Polyphenol-mediated high-internal-phase Pickering emulsion as well as preparation method and application thereof

Abstract

The invention discloses a polyphenol-mediated high-internal-phase Pickering emulsion as well as a preparation method and application thereof, and belongs to the crossing field of food science and biological medicine application. The preparation method of the high-internal-phase Pickering emulsion comprises the following steps: respectively dissolving polyphenol, food-grade aldehydes, proteins, peptides or amino acids in water, then mixing the three solutions, and carrying out magnetic stirring under a normal-temperature dark condition to obtain a mixed system; then centrifuging, and freeze-drying the precipitate to obtain polyphenol composite nanoparticles; and dissolving the polyphenol composite nanoparticles in water to obtain a nanoparticle suspension, mixing the nanoparticle suspension with vegetable oil, and performing high-speed dispersion to obtain the polyphenol-mediated high-internal-phase Pickering emulsion. The polyphenol-mediated high-internal-phase Pickering emulsion has good biocompatibility, is stable in digestion of gastrointestinal tracts, and has a very excellent application prospect in the industries of medicines and health care products.

Description

Technical Field

[0001] The invention relates to the intersecting field of food science and biomedical application, and relates to a polyphenol-mediated high internal phase Pickering emulsion and a preparation method and application thereof. Background Art

[0002] With the development of society and the continuous improvement of living standards, food that only meets basic needs can no longer meet people's basic living needs. Nutritional diet has gradually become the dietary concept of modern people, among which food functional factors play a huge role. Food functional factors include amino acids, peptides, proteins, functional lipids, polysaccharides, oligosaccharides, terpenes, polyphenols, carotenoids, flavonoids, probiotics, minerals, vitamins, etc. They are substances that regulate human functions by activating enzyme activity or other pathways, and are the ingredients in functional foods that truly play a physiological role. However, due to their wide variety, different chemical compositions and functions, they can not only provide basic nutrition for the human body, but also provide additional health benefits. They also play an important role in improving food quality, regulating physiological functions of the body, preventing diseases and regulating human health. Therefore, the research and application of food functional factors are receiving more and more attention.

[0003] However, there are many problems in the application of food functional factors, such as poor solubility, poor stability, low bioavailability, and susceptibility to factors such as oxygen, pH, enzymes, and metal ions. Therefore, although food functional factors have great potential in promoting health, they still face many challenges in practical application.

[0004] Emulsion delivery systems can be used to encapsulate, transport and release bioactive substances, and are widely used in the fields of food and medicine. Studies have found that high internal phase Pickering emulsions have higher loading and bioaccessibility than traditional emulsions in delivering food functional factors, and have broad prospects in the development of drugs and fortified foods containing nutrients. As a new type of emulsion, it has the advantages of high stability, good safety, good biocompatibility, and resistance to Oswald ripening.

[0005] However, the stability of high internal phase Pickering emulsion depends on the type and properties of the particle stabilizer. The existing solid particles are divided into inorganic solid particles and organic solid particles. Inorganic particles mainly include modified silica, hydroxyapatite, titanium dioxide particles, graphene and zinc oxide. Although the stability of high internal phase Pickering emulsion stabilized by inorganic particles is improved, the biocompatibility is poor, which limits its application in biomedicine, food and other fields. Solid particles of biological origin have the advantages of environmental friendliness, renewability and biodegradability, and are mainly divided into: polysaccharide particles, protein particles, composite particles, etc. Composite particles have better emulsification properties and stronger spatial stability than single particles; for example, a tea polyphenol nanoparticle-stabilized Pickering emulsion and its preparation method disclosed in Chinese patent CN114672318A use polyphenol as a stabilizer to prepare Pickering emulsion, but polyphenol will be destroyed by high temperature, oxygen and radiation during processing or storage, and the stability is poor, which limits its application in food, biomedicine and other fields. Generally, the stability of polyphenols is improved by covalently or non-covalently combining polyphenols with polysaccharides or proteins to form stable particles. However, non-covalent bonding may affect the biological activity of polyphenols, proteins and polysaccharides, causing them to lose their original functions; most covalent bonding methods require high reaction conditions, such as high temperature, high pressure, strong acid, strong alkali, etc., which may damage the structure and function of proteins or polysaccharides. If the reaction is not appropriate during the covalent bonding process, toxic substances may even be produced, posing potential risks to human health. Summary of the invention

[0006] [Technical issues]

[0007] Nanoparticle solids that stabilize high internal phase Pickering emulsions have limitations such as safety issues and poor biocompatibility. Polyphenols as high internal phase Pickering emulsion stabilizers are easily decomposed by heat, oxygen and light, have poor chemical stability and low solubility in gastrointestinal fluid, resulting in extremely low oral bioavailability of only about 1%-5%. Therefore, a method is needed to enhance the functional properties of polyphenols and prepare safe and effective high internal phase Pickering emulsion stabilizers by complexing polyphenols with other biopolymers, so as to effectively improve the stability and bioavailability of high internal phase Pickering emulsions.

[0008] [Technical solution]

[0009] In view of the above technical problems, the present invention provides a polyphenol-mediated high internal phase Pickering emulsion and a preparation method and application thereof. The high internal phase Pickering emulsion prepared by the present invention uses polyphenol composite nanoparticles as a stabilizer to stabilize the high internal phase Pickering emulsion, improve the stability of the emulsion, prepare an emulsion carrier system, and promote the bioavailability of food functional factors.

[0010] Among them, the polyphenol composite nanoparticles are prepared by a one-pot method, specifically using polyphenols, food-grade aldehydes, proteins, peptides or amino acids as raw materials; the phenolic hydroxyl groups of polyphenols, the aldehyde groups of food-grade aldehydes and the amino groups of proteins, peptides and amino acids can undergo Mannich reactions to self-assemble into polyphenol composite nanoparticles, thereby stabilizing the high internal phase Pickering emulsion and preparing the emulsion carrier system.

[0011] In order to achieve the above purpose, the technical solutions provided are as follows:

[0012] A method for preparing a polyphenol-mediated high internal phase Pickering emulsion, the method comprising the following steps:

[0013] (1) dissolving polyphenols, food-grade aldehydes, proteins, peptides or amino acids in water to form a polyphenol solution, a food-grade aldehyde solution and a protein, peptide or amino acid solution respectively, then mixing the three solutions and magnetically stirring them at room temperature and in the dark to obtain a mixed system;

[0014] (2) centrifuging the mixed system of step (1), washing the precipitate obtained by centrifugation, and then freeze-drying the precipitate to obtain polyphenol composite nanoparticles;

[0015] (3) dissolving the polyphenol composite nanoparticles obtained in step (2) in water to obtain a nanoparticle suspension, and mixing it with vegetable oil and dispersing it at high speed to obtain a polyphenol-mediated high internal phase Pickering emulsion.

[0016] In one embodiment, the polyphenols in step (1) include one or more of tea polyphenols, catechins, brown algae polyphenols, and proanthocyanidins.

[0017] In one embodiment, the food-grade aldehydes in step (1) include one or more of vanillin, cinnamaldehyde, and benzaldehyde.

[0018] In one embodiment, the protein in step (1) includes one or more of whey protein isolate, casein, zein, and soy protein.

[0019] In one embodiment, the peptide in step (1) comprises one or both of sea cucumber peptide and sea ascidian polypeptide.

[0020] In one embodiment, the amino acids in step (1) include one or more of lysine, arginine, glutamic acid, and aspartic acid.

[0021] In one embodiment, the polyphenol concentration in the solution after mixing in step (1) is 1.25-7.50 mg / mL.

[0022] In one embodiment, the concentration of the food-grade aldehyde solution in the solution after mixing in step (1) is 1.25-7.50 mg / mL.

[0023] In one embodiment, the concentration of the protein, peptide or amino acid solution in the solution after mixing in step (1) is 0.5-2.5 mg / mL.

[0024] In one embodiment, the magnetic stirring speed in step (1) is 500-1000 rpm, and the stirring time is 20-24 h.

[0025] In one embodiment, the centrifugation parameters in step (2) are: 10000-12000 rpm, and the time is 5-10 min.

[0026] In one embodiment, the washing in step (2) is performed using deionized water for 3 to 5 times.

[0027] In one embodiment, the freeze drying in step (2) is performed using a freeze dryer under the following conditions: vacuum pressure of 1-20 Pa and temperature of -50--80°C.

[0028] In one embodiment, the concentration of the nanoparticle suspension in step (3) is 0.5% to 4.5%, w / v, mg / ml; preferably 4% to 4.5%, w / v, mg / ml.

[0029] In one embodiment, the volume proportion of the vegetable oil phase in step (3) is 75-80%, preferably 75-78%.

[0030] In one embodiment, the vegetable oil in step (3) includes one or more of soybean oil, rapeseed oil, and corn oil.

[0031] In one embodiment, the speed of the high-speed disperser for high-speed dispersion in step (3) is 10000-12000 rpm, and the dispersion time is 2-5 min.

[0032] Another object of the present invention is to provide a polyphenol-mediated high internal phase Pickering emulsion prepared by the method described above.

[0033] The third object of the present invention is to provide an application of the polyphenol-mediated high internal phase Pickering emulsion in the preparation of food, medicine and health care products.

[0034] A fourth object of the present invention is to provide a method for preparing a high internal phase Pickering emulsion loaded with a fat-soluble active substance, the method comprising the following steps:

[0035] (1) dissolving polyphenol powder, food-grade aldehydes, proteins, peptides or amino acids in water to form polyphenol solutions, food-grade aldehyde solutions and protein, peptide or amino acid solutions respectively, then mixing the three solutions and magnetically stirring them at room temperature and in the dark to obtain a mixed system;

[0036] (2) centrifuging the mixed system of step (1), washing the precipitate obtained by centrifugation, and then freeze-drying the precipitate to obtain polyphenol composite nanoparticles;

[0037] (3) dissolving the fat-soluble active substance in vegetable oil to obtain an oil phase;

[0038] (4) dissolving the polyphenol composite nanoparticles obtained in step (2) in water to obtain a nanoparticle suspension, and mixing it with the oil phase obtained in step (3), and dispersing it at high speed to obtain a high internal phase Pickering emulsion loaded with fat-soluble active substances.

[0039] In one embodiment, the fat-soluble active substance in step (3) includes one or more of astaxanthin, lutein and curcumin.

[0040] In one embodiment, the concentration of the fat-soluble active substance in the oil phase in step (3) is 1-5 mg / mL.

[0041] In one embodiment, the volume proportion of the oil phase in the high internal phase Pickering emulsion loaded with fat-soluble active substances in step (4) is 75%-80%, preferably 78%.

[0042] The fifth object of the present invention is to provide a high internal phase Pickering emulsion loaded with fat-soluble active substances prepared by the above preparation method.

[0043] In one embodiment, the loading amount of the high internal phase Pickering emulsion loaded with the fat-soluble active substance is 0.778-0.804 mg / mL; and the encapsulation efficiency is 80-85%.

[0044] The sixth object of the present invention is to provide an application of the above-mentioned high internal phase Pickering emulsion loaded with fat-soluble active substances in the field of food, medicine and health care product preparation.

[0045] Beneficial Effects

[0046] (1) The polyphenol-mediated high internal phase Pickering emulsion of the present invention is prepared under mild conditions and effectively maintains the activity of the polyphenol active substance. The preparation process is simple to operate, low in energy consumption, and green and environmentally friendly. In addition, the high internal phase Pickering emulsion has good safety, biocompatibility, semi-solid gel stability, etc.;

[0047] (2) The present invention uses a polyphenol-mediated high internal phase Pickering emulsion to load fat-soluble food functional factors, which significantly improves the poor water solubility and stability of the fat-soluble functional factors, improves their bioavailability, and has good biocompatibility. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 This is a macroscopic image of proanthocyanidin nanoparticles without adding vanillin to the mixed system in Comparative Example 1 of the present invention;

[0049] Figure 2 This is a SEM scanning electron microscope image of proanthocyanidin nanoparticles in the mixed system in Example 1 of the present invention;

[0050] Figure 3 This is a macroscopic inverted image of a high internal phase Pickering emulsion with different concentrations of proanthocyanidin nanoparticles stabilizing 75% oil phase in Example 2 of the present invention;

[0051] Figure 4 It is a macroscopic inverted image of high internal phase Pickering emulsions with different oil phase fractions mediated by 4.5% proanthocyanidin nanoparticles in Example 3 of the present invention;

[0052] Figure 5 This is a SEM scanning electron micrograph of a high internal phase Pickering emulsion with different oil phase fractions mediated by 4.5% proanthocyanidin nanoparticles in Example 3 of the present invention;

[0053] Figure 6 This is a data diagram of the loading rate and encapsulation rate of astaxanthin in a high internal phase Pickering emulsion mediated by proanthocyanidin nanoparticles in Example 4 of the present invention;

[0054] Figure 7 The storage effect and astaxanthin retention rate of the high internal phase Pickering emulsion loaded with astaxanthin prepared in Example 4 of the present invention are broken line data graphs; (A) storage effect macrograph, (B) retention rate broken line data graph;

[0055] Figure 8 This is a digestion microscopic image of astaxanthin loaded in a high internal phase Pickering emulsion with 78% oil phase prepared in Example 4 of the present invention. DETAILED DESCRIPTION

[0056] The preferred embodiments of the present invention are described in detail below. The protection scope of the present invention is not limited to the specific conditions and details in the following embodiments.

[0057] Example 1

[0058] A method for preparing a polyphenol-mediated high internal phase Pickering emulsion, the method comprising the following steps:

[0059] (1) Dissolving proanthocyanidin powder, vanillin, and arginine in deionized water respectively to completely dissolve them, then mixing the three solutions and magnetically stirring them for 24 hours at room temperature and in the dark to obtain a mixed system; wherein the concentration of the proanthocyanidin solution after mixing is 2.5 mg / mL, the concentration of the vanillin solution is 5 mg / mL, and the concentration of the arginine solution is 2.5 mg / mL; the mixed system solution is imaged by SEM scanning electron microscopy, and the results are shown in FIG. Figure 1 ;

[0060] (2) centrifuging the mixed system of step (1), washing the precipitate obtained by centrifugation, and then freeze-drying the precipitate for 24 hours using a freeze dryer to obtain proanthocyanidin nanoparticle solids; wherein the centrifugal force is 10000 rpm, the centrifugation is performed twice, each time for 5 minutes; the washing condition is washing 3 times with deionized water; the freeze-drying condition is a vacuum pressure of 15 Pa and a temperature of -50°C;

[0061] (3) adding deionized water to the proanthocyanidin nanoparticle solid obtained in step (2) to obtain a proanthocyanidin nanoparticle suspension, and mixing it with soybean oil, dispersing the sample at 10000 rpm for 2 min under a high-speed disperser to obtain a high internal phase Pickering emulsion; wherein the concentration of the proanthocyanidin nanoparticle suspension is 4.5% (w / v, mg / ml), and the soybean oil accounts for 75% of the oil phase volume.

[0062] Example 2

[0063] The only difference from Example 1 is that the concentration of the proanthocyanidin nanoparticle suspension in step (3) is adjusted to 0.5% (w / v, mg / ml), 1.0% (w / v, mg / ml), 2.0% (w / v, mg / ml), 3.0% (w / v, mg / ml), and 4.0% (w / v, mg / ml), respectively, to obtain a proanthocyanidin-mediated high internal phase Pickering emulsion.

[0064] Example 3

[0065] The only difference from Example 1 is that the oil phase volume proportion of soybean oil in step (3) is adjusted to 76%, 77%, 78%, 79% and 80%, respectively, to prepare high internal phase Pickering emulsions with different oil phase volume proportions mediated by proanthocyanidins.

[0066] Comparative Example 1

[0067] The only difference from Example 1 is that vanillin is omitted in step (1), and the other parameters and conditions are the same as those in Example 1. Figure 1 ;

[0068] Depend on Figure 1The results show that the high internal phase Pickering emulsion stabilized by proanthocyanidin nanoparticles without vanillin has poor stability and oil-water separation. It is obvious that the nanoparticles without vanillin are not sufficient to stabilize the high internal phase Pickering emulsion.

[0069] Results Analysis

[0070] 1. The mixed system prepared in Example 1 was imaged by SEM scanning electron microscope ( Figure 2 ). The SEM scanning electron microscope image results show that the proanthocyanidin nanoparticles are tightly combined and present a spherical shape, and the surface is mostly smooth, indicating that the proanthocyanidin nanoparticles were successfully prepared.

[0071] 2. Macroscopic analysis of the high internal phase Pickering emulsion stabilized by proanthocyanidin nanoparticles of different concentrations before and after inversion. The results are shown in Figure 3 ;

[0072] It can be seen from the results in the figure that the concentration of proanthocyanidin nanoparticles determines the semi-solid gel stability of the high internal phase Pickering emulsion. Obviously, too low a concentration of nanoparticles is not enough to stabilize the high internal phase Pickering emulsion. When the concentration of proanthocyanidin nanoparticles is 4-4.5% (w / v, mg / ml), it can maintain good stability; especially when the concentration of proanthocyanidin nanoparticles is 4.5% (w / v, mg / ml), the stability is better.

[0073] 3. The macroscopic analysis and SEM scanning electron microscope images of the high internal phase Pickering emulsions mediated by proanthocyanidins with different oil phase volume ratios before and after inversion are as follows: Figure 4 and Figure 5 ;

[0074] Depend on Figure 4 The results show that the volume ratio of the oil phase affects the semisolid gel stability of the high internal phase Pickering emulsion. Figure 4 It can be seen that when the volume proportion of the oil phase is equal to 80%, the emulsion system undergoes obvious macroscopic phase separation and cannot successfully construct a stable high internal phase Pickering emulsion. Figure 5 It can be seen that in the 75%-77% oil phase volume ratio, the solid particles do not completely wrap the emulsion droplets, and a small part of the particles are free in the dispersed phase; in the 78%-80% oil phase volume ratio, the emulsion droplets are rough in appearance, indicating that they are completely wrapped by solid particles, but the particle size of the 79% and 80% emulsion droplets is larger than that of 78%.

[0075] Example 4

[0076] A method for preparing a high internal phase Pickering emulsion loaded with food functional factors, the method comprising the following steps:

[0077] (1) dissolving proanthocyanidin powder, vanillin, and arginine in deionized water respectively to completely dissolve them, then mixing the three solutions and magnetically stirring them for 24 hours at room temperature and in the dark to obtain a mixed system; wherein the concentration of the proanthocyanidin solution after mixing is 2.5 mg / mL, the concentration of the vanillin solution is 5 mg / mL, and the concentration of the arginine solution is 2.5 mg / mL;

[0078] (2) centrifuging the mixed system of step (1), washing the precipitate obtained by centrifugation, and then freeze-drying the precipitate for 24 hours using a freeze dryer to obtain proanthocyanidin nanoparticle solids; wherein the centrifugal force is 10000 rpm, the centrifugation is performed twice, each time for 5 minutes; the washing condition is washing 3 times with deionized water; the freeze-drying condition is a vacuum pressure of 15 Pa and a temperature of -50°C;

[0079] (3) dissolving astaxanthin in soybean oil, treating it with magnetic stirring overnight at room temperature and in the dark to obtain soybean oil containing astaxanthin, and centrifuging to remove insoluble impurities; wherein the astaxanthin concentration is 1 mg / mL; the magnetic stirring speed is 500 rpm, the centrifugal condition is a centrifugal force of 10000 rpm, and the centrifugation condition is 10 min;

[0080] (4) adding deionized water to the proanthocyanidin nanoparticle solid obtained in step (2) to obtain a proanthocyanidin nanoparticle suspension, and mixing it with the soybean oil containing astaxanthin obtained in step (3), dispersing the sample at 10000 rpm for 2 min under a high-speed disperser to obtain a high internal phase Pickering emulsion loaded with astaxanthin; wherein the concentration of the proanthocyanidin nanoparticle suspension is 45 mg / mL, and the volume proportion of the soybean oil containing astaxanthin is 75% to 80%.

[0081] Results Analysis

[0082] 1. The performance of the high internal phase Pickering emulsion loaded with astaxanthin prepared in Example 4 was analyzed. The results are as follows: Figure 6 As shown. Figure 6 The results show that the loading rate and encapsulation rate of astaxanthin are best when the soybean oil phase accounts for 78% by volume, the loading amount is 0.804 mg / mL, and the encapsulation rate is 83.33%, while when it exceeds 78%, a downward trend appears. The high loading rate and encapsulation rate of the high internal phase Pickering emulsion are attributed to the high viscosity of the high internal phase Pickering emulsion with a volume ratio of 78% oil phase, and a small amount of emulsion can load and encapsulate a large amount of astaxanthin.

[0083] 2. The stability of the astaxanthin-loaded high internal phase Pickering emulsion prepared in Example 4 was analyzed. The results are as follows: Figure 7 As shown;

[0084] Astaxanthin loaded in soybean oil was used as the blank control group. Figure 7 A. The color of soybean oil in which astaxanthin is dissolved is reddish brown, while the color of the high internal phase Pickering emulsion mediated by proanthocyanidin nanoparticles loaded with astaxanthin is light brown. This is because the water phase in the emulsion dilutes the color of astaxanthin, causing its color to become lighter. After storage for 28 days, the color of astaxanthin in pure soybean oil becomes lighter, indicating that astaxanthin has been degraded, while the color of astaxanthin loaded in the high internal phase Pickering emulsion remains basically unchanged in the oil phase of 75%-78%. After exceeding 78% of the oil phase, the color of the emulsion changes significantly, indicating that the high internal phase Pickering emulsion mediated by proanthocyanidins with an oil phase below 78% (including 78%) has a good protective effect on astaxanthin. Figure 7 B and Table 1 show that the retention rate of the 78% high internal phase Pickering emulsion loaded with astaxanthin is about 1.5 times that of the blank control group after storage for 28 days. These results indicate that the 78% high internal phase Pickering emulsion mediated by proanthocyanidins can significantly enhance the photostability and storage stability of astaxanthin.

[0085] Table 1. Astaxanthin retention rate

[0086]

[0087] 3. In vitro digestion analysis of the high internal phase Pickering emulsion loaded with astaxanthin with an oil phase volume ratio of 78% prepared in Example 4

[0088] The emulsion, artificial saliva, simulated gastric fluid, and simulated intestinal fluid were preheated to 37°C before the experiment, then exposed to artificial saliva for 10 min, added with simulated gastric fluid, adjusted to pH 2.0 with HCl, and incubated for 2 h. After 2 h, the pH was quickly adjusted to 7.0 with NaOH and placed in simulated intestinal fluid for 2 h. After simulating the in vitro digestion stage, optical microscopy images were obtained. Figure 8 It was found that the emulsion droplets with an oil phase fraction of 78% underwent significant changes. In simulated saliva and gastric fluid, the emulsion droplets maintained a high stability despite a slight increase in size. The digestion and absorption of the emulsion mainly occurred in simulated intestinal fluid. Under simulated intestinal fluid conditions, a large number of emulsion droplets aggregated after 1 hour, and the number of emulsion droplets decreased significantly after 2 hours, but did not completely disappear at the end of digestion. The above results indicate that the high internal phase Pickering emulsion mediated by proanthocyanidin nanoparticles is a good carrier for better delivery of astaxanthin.

[0089] The embodiments provided above are not intended to limit the scope of the present invention, and the steps described are not intended to limit the execution order thereof. Those skilled in the art may make obvious improvements to the present invention in combination with existing common knowledge, which also fall within the scope of protection defined by the claims of the present invention.

Claims

1. A method for preparing a polyphenol-mediated high internal phase Pickering emulsion, characterized in that: The method comprises the following steps: (1) dissolving polyphenols, food-grade aldehydes, proteins, peptides or amino acids in water to form a polyphenol solution, a food-grade aldehyde solution and a protein, peptide or amino acid solution respectively, then mixing the three solutions and magnetically stirring them at room temperature and in the dark to obtain a mixed system; (2) centrifuging the mixed system of step (1), washing the precipitate obtained by centrifugation, and then freeze-drying the precipitate to obtain polyphenol composite nanoparticles; (3) dissolving the polyphenol composite nanoparticles obtained in step (2) in water to obtain a nanoparticle suspension, and mixing it with vegetable oil and dispersing it at high speed to obtain a polyphenol-mediated high internal phase Pickering emulsion.

2. The preparation method according to claim 1, characterized in that: The polyphenols in step (1) include one or more of tea polyphenols, catechins, brown algae polyphenols, and proanthocyanidins.

3. The preparation method according to claim 1, characterized in that: The food-grade aldehydes in step (1) include one or more of vanillin, cinnamaldehyde, and benzaldehyde.

4. The preparation method according to claim 1, characterized in that: The concentration of the nanoparticle suspension in step (3) is 0.5% to 4.5%, w / v, mg / ml.

5. The preparation method according to claim 1, characterized in that: In step (3), the volume proportion of the vegetable oil phase is 75-80%.

6. A polyphenol-mediated high internal phase Pickering emulsion prepared by the preparation method according to any one of claims 1 to 5.

7. Use of the polyphenol-mediated high internal phase Pickering emulsion according to claim 6 in the preparation of foods, medicines and health products.

8. A method for preparing a high internal phase Pickering emulsion loaded with a fat-soluble active substance, characterized in that: The method comprises the following steps: (1) dissolving polyphenol powder, food-grade aldehydes, proteins, peptides or amino acids in water to form polyphenol solutions, food-grade aldehyde solutions and protein, peptide or amino acid solutions respectively, then mixing the three solutions and magnetically stirring them at room temperature and in the dark to obtain a mixed system; (2) centrifuging the mixed system of step (1), washing the precipitate obtained by centrifugation, and then freeze-drying the precipitate to obtain polyphenol composite nanoparticles; (3) dissolving the fat-soluble active substance in vegetable oil to obtain an oil phase; (4) dissolving the polyphenol composite nanoparticles obtained in step (2) in water to obtain a nanoparticle suspension, and mixing it with the oil phase obtained in step (3), and dispersing it at high speed to obtain a high internal phase Pickering emulsion loaded with fat-soluble active substances.

9. A high internal phase Pickering emulsion loaded with fat-soluble active substances prepared by the method according to any one of claims 8.

10. Use of the high internal phase Pickering emulsion loaded with fat-soluble active substances according to claim 9 in the preparation of foods, medicines and health products.

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