LWT - Food Science and Technology 153 (2022) 112454

Contents lists available at ScienceDirect

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

Impact of composite gelators on physicochemical properties of oleogels and

astaxanthin delivery of oleogel-based nanoemulsions
Tianhang Xia a, Zihao Wei a, *, Changhu Xue a, b
a
College of Food Science and Engineering, Ocean University of China, Qingdao, 266003, China
b
Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266235, China

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

Keywords: In this study, binary systems of carnauba wax (CW) and 1-docosanol (DS) were first explored to enhance the
Carnauba wax properties of oleogels. Compared to the corresponding single component oleogels (0:5 and 5:0), a synergistic
1-Docosanol effect of oleogel strength was observed at CW:DS ratios of 1:4 (C1D4). At this ratio, the formed oleogel showed
Rheology
the highest hardness, rheological property and oil binding ability. Meanwhile, the gel-sol melting temperature of
Oil binding ability
Astaxanthin bioaccessibility
this oleogel at this ratio was lower than most of others. Subsequently, the effect of composite gelators on the
astaxanthin delivery of oleogel-based nanoemulsion was explored. The oleogels at CW:DS ratios of 1:4, 0:5 and
5:0 were chosen to prepare oleogel-based nanoemulsions. The oleogel-based nanoemulsion formulated with
C1D4 had the largest droplet size. In vitro digestion study revealed that although C1D4 oleogel-based nano­
emulsion could not improve the extent of lipolysis, it could significantly improve astaxanthin bioaccessibility
when compared with the other two oleogel-based nanoemulsions. This study developed a new formula of
oleogel-based nanoemulsion with improved astaxanthin bioaccessibility. This novel formulation approach might
also be suitable for other hydrophobic nutraceuticals and provided a new idea for the preparation of oleogel-
based nanoemulsions with improved nutraceutical bioaccessibility.

1. Introduction Marangoni, 2017). Moreover, there are some potential problems of CW

to be considered, such as shear sensitivity (Tavernier, Doan, Van der
Fat, which is the ingredient and energy storage material of organism, Meeren, Heyman, & Dewettinck, 2018). To improve the properties of
plays a very important role in the texture and flavor of food. There are CW-based oleogel, the use of composite gelators can be a logical
two main sources of solid fat: animal fats and hydrogenated vegetable approach (Okuro et al., 2018). Many composite gelators have been used
oil. The former has a higher content of saturated fatty acids, while the in oleogels, such as behenyl alcohol and behenic acid (Callau, Sow-Kebe,
latter may lead to the production of trans fats, both of which increase the Nicolas-Morgantini, & Fameau, 2020), sucrose esters and lecithin (Bin
risk of metabolic syndrome and cardiovascular disease (Dassanayake, Sintang et al., 2017), lecithin and fruit wax (Okuro et al., 2018), phy­
Kodali, & Ueno, 2011). Therefore, a large amount of animal fats and tosterols and γ-oryzanol (Sawalha et al., 2015), and so on.
hydrogenated vegetable oil intake will have adverse effect on human Fatty alcohols are good gelators for vegetable oil and can be used in
health (Oh, Amoah, Lim, Jeong, & Lee, 2017). Developing oleogel with the field of food (Callau, Sow-Kebe, Nicolas-Morgantini, & Fameau,
good plasticity is an effective way to replace animal fats and hydroge­ 2020). Fatty alcohols with long chain length can lead to the formation of
nated vegetable oil. Oleogel is formed by blocking liquid oil through the oleogels with stronger structure and physical properties (Valoppi, Cal­
three-dimensional network structure of gelator (Chaves, ligaris, & Marangoni, 2016). 1-Docosanol (DS), which is a kind of fatty
Barrera-Arellano, & Ribeiro, 2018). Considering that carnauba wax alcohol, can play an excellent role in the preparation of oleogel. So far,
(CW) is one of the most efficient gelators due to its ability to improve there is no report on the combination of CW and DS to prepare oleogel.
texture and oil binding capacity of obtained oleogels, CW is often Both CW and DS are low-molecular weight gelators, and the major
applied in food industry (Doan, Tavernier, Okuro, & Dewettinck, 2018). gelation mechanism of small molecular weight gelators is to form a
However, the higher melting point of CW has a negative effect on the crystallization network through self-assembly to bind oil (Hwang,
structure properties of oleogels (Gravelle, Blach, Weiss, Barbut, & 2020). Due to the lower melting point of DS compared with CW, the

* Corresponding author. College of Food Science and Engineering, Ocean University of China, 5 Yushan Road, Qingdao, Shandong Province, 266003, China.
E-mail address: weizihao@ouc.edu.cn (Z. Wei).

https://doi.org/10.1016/j.lwt.2021.112454
Received 21 June 2021; Received in revised form 11 August 2021; Accepted 10 September 2021
Available online 12 September 2021
0023-6438/© 2021 Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
T. Xia et al. LWT 153 (2022) 112454

formation of assemblies with the help of DS is expected to reduce the was obtained by combining CW and DS at different mass ratios of 5:0,
melting point of the whole oleogel system. 4:1, 3:2, 2:3, 1:4, 0:5 and named as C5D0, C4D1, C3D2, C2D3, C1D4,
In addition to replacing animal fats and hydrogenated vegetable oil, C0D5, respectively. Afterwards, 10% (w/w) of the gelator was added
oleogel can also serve as a nutrient delivery system (Calligaris, Alongi, into astaxanthin-loaded corn oil, and the system was heated at 90 ◦ C
Lucci, & Anese, 2020). Besides, emulsions are common nutraceutical with a stirring speed of 300 rpm for 3 min. Then, the samples were
delivery systems in food systems (Salvia-Trujillo, Artiga-Artigas, Mole­ cooled at room temperature. Typical tube inversion method was used to
t-Rodríguez, Turmo-Ibarz, & Martín-Belloso, 2018; Xia, Xue, & Wei, inspect whether the oleogel was formed (Chung, Simmons, Gutowska, &
2021). The dispersity of oleogel in water is poor, but the emulsion has Jeong, 2002). After the oleogel was formed, the sample was stored at
good dispersibility. Therefore, oleogel-based nanoemulsion was devel­ 4 ◦ C for 24 h. Finally, all the measurements were performed after the
oped to improve delivery capability. Oleogel-based nanoemulsion was oleogel samples were placed at room temperature for 1 h.
first developed by Yu and Huang (Yu & Huang, 2012). Previous studies
have shown that oleogel-based nanoemulsion can improve the bio­ 2.3. Characterization of oleogel
accessibility of nutraceutical when compared with oleogel (Wei &
Huang, 2019). Nanoemulsions are usually stabilized by chemical sur­ 2.3.1. Hardness
factants (e.g., Tween), some of which have adverse effect on the gut The hardness of oleogel was measured with a texture analyzer (TMS-
microbiota and limit their application in food industry (Chassaing et al., TOUCH, Food Technology Corporation, USA) by using a cylindrical
2015; Lu, Cao, Ho, & Huang, 2016; Xia et al., 2021). In this study, to probe with a 30 mm diameter. The oleogel samples were placed in the
avoid adverse biosafety issues, fish collagen as an amphiphilic macro­ 100 mL beaker. All the oleogel samples were extruded to 30% defor­
molecule with good solubility was selected as emulsifier. mation once at 30 mm/min. For each sample, every moment of force (N)
Astaxanthin is a carotenoid that widely exists in the biological world, was recorded and the maximum force was considered as hardness.
especially in aquatic animals such as shrimp, crab and fish. Astaxanthin
has 13 conjugated double bonds, which endows it stronger antioxidant 2.3.2. Rheological analysis
properties than other antioxidants (Wan et al., 2020). Astaxanthin also A stress-controlled rheometer (Anton Paar Physica MCR 301, Graz,
has anti-inflammatory, antiapoptotic and antitumor effects (Zhou et al., Austria), which was equipped with a Peltier plate temperature controller
2021). In addition, astaxanthin is a lipophilic compound and has small (Julabo, Seelbach, Germany), was used to monitor the rheological
molecule size, which can easily cross the blood-brain barrier and play properties of oleogels. All tests were carried out with a 50 mm parallel
the strong role of neuroprotective effect (Galasso et al., 2018). But plate (PP50) at a constant temperature of 25 ◦ C and the measuring gap
similar to other carotenoids, astaxanthin has low bioaccessibility (Yang was set at 1 mm. Stress sweep measurement was performed at 1 Hz
et al., 2021). Thus, improving the bioaccessibility of astaxanthin is the frequency in the range of 0.001%–10% to determine the linear visco­
key to expand its application in food. So far, many emulsion-based de­ elastic region. Frequency sweep was performed at a constant low stress
livery systems have been developed. The oleogel-based nanoemulsion level in the linear viscoelastic region (0.1%) and the frequency range
has been proven to improve the bioaccessibility of capsaicin (Lu et al., was from 5 Hz to 20 Hz.
2016). Because astaxanthin has similar properties to capsaicin and there
is no study on the effect of composite gelators on oleogel-based nano­ 2.3.3. Oil binding capacity (OBC)
emulsion, oleogel-based nanoemulsion with improved properties is ex­ The measurement of oil binding capacity of oleogel was carried out
pected to be the delivery system of astaxanthin and improve the by centrifugation according to the method proposed by Fayaz et al. with
bioaccessibility of astaxanthin. some modifications (Fayaz et al., 2017). Briefly, 5 g of each oleogel
This study was started with the preparation of astaxanthin-loaded sample was placed in a 10 mL centrifuge tube and centrifuged (TGL20A,
oleogels using different ratios of CW and DS. Afterwards, the oleogel Changsha Yingtai Instrument Co., Ltd, Changsha, China) at 16050×g for
was characterized by texture, rheology, oil binding capacity and gel-sol 30 min. Then, the sample tube was inverted on the filter paper for 1 h to
melting temperature. Then several representative samples of oleogels separate the free oil. Finally, the total weight of the remaining sample
were selected to prepare oleogel-based nanoemulsions and the basic and tube was weighed. The OBC of oleogel was calculated according to
property of nanoemulsion was investigated. Finally, in vitro lipolysis and the following equation:
astaxanthin bioaccessibility in oleogel-based nanoemulsions were ( )
examined. Hopefully, this study may help to understand impact of OBC(%) = 1－
b－c
× 100% (1)
composite gelators on physicochemical properties of oleogels and nu­ a
traceutical delivery of oleogel-based nanoemulsions.
where a is the total mass of oleogel sample, b is the mass of tube and
2. Materials and methods sample before centrifugation, c is the mass of tube and sample after
centrifugation.
2.1. Materials
2.3.4. Gel-sol melting temperature of oleogel
Carnauba wax (CW) was bought from Aladdin Industrial Corporation The gel-sol melting temperature was measured by tube inversion
(Shanghai, China). 1-Docosanol (DS) was purchased from Shanghai method (Jadhav, Hwang, Huang, & John, 2013). Oleogel was first
Macklin Biochemical Co., Ltd (China). Astaxanthin was provided by packed in vial, and then the vial was inverted and placed in water-bath.
Shanxi Kangsheng Biotechnology Co., Ltd (China). Corn oil was pur­ Subsequently, raised the water temperature at a slow rate while the flow
chased from a local market. Fish collagen was obtained from Qingdao of oleogel samples was observed. The temperature in which the oleogel
Dongyi Technology Development Co., Ltd (China). Pig bile salt, pepsin, melted and flowed was considered to be the gel-sol melting temperature.
pancreatin and Tris was purchased from Beijing Solarbio Science &
Technology Co., Ltd (China). All other chemicals were bought from 2.4. Preparation of oleogel-based nanoemulsion
Sinopharm Chemical Reagent Co., Ltd (China), unless otherwise stated.
Fish collagen (50 mg/mL) was dispersed in Milli-Q water and stirred
2.2. Preparation of astaxanthin-loaded oleogels until dissolved. NaOH and HCI solutions were used to adjust the pH of
fish collagen solution to 3.0. Three oleogel samples (C5D0, C1D4, C0D5)
Astaxanthin (1 mg/g) was dispersed and dissolved in corn oil under were used to prepare oleogel-based nanoemulsions. Astaxanthin-loaded
heating (120 ◦ C) to prepare astaxanthin-loaded oil. Composite gelator oleogels were mixed with fish collagen solutions to obtain mixtures with

2
T. Xia et al. LWT 153 (2022) 112454

10% oleogel (w/v). Then, the mixtures were sonicated by an intelligent analysis of astaxanthin in this study was modified according to the
ultrasonic processor (SM-900A, Nanjing Shunma Instrument Equipment method described by Biehler, Mayer, Hoffmann, Krause, and Bohn
Co., Ltd, Nanjing, China) at 70% pressure amplitude for 10 min in a (2010). Methanol and tert-butyl methyl ether (1:1, v/v) was added into
pulse mode (4 s on and 6 s off). 5 mL of the intermediate micelle phase for oscillation, followed by
centrifugation at 11150×g for 2 min, and the upper solution was
2.5. Droplet size of nanoemulsion collected. Repeating above steps for the lower solution until the upper
solution was colorless. Then, all the upper solutions were collected and
The droplet size was measured using a Nano ZS90 Zetasizer (Malvern dried using a vacuum concentrator. The residue was redissolved with 1
Instruments Ltd., U.K.) with Zetasizer software v7.13 (Malvern In­ mL methanol/tert-butyl methyl ether (1:1, v/v). filtered through a
struments). The instrument was used to measure the droplet size of 0.22-μm nylon membrane filter and then analyzed by HPLC.
nanoemulsion by dynamic light scattering (DLS) technology. Each An Agilent 1260 HPLC system (Agilent Technologies Inc., California,
sample was diluted 100 times by Milli-Q water (pH 3) to avoid the effect USA) with a YMC-C30 (YMC CO., LTD, Japan) column (4.6 mm × 250
of multiple scattering before measurement. Then the measurements mm, 5 μm) was applied to measure the content of astaxanthin. The
were carried out at 25 ◦ C. The surface average diameter D[3,2] of the mobile phase was consisted with (A) methanol and (B) tert-butyl methyl
droplet size could be estimated: ether. A gradient elution procedure with a flow rate of 1.0 mL/min was
∑ carried out. Elution condition was programmed as follows: 0–10 min,
Ni D3i 90% A and 10% B; 10–20 min, A went from 90% to 40% linearly; 20–30
D[3,2] = ∑i (2)
min, 40% A and 60% B; 30–35 min, linear gradient from 40% to 60% A;
2
i Ni Di

35–40 min, 90% A and 10% B. The detection wavelength was set at 476
Where Ni was the total number of droplets with diameter Di . The re­
nm using a G7114A UV detection system (Agilent Technologies Inc.,
ported values were average of three different batches of each
California, USA), and the injection volume were 30 μL.
nanoemulsion.
After using the HPLC quantification of astaxanthin in the micelle
phase, astaxanthin bioaccessibility of three oleogel-based nanoemulsion
2.6. In vitro gastrointestinal digestion of astaxanthin in oleogel-based samples was calculated as follows:
nanoemulsion
astaxanthin content in the micelle phase
bioaccessibility = (4)
Simulated gastric digestion: Simulated gastric fluid (SGF) without total astaxanthin content in the formulations
pepsin was obtained by adjusting the pH of sodium chloride solutions (2
g/L) to 1.2 with HCl. The oleogel-based nanoemulsion sample contain­ 2.9. Statistical analysis
ing 2 g of oleogel was mixed with 16 mL SGF and cultured in a
temperature-controlled oil bath (37 ◦ C) under magnetic stirring (210 Each experiment was performed at least in triplicate. The data were
rpm). Afterwards, dissolved 32 mg of pepsin in 4 mL of SGF and added to processed by a one-way ANOVA analysis of variance and Duncan’s
the mixture to commence the digestion process. After 2 h of digestion, multiple range test. Statistical analysis was performed by SPSS 20.0
the pH of the system was adjusted to 7.5 to inactive the pepsin and (SPSS Inc., Chicago, IL, USA). A p-value of less than 0.05 was considered
closure gastric digestion. as statistically significant. Origin 2017 software was applied to perform
Simulated intestinal digestion: Simulated intestinal fluid (SIF) was all of the statistical analysis.
obtained by dissolving bile salt (10 mg/mL), CaCl2 (10 mM), Tris (50
mM) into water, followed by adjusting the pH of the system to 7.5 with 3. Results and discussion
HCl. Then, 64 mg of pancreatin was dissolved in 20 mL SIF. Afterwards,
the SIF was added to gastric digesta under continuous stirring in 37 ◦ C 3.1. Preparation of astaxanthin-loaded oleogel with composite gelators
oil bath to start intestinal digestion. During the digestion process, the pH
of the system should be monitored at all times, and 0.25M NaOH was Corn oil has high nutritional value due to high content of phytos­
adding to the system to keep the pH at 7.5. The volume and time of terols, and corn oil is widely consumed in the world. In this study, corn
added NaOH solution in the whole process of intestinal digestion were oil-based oleogel was developed to investigate influence of composite
recorded to calculate extent and speed of lipolysis. The whole process of gelators on oleogels and oleogel-based nanoemulsions. Many nutra­
simulated intestinal digestion lasted for 2 h. ceuticals such as curcumin, hesperidin and astaxanthin were difficult to
be incorporated in food because of their hydrophobicity, and dissolving
2.7. Extent of lipolysis in oleogel-based nanoemulsion them into oil could broaden their application in food. In this study, non-
dissolved astaxanthin was not found in astaxanthin-loaded corn oil after
The degree of lipolysis can be analyzed by the release of free fatty heating. This proved that astaxanthin could be completely dissolved in
acids (FFA). It was assumed that the digestion of 1 mol of triglycerides corn oil at the concentration of 1 mg/g. According to the non-flow state
released 2 mol of FFA and consumed 2 mol of NaOH. For this reason, the of the samples after inversion, all samples could form oleogels under the
fraction of released FFA can be calculated as: gelator concentration of 10%.
Mcorn oil × VNaOH × mNaOH
%FFA = 100 × (3) 3.2. Characterization of oleogel
wcorn oil × 2

where Mcorn oil was molecular mass of the corn oil (in g/mol), VNaOH was 3.2.1. Hardness
the amount of 0.25m NaOH added to the system (in L), mNaOH was the The hardness of oleogel is the main factor affecting its practical ap­
molarity of NaOH solution (in mol/L), and wcorn oil was the total mass of plications such as baked food. Fig. 1a showed the relationship between
initial present corn oil (in g). the hardness of oleogels and the ratio of CW to DS. It exhibited that the
combination of CW and DS could produce either harder oleogels (e.g.
2.8. Astaxanthin bioaccessibility in oleogel-based nanoemulsion formulation 1:4) or softer oleogels (e.g. formulation 3:2) compared to
oleogels prepared by a single gelator. The maximum hardness value was
When gastrointestinal digestion ended, the intestinal digesta were found in C1D4 oleogel, followed by C4D1 oleogel. The hardness of C3D2
centrifuged at 16050×g for 45 min. The intermediate micelle phase was oleogel was the lowest. The hardness of oleogels prepared by a single
collected for bioaccessibility analysis of astaxanthin. Method for the gelator was at the intermediate level, which revealed that the

3
T. Xia et al. LWT 153 (2022) 112454

70 2020). In this study, one of the main components of CW was long chain
fatty acid, and DS was a kind of long chain fatty alcohol (Soleimanian,
60 Goli, Shirvani, Elmizadeh, & Marangoni, 2020). When CW was mixed
with DS in a certain proportion, DS and the fatty acids in CW reached the
appropriate ratio, resulting in synergistic effect and enhancing the
50
hardness of the oleogel.
Hardness (N)

40 3.2.2. Rheological analysis

Rheological tests were performed to characterize viscoelastic prop­
30 erties of the oleogel. According to the results of strain sweep (data not
shown), storage modulus (G′ ) and loss modulus (G′′ ) of oleogel were
20 stable in the 0.01%–0.5% strain region, which was defined as the linear
viscoelastic region (LVE) for this oleogel. Frequency sweep test was
performed in a constant strain of 0.1%. Fig. 2 showed the variation of G′
10
and G′′ with frequency. For viscous materials, G′ and G′′ could be used to
measure the solid-like and liquid-like properties of materials, respec­
0 tively (Patel, Babaahmadi, Lesaffer, & Dewettinck, 2015). In this study,
5:0 4:1 3:2 2:3 1:4 0:5 all the results had the tendency that with the increase of frequency, the
CW:DS radio (w/w) slope of G′ was smaller. It showed that they had low frequency depen­
dence, which was symbol of elastic material. Since G′ and G′′ repre­
(a)
sented elastic and viscous part of oleogel, G′ of all oleogel samples was
much larger than G′′ , which indicated that all samples had a gel-like
structure (Wei, Zhang, & Huang, 2019). The oleogel produced with
100
C1D4 showed the utmost G′ , suggesting that a more elastic structure was
formed. In particular, when the storage modulus and shear modulus of
98
the gels meet G’’/G’ (ω) ≤ 0.1, they are defined as “strong gel” (Tav­
ernier et al., 2018). In these samples, only C1D4 oleogels could be
96
defined as “strong gel”. The reason of this phenomenon could be related
to a strong interaction between CW and DS under the radio of 1:4, which
OBC (%)

94 helped to form a more compact three-dimensional network. The

cross-linking of the three-dimensional network could be explained by
92 the formation mechanism of the oleogel. Oleogel formed by low mo­
lecular weight gelator was usually achieved through competitive in­
90 teractions between fibrous structure formation and crystallization
(Vintiloiu et al., 2008). Both CW and DS crystallized and blocked oil to
88 form oleogels (Vintiloiu et al., 2008). The crystalline properties mainly
depended on the composition of the mixture. When the ratio of CW to DS
86 changed, the crystal morphology also changed. When CW and DS were
5:0 4:1 3:2 2:3 1:4 0:5 mixed at the ratio of 1:4, the mixture formed the smallest needle-like
CW:DS radio (w/w) shape crystal. Small and numerous crystals made the overall structure
more compact, so the oil phase was difficult to escape, thus forming
(b) “strong gel”. This could also explain the change of the hardness of the
oleogel.
Fig. 1. (a) The hardness of oleogel at different ratios of carnauba wax (CW) and
1-docosanol (DS). (b) Oil binding capacity (OBC) of oleogel at different ratios of
CW and DS.

combination of CW and DS could produce both synergistic effect and

antagonistic effect with the change of proportion. The synergistic effect 300000
was produced when the ratio of CW to DS was far from 1, resulting in C5D0 G'
C5D0 G''
increased hardness of oleogel. The increase of hardness might be C4D1 G'
250000
enhanced by intermolecular hydrogen bonds between the three- C4D1 G''
dimensional reticular structures of oleogel. Conversely, when the ratio C3D2 G'
200000 C3D2 G''
of CW to DS was close to 1, antagonism effect appeared and led to the
G', G'' (Pa)

C2D3 G'
decrease of hardness. With the change of gelator ratio, the phenomenon C2D3 G''
of synergistic effect and antagonistic effect transforming into each other 150000 C1D4 G'
C1D4 G''
also appeared in other studies (Blach et al., 2016; Callau, Sow-Kebe, C0D5 G'
Jenkins, & Fameau, 2020). It was confirmed that the combination of 100000 C0D5 G''
long chain fatty alcohols and long chain fatty acids under a certain
proportion could produce synergistic effect to improve the hardness and 50000
oil binding ability of oleogel. For example, the oleogel formed by stearyl
alcohol and stearic acid at the radio of 8:2 exhibited the greatest hard­ 0
ness. But when their proportions changed to 4:6, they produced antag­
onism and reduced the hardness of the oleogel (Blach et al., 2016). 0 5 10 15 20 25
Similar results also occurred in the oleogel prepared by behenyl alcohol Frequency (Hz)
and behenic acid (Callau, Sow-Kebe, Nicolas-Morgantini, & Fameau,
Fig. 2. Rheological properties of oleogel during frequency sweep.

4
T. Xia et al. LWT 153 (2022) 112454

3.2.3. Oil binding capacity (OBC) It was also interesting to explore whether the synergistic effect of gela­
Oil binding capacity is an important parameter for evaluating the tors would also be advantageous in preparation of oleogel-based
quality of oleogel and predicting the application of oleogel in food nanoemulsion.
products. OBC represents not only the oil structuring capabilities of
oleogel network but also the stability of the oleogel (Yang, Yang, Chen, 3.3. Preparation of oleogel-based nanoemulsion
Chen, & Liu, 2020). In food manufacturing, the ideal oleogel is one that
oil oozes out little and slowly (Tavernier et al., 2018). Fig. 1b showed the Since the effect of composite gelators on oleogel-based nano­
results of OBC measurement. In general, the OBC of all the oleogel emulsions was not reported before, this paper was the first to investigate
samples was above 90%. This indicated that CW, DS and their complexes it. C1D4 oleogel had the optimal performance in the analysis results of
had strong gel properties. The results also showed that the combination texture, rheology and OBC. In the gel-sol melting temperature, C1D4
of CW and DS had perceptible effect on OBC of the oleogel. With the oleogel also had a good performance. In order to simplify the experi­
increase of DS content, the OBC of the oleogels showed a M-shaped ment, oleogel samples prepared by C5D0, C1D4, C0D5 were selected for
change trend. In particular, it could be found that the OBC results were subsequent experiments. Protein fiber is an excellent emulsifier, while
consistent with the hardness of oleogels. When one of the two kinds of collagen is a natural fibrous protein (Wei, Cheng, & Huang, 2019; de
gelators was slightly substituted by the other (e.g. C1D4 oleogel and Castro Santana, Kawazoe Sato, & Lopes da Cunha, 2012; Wei, Zhu,
C4D1 oleogel), it could significantly increase its OBC of oleogel. The Cheng, & Huang, 2019). Compared with terrestrial mammalian
reason for the increase of OBC might be that a small amount of gelator collagen, the possibility of zoonosis and religious restrictions of using
replacement improved the nucleation and crystallization process. Then fish collagen is lower in most cases (Luo, Zhou, Yao, & Fu, 2020).
the gelators entrapped of large volumes of corn oil between the crys­ Therefore, fish collagen has great potential as an emulsifier. The iso­
talline strands. Therefore, the oil loss of the oleogel samples was reduced electric point of collagen was near pH 5.0 (de Castro Santana, Kawazoe
during centrifugal process. Another reason was that OBC of oleogel was Sato, & Lopes da Cunha, 2012). When the pH of the emulsifiers was close
related to rheology. Due to higher G′ and yield stress, low value of corn to the isoelectric point, the electrostatic repulsion between emulsifiers
oil was discharged during centrifugation. Similar results were found in decreased, resulting in aggregation and sedimentation of emulsifiers,
other oleogel prepared by mixtures of gelators. With the change of the which led to poor emulsifying property. Thus, in this study, the pH of
proportion of the two kinds of gelator, the OBC of the oleogel might fish collagen was adjusted to 3.0 to endow it better emulsifying prop­
become larger or smaller (Blach et al., 2016; Yang et al., 2020). erty. All the oleogel-based nanoemulsions stabilized by fish collagen had
good stability in this study.
3.2.4. Gel-sol melting temperature of oleogel
Since oleogel is thermoreversible and gel-sol transition temperature 3.4. Droplet size of oleogel-based nanoemulsion
plays an important role in food applications, gel-sol melting temperature
of oleogel was investigated in this study. Higher gel-sol melting tem­ The droplet size and polymer dispersity index (PdI) of the nano­
perature could lead to an undesirable waxy mouthcoating. Generally, emulsion were shown in Table 2. The nanoemulsion prepared by C1D4
the lower the gel-sol melting temperature of oleogel is, the easier it is to had the maximum droplet size. This phenomenon might be related to the
be applied in food. It could be seen from Table 1 that the gel-sol melting hardness of oleogel. It was difficult to break the oleogel with high
temperature of oleogel decreased gradually with the increase of DS hardness during ultrasonic process, resulting in larger oleogel particles
content. This phenomenon could be explained from the melting point of wrapped in fish collagen. In addition, the combination of CW and DS
the two kinds of gelators. The melting point of CW and DS was about enhanced the gel network of the oleogel, so it was difficult to disperse
83 ◦ C and 67 ◦ C, respectively. Therefore, the higher the content of CW, the oleogel into small droplets when prepared the oleogel-based nano­
the more heat it needed to melt. On the other hand, the combination of emulsion. The smallest droplet size of nanoemulsion was obtained at
CW and DS changed the crystallization state of the oleogel. Different C5D0. One explanation could support this result. As mentioned above,
crystal structures required different melting temperatures. There was no the mechanical strength of the oleogel prepared by a single CW was
doubt that the combine of CW and DS provided a solution for reducing relatively low. The oleogel could break easily when the system under­
the gel-sol melting temperature of wax-based oleogel. This strategy not went ultrasonic treatment, and more broken oleogel might contribute to
only reduced the gel-sol melting temperature of wax-based oleogel, but the formation of smaller droplet.
also preserved and even enhanced the other properties of wax-based PdI is an important factor affecting the properties of emulsion (Jiang
oleogel such as hardness and rheological behavior. et al., 2019), and PdI stands for the width of the droplet size (Cheong,
After a comprehensive discussion of the above characterization re­ Tan, Tan, & Nyam, 2016). The PdI of C5D0 and C0D5 oleogel-based
sults, it could be seen that CW and DS had strong synergistic effect at the nanoemulsions was less than 0.2, which indicated a narrow size distri­
ratio of 1:4. When the ratio of CW to DS was 2:3 or 3:2, it had antago­ bution and displayed that the nanoemulsions had better dispersibility as
nistic effect, which was manifested in the reduction of hardness, OBC well as stability (Li et al., 2019; Wei, Yang, Fan, Yuan, & Gao, 2015). The
and other properties. This provided a new way to prepare oleogels with PdI of nanoemulsion prepared by C1D4 oleogel was more than 0.5, it
distinctive performance. In practical application, the suitable proportion suggesting that the droplet size distribution was wide and the nano­
of gelators could be selected to prepare oleogels according to the needs. emulsion was easy to gather and destabilize.

Table 1
Gel-sol melting temperature of oleogels.
CW:DS radio (w/w) Gel-sol melting temperature (◦ C) Table 2
5:0 85.0 ± 1.4d Droplet size and PdI (polymer dispersity index) of oleogel-based nanoemulsion.
4:1 73.0 ± 1.9c
CW:DS radio (w/w) Droplet size (nm) PdI
3:2 71.0 ± 4.0bc
2:3 65.0 ± 3.0ab 5:0 227.2 ± 2.4a 0.118 ± 0.021a
1:4 63.0 ± 2.6a 1:4 392.9 ± 26.7c 0.596 ± 0.032c
0:5 61.0 ± 3.2a 0:5 298.8 ± 6.0b 0.200 ± 0.020b

Note: CW and DS refer to carnauba wax and 1-docosanol, respectively. Note: CW and DS refer to carnauba wax and 1-docosanol, respectively.
Values are means ± SD (n = 3). Different superscript letters in the table Values are means ± SD (n = 3). Different superscript letters in the table indicate
indicate significant differences (p < 0.05). significant differences (p < 0.05).

5
T. Xia et al. LWT 153 (2022) 112454

3.5. Lipolysis in oleogel-based nanoemulsion network structure which was denser than the other two, so its void
spaces were smaller than that of the other two. This prevented lipase
Analysis of lipolysis could determine the digestibility of lipid during contact with oil directly through void spaces.
in vitro digestion. Fig. 3a showed that fraction of FFA released in C5D0
oleogel-based nanoemulsion was the highest (41.0%), suggesting higher 3.6. Bioaccessibility of astaxanthin in oleogel-based nanoemulsion
extent of lipolysis. C1D4 oleogel-based nanoemulsion had the lowest
lipid digestibility (26.4%), indicating that the degree of lipolysis was the The bioaccessibility of nutrients is mainly affected by solubilization,
lowest. Considering that the digestion rate and degree of lipid were permeation and metabolism (Yu et al., 2012). This study mainly
related to the droplet size and sufficient contact of lipase with oil, dif­ explored the bioaccessibility of astaxanthin from the aspect of solubili­
ference of lipid digestion among the three samples might be accounted zation. Nanoemulsion can transform the oil phase into free fatty acids
for several factors. First, since the droplet size of C5D0 oleogel-based and monoacylglycerols in the process of digestion, both of which
nanoemulsion was the smallest, C5D0 oleogel-based nanoemulsion contribute to the formation of mixed micelles. With the progress of
had a much larger interfacial area than the other two oleogel-based lipolysis, astaxanthin, which was originally dissolved in lipid, was
nanoemulsions, which was beneficial to increase the lipase-oil inter­ encapsulated into mixed micelles. Subsequently, astaxanthin entered
face and accelerate the speed of lipolysis (Salvia-Trujillo, Qian, the systemic circulation through passive diffusion and became
Martin-Belloso, & McClements, 2013). Second, void spaces might exist bioavailable (Wei, Cheng, Zhu, & Huang, 2019). Therefore, it was
among gel network, and lipase might directly enter the oil phase through believed that astaxanthin in mixed micelles was bioaccessible. As shown
the void spaces for lipolysis (Tan, Sun, Lin, Mu, & Ngai, 2014; Wei & in Fig. 3b, astaxanthin bioaccessibility was the highest in the C1D4
Huang, 2019). The oleogel formed by C1D4 had a three-dimensional oleogel-based nanoemulsion, followed by C0D5 oleogel-based nano­
emulsion. Astaxanthin bioaccessibility in the C5D0 oleogel-based
nanoemulsion was the lowest, which was opposite to the digestibility
60
of lipid. The release of more fatty acids helped to produce more micelles.
5:0 Hydrophobic astaxanthin should be dissolved in the core of micelles
50 1:4 before it becomes bioaccessible. In general, the higher the release of
0:5 FFA, the higher the bioaccessibility of astaxanthin (Wei & Huang, 2019;
Wei, Cheng, et al., 2019). However, the extent of lipolysis and astax­
40 anthin bioaccessibility had a negative correlation in this oleogel-based
% FFA Released

system. Several reasons could be used to explain the opposite phenom­

enon. As shown in Fig. 4, the mixed micelles formed after the digestion
30
of the C1D4 oleogel-based nanoemulsion could dissolve more astax­
anthin. After digestion, the oil phase was decomposed into micelles,
20 while CW and DS continued to adsorb on the surface of micelles because
they were hard to digest (de Freitas et al., 2019). Both CW and DS were
food-grade surfactants in nature. They attached to the surface of micelles
10
and produced synergistic effect, which made more astaxanthin dissolve
in micelles and became the core of micelles. Therefore, the bio­
0 accessibility of astaxanthin was improved. Previous studies have sug­
0 20 40 60 80 100 120 gested that the bioaccessibility of astaxanthin encapsulated in
Digestion time (min) nanoemulsion was 33% (Khalid et al., 2017). In this study, the bio­
accessibility of astaxanthin reached 55% and increased by 22%, which
(a) was a very exciting result. These results provided a new idea for the high
value utilization of astaxanthin.

(b)

Fig. 3. (a) Release of free fatty acids (FFA) in oleogel-based nanoemulsion

stabilized by fish collagen. (b) The bioaccessibility of astaxanthin in oleogel- Fig. 4. Morphology of astaxanthin-loaded oleogel-based nanoemulsion before
based nanoemulsion after in vitro digestion. and after digestion.

6
T. Xia et al. LWT 153 (2022) 112454

4. Conclusion Cheong, A. M., Tan, K. W., Tan, C. P., & Nyam, K. L. (2016). Kenaf (Hibiscus cannabinus
L.) seed oil-in-water Pickering nanoemulsions stabilised by mixture of sodium
caseinate, Tween 20 and β-cyclodextrin. Food Hydrocolloids, 52, 934–941. https://
In conclusion, after CW and DS combined at the ratio of 1:4, the doi.org/10.1016/j.foodhyd.2015.09.005
physicochemical properties of composite oleogel such as texture, Chung, Y., Simmons, K., Gutowska, A., & Jeong, B. (2002). Sol-gel transition temperature
rheology and OBC were improved. Therefore, these two kinds of gelators of PLGA-g-PEG aqueous solutions. Biomacromolecules, 3, 511–516. https://doi.org/
10.1021/bm0156431
could produce synergistic effect at the ratio of 1:4. Compared with CW- Dassanayake, L. S. K., Kodali, D. R., & Ueno, S. (2011). Formation of oleogels based on
based oleogel, the decrease of gel-sol melting temperature endowed edible lipid materials. Current Opinion in Colloid & Interface Science, 16, 432–439.
composite oleogel with more application opportunities. In the prepa­ https://doi.org/10.1016/j.cocis.2011.05.005
Doan, C. D., Tavernier, I., Okuro, P. K., & Dewettinck, K. (2018). Internal and external
ration of nanoemulsion, the performance of composite oleogel-based factors affecting the crystallization, gelation and applicability of wax-based oleogels
nanoemulsion was different from single components oleogel-based in food industry. Innovative Food Science & Emerging Technologies, 45, 42–52. https://
nanoemulsions. It is mainly manifested in larger droplet size and doi.org/10.1016/j.ifset.2017.09.023
Fayaz, G., Goli, S. A. H., Kadivar, M., Valoppi, F., Barba, L., Calligaris, S., et al. (2017).
wider PdI. Although the degree of lipolysis of composite oleogel-based Potential application of pomegranate seed oil oleogels based on monoglycerides,
nanoemulsion was lower than that of single gelator oleogel-based beeswax and propolis wax as partial substitutes of palm oil in functional chocolate
nanoemulsion, the bioaccessibility of astaxanthin was significantly spread. Lebensmittel-Wissenschaft und -Technologie- Food Science and Technology, 86,
523–529. https://doi.org/10.1016/j.lwt.2017.08.036
improved. There is no doubt that composite oleogel-based nano­ de Freitas, C. A. S., de Sousa, P. H. M., Soares, D. J., da Silva, J. Y. G., Benjamin, S. R., &
emulsion has profound significance in improving the bioaccessibility of Guedes, M. I. F. (2019). Carnauba wax uses in food - a review. Food Chemistry, 291,
nutrients. The obtained results will provide reference for the application 38–48. https://doi.org/10.1016/j.foodchem.2019.03.133
Galasso, C., Orefice, I., Pellone, P., Cirino, P., Miele, R., Ianora, A., et al. (2018). On the
of oleogel-based nanoemulsion in the delivery of nutraceutical.
neuroprotective role of astaxanthin: New perspectives? Marine Drugs, 16, 247.
https://doi.org/10.3390/md16080247
CRediT authorship contribution statement Gravelle, A. J., Blach, C., Weiss, J., Barbut, S., & Marangoni, A. G. (2017). Structure and
properties of an ethylcellulose and stearyl alcohol/stearic acid (EC/SO:SA) hybrid
oleogelator system. European Journal of Lipid Science and Technology, 119, 1700069.
Tianhang Xia: Investigation, Software, Data curation, Writing – https://doi.org/10.1002/ejlt.201700069
original draft. Zihao Wei: Conceptualization, Methodology, Resources, Hwang, H. (2020). A critical review on structures, health effects, oxidative stability, and
sensory properties of oleogels. Biocatalysis and Agricultural Biotechnology, 26,
Writing – review & editing, Project administration, Funding acquisition,
101657. https://doi.org/10.1016/j.bcab.2020.101657
Supervision. Changhu Xue: Funding acquisition, Supervision. Jadhav, S. R., Hwang, H., Huang, Q., & John, G. (2013). Medium-chain sugar
amphiphiles: A new family of healthy vegetable oil structuring agents. Journal of
Agricultural and Food Chemistry, 61, 12005–12011. https://doi.org/10.1021/
Declaration of competing interest jf401987a
Jiang, Y., Zhang, C., Yuan, J., Wu, Y., Li, F., Li, D., et al. (2019). Effects of pectin
polydispersity on zein/pectin composite nanoparticles (ZAPs) as high internal-phase
The authors declare no conflict of interest and no competing finan­ Pickering emulsion stabilizers. Carbohydrate Polymers, 219, 77–86. https://doi.org/
cial interest. 10.1016/j.carbpol.2019.05.025
Khalid, N., Shu, G., Holland, B. J., Kobayashi, I., Nakajima, M., & Barrow, C. J. (2017).
Formulation and characterization of O/W nanoemulsions encapsulating high
Acknowledgments concentration of astaxanthin. Food Research International, 102, 364–371. https://doi.
org/10.1016/j.foodres.2017.06.019
This work was supported by the start-up fund from Ocean University Li, X. M., Zhu, J., Pan, Y., Meng, R., Zhang, B., & Chen, H. Q. (2019). Fabrication and
characterization of pickering emulsions stabilized by octenyl succinic anhydride
of China (grant No. 862001013134). This work was also funded by the -modified gliadin nanoparticle. Food Hydrocolloids, 90, 19–27. https://doi.org/
National Key R&D Program of China (grant No. 2018YFC0311201), 10.1016/j.foodhyd.2018.12.012
China Postdoctoral Science Foundation funded project (2021M693025) Lu, M., Cao, Y., Ho, C. T., & Huang, Q. (2016). Development of organogel-derived
capsaicin nanoemulsion with improved bioaccessibility and reduced gastric mucosa
and Qingdao Postdoctoral Application Foundation funded project.
irritation. Journal of Agricultural and Food Chemistry, 64, 4735–4741. https://doi.
org/10.1021/acs.jafc.6b01095
References Luo, J., Zhou, Z., Yao, X., & Fu, Y. (2020). Mineral-chelating peptides derived from fish
collagen: Preparation, bioactivity and bioavailability. Lebensmittel-Wissenschaft &
Technologie, 134, 110209. https://doi.org/10.1016/j.lwt.2020.110209
Biehler, E., Mayer, F., Hoffmann, L., Krause, E., & Bohn, T. (2010). Comparison of 3
Oh, I. K., Amoah, C., Lim, J., Jeong, S., & Lee, S. (2017). Assessing the effectiveness of
spectrophotometric methods for carotenoid determination in frequently consumed
wax-based sunflower oil oleogels in cakes as a shortening replacer. Lebensmittel-
fruits and vegetables. Journal of Food Science, 75, C55–C61. https://doi.org/
Wissenschaft und -Technologie- Food Science and Technology, 86, 430–437. https://doi.
10.1111/j.1750-3841.2009.01417.x
org/10.1016/j.lwt.2017.08.021
Bin Sintang, M. D., Danthine, S., Patel, A. R., Rimaux, T., Van De Walle, D., &
Okuro, P. K., Tavernier, I., Bin Sintang, M. D., Skirtach, A. G., Vicente, A. A.,
Dewettinck, K. (2017). Mixed surfactant systems of sucrose esters and lecithin as a
Dewettinck, K., et al. (2018). Synergistic interactions between lecithin and fruit wax
synergistic approach for oil structuring. Journal of Colloid and Interface Science, 504,
in oleogel formation. Food & Function, 9, 1755–1767. https://doi.org/10.1039/
387–396. https://doi.org/10.1016/j.jcis.2017.05.114
c7fo01775h
Blach, C., Gravelle, A. J., Peyronel, F., Weiss, J., Barbut, S., & Marangoni, A. G. (2016).
Patel, A. R., Babaahmadi, M., Lesaffer, A., & Dewettinck, K. (2015). Rheological profiling
Revisiting the crystallization behavior of stearyl alcohol : stearic acid (SO : SA)
of organogels prepared at critical gelling concentrations of natural waxes in a
mixtures in edible oil. RSC Advances, 6, 81151–81163. https://doi.org/10.1039/
triacylglycerol solvent. Journal of Agricultural and Food Chemistry, 63, 4862–4869.
c6ra15142f
https://doi.org/10.1021/acs.jafc.5b01548
Callau, M., Sow-Kebe, K., Jenkins, N., & Fameau, A. L. (2020). Effect of the ratio between
Salvia-Trujillo, L., Artiga-Artigas, M., Molet-Rodríguez, A., Turmo-Ibarz, A., & Martín-
fatty alcohol and fatty acid on foaming properties of whipped oleogels. Food
Belloso, O. (2018). Emulsion-based nanostructures for the delivery of active
Chemistry, 333, 127403. https://doi.org/10.1016/j.foodchem.2020.127403
ingredients in foods. Frontiers in Sustainable Food Systems, 2, 79. https://doi.org/
Callau, M., Sow-Kebe, K., Nicolas-Morgantini, L., & Fameau, A. L. (2020). Effect of the
10.3389/fsufs.2018.00079
ratio between behenyl alcohol and behenic acid on the oleogel properties. Journal of
Salvia-Trujillo, L., Qian, C., Martin-Belloso, O., & McClements, D. J. (2013). Influence of
Colloid and Interface Science, 560, 874–884. https://doi.org/10.1016/j.
particle size on lipid digestion and beta-carotene bioaccessibility in emulsions and
jcis.2019.10.111
nanoemulsions. Food Chemistry, 141, 1472–1480. https://doi.org/10.1016/j.
Calligaris, S., Alongi, M., Lucci, P., & Anese, M. (2020). Effect of different oleogelators on
foodchem.2013.03.050
lipolysis and curcuminoid bioaccessibility upon in vitro digestion of sunflower oil
Sawalha, H., Venema, P., Bot, A., Floter, E., Adel, R. D., & van der Linden, E. (2015). The
oleogels. Food Chemistry, 314, 126146. https://doi.org/10.1016/j.
phase behavior of gamma-oryzanol and beta-sitosterol in edible oil. Journal of
foodchem.2019.126146
American Oil Chemical Society, 92, 1651–1659. https://doi.org/10.1007/s11746-
de Castro Santana, R., Kawazoe Sato, A. C., & Lopes da Cunha, R. (2012). Emulsions
015-2731-3
stabilized by heat-treated collagen fibers. Food Hydrocolloids, 26, 73–81. https://doi.
Soleimanian, Y., Goli, S. A. H., Shirvani, A., Elmizadeh, A., & Marangoni, A. G. (2020).
org/10.1016/j.foodhyd.2011.04.006
Wax-based delivery systems: Preparation, characterization, and food applications.
Chassaing, B., Koren, O., Goodrich, J. K., Poole, A. C., Srinivasan, S., Ley, R. E., et al.
Comprehensive Reviews in Food Science and Food Safety, 19, 2994–3030. https://doi.
(2015). Dietary emulsifiers impact the mouse gut microbiota promoting colitis and
org/10.1111/1541-4337.12614
metabolic syndrome. Nature, 519, 92–96. https://doi.org/10.1038/nature14232
Tan, H., Sun, G., Lin, W., Mu, C., & Ngai, T. (2014). Gelatin particle-stabilized high
Chaves, K. F., Barrera-Arellano, D., & Ribeiro, A. P. B. (2018). Potential application of
internal phase emulsions as nutraceutical containers. ACS Applied Materials &
lipid organogels for food industry. Food Research International, 105, 863–872.
Interfaces, 6, 13977–13984. https://doi.org/10.1021/am503341j
https://doi.org/10.1016/j.foodres.2017.12.020

7
T. Xia et al. LWT 153 (2022) 112454

Tavernier, I., Doan, C. D., Van der Meeren, P., Heyman, B., & Dewettinck, K. (2018). The model β-carotene emulsion. Food Hydrocolloids, 45, 337–350. https://doi.org/
potential of waxes to alter the microstructural properties of emulsion-templated 10.1016/j.foodhyd.2014.12.008
oleogels. European Journal of Lipid Science and Technology, 120, 1700393. https:// Wei, Z., Zhang, H., & Huang, Q. (2019). Curcumin-loaded Pickering emulsion stabilized
doi.org/10.1002/ejlt.201700393 by insoluble complexes involving ovotransferrin-gallic acid conjugates and
Valoppi, F., Calligaris, S., & Marangoni, A. G. (2016). Structure and physical properties of carboxymethyldextran. Food & Function, 10, 4911–4923. https://doi.org/10.1039/
oleogels containing peanut oil and saturated fatty alcohols. European Journal of Lipid c9fo01162e
Science and Technology, 119, 1600252. https://doi.org/10.1002/ejlt.201600252 Wei, Z., Zhu, J., Cheng, Y., & Huang, Q. (2019). Ovotransferrin fibril-stabilized Pickering
Vintiloiu, A., & Leroux, J. C. (2008). Organogels and their use in drug delivery–a review. emulsions improve protection and bioaccessibility of curcumin. Food Research
Journal of Controlled Release, 125, 179–192. https://doi.org/10.1016/j. International, 125, 108602. https://doi.org/10.1016/j.foodres.2019.108602
jconrel.2007.09.014 Xia, T., Xue, C., & Wei, Z. (2021). Physicochemical characteristics, applications and
Wan, X., Zhou, X. R., Moncalian, G., Su, L., Chen, W. C., Zhu, H. Z., et al. (2020). research trends of edible Pickering emulsions. Trends in Food Science & Technology,
Reprogramming microorganisms for the biosynthesis of astaxanthin via metabolic 107, 1–15. https://doi.org/10.1016/j.tifs.2020.11.019
engineering. Progress in Lipid Research, 81, 101083. https://doi.org/10.1016/j. Yang, L., Qiao, X., Gu, J., Li, X., Cao, Y., Xu, J., et al. (2021). Influence of molecular
plipres.2020.101083 structure of astaxanthin esters on their stability and bioavailability. Food Chemistry,
Wei, Z., Cheng, J., & Huang, Q. (2019). Food-grade Pickering emulsions stabilized by 343, 128497. https://doi.org/10.1016/j.foodchem.2020.128497
ovotransferrin fibrils. Food Hydrocolloids, 94, 592–602. https://doi.org/10.1016/j. Yang, S., Yang, G., Chen, X., Chen, J., & Liu, W. (2020). Interaction of monopalmitate
foodhyd.2019.04.005 and carnauba wax on the properties and crystallization behavior of soybean oleogel.
Wei, Z., Cheng, Y., Zhu, J., & Huang, Q. (2019). Genipin-crosslinked ovotransferrin Grain & Oil Science and Technology, 3, 49–56. https://doi.org/10.1016/j.
particle-stabilized Pickering emulsions as delivery vehicles for hesperidin. Food gaost.2020.05.001
Hydrocolloids, 94, 561–573. https://doi.org/10.1016/j.foodhyd.2019.04.008 Yu, H., & Huang, Q. (2012). Improving the oral bioavailability of curcumin using novel
Wei, Z., & Huang, Q. (2019). Developing organogel-based Pickering emulsions with organogel-based nanoemulsions. Journal of Agricultural and Food Chemistry, 60,
improved freeze-thaw stability and hesperidin bioaccessibility. Food Hydrocolloids, 5373–5379. https://doi.org/10.1021/jf300609p
93, 68–77. https://doi.org/10.1016/j.foodhyd.2019.01.050 Zhou, X., Zhang, J., Li, Y., Cui, L., Wu, K., & Luo, H. (2021). Astaxanthin inhibits
Wei, Z., Yang, W., Fan, R., Yuan, F., & Gao, Y. (2015). Evaluation of structural and microglia M1 activation against inflammatory injury triggered by lipopolysaccharide
functional properties of protein–EGCG complexes and their ability of stabilizing a through down-regulating miR-31-5p. Life Sciences, 267, 118943. https://doi.org/
10.1016/j.lfs.2020.118943