) particles size distribution of oat slurries (before centrifugation), and (B) D43. Control
(0 min) was the sample without ultrasound treatment. Different letters above column indicate significant differences (p < 0.05).

2.7. β-glucan characterization

2.6.2. SDS-PAGE
SDS-PAGE was performed under reducing and non-reducing condi­
2.7.1. β-glucan quantification
tions (Zhang, Huang, Roopesh, & Chen, 2022). The samples (3 mg
β-glucan content in lyophilized oat beverages was determined using
protein/mL) were mixed with 2 × sample buffers (Bio-Rad Laboratories
the Megazyme β-glucan (AOAC 992.28) method. The absorption read­
Inc.) with or without 2-mercaptoethanol and boiled for 5 min. After
ings were measured at 510 nm using the same Spectra Max
centrifugation (9703 rpm for 5 min), 12 μL of supernatants were loaded
spectrophotometer.
on each lane of 4% stacking gel and 12% separating gel. The electro­
phoresis was run at a constant voltage of 80 V. After electrophoresis, the
2.7.2. Determination of β-glucan molecular weight (Mw)
gels were stained with Coomassie Brilliant Blue R-250 and destained in
The Mw of β-glucan in the ultrasound treated and untreated oat
water: methanol: and acetic acid solution was at the ratio of 8:1:1
beverage samples was determined by SE-HPLC using an Agilent 1100
(v/v/v).
series HPLC system equipped an Ultrahydrogel™ Linear (7.8 × 300 mm,
Waters Corp., Mass., USA) according to Åman, Rimsten, & Andersson, R.
2.6.3. Fourier-transform infrared spectroscopy (FT-IR)
(2004) with modifications. The lyophilized oat beverages (200 mg) were
The conformation of protein in the oat beverages was characterized
mixed with 10 mL Milli-Q water and 70 μL 100% ethanol, and then
by FT-IR. Lyophilized oat beverages were mixed and pressed with po­
hydrolyzed with 50 μL thermostable α-amylase from Bacillus spp. (EC
tassium bromide in 1:100 ratio. The spectra were recorded by a Nicolet
3.2.1.1., 3000 U/mL) (Megazyme International Ireland Ltd) at 90 ◦ C for
6700 spectrophotometer (Thermo Fisher scientific Inc., MA, US) at 400
2 h to remove starch and polysaccharides. Then, the mixtures were
to 4000 cm− 1 with 128 scans at a 4 cm− 1 resolution. Fourier self-
centrifugated at 2500 rpm for 15 min. The supernatants filtered with
deconvolution for amide I region (1700-1600 cm− 1) was analyzed at a
0.45 μm membrane prior to SE-HPLC analysis. The column was kept at
bandwidth of 24 cm− 1 and an enhancement factor of 2.5 using Omnic
40 ± 0.5 ◦ C, and the flow rate of mobile phase was at 0.5 mL/min.
8.1 software (Byler & Susi, 1986).
Signals were detected by a refractive index detector (RID). Pullulans of
different Mw (10,000, 21,700, 48,800, 113,000, 200,000, and 348,000
2.6.4. Intrinsic fluorescence
Da) were used as standards to estimate the β-glucan Mw.
The intrinsic fluorescence spectra of the oat beverages were
measured using a spectrophotometer (SpectraMax M3, Molecular De­
vices, Sunnyvale, US). Lyophilized samples were prepared in 1 mg 2.8. Confocal laser scanning microscopy (CLSM)
protein/mL with distilled water before testing. The excitation wave­
length was set at 295 nm and emission wavelength was in the range of The ultrasound treated and untreated residual samples were
315–500 nm (slit = 2 nm). observed by CLSM. Acid Fuchsin and Calcofluor White were used to
highlight the endosperm protein and plant cell wall, respectively (Dor­
nez et al., 2011). Specifically, the residual samples were stained with

3
E. Kwok et al. LWT 202 (2024) 116303

Acid Fuchsin (0.1 mg/mL) for 2 h, followed by Calcofluor White (0.01

mg/mL) for 2 h. After each staining, the samples were washed with
Milli-Q water twice through centrifugation for 30 s at 500 rpm to remove
excess stain. When the stained samples were imaged with a confocal
laser scanning microscope (Zeiss LSM 710, Zeiss AxioObserver, Jena,
Germany) with 10 × and 20 × objectives. The images were analyzed at
the wavelength of 405 and 561 nm for Calcofluor White and Acid
Fuchsin, respectively. The ZEN 2011 software (Carl Zeiss AG, Oberko­
chen, Germany) was used to process the images.

2.9. Foaming properties of oat beverage

The foaming properties of the oat beverages were measured ac­

cording to Xiong et al. (2018) with modifications. 5 mL of oat beverage
samples (stored at 4 ◦ C) were whipped at a high speed (24,000 rpm, 1
min) using an Ultra-Turrax ® homogenizer (T18 Ultra Turrax, IKA,
Wilmington, US). The foaming capacity was determined by comparing
the foam volume at 1 min to the initial liquid volume. The foaming
capacity (FC) was calculated as follows (Eq. (4)):
V1 − V
FC = × 100% (4)
V

where V is the volume of the oat beverages, and V1 is the volume of foam
1 min after whipping, respectively.
The foam volume was measured after storage at room temperature
for 30 min. The foam stability (FS) was determined as the remained
foam volume percentage using (Eq. (5)),
V2
FS = × 100% (5)
V1

where V2 and V1 represent the volume of foam after storage for 30 min
and the initial volume of foam after whipping.

2.10. Statistical analysis

The experiments were preformed triplicate, and the data were pre­
sented as mean ± standard deviation. The statistical evaluations were
computed by one-way analysis of variance (ANOVA) using Origin 2022
(Hampton, MA, USA). The statistical difference between samples was
performed by the Tukey test with a p-value<0.05.

3. Results & discussion

3.1. Particle size distribution

Ultrasound can rupture plant cell walls through the generated high
shear rate and microbubbles collapse in a liquid medium. Thus, the
impact of ultrasound power levels (60, 240, and 960 W) and treatment
durations (1, 5, and 9 min) on particle size change of the oat slurries
before centrifugation and the oat beverages were studied. As shown in
Fig. 1A, the oat slurries were a polydispersed system, which could be
divided into two regions including one with small particle size (1–70
Fig. 2. Confocal micrographs of oat grain fibrous materials in the oat beverage
μm) and the other one with larger particle size (100–1600 μm). The residues undergo different ultrasound conditions. Protein bodies presented in
ultrasound showed some impacts on the larger particle region in particle red channel and polysaccharides (cell wall materials) are highlighted in blue.
size distribution curves, leading to a left shifting of the size distribution. (For interpretation of the references to colour in this figure legend, the reader is
The more left shifting indicates the smaller size of the large particles. referred to the Web version of this article.)
The pronounced peak left shifted from 709.68 to 329.93 μm as the ul­
trasound power level increased to 960 W for 9 min. In the small particle treatment (Fig. 1B). As the ultrasound power level was raised to 960 W
region, ultrasound treated oat slurries had distribution curves similar to with 9 min treatment, the lowest D43 of 126.33 μm was achieved. From
that of the control.The effect of ultrasound on the large particle size the optic microscopic observations, the larger particles were oat grain
reduction was also supported by the D43 (volume-based diameter) fibrous materials, and their size reduction was clearly observed when
measurement, which is highly sensitive to any larger particles (Zhu the ultrasound power levels were increased to 240 and 960 W with 9
et al., 2018). The ultrasound treated oat slurries also exhibited a min. This indicated the sizes of the fibrous materials had more impact at
reduction in D43 as the ultrasound exposure time increased to 9 min the higher ultrasound power with longer exposure time (Fig. S1). The
across all power levels when compared to the control without the

4
E. Kwok et al. LWT 202 (2024) 116303

Fig. 3. Effect of ultrasound treatment (60, 240, and 960 W for 1, 5, and 9 min) on the (A) β-glucan content and (B) Mw in oat beverages. Control (0 min) was the
sample without ultrasound treatment. Different letters above column denote significant differences (p < 0.05).

smaller particles were protein, polysaccharides and lipid (He & Xu, shock wave can damage the cell structure including breakages and
2024). The size reduction of fibrous materials was due to cavitation loosening of the cell structure (Yusoff et al., 2022). Although the cell
resulting from bubble collapse, which generated micro-turbulence and walls were damaged, the overall shape of the cell walls was retained.
hydrodynamic shockwaves, leading to the fibrous materials disintegra­ Whereas several studies demonstrated completely rupture of cell walls
tion (Yusoff et al., 2022). The fibrous materials are in larger size, when fruit juices were treated by a high-power level of ultrasound with a
allowing more contact with the cavitation bubbles, thus resulting in longer duration (Rojas, Leite, Cristianini, Alvim, & Augusto, 2016;
greater hydrodynamic forces for the fragmentation. Furthermore, Campoli, Rojas, do Amaral, Canniatti-Brazaca, & Augusto, 2018). Due to
similar particle size distribution curves and no significant D43 was the difference in rigidity of the plant cell walls, ultrasound had less
observed among the ultrasound treated oat beverages (Fig. S2). This destructive effects on oat cell walls than those of fruits. The mechanism
indicated that ultrasound primarily affected the fibrous materials, of intercellular materials extraction through ultrasound involves cell
potentially improving the embedded nutrient compounds to be available disruption, resulted in increasing the contact area of intracellular ma­
for extraction. Meanwhile, less impact on the small particles may allow terials and extraction solvent (water) to promote mass transfer (Yusoff
better preservation of protein and β-glucan. et al., 2022). Since the oat cells were not fully disrupted, the improve­
ment of mass transfer of proteins into the water was limited. It was
observed that some of the cells still contained fully packed protein
3.2. CLSM
bodies, even after treatment with 960 W for 9 min. Similar result was
observed from the study of Preece, Hooshyar, Krijgsman, Fryer, and
The oat beverage residues were observed by the CLSM to further
Zuidam (2017) in which protein bodies remained within the cells when
study the impact of ultrasound on the oat grain fibrous materials, as
the soy slurry was treated with 1700 W for 2.3 min.
shown in Fig. 2. The protein bodies were stained with Acid Fuchsin
(red), and the cell wall was stained with Calcofluor White (blue).
Without the ultrasound treatment, large pieces of intact cell wall 3.3. The effect of ultrasound treatment on β-glucan content and Mw in oat
structure remained with protein bodies embedded in the cells. When 60 beverage
W was applied, the cells adjacent to the disintegrated edge became less
packed, but the centre of the cells was remained intact and fully packed The β-glucan content of oat beverages by ultrasound treatments is
with protein bodies. When the higher ultrasound power level was shown in Fig. 3A. The oat beverages treated with ultrasound of 60 and
applied, the destruction of cell walls not only limited to the edge of the 240 W showed gradually increased β-glucan content. At 240 W for 9 min
residue fragment, with the observable damage towards its centre. This the highest content of β-glucan of 7.45 mg/mL was observed in the oat
phenomenon was attributed to the greater exposure of cavitation effect beverage which increased by almost 2 folds compared to the control
on these cell walls. Prolonged treatment at 960 W for 9 min further (4.58 mg/mL). The explosions of bubbles generate energy to loosen the
intensified this impact, resulting in more damage in the centre of cell outer layer of oat cell walls, allowing more water to penetrate to the cell
walls as indicated by the arrows in Fig. 2. The generated hydrodynamic wall inner layer where β-glucan is located (Hematian Sourki et al.,

5
E. Kwok et al. LWT 202 (2024) 116303

Fig. 4. Effect of ultrasound (60, 240, and 960 W) for 1, 5, and 9 min on the protein content of oat beverages. Control (0 min) was the sample without ultrasound
treatment. Different letters above column denote significant differences (p < 0.05).

2017). However, when the ultrasound power level was further raised to suggested high contents of carbohydrates and lipids lowered the protein
960 W and time was prolonged to 9 min, the β-glucan content was yield in the products (Byanju et al., 2020). An increase in water ratio
reduced to 5.49 mg/mL, even no significant difference compared to the might increase protein content in the oat beverages as water allowed
control sample (p > 0.05). This reduction could be attributed to more more cavitation effects. However, from the industrial perspective, in­
insoluble materials such as cellulose and arabinoxylan were also crease in water usage is not desirable because the oat beverage will be
released and suspend in the solvent (water), which possibly lower the diluted, which also leads to diminish of its nutritive quality and desir­
permeability of water to extract β-glucan (Prakash Maran, Manikandan, able characteristics such as mouthfeel.
Thirugnanasambandham, VignaNivetha, & Dinesh, 2013). The pro­
longed exposure time (9 min) may cause excessive release of these
insoluble materials, leading to the further reduction of the β-glucan 3.5. The effect of ultrasound on protein structure
content in the oat beverage. Similar results were observed by the work of
Hu et al. (2022), in which reduced extraction yield of polysaccharides 3.5.1. SDS-PAGE
from coix seeds was observed when ultrasound time was extended to 15 SDS-PAGE profiles of protein in control and oat beverages treated by
min at the high power level (480 W). Meanwhile, only a minor change ultrasound are shown in Fig. 5A. In oat protein, the major protein is 12S
was observed for the β-glucan Mw. The samples showed β-glucan Mw globulin. The former is the 320 kDa hexamer is comprised of 54 kDa
range of 180.51–204.23 kDa. Even with the highest ultrasound power subunits that bind non-covalently. Each subunit is disulfide bonded,
level with prolonged treatment time (960 W for 9 min), only approxi­ which separated into acidic (32 kDa) and basic (22 kDa) subunits by 2-
mately 4.2% decrease of β-glucan Mw when ultrasound timing was mercaptoethanol (Li & Xiong, 2021). Other bands at 50–75 and 15–17
extended from 0 to 9 min at 960 W. Since the large amount of micro­ kDa correspond to 7S and 3S subunit of oat globulin, respectively
bubble formation at a higher ultrasound power level which could act as a (Nieto-Nieto, Wang, Ozimek, & Chen, 2014). Comparison with the un­
barrier of energy transmission within the system. These bubbles may not treated oat beverage showed that ultrasound did not induce major
effectively to produce the required energy to further decrease the Mw changes in oat protein subunits or break down the disulfide bonds in
(Muñoz-Almagro, Montilla, Moreno, & Villamiel, 2017). Comparing to protein structure. Similar results were observed when using ultrasound
ultrasound, multiple enzymatic (e.g. the combination of α-amylase, to hemp protein (Liu, Wang, Xue, & Adhikari, 2022) and quinoa protein
protease and xylanase) and chemical (alkaline and acidic solutions) (Li et al., 2023). In other studies, modification of extracted plant-based
extraction methods can lead to extensively fragmentation of β-glucan proteins subunits by ultrasound were observed. For example, bitter
Mw into shorter chains, which may interfere its blood-glucose and melon seed and olive leaf proteins were broken down into smaller
blood-cholesterol lowering abilities (Bozbulut & Sanlier, 2019; Goudar, structures through extra bands were revealed at the lower molecular
Sharma, Janghu, & Longvah, 2020). Significantly increase in extraction weight ladder (Naik, Natarajan, Modupalli, Thangaraj, & Rawson, 2022;
of β-glucan content by the ultrasound treatment (240 W for 9 min) Ortega et al., 2024). This result suggests that the modification of protein
without extensively reducing its Mw makes this technology favorable for molecular weight using ultrasound extraction is contingent on the spe­
improving the nutritional quality of oat beverage. cific plant sources employed.

3.5.2. Fourier transform infrared spectroscopy (FT-IR)

3.4. The effect of ultrasound on protein content in oat beverage FT-IR was used to understand the secondary structure of protein. The
absorption changes in amide region (amide I) were mainly studied and
The effect of ultrasound on protein content in the oat beverages is the results are shown in Fig. 5B. The amid I band with and without ul­
presented in Fig. 4. Comparing with the control, the ultrasound treated trasound treatment showed 7 bands, including 1691 cm− 1 (β-sheets/
oat beverages showed either no significant difference or a reduction in turns), 1670 cm1 (β-turns), 1656 cm− 1 (α-helix), 1641 cm− 1 (random
the protein content. One explanation could be due to the rigid cell wall coil), 1629 cm− 1 (β-sheet), 1618 cm− 1 (intermolecular β-sheet), and
protected the intercellular proteins from cavitation effects, which 1609 cm− 1 (vibration of amino acid residues) (Kong & Yu, 2007; Liu
limiting the extraction of cellular proteins (Karabulut, Yildiz, Karaca, & et al., 2009; Nieto-Nieto, Wang, Ozimek, & Chen, 2015). When oat
Yemiş, 2023). Another explanation was ultrasound not only released the grains underwent ultrasound treatment, no changes in amid I region
protein, other components including dietary fibers such as β-glucan and were observed. This result suggests that ultrasound leave the secondary
oil were also being released in the oat beverages. These released com­ structure of oat protein intact.
ponents might interact with the oat protein which inhibited the disso­
lution of protein during extraction, leading to its precipitation by 3.5.3. Intrinsic fluorescence spectroscopy
centrifugation (Byanju, Rahman, Hojilla-Evangelista, & Lamsal, 2020; Intrinsic fluorescence can be applied to characterize the tertiary
Jaramillo, Roberts, & Coupland, 2011). Similar findings were found structure changes of protein, mainly due to the presence of chromo­
when using ultrasound to extract protein from chickpea as the authors phores (tryptophan, tyrosine, and phenylalanine residues). The

6
E. Kwok et al. LWT 202 (2024) 116303

Fig. 5. Effect of ultrasound treatment (60, 240, and 960 W for 1, 5, and 9 min) on the (A) SDS-PAGE profiles (Marker, M; Control, lane 1; 60 W–1 min, lane 2; 60
W–5 min, lane 3; 60 W–9 min, lane 4; 240 W–1 min, lane 5; 240 W–5 min, lane 6; 240 W–9 min, lane 7; 960 W–1 min, lane 8; 960 W–5 min, lane 9; 960 W–9 min,
lane 10), (B) deconvoluted FT-IR spectra, (C) intrinsic fluorescent spectra of oat beverages.

fluorescence spectra particularly reflect tryptophan as its intensity is (red shift) indicates that tryptophan side chains in protein were exposed
greater than tyrosine and phenylalanine residues (Li & Xiong, 2021). to the polar environment. This was aligned with the results of surface
The position of tryptophan in a protein can be analyzed through study of hydrophobicity (H0) measurement, as shown in Fig. S3. H0 indicates the
the λmax emission; if the λmax is less than 330 nm, tryptophan is buried in number of hydrophobic groups exposed to the protein molecular sur­
a non-polar region in a protein; if λmax is greater than 330 nm, trypto­ face. Treatment by 240 W for 9 min resulted the significant increased in
phan is exposure to a polar environment (Vivian & Callis, 2001). As H0 when compared to the samples treated by 1 min and the control. Both
presented in Fig. 5C, the λmax of control was 334 nm and ultrasound results indicates that ultrasound partially unfolded oat protein, led to
treated oat beverages were between 334 and 340 nm. When the ultra­ exposure of buried hydrophobic groups to the protein surface (Arranz
sound duration was increased to 9 min at 60 W, the λmax was shifted to et al., 2023). When the power level increased to 960 W, a reduction in H0
336 nm. Further peak shift was observed as the oat beverage was treated was observed when the treatment duration was prolonged to 9 min
with 240 W for 9 min. The shift of λmax to the longer wavelength wave (Fig. S3). The reduction in H0 was also observed for back bean and hemp

7
E. Kwok et al. LWT 202 (2024) 116303

Fig. 6. Effect of ultrasound (60, 240, and 960 W) for 1, 5, and 9 min on the protein solubility of oat beverages. Control (0 min) was the sample without ultrasound
treatment. Different letters above column denote significant differences (p < 0.05).

Fig. 7. Effect of ultrasound treatments on the apparent viscosity of oat beverages. Control (0 min) was the sample without ultrasound treatment.

proteins when treated by a high power ultrasound for a longer exposure et al., 2023). As displayed in Fig. 6, the protein solubility showed no
time. This could be related to the aggregation of partially unfolded statistical change compared with the control at low power ultrasound
proteins through hydrophobic and other interactions, and thus the treatment. As the power level was increased to 240 W and treated for 9
exposed hydrophobic groups were buried inside the protein aggregates min, the protein solubility was significantly enhanced from 24.7 to
(Liu et al., 2022; Mir, Riar, & Singh, 2019). Consequently, a blue shift 52.8%. This could be attributed to partial unfolding structure of oat
occurred as the λmax shifted back from 338 to 336 nm when the ultra­ protein, increasing the interaction between the protein and water mol­
sound treatment was prolonged from 1 to 9 min (Fig. 5C). ecules. When the ultrasound power level and treatment time increased,
more hydrophobic groups were exposed that could caused protein to
reaggregate through non-covalent interactions and thus, reduce its sol­
3.6. The effect of ultrasound treatments on protein solubility ubility (Li & Xiong, 2021; Mir et al., 2019). Despite an increase in the
solubility at 240 W, the protein recovery did not improve. This contracts
Protein solubility is a critical factor in plant-based beverages as with the expectations, as higher water solubility usually correlates with
having a good protein solubility will improve physiochemical proper­ a better recovery (Karabulut et al., 2023; Li et al., 2023). Further studies
ties, including foaming properties of plant-based beverages. In this are needed to understand this phenomenon.
study, the oat beverage samples have a range of pH from 6.26 to 6.40,
and oat protein is known to have limited solubility in such pH range (Yu
3.7. Rheological properties

Table 1 The rheological properties of oat beverage are important for its
Consistency index (k) and flow behavior index (n) of untreated and ultrasound processing and sensory quality. Too high viscosity can result in in­
treated oat beverages using the Power Law model fitting.
efficiency in operation such as low extraction yield of oat beverage and
Samples Duration Consistency coefficient Flow behavior R2 high energy usage. In addition, oat beverage with a high viscosity will
(min) (k, Pa.sn) index (n)
have adverse impacts on its sensory properties such as mouthfeel,
60 W 0 0.333 ± 0.009a 0.679 ± 0.013a 0.988 resulting in a distinct difference from bovine milk. Consumers often
1 0.313 ± 0.012ab 0.641 ± 0.020a 0.984 anticipate plant-based beverages to exhibit sensory characteristics
5 0.277 ± 0.007a 0.663 ± 0.011a 0.999
similar to bovine milk, as they are perceived as beverage substitutes. The
9 0.299 ± 0.014a 0.644 ± 0.008a 0.986
apparent viscosities of untreated and ultrasound treated oat beverages
240 W 0 0.333 ± 0.009ab 0.679 ± 0.013a 0.988
were presented in Fig. 7. At 60 W, the ultrasound treated oat beverages
1 0.266 ± 0.002b 0.685 ± 0.012a 0.992
5 0.289 ± 0.020b 0.666 ± 0.087a 0.974 showed a reduction in apparent viscosity than that of the control. This
9 0.358 ± 0.038a 0.637 ± 0.066a 0.979 could be explained by the collapse of the cavitation bubbles that caused
960 W 0 0.333 ± 0.009a 0.679 ± 0.013a 0.988
physical disintegration of starch granules (Falsafi et al., 2019), thus
1 0.263 ± 0.020bc 0.664 ± 0.027a 0.983 allowing easier access of α-amylase to starch granules for a more
5 0.287 ± 0.008b 0.654 ± 0.004a 0.991 completed hydrolysis. When the treatment time was extended to 9 min,
9 0.248 ± 0.010c 0.774 ± 0.011b 0.995 a slight increase of the viscosity was observed, which was due to the
Data were expressed as mean ± standard deviation. 0 min indicated the oat increase of β-glucan content. As the ultrasound power level and timing
beverage was not treated with ultrasound and only enzymatic hydrolysis. increased to 240 W and 9 min, the apparent viscosity was even slightly
Different superscripted letters indicate significant differences (p < 0.05). higher compared to the control due to the highest β-glucan content in

8
E. Kwok et al. LWT 202 (2024) 116303

movement (Papageorgiou, Lakhdara, Lazaridou, Biliaderis, & Izy­

dorczyk, 2005). However, the viscosity of ultrasound treated oat
beverage under 240 W for 9 min exhibited a different trend. A higher
viscosity compared to the control and other samples was observed
especially at high shear rates (100–1000 1/s). This could be related to
the increased amount of β-glucan extracted into the oat beverage under
such conditions, leading to formation of stronger entanglements that
were more resistant to deformation by the imposed movement (Yuan,
Ritzoulis, & Chen, 2019). Nevertheless, its consistency coefficient (k)
was not significantly different than that of the control (p > 0.05)
(Table 1), indicating their viscosities were still similar and the increase
in β-glucan in this study would not necessarily change its viscosity to a
level that can impact the sensory properties (Rao, 2014). At 960 W, the
apparent viscosities were further reduced compared with other ultra­
sound treated samples. Moreover, their consistency coefficients (k) were
significantly lower compared to the control (p < 0.05) (Table 1), indi­
cating the oat beverages became less viscous. The ultrasound with the
highest power level (960 W) caused more destructive effects on starch
granules, allowing enhanced accessibility of α-amylase to hydrolysis
starch and hence, significantly decreased in viscosity of oat beverages.
Starch hydrolysis and β-glucan play important roles to determine the
viscosity of oat beverage, that subsequently affect the operation pro­
cessing of oat beverage and the sensory quality of the final products.
In Table 1, flow behaviour index reflects the rheology of a fluid
system; when n = 1 indicates the fluid has a Newtonian, the fluid with n
< 1 is shear-thinning (pseudoplastic) and n > 1 is shear thickening
(dilatant) in nature (Rao, 2014). The control and samples by ultrasound
treatment showed n < 1, indicating ultrasound did not alter the fluid
types of oat beverage as all the samples exhibited shear-thinning
behaviour. The positive impacts of shear thinning behavior on opera­
tional processing including ease of mixing and pumping when subjected
to mechanical forces, and product sensory quality as desirable mouthfeel
in the oat beverage development.

3.8. Foaming properties

The values for foaming capacity and stability are presented in Fig. 8.
Notably, the foaming capacity of oat beverages showed an improvement
after ultrasound treatments. This could be attributed to the reduction in
larger particle size as larger particles in wet foam systems can decrease
in foaming capacity. It was observed in a foam system with silica par­
ticles of different sizes, larger particles became pinned to the well of film
between air bubbles, limiting their diffusivity, while the smaller parti­
cles continued to diffuse freely (Dickinson, 2015). This creates an os­
motic pressure difference, pulling water from regions with larger
particles, thinning the film in those areas and causing rupture. More­
over, the foaming capacity can be impacted by the viscosity of oat
beverages. For example, the oat beverages treated at 960 W showed a
significant decrease in viscosity, making proteins easier to absorb to the
air-water interfaces (Sheng et al., 2018). Furthermore, the foaming
stability was also significantly improved when ultrasound was applied
than that of the control, typically at 240 and 960 W (p < 0.05). The
increase of foaming stability could be due to the enhanced β-glucan
content, providing a certain level of viscosity to lower the mobility of air
bubbles (Eghbaljoo et al., 2022). More importantly, ultrasound induced
Fig. 8. Impact of ultrasound treatment (60, 240, and 960 W for 1, 5, and 9 min)
the partial unfolding of oat proteins, increasing the number of hydro­
on the foaming properties of oat beverages. Control represented no ultrasound
phobic groups exposure to surface, which enhanced their ability to
treatment. Different letters above column indicate significant differences in the
same characteristic (p < 0.05). absorb at the air-water interface and formation of a cohesive layer
around the air bubbles to prevent coalescence and thereby increase the
foaming ability (Mir et al., 2019). The improved foaming capacity and
the oat beverage (Fig. 3A). β-Glucan has steric hindrance effects on
stability of ultrasound treated oat beverages will be desirable when
water molecules and reduces its mobility, leading to a higher viscosity
using as an ingredient in such as coffee-based beverages and desserts.
(Hematian Sourki et al., 2017). For the oat beverages treated by the
ultrasound under 60 W for 1–9 min and 240 W for 1–5 min, the vis­
4. Conclusion
cosities were reduced with the increasing shear rate because the en­
tanglements formed by β-glucan were disrupted by the imposed
This research has provided scientific insight for a fundamental

9
E. Kwok et al. LWT 202 (2024) 116303

understanding of the impact of ultrasound as a pre-treatment technique barrier biomarkers. Food Research International, 169, Article 112815. https://doi.
org/10.1016/j.foodres.2023.112815
on the nutritive quality and physicochemical properties of oat beverage.
Bozbulut, R., & Sanlier, N. (2019). Promising effects of β-glucans on glyceamic control in
Ultrasound effectively reduced the fibrous materials into the smaller diabetes. Trends in Food Science & Technology, 83, 159–166. https://doi.org/
size, and partially disrupted oat grain cell wall structure. This resulted in 10.1016/j.tifs.2018.11.018
up to 39% enhancement in β-glucan recovery when treated at the ul­ Byanju, B., Rahman, M. M., Hojilla-Evangelista, M. P., & Lamsal, B. P. (2020). Effect of
high-power sonication pretreatment on extraction and some physicochemical
trasound power level at 240 W for 9 min, but the content was reduced properties of proteins from chickpea, kidney bean, and soybean. International Journal
when the greater power level was implied (960 W). Interestingly, ul­ of Biological Macromolecules, 145, 712–721. https://doi.org/10.1016/j.
trasound exhibited minimal impact on the β-glucan Mw that could pre­ ijbiomac.2019.12.118
Byler, D. M., & Susi, H. (1986). Examination of the secondary structure of proteins by
served its inherent health benefits. Conversely, the protein recovery in deconvolved FTIR spectra. Biopolymers, 25(3), 469–487. https://doi.org/10.1002/
oat beverage was not improved by ultrasound treatment, instead, ul­ bip.360250307
trasound treatment led to increased hydrophobic amino acid side chains Campoli, S. S., Rojas, M. L., do Amaral, J. E. P. G., Canniatti-Brazaca, S. G., &
Augusto, P. E. D. (2018). Ultrasound processing of guava juice: Effect on structure,
exposure to the surface. This led to enhanced foaming properties, which physical properties and lycopene in vitro accessibility. Food Chemistry, 268,
would be advantageous for developing coffee-based beverages and 594–601. https://doi.org/10.1016/j.foodchem.2018.06.127
desserts. Furthermore, shear-thinning behavior was retained in ultra­ Chen, C., Wang, L., Wang, R., Luo, X., Li, Y., Li, J., et al. (2018). Ultrasound-assisted
extraction from defatted oat (Avena sativa L.) bran to simultaneously enhance
sound treated oat beverages, offering advantages for operational effi­ phenolic compounds and β-glucan contents: Compositional and kinetic studies.
ciency, and maintaining desirable mouthfeel. Therefore, ultrasound is a Journal of Food Engineering, 222, 1–10. https://doi.org/10.1016/j.
potential pre-treatment technology to improve the nutritive values and jfoodeng.2017.11.002
Cui, L., Jia, Q., Zhao, J., Hou, D., & Zhou, S. (2023). A comprehensive review on oat milk:
physicochemical properties of oat beverage. To further enhance the
From oat nutrients and phytochemicals to its processing technologies, product
extractability of protein in oat beverage, it will be worthy of investiga­ features, and potential applications. Food & Function, 14(13), 5858–5869. https://
tion to combine ultrasound with other technologies such as cell wall doi.org/10.1039/D3FO00893B
degrading enzymes for a better disruption of the rigid cell walls, thereby Dickinson, E. (2015). Structuring of colloidal particles at interfaces and the relationship
to food emulsion and foam stability. Journal of Colloid and Interface Science, 449,
recovering more nutritious components to improve the overall nutri­ 38–45. https://doi.org/10.1016/j.jcis.2014.09.080
tional quality of oat beverage. Dornez, E., Holopainen, U., Cuyvers, S., Poutanen, K., Delcour, J. A., Courtin, C. M., et al.
(2011). Study of grain cell wall structures by microscopic analysis with four different
staining techniques. Journal of Cereal Science, 54(3), 363–373. https://doi.org/
CRediT authorship contribution statement 10.1016/j.jcs.2011.07.003
Eckert, E., Han, J., Swallow, K., Tian, Z., Jarpa-Parra, M., & Chen, L. (2019). Effects of
Esther Kwok: Writing – original draft, Validation, Methodology, enzymatic hydrolysis and ultrafiltration on physicochemical and functional
properties of faba bean protein. Cereal Chemistry, 96(4), 725–741. https://doi.org/
Data curation, Conceptualization. Jingqi Yang: Methodology, Concep­ 10.1002/cche.10169
tualization. Parinaz Taheri: Writing – review & editing, Resources. Eghbaljoo, H., Sani, I. K., Sani, M. A., Rahati, S., Mansouri, E., Molaee-Aghaee, E., et al.
Lingyun Chen: Writing – review & editing, Supervision, Resources, (2022). Advances in plant gum polysaccharides; Sources, techno-functional
properties, and applications in the food industry - a review. International Journal of
Project administration, Funding acquisition, Conceptualization. Biological Macromolecules, 222, 2327–2340. https://doi.org/10.1016/j.
ijbiomac.2022.10.020
Declaration of competing interest Falsafi, S. R., Maghsoudlou, Y., Rostamabadi, H., Rostamabadi, M. M., Hamedi, H., &
Hosseini, S. M. H. (2019). Preparation of physically modified oat starch with
different sonication treatments. Food Hydrocolloids, 89, 311–320. https://doi.org/
The authors declare that they have no known competing financial 10.1016/j.foodhyd.2018.10.046
interests or personal relationships that could have appeared to influence Goudar, G., Sharma, P., Janghu, S., & Longvah, T. (2020). Effect of processing on barley
β-glucan content, its molecular weight and extractability. International Journal of
the work reported in this paper. Biological Macromolecules, 162, 1204–1216. https://doi.org/10.1016/j.
ijbiomac.2020.06.208
Data availability He, A., & Xu, B. (2024). High-pressure homogenisation improves food quality of plant-
based milk alternatives. International Journal of Food Science and Technology, 59(1),
399–407. https://doi.org/10.1111/ijfs.16822
Data will be made available on request. Hematian Sourki, A., Koocheki, A., & Elahi, M. (2017). Ultrasound-assisted extraction of
β-d-glucan from hull-less barley: Assessment of physicochemical and functional
properties. International Journal of Biological Macromolecules, 95, 462–475. https://
Acknowledgment
doi.org/10.1016/j.ijbiomac.2016.10.111
Hu, X., Xu, F., Li, J., Li, J., Mo, C., Zhao, M., et al. (2022). Ultrasonic-assisted extraction
The authors are grateful to the Natural Sciences and Engineering of polysaccharides from coix seeds: Optimization, purification, and in vitro
digestibility. Food Chemistry, 374, Article 131636. https://doi.org/10.1016/j.
Research Council of Canada (NSERC-Discovery and CREATE program),
foodchem.2021.131636
Alberta Innovates, Results Driven Agriculture Research (RDAR) and Hughes, J., & Grafenauer, S. (2021). Oat and barley in the food supply and use of beta
Prairie Oat Growers Association for financial support. Professor Lingyun glucan health claims. Nutrients, 13(8), 2556. https://doi.org/10.3390/nu13082556
Chen would like to thank the Natural Sciences and Engineering Research Jaramillo, D. P., Roberts, R. F., & Coupland, J. N. (2011). Effect of pH on the properties of
soy protein–pectin complexes. Food Research International, 44(4), 911–916. https://
Council of Canada (NSERC)-Canada Research Chairs Program for its doi.org/10.1016/j.foodres.2011.01.057
financial support. Karabulut, G., Yildiz, S., Karaca, A. C., & Yemiş, O. (2023). Ultrasound and enzyme-
pretreated extraction for the valorization of pea pod proteins. Journal of Food Process
Engineering, 46(12), Article e14452. https://doi.org/10.1111/jfpe.14452
Appendix A. Supplementary data Kong, J., & Yu, S. (2007). Fourier transform infrared spectroscopic analysis of protein
secondary structures. Acta Biochimica et Biophysica Sinica, 39(8), 549–559. https://
Supplementary data to this article can be found online at https://doi. doi.org/10.1111/j.1745-7270.2007.00320.x
Li, Y., Wang, W., Wu, T., You, H., Liu, H., Liu, X., et al. (2023). Preparation of quinoa
org/10.1016/j.lwt.2024.116303. protein with ultrasound pretreatment and its effects on the physicochemical
properties, structural and digestion characterizations. International Journal of
References Biological Macromolecules, 238, Article 124202. https://doi.org/10.1016/j.
ijbiomac.2023.124202
Li, R., & Xiong, Y. L. (2021). Ultrasound-induced structural modification and thermal
Åman, P., Rimsten, L., & Andersson, R. (2004). Molecular weight distribution of β-glucan
properties of oat protein. Lebensmittel-Wissenschaft & Technologie, 149, Article
in oat-based foods. Cereal Chemistry, 81(3), 356–360. https://doi.org/10.1094/
111861. https://doi.org/10.1016/j.lwt.2021.111861
CCHEM.2004.81.3.356
Liu, G., Li, J., Shi, K., Wang, S., Chen, J., Liu, Y., et al. (2009). Composition, secondary
Amoura, H., Mokrane, H., & Nadjemi, B. (2020). Effect of wet and dry milling on the
structure, and self-assembly of oat protein isolate. Journal of Agricultural and Food
functional properties of whole sorghum grain flour and kafirin. Journal of Food
Chemistry, 57(11), 4552–4558. https://doi.org/10.1021/jf900135e
Science and Technology, 57(3), 1100–1109. https://doi.org/10.1007/s13197-019-
Liu, X., Wang, M., Xue, F., & Adhikari, B. (2022). Application of ultrasound treatment to
04145-2
improve the technofunctional properties of hemp protein isolate. Future Foods, 6,
Arranz, E., Segat, A., Velayos, G., Flynn, C., Brodkorb, A., & Giblin, L. (2023). Dairy and
Article 100176. https://doi.org/10.1016/j.fufo.2022.100176
plant based protein beverages: In vitro digestion behaviour and effect on intestinal

10
E. Kwok et al. LWT 202 (2024) 116303

Lu, X., Chen, J., Zheng, M., Guo, J., Qi, J., Chen, Y., et al. (2019). Effect of high-intensity Preece, K. E., Hooshyar, N., Krijgsman, A., Fryer, P. J., & Zuidam, N. J. (2017).
ultrasound irradiation on the stability and structural features of coconut-grain milk Intensified soy protein extraction by ultrasound. Chemical Engineering and Processing:
composite systems utilizing maize kernels and starch with different amylose Process Intensification, 113, 94–101. https://doi.org/10.1016/j.cep.2016.09.003
contents. Ultrasonics Sonochemistry, 55, 135–148. https://doi.org/10.1016/j. Rao, M. A. (2014). Rheology of fluid, semisolid, and solid foods : Principles and applications
ultsonch.2019.03.003 (3rd ed.). Springer. https://doi.org/10.1007/978-1-4614-9230-6 [eBook]. Imprint.
Mir, N. A., Riar, C. S., & Singh, S. (2019). Physicochemical, molecular and thermal Rojas, M. L., Leite, T. S., Cristianini, M., Alvim, I. D., & Augusto, P. E. D. (2016). Peach
properties of high-intensity ultrasound (HIUS) treated protein isolates from album juice processed by the ultrasound technology: Changes in its microstructure improve
(Chenopodium album) seed. Food Hydrocolloids, 96, 433–441. https://doi.org/ its physical properties and stability. Food Research International, 82, 22–33. https://
10.1016/j.foodhyd.2019.05.052 doi.org/10.1016/j.foodres.2016.01.011
Muñoz-Almagro, N., Montilla, A., Moreno, F. J., & Villamiel, M. (2017). Modification of Sheng, L., Wang, Y., Chen, J., Zou, J., Wang, Q., & Ma, M. (2018). Influence of high-
citrus and apple pectin by power ultrasound: Effects of acid and enzymatic intensity ultrasound on foaming and structural properties of egg white. Food
treatment. Ultrasonics Sonochemistry, 38, 807–819. https://doi.org/10.1016/j. Research International, 108, 604–610. https://doi.org/10.1016/j.
ultsonch.2016.11.039 foodres.2018.04.007
Naik, M., Natarajan, V., Modupalli, N., Thangaraj, S., & Rawson, A. (2022). Pulsed Siddiqui, S. A., Mehany, T., Schulte, H., Pandiselvam, R., Nagdalian, A. A., Golik, A. B.,
ultrasound assisted extraction of protein from defatted Bitter melon seeds et al. (2023). Plant-based milk – thoughts of researchers and industries on what
(Momardica charantia L.) meal: Kinetics and quality measurements. Lebensmittel- should be called as -milk. Food Reviews International, 1–28. https://doi.org/10.1080/
Wissenschaft & Technologie, 155, Article 112997. https://doi.org/10.1016/j. 87559129.2023.2228002
lwt.2021.112997 Triantafyllou, A. (2014). A liquid oat base and a process for preparing it using protein-
Nieto-Nieto, T. V., Wang, Y. X., Ozimek, L., & Chen, L. (2014). Effects of partial deamidase. (Sweden Patent No. WO 2014/123466 A1). World Intellectual Property
hydrolysis on structure and gelling properties of oat globular proteins. Food Research Organization. https://patents.google.com/patent/WO2014123466A1/en.
International, 55, 418–425. https://doi.org/10.1016/j.foodres.2013.11.038 Vallath, A., & Shanmugam, A. (2022). Study on model plant based functional beverage
Nieto-Nieto, T. V., Wang, Y. X., Ozimek, L., & Chen, L. (2015). Inulin at low emulsion (non-dairy) using ultrasound - a physicochemical and functional
concentrations significantly improves the gelling properties of oat protein – a characterization. Ultrasonics Sonochemistry, 88, Article 106070. https://doi.org/
molecular mechanism study. Food Hydrocolloids, 50, 116–127. https://doi.org/ 10.1016/j.ultsonch.2022.106070
10.1016/j.foodhyd.2015.03.031 Vivian, J. T., & Callis, P. R. (2001). Mechanisms of tryptophan fluorescence shifts in
Ortega, M. L. S., Orellana-Palacios, J. C., Garcia, S. R., Rabanal-Ruiz, Y., Moreno, A., & proteins. Biophysical Journal, 80(5), 2093–2109. https://doi.org/10.1016/S0006-
Hadidi, M. (2024). Olive leaf protein: Extraction optimization, in vitro digestibility, 3495(01)76183-8
structural and techno-functional properties. International Journal of Biological Xiong, T., Xiong, W., Ge, M., Xia, J., Li, B., & Chen, Y. (2018). Effect of high intensity
Macromolecules, 256, Article 128273. https://doi.org/10.1016/j. ultrasound on structure and foaming properties of pea protein isolate. Food Research
ijbiomac.2023.128273 International, 109, 260–267. https://doi.org/10.1016/j.foodres.2018.04.044
Ozturk, O. K., Kaasgaard, S. G., Palmén, L. G., Vidal, B. C., & Hamaker, B. R. (2021). Yu, Y., Li, X., Zhang, J., Li, X., Wang, J., & Sun, B. (2023). Oat milk analogue versus
Enzyme treatments on corn fiber from wet-milling process for increased starch and traditional milk: Comprehensive evaluation of scientific evidence for processing
protein extraction. Industrial Crops and Products, 168, Article 113622. https://doi. techniques and health effects. Food Chemistry X, 19, Article 100859. https://doi.org/
org/10.1016/j.indcrop.2021.113622 10.1016/j.fochx.2023.100859
Papageorgiou, M., Lakhdara, N., Lazaridou, A., Biliaderis, C. G., & Izydorczyk, M. S. Yuan, B., Ritzoulis, C., & Chen, J. (2019). Rheological investigations of beta glucan
(2005). Water extractable (1→3,1→4)-β-d-glucans from barley and oats: An functionality: Interactions with mucin. Food Hydrocolloids, 87, 180–186. https://doi.
intervarietal study on their structural features and rheological behaviour. Journal of org/10.1016/j.foodhyd.2018.07.049
Cereal Science, 42(2), 213–224. https://doi.org/10.1016/j.jcs.2005.03.002 Yusoff, I. M., Mat Taher, Z., Rahmat, Z., & Chua, L. S. (2022). A review of ultrasound-
Perez-Vega, S., Ayokanmbi, S., Kwofie, E. M., Mba, O., Salmeron, I., Gutierrez, N., et al. assisted extraction for plant bioactive compounds: Phenolics, flavonoids, thymols,
(2023). Early-stage analysis on developing different oat (Avena sativa L.) added value saponins and proteins. Food Research International, 157, Article 111268. https://doi.
fractions by ultrasound-assisted three-phase partition (UATPP): Effect of variables on org/10.1016/j.foodres.2022.111268
yield and environmental impacts. Journal of Cleaner Production, 430, Article 139624. Zhang, S., Huang, W., Roopesh, M. S., & Chen, L. (2022). Pre-treatment by combining
https://doi.org/10.1016/j.jclepro.2023.139624 atmospheric cold plasma and pH-shifting to prepare pea protein concentrate
Prakash Maran, J., Manikandan, S., Thirugnanasambandham, K., Vigna Nivetha, C., & powders with improved gelling properties. Food Research International, 154, Article
Dinesh, R. (2013). Box–Behnken design based statistical modeling for ultrasound- 111028. https://doi.org/10.1016/j.foodres.2022.111028
assisted extraction of corn silk polysaccharide. Carbohydrate Polymers, 92(1), Zhu, Z., Zhu, W., Yi, J., Liu, N., Cao, Y., Lu, J., et al. (2018). Effects of sonication on the
604–611. https://doi.org/10.1016/j.carbpol.2012.09.020 physicochemical and functional properties of walnut protein isolate. Food Research
International, 106, 853–861. https://doi.org/10.1016/j.foodres.2018.01.060

11