CN113693225B - Ovalbumin-ferulic acid-polysaccharide compound emulsion and preparation method and application thereof - Google Patents

Ovalbumin-ferulic acid-polysaccharide compound emulsion and preparation method and application thereof Download PDF

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CN113693225B
CN113693225B CN202110899371.8A CN202110899371A CN113693225B CN 113693225 B CN113693225 B CN 113693225B CN 202110899371 A CN202110899371 A CN 202110899371A CN 113693225 B CN113693225 B CN 113693225B
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ovalbumin
ferulic acid
polysaccharide
solution
stirring
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CN113693225A (en
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苏东晓
黄贞贞
杨欣禧
梁思月
陈乐祺
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Guangzhou University
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Abstract

The invention discloses an ovalbumin-ferulic acid-polysaccharide compound emulsion and a preparation method and application thereof. The method comprises the following steps: (1) Adding egg albumin into water to fully hydrate the protein to obtain egg albumin dispersion liquid; (2) Adding ferulic acid into water to obtain ferulic acid solution; (3) Adding ferulic acid solution into ovalbumin dispersion to obtain ovalbumin-ferulic acid solution; (4) adding polysaccharide into water to obtain a polysaccharide solution; (5) Adding the ovalbumin-ferulic acid solution into the polysaccharide solution to obtain an ovalbumin-ferulic acid-polysaccharide complex solution; (6) Medium chain triglyceride is added into the ovalbumin-ferulic acid-polysaccharide compound solution drop by drop to obtain the ovalbumin-ferulic acid-polysaccharide compound emulsion. The composite emulsion has good viscoelasticity and storage stability, has the characteristic of high embedding rate, and can be used as a carrier for conveying fat-soluble substances.

Description

Ovalbumin-ferulic acid-polysaccharide compound emulsion and preparation method and application thereof
Technical Field
The invention relates to the technical field of food processing, in particular to an ovalbumin-ferulic acid-polysaccharide compound emulsion and a preparation method and application thereof.
Background
Ovalbumin (OVA) is the most abundant protein in egg white, has excellent emulsifying and gelling properties, and is widely used for preparing nano particles or nano emulsion. Some researches show that Ferulic Acid (FA) is a cross-linking agent of ovalbumin and the production of an ovalbumin coupled ferulic acid reagent is available, but the reagent is formed by using chemical cross-linking of the ovalbumin and the ferulic acid reagent, and the process is complex. In addition, it has been reported that the stability of the emulsion is improved by using a polyphenol-protein interface layer formed by competitive adsorption of natural polyphenol compounds. However, complexing polyphenols with proteins may affect the availability of certain amino acids and also alter the structure of the protein, thus affecting the functionality of the protein, and complex stabilized emulsions have a significant impact on environmental changes. Therefore, there is a need to develop an emulsion which is natural and healthy and has good stability, and an effective formulation which can be used for health products, cosmetics or functional foods.
Proteins and Polysaccharides (PS) form polymers through electrostatic, hydrophobic and hydrogen bonding interactions, whereas aggregated polysaccharides can stabilize emulsions by attaching to emulsion droplets, thereby controlling colloidal interactions, increasing viscosity to reduce droplet aggregation, or creating yield stress to the movement of stationary particles. Researchers find that an antibacterial film (CN 103554532) with good barrier property and higher mechanical strength can be prepared by using collagen, ferulic acid and sodium alginate, and researches find that the addition of polysaccharide can obviously change the rheological behavior of emulsion and plays a key role in the adsorption of protein from a bulk phase to an interface. However, under high shear conditions, the emulsion structure prepared by the protein-polysaccharide complex is broken, resulting in serious instability of the emulsion.
Currently, studies have shown that hydrogen bonds are formed between the hydroxyl groups of polyphenols and the oxygen atoms of the polysaccharide glycosidic linkages, non-covalent bonds can be formed between phenolic acids and polysaccharides, and different polysaccharides can influence these interactions by protecting or destroying the charge, and can even be used as additives to stabilize colloidal solutions. However, no report has been published on the fortification of ovalbumin-ferulic acid (OVA-FA) emulsion with polysaccharide. The invention expands the application of eggs to a certain extent, expands the cognition of people on the efficacy of the ovalbumin-ferulic acid nano particles, solves the problem of poor emulsion storage stability, and increases social and economic values.
Disclosure of Invention
The primary aim of the invention is to overcome the defects and shortcomings of the prior art and provide a preparation method of an ovalbumin-ferulic acid-polysaccharide compound emulsion.
Another object of the present invention is to provide an ovalbumin-ferulic acid-polysaccharide complex emulsion prepared by the method.
It is a further object of the present invention to provide the use of said ovalbumin-ferulic acid-polysaccharide complex emulsion.
The aim of the invention is achieved by the following technical scheme:
a preparation method of an ovalbumin-ferulic acid-polysaccharide complex emulsion comprises the following steps:
(1) Adding egg albumin into water, stirring uniformly, regulating the pH value to 7.0+/-0.1, and standing to fully hydrate the protein to obtain an egg albumin dispersion liquid;
(2) Adding ferulic acid into water, stirring uniformly, and regulating pH value to 7.0+ -0.1 to obtain ferulic acid solution;
(3) Adding the ferulic acid solution obtained in the step (2) into the ovalbumin dispersion liquid obtained in the step (1), stirring and mixing uniformly, and regulating the pH value to 6.0+/-0.1 to obtain ovalbumin-ferulic acid solution;
(4) Adding Polysaccharide (PS) into water, stirring uniformly, and regulating the pH value to 6.0+/-0.1 to obtain a polysaccharide solution; wherein the polysaccharide is at least one of Sodium Alginate (SA), carrageenan (KC), hyaluronic Acid (HA) and Agar (Agar);
(5) Adding the ovalbumin-ferulic acid solution obtained in the step (3) into the polysaccharide solution obtained in the step (4), and uniformly stirring and mixing to obtain an ovalbumin-ferulic acid-polysaccharide compound solution;
(6) And (3) dropwise adding Medium Chain Triglyceride (MCT) into the ovalbumin-ferulic acid-polysaccharide compound solution obtained in the step (5), stirring and uniformly mixing, and homogenizing to obtain the ovalbumin-ferulic acid-polysaccharide compound emulsion.
The conditions for sufficient hydration of the protein by standing as described in step (1) are: standing for 12-24 h at a low temperature of 4 ℃; preferably, it is: standing at a low temperature of 4 ℃ for 24 hours.
The water described in steps (1), (2) and (4) is preferably deionized water.
The stirring conditions in the steps (1), (2) and (4) are as follows: stirring is continuously carried out for more than 2 hours at room temperature.
The adjusting agent for adjusting the pH value described in steps (1), (2), (3) and (4) is preferably a solution of 0.1 to 1mol/L of HCl and 0.1 to 1mol/L of NaOH.
The mass concentration of the ovalbumin dispersion liquid in the step (1) is 0.1% -20%; preferably 1%.
The mass concentration of the ferulic acid solution in the step (2) is 0.01-10%; preferably 0.5%.
The volume ratio of the ovalbumin dispersion liquid to the ferulic acid solution in the step (3) is 4:1.
The stirring conditions in the step (3) are as follows: stirring at 500-1400 rpm for 30-120 min; preferably, it is: stirring at 800-1400 rpm for 60-120 min.
The polysaccharide in the step (4) is preferably at least one of Sodium Alginate (SA) and carrageenan (KC); more preferably carrageenan (KC).
The mass concentration of the polysaccharide solution in the step (4) is 0.01% -10%; further preferably 0.2% -10%; still more preferably 0.5% to 10%; still more preferably 0.5%.
The volume ratio of the ovalbumin-ferulic acid solution to the polysaccharide solution in the step (5) is 1-12:1-8; preferably 1-12:1; more preferably 1:1.
The stirring conditions in the step (5) are as follows: stirring at 400-1200 rpm for 10-120 min; preferably, it is: stirring at 500-1200 rpm for 30-120 min.
The volume ratio of the ovalbumin-ferulic acid-polysaccharide complex solution and Medium Chain Triglyceride (MCT) in the step (6) is 5:1 to 15; preferably 5:5 to 9; more preferably 5:7 to 9.
The stirring conditions in the step (6) are as follows: stirring at 200-1500 rpm for 10-100 min; preferably 600rpm to 1200rpm, for 30min to 60min.
The homogenization conditions described in step (6) are: homogenizing at 2000-20000 rpm for 1-20 min; preferably, it is: homogenizing at 6000-16000 rpm for 1-10 min; more preferably: homogenizing at 12000rpm for 2min.
An ovalbumin-ferulic acid-polysaccharide complex emulsion prepared by the method of any one of the above.
The particle size of the ovalbumin-ferulic acid-polysaccharide composite emulsion is 1-10 um (measured by Dynamic Light Scattering (DLS)).
The ovalbumin-ferulic acid-polysaccharide complex emulsion is applied to the fields of foods, medicines, health care products or cosmetics.
The application of the ovalbumin-ferulic acid-polysaccharide complex emulsion in preparing a drug delivery system.
The ovalbumin-ferulic acid-polysaccharide complex emulsion can be used for conveying fat-soluble polyphenol compounds, fat-soluble vitamin substances, fat-soluble medicaments and medicaments interacted with proteins and/or polyphenols.
A preparation method of an ovalbumin-ferulic acid-polysaccharide complex emulsion loaded with a drug, which comprises the steps (1) to (5) in the preparation method of the ovalbumin-ferulic acid-polysaccharide complex emulsion, and the following steps:
(6) Adding the medicine into Medium Chain Triglyceride (MCT), and stirring and mixing uniformly to obtain medium chain triglyceride loaded with the medicine;
(7) Dropwise adding the medium chain triglyceride loaded with the drug obtained in the step (6) into the ovalbumin-ferulic acid-polysaccharide compound solution obtained in the step (5), uniformly stirring and mixing, and homogenizing to obtain the ovalbumin-ferulic acid-polysaccharide compound emulsion loaded with the drug.
The medicine in the step (6) is at least one of fat-soluble polyphenol compounds, fat-soluble vitamin substances and fat-soluble medicines; rutin is preferred.
The dosage of the medicine in the step (6) is calculated according to the proportion of 85-90 mL medium chain triglyceride in each gram of medicine; preferably 87.5mL medium chain triglycerides per gram of drug.
The volume ratio of the ovalbumin-ferulic acid-polysaccharide complex solution and the drug-loaded Medium Chain Triglyceride (MCT) in the step (7) is 5:5 to 9; further preferably 5: 7-9; still more preferably 5:9.
compared with the prior art, the invention has the following advantages and effects:
(1) The ovalbumin-ferulic acid is prepared by forming stable nano particles through hydrophobic bonds and hydrogen bonds between the ovalbumin-ferulic acid and the polysaccharide, strengthening the structure of the ovalbumin-ferulic acid emulsion through polysaccharide, regulating and controlling the reaction between an ovalbumin-ferulic acid complex and the polysaccharide through changing the mixing proportion, and obtaining the ovalbumin-ferulic acid-polysaccharide emulsion with good viscoelasticity and stability through regulating the mixing proportion of the ovalbumin-ferulic acid-polysaccharide complex and medium chain triglyceride; in addition, when the OVA-FA-polysaccharide and the triglyceride are mixed in a volume ratio of 5:9 to prepare the emulsion, the OVA-FA-polysaccharide can embed 9mL (namely 9 g) of the triglyceride only by 5mL (with a mass of 0.035 g), namely the ovalbumin-ferulic acid-polysaccharide emulsion has the characteristic of high embedding rate.
(2) The raw materials adopted in the process of the invention are natural biological macromolecules, can be degraded, are cheap and easy to obtain, have mild preparation process, do not use toxic and harmful reagents, have easy control of reaction process, short production period and low equipment investment and production cost, and can be used in various fields of foods, health care products, cosmetics and the like.
(3) The ovalbumin-ferulic acid-polysaccharide compound stable emulsion prepared by the invention has good viscoelasticity and storage stability, and compared with the ovalbumin-ferulic acid nanoparticle stable emulsion, the addition of polysaccharide obviously improves the problem of poor effect of the ovalbumin-ferulic acid nanoparticle stable emulsion, and avoids layering phenomenon.
(4) The ovalbumin-ferulic acid-polysaccharide emulsion prepared by the invention not only can transmit fat-soluble polyphenol compounds, fat-soluble vitamin substances and the like, but also can transmit fat-soluble or medicines interacted with proteins and polyphenols, thereby realizing the common transmission of various medicines.
Drawings
FIG. 1 is a graph showing the transmittance of an ovalbumin-ferulic acid-polysaccharide composite solution (in the graph, OVA: ovalbumin, FA: ferulic acid, PS: polysaccharide, SA: sodium alginate, KC: carrageenan, HA: hyaluronic acid, agar: agar) obtained when the mass concentration of ovalbumin-ferulic acid is 1% and the mass concentration of polysaccharide is 0.5% and the mixing ratio is 12:1-1:8, respectively, at pH 6.0.+ -. 0.1.
FIG. 2 is an ultraviolet spectrum of an ovalbumin-ferulic acid-polysaccharide composite solution obtained when the mass concentration of the ovalbumin-ferulic acid is 1%, the mass concentration of the polysaccharide is 0.5%, and the pH value of the ovalbumin-ferulic acid-polysaccharide composite solution is 6.0+/-0.1 and the mixing ratio is (12:1) - (1:8); wherein A is Sodium Alginate (SA); b is carrageenan (KC); c is Agar (Agar); d is Hyaluronic Acid (HA).
FIG. 3 is an infrared spectrum of an ovalbumin-ferulic acid-polysaccharide composite solution obtained when the mass concentration of ovalbumin-ferulic acid is 1%, the mass concentration of polysaccharide is 0.5%, and the pH value of the ovalbumin-ferulic acid-polysaccharide composite solution is 6.0+/-0.1 and the mixing ratio of the ovalbumin-ferulic acid-polysaccharide composite solution is (12:1) - (1:8).
FIG. 4 is an external view showing that ovalbumin-ferulic acid-polysaccharide composite solution, which is obtained by mixing ovalbumin-ferulic acid-polysaccharide composite solution with medium-chain triglycerides in a volume ratio of (5:1) to (5:15), has a mass concentration of 1% and a mass concentration of polysaccharide of 0.5%, both of which are at pH 6.0.+ -. 0.1, and a mixing ratio of 1:1.
FIG. 5 is a rheological diagram of an ovalbumin-ferulic acid-polysaccharide composite solution obtained by mixing ovalbumin-ferulic acid-polysaccharide composite solution with medium-chain triglycerides in a volume ratio of (5:5) - (5:9) at a mass concentration of 1% and a mass concentration of polysaccharide of 0.5%, both at pH 6.0.+ -. 0.1, and a mixing ratio of 1:1.
FIG. 6 is a microstructure of an ovalbumin-ferulic acid-polysaccharide composite solution obtained by mixing ovalbumin-ferulic acid-polysaccharide composite solution with medium chain triglycerides in a volume ratio of (5:7) - (5:9) at a mass concentration of 1% and a mass concentration of polysaccharide of 0.5%, both at pH 6.0.+ -. 0.1, and a mixing ratio of 1:1.
FIG. 7 is a graph showing changes in the stable emulsions of OVA-FA-SA and OVA-FA-KC at volume ratios of 5:7 and 5:9 during 14 days of storage at 37 ℃; wherein A is an emulsion appearance diagram; b is a particle size diagram of the emulsion; c is the absolute value diagram of the emulsion potential.
FIG. 8 is a rheogram showing that ovalbumin-ferulic acid mass concentration is 1%, polysaccharide mass concentration is 0.5%, both of which are at pH 6.0.+ -. 0.1, and when the mixing ratio is 1:1, the obtained ovalbumin-ferulic acid-polysaccharide composite solution is mixed with rutin-loaded medium chain triglyceride in a volume ratio of 5:9; wherein A is storage modulus (also called elastic modulus; G'); b is the loss modulus (also known as the viscous modulus; G ").
Detailed Description
The present invention will be described in further detail with reference to examples, but embodiments of the present invention are not limited thereto. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art. The test methods for specific experimental conditions are not noted in the examples below, and are generally performed under conventional experimental conditions or under experimental conditions recommended by the manufacturer. The reagents and starting materials used in the present invention are commercially available unless otherwise specified.
The acid-alkali solution used for adjusting the pH value in the invention is a pH value adjusting agent which is conventional in the art, such as 0.1 mol/L-1 mol/L hydrochloric acid, sodium hydroxide and the like.
The examples of the present invention relate to ovalbumin (biotechnological grade) and ferulic acid purchased from Shanghai microphone Biochemical technology Co.Ltd (China, shanghai); sodium alginate, carrageenan, hyaluronic acid, agar and medium chain triglycerides were purchased from Shanghai Yuan leaf Biotechnology Co.Ltd (China, shanghai).
Example 1
(1) Accurately weighing 2g of Ovalbumin (OVA) powder, dispersing the powder in 200g of deionized water, continuously stirring at room temperature for 2 hours, adjusting the pH to 7.0+/-0.1 by using acid-base solution, and then placing the powder at a low temperature of 4 ℃ for 24 hours to fully hydrate the protein so as to obtain the ovalbumin storage solution with the mass concentration of 1%.
(2) 1g of Ferulic Acid (FA) powder is weighed and added into 200g of deionized water, and is continuously stirred for 2 hours at room temperature, the pH is adjusted to 7.0+/-0.1 by alkali liquor, and the ferulic acid is fully and uniformly mixed to obtain the ferulic acid solution with the mass concentration of 0.5 percent.
(3) Mixing the ovalbumin solution in the step (1) and the ferulic acid solution in the step (2) according to the volume ratio of 4:1, namely dropwise adding the ferulic acid solution into the ovalbumin solution under magnetic stirring, wherein the magnetic stirring speed is 800-1400 rpm, the stirring time is 60-120 min, and the pH value of the ovalbumin-ferulic acid mixed solution is regulated to 6.0+/-0.1 to obtain the ovalbumin-ferulic acid nanoparticle solution.
(4) 1g of Sodium Alginate (SA), carrageenan (KC), hyaluronic Acid (HA) and Agar (Agar) powder are respectively weighed, then are respectively added into 200g of deionized water, are continuously stirred for 2 hours at room temperature, and are adjusted to have pH value of 6.0+/-0.1 by alkali liquor, so that polysaccharide is fully and uniformly mixed, and a polysaccharide solution with mass concentration of 0.5% is obtained.
(5) Mixing the ovalbumin-ferulic acid nanoparticle solution (OVA-FA) obtained in the step (3) with the Polysaccharide Solution (PS) obtained in the step (4) according to the volume ratio of 12:1, 8:1, 4:1, 1:4 and 1:8 respectively, namely dropwise adding the polysaccharide solution into the ovalbumin-ferulic acid nanoparticle solution under magnetic stirring, wherein the magnetic stirring speed is 400-1200 rpm, and the stirring time is 30-120 min, so as to obtain an ovalbumin-ferulic acid-polysaccharide compound solution.
The present example examined the light transmittance change of ovalbumin-ferulic acid nanoparticle solution and polysaccharide solution mixed according to different volume ratios. The light transmittance and ultraviolet spectra of the ovalbumin-ferulic acid-polysaccharide composite solutions with different volume ratios are shown in fig. 1 and 2 respectively:
when the mixing ratio of sodium alginate, carrageenan and hyaluronic acid to the ovalbumin-polyphenol (ferulic acid) composite liquid is larger than 1:1, the light transmittance of the formed solution is reduced along with the increase of the proportion of the ovalbumin-polyphenol composite liquid, and the formed solution has similar trend to that of the ovalbumin-polyphenol composite liquid. However, the transmittance of the complex solutions of OVA-FA-polysaccharide (OVA-FA-SA, OVA-FA-KC, OVA-FA-HA, OVA-FA-Agar) was always lower than that of the ovalbumin-polyphenol complex solution, indicating that the polysaccharide interacted with the ovalbumin-polyphenol complex solution to form a complex. The low light transmittance is due to the formation of aggregates between protein and polysaccharide, while when light transmittance is minimized, it is indicated that the OVA-FA complex with polysaccharide is saturated. Thus, the optimal mixing ratio of polysaccharide to OVA-FA complex was 1:1 (FIG. 1).
Different mixing volume ratios of polysaccharide to OVA-FA have different effects on the uv spectrum of OVA. As the proportion of polysaccharide added increases, the absorption peak of OVA-FA decreases in turn. This is probably due to structural changes caused by interactions of proteins with polysaccharides, which change the conformation of protein molecules by exposing aromatic heterocyclic hydrophobic groups in tryptophan and tyrosine residues, resulting in reduced protein absorption peaks (fig. 2).
Example 2
(1) Accurately weighing 2g of Ovalbumin (OVA) powder, dispersing the powder in 200g of deionized water, continuously stirring at room temperature for 2 hours, adjusting the pH to 7.0+/-0.1 by using acid-base solution, and then placing the powder at a low temperature of 4 ℃ for 24 hours to fully hydrate the protein so as to obtain the ovalbumin storage solution with the mass concentration of 1%.
(2) 1g of Ferulic Acid (FA) powder is weighed and added into 200g of deionized water, and is continuously stirred for 2 hours at room temperature, the pH is adjusted to 7.0+/-0.1 by alkali liquor, and the ferulic acid is fully and uniformly mixed to obtain the ferulic acid solution with the mass concentration of 0.5 percent.
(3) Mixing the ovalbumin solution in the step (1) and the ferulic acid solution in the step (2) according to the volume ratio of 4:1, namely dropwise adding the ferulic acid solution into the ovalbumin solution under magnetic stirring, wherein the magnetic stirring speed is 800-1400 rpm, the stirring time is 60-120 min, and the pH value of the ovalbumin-ferulic acid mixed solution is regulated to 6.0+/-0.1 to obtain the ovalbumin-ferulic acid nanoparticle solution.
(4) 1g of Sodium Alginate (SA), carrageenan (KC), hyaluronic Acid (HA) and Agar (Agar) powder are respectively weighed and added into 200g of deionized water, and the mixture is continuously stirred for 2 hours at room temperature, and the pH is adjusted to 6.0+/-0.1 by alkali liquor, so that polysaccharide is fully and uniformly mixed, and a polysaccharide solution with the mass concentration of 0.5% is obtained.
(5) Mixing the ovalbumin-ferulic acid nanoparticle solution obtained in the step (3) with the polysaccharide solution obtained in the step (4) according to the volume ratio of 1:1, namely, dropwise adding the polysaccharide solution into the ovalbumin-ferulic acid nanoparticle solution under magnetic stirring, wherein the magnetic stirring speed is 400-1200 rpm, and the stirring time is 30-120 min, so as to obtain an ovalbumin-ferulic acid-polysaccharide compound solution.
The present example examined the change in infrared spectrum of an ovalbumin-ferulic acid-polysaccharide complex solution. The infrared spectrum of the ovalbumin-ferulic acid-polysaccharide composite solution is shown in figure 3. FIG. 3 shows respective infrared amide I band Gaussian fits for OVA, OVA-FA-SA, OVA-FA-KC, OVA-FA-Agar and OVA-FA-HA complexes. The different regions of the amide I band were used to calculate the secondary structure of the protein. The addition of ferulic acid resulted in an increase in the alpha-helix (alpha-helix) and beta-sheet (beta-sheet) content of the OVA and a decrease in the beta-turn (beta-turn) and random coil (random coil) content. This is probably mainly due to the formation of ordered hydrogen bonds by the interaction of ferulic acid and OVA. However, the addition of polysaccharide to the OVA-FA complex resulted in a significant reduction in the alpha-helix content, with the exception of agar, which resulted in a significant increase in the beta-sheet content. On the one hand, studies have shown that an increase in β -sheet content results in a decrease in interfacial tension and an increase in emulsion stability, mainly due to the disordered structure leading to a rapid conformational change of the protein at the oil-water interface, exposing hydrophobic amino acids in the internal structure, thus lowering the interfacial tension and producing a lower equilibrium interfacial tension over time. On the other hand, β -sheet favors the formation of hydrogen bonds, while hydrogen bond micelles favors the formation of oil-water interface structures, confirming our findings on the ability of OVA-FA-polysaccharide to stabilize emulsions.
Example 3
(1) Accurately weighing 2g of Ovalbumin (OVA) powder, dispersing the powder in 200g of deionized water, continuously stirring at room temperature for 2 hours, adjusting the pH to 7.0+/-0.1 by using acid-base solution, and then placing the powder at a low temperature of 4 ℃ for 24 hours to fully hydrate the protein so as to obtain the ovalbumin storage solution with the mass concentration of 1%.
(2) 1g of Ferulic Acid (FA) powder is weighed and added into 200g of deionized water, and is continuously stirred for 2 hours at room temperature, the pH is adjusted to 7.0+/-0.1 by alkali liquor, and the ferulic acid is fully and uniformly mixed to obtain the ferulic acid solution with the mass concentration of 0.5 percent.
(3) Mixing the ovalbumin solution in the step (1) and the ferulic acid solution in the step (2) according to the volume ratio of 4:1, namely dropwise adding the ferulic acid solution into the ovalbumin solution under magnetic stirring, wherein the magnetic stirring speed is 800-1400 rpm, the stirring time is 60-120 min, and the pH value of the ovalbumin-ferulic acid mixed solution is regulated to 6.0+/-0.1 to obtain the ovalbumin-ferulic acid nanoparticle solution.
(4) 1g of Sodium Alginate (SA), carrageenan (KC), hyaluronic Acid (HA) and Agar (Agar) powder are respectively weighed, then are respectively added into 200g of deionized water, are continuously stirred for 2 hours at room temperature, and are adjusted to have pH value of 6.0+/-0.1 by alkali liquor, so that polysaccharide is fully and uniformly mixed, and a polysaccharide solution with mass concentration of 0.5% is obtained.
(5) Mixing the ovalbumin-ferulic acid nanoparticle solution (OVA-FA) obtained in the step (3) with the polysaccharide solution obtained in the step (4) according to the volume ratio of 1:1, namely adding the polysaccharide solution into the ovalbumin-ferulic acid nanoparticle solution dropwise under magnetic stirring, wherein the magnetic stirring speed is 400-1200 rpm, and the stirring time is 30-120 min, so as to obtain the ovalbumin-ferulic acid-polysaccharide composite solution.
(6) Mixing the ovalbumin-ferulic acid-polysaccharide composite solution obtained in the step (5) with medium chain triglyceride according to the volume ratio of 5:1, 5:3, 5:5,5:7,5:9, 5:11, 5:13 and 5:15 respectively, namely dropwise adding the Medium Chain Triglyceride (MCT) into the ovalbumin-ferulic acid-polysaccharide composite solution under magnetic stirring, wherein the magnetic stirring speed is 200-1500 rpm, and the stirring time is 30min.
(7) Homogenizing the mixed solution obtained in the step (6) for a certain time (about 2 min) under the condition of 12000rpm, thus obtaining the stable emulsion of the ovalbumin-ferulic acid-polysaccharide ternary system.
In this example, the appearance of stable emulsion of ovalbumin-ferulic acid-polysaccharide complex liquid (standing at room temperature for 120 min) was examined by the volume fraction of the oil phase (the emulsion prepared by OVA-FA was used as a control, i.e., the OVA-FA emulsion was prepared directly without adding polysaccharide solution according to the above method). The appearance of the stable emulsion of ovalbumin-ferulic acid-polysaccharide complex is shown in figure 4. The figure 4 shows that the ovalbumin-ferulic acid nanoparticle stable emulsion HAs poor effect, layering phenomenon is generated in the emulsion under all oil phase volume fractions, compared with the ovalbumin-ferulic acid nanoparticle stable emulsion, after polysaccharide is added, the emulsion can be better stabilized at a certain oil phase volume fraction, the problem of poor emulsion stabilizing effect is not improved due to the addition of agar, and obvious layering phenomenon still occurs, and the figure 4 also shows that the emulsion can be better stabilized when the mixing volume ratio of OVA-FA-SA, OVA-FA-KC and OVA-FA-HA to MCT is 5:5,5:7 and 5:9.
Example 4
(1) Accurately weighing 2g of Ovalbumin (OVA) powder, dispersing the powder in 200g of deionized water, continuously stirring at room temperature for 2 hours, adjusting the pH to 7.0+/-0.1 by using acid-base solution, and then placing the powder at a low temperature of 4 ℃ for 24 hours to fully hydrate the protein so as to obtain the ovalbumin storage solution with the mass concentration of 1%.
(2) 1g of Ferulic Acid (FA) powder is weighed and added into 200g of deionized water, and is continuously stirred for 2 hours at room temperature, the pH is adjusted to 7.0+/-0.1 by alkali liquor, and the ferulic acid is fully and uniformly mixed to obtain the ferulic acid solution with the mass concentration of 0.5 percent.
(3) Mixing the ovalbumin solution in the step (1) and the ferulic acid solution in the step (2) according to the volume ratio of 4:1, namely dropwise adding the ferulic acid solution into the ovalbumin solution under magnetic stirring, wherein the magnetic stirring speed is 800-1400 rpm, the stirring time is 60-120 min, and the pH value of the ovalbumin-ferulic acid mixed solution is regulated to 6.0+/-0.1 to obtain the ovalbumin-ferulic acid nanoparticle solution.
(4) 1g of Sodium Alginate (SA), carrageenan (KC), hyaluronic Acid (HA) and Agar (Agar) powder are respectively weighed, then are respectively added into 200g of deionized water, are continuously stirred for 2 hours at room temperature, and are adjusted to have pH value of 6.0+/-0.1 by alkali liquor, so that polysaccharide is fully and uniformly mixed, and a polysaccharide solution with mass concentration of 0.5% is obtained.
(5) Mixing the ovalbumin-ferulic acid nanoparticle solution obtained in the step (3) with the polysaccharide solution obtained in the step (4) according to the volume ratio of 1:1, namely, dropwise adding the polysaccharide solution into the ovalbumin-ferulic acid nanoparticle solution under magnetic stirring, wherein the magnetic stirring speed is 400-1200 rpm, and the stirring time is 30-120 min, so as to obtain the ovalbumin-ferulic acid-polysaccharide composite solution.
(6) Mixing the ovalbumin-ferulic acid-polysaccharide composite solution obtained in the step (5) with medium-chain triglyceride according to the volume ratio of 5:5,5:7 and 5:9 respectively, namely adding the medium-chain triglyceride into the ovalbumin-ferulic acid-polysaccharide composite solution dropwise under magnetic stirring, wherein the magnetic stirring speed is 200-1500 rpm, and the stirring time is 30min.
(7) Homogenizing the mixed solution obtained in the step (6) for a certain time (about 2 min) under the condition of 12000rpm, thus obtaining the stable emulsion of the ovalbumin-ferulic acid-polysaccharide ternary system.
The rheological property of the emulsion is researched, and the influence of different oil phase volume ratios on the mechanical property of the emulsion is discussed. After being placed at 4 ℃ for 12 hours, the mixture is measured by a dynamic shear rheometer, and the specific measuring steps are as follows: the sample was placed between two parallel plates and the gap height was set to 1mm; performing an amplitude sweep to determine a Linear Viscoelastic Region (LVR); at a constant frequency of 1hz, the strain increases logarithmically from 0.1% to 100%; the frequency sweep was from 0.1 to 100rad/s with a fixed strain of 0.5% (within LVR).
The results are shown in FIG. 5: with the increase of the frequency, the intersection point of the G 'and the G' of the OVA-FA-HA emulsion shows that the frequency exceeds a certain range, the structure of the sample changes irreversibly, and the structure of the sample is damaged to a certain extent. However, when the oil phase volume ratio of the OVA-FA-sodium alginate or the OVA-FA-carrageenan is 5:7 and 5:9, the G 'of the sample is always larger than G' (G 'is the storage modulus and also called the elastic modulus, G' is the loss modulus and also called the viscous modulus, and the viscoelasticity is the sum of G 'and G'), the main elastic behavior is shown, and the stability is good. Meanwhile, at high frequencies, the intersection point of the G 'and G' of the sample still does not appear, which indicates that the stable emulsion of the OVA-FA-SA or the OVA-FA-KC at the oil phase volume ratio of 5:7 and 5:9 does not generate structural damage under high-rate deformation.
Example 5
(1) Accurately weighing 2g of Ovalbumin (OVA) powder, dispersing the powder in 200g of deionized water, continuously stirring at room temperature for 2 hours, adjusting the pH to 7.0+/-0.1 by using acid-base solution, and then placing the powder at a low temperature of 4 ℃ for 24 hours to fully hydrate the protein so as to obtain the ovalbumin storage solution with the mass concentration of 1%.
(2) 1g of Ferulic Acid (FA) powder is weighed and added into 200g of deionized water, and is continuously stirred for 2 hours at room temperature, the pH is adjusted to 7.0+/-0.1 by alkali liquor, and the ferulic acid is fully and uniformly mixed to obtain the ferulic acid solution with the mass concentration of 0.5 percent.
(3) Mixing the ovalbumin solution in the step (1) and the ferulic acid solution in the step (2) according to the volume ratio of 4:1, namely dropwise adding the ferulic acid solution into the ovalbumin solution under magnetic stirring, wherein the magnetic stirring speed is 800-1400 rpm, the stirring time is 60-120 min, and the pH value of the ovalbumin-ferulic acid mixed solution is regulated to 6.0+/-0.1 to obtain the ovalbumin-ferulic acid nanoparticle solution.
(4) 1g of Sodium Alginate (SA), carrageenan (KC), hyaluronic Acid (HA) and Agar (Agar) powder are respectively weighed, then are respectively added into 200g of deionized water, are continuously stirred for 2 hours at room temperature, and are adjusted to have pH value of 6.0+/-0.1 by alkali liquor, so that polysaccharide is fully and uniformly mixed, and a polysaccharide solution with mass concentration of 0.5% is obtained.
(5) Mixing the ovalbumin-ferulic acid nanoparticle solution (OVA-FA) obtained in the step (3) with the polysaccharide solution obtained in the step (4) according to the volume ratio of 1:1, namely respectively dropwise adding the polysaccharide solution into the ovalbumin-ferulic acid nanoparticle solution under magnetic stirring, wherein the magnetic stirring speed is 400-1200 rpm, and the stirring time is 30-120 min, so as to obtain the ovalbumin-ferulic acid-polysaccharide composite solution.
(6) Mixing the ovalbumin-ferulic acid-polysaccharide composite solution obtained in the step (5) with Medium Chain Triglyceride (MCT) according to the volume ratio of 5:7 and 5:9, namely adding the medium chain triglyceride into the ovalbumin-ferulic acid-polysaccharide composite solution dropwise under magnetic stirring, wherein the magnetic stirring speed is 200-1500 rpm, and the stirring time is 30min.
(7) Homogenizing the mixed solution obtained in the step (6) for a certain time (2 min) under the condition of 12000rpm, thus obtaining the stable emulsion of the ovalbumin-ferulic acid-polysaccharide ternary system.
In this example, the microstructure of the stable emulsion of OVA-FA-SA or OVA-FA-KC and MCT at 5:7 or 5:9 (i.e. OVA-FA-PS: oil=5:7 or 5:9) was observed by a laser confocal scanning microscope (CLSM), i.e. after homogenization, the stable emulsion was left at 4℃for 12 hours, and then observed by a laser confocal microscope. The results are shown in FIG. 6: the result shows that compared with emulsion prepared by OVA-SA or OVA-KC (under the same condition, the polysaccharide solution is replaced by distilled water as a control), a layer of red film is adsorbed around spherical oil drops in the emulsion stabilized by the OVA-FA-SA or OVA-FA-KC compound, so that aggregation of the emulsion can be effectively prevented, the stability of the emulsion is improved, and the spherical oil drops are more compact; as the volume ratio of the oil phase increases, the voids between the emulsion droplets gradually decrease and the droplets become more compact. However, by comparing the stable emulsions of OVA-FA-SA and OVA-FA-KC with MCT in a volume ratio of 5:9, it was found that the stable emulsion of OVA-FA-KC was more compact, the droplets were smaller, the distribution was more uniform, and a layer of red film was adsorbed around the droplets of the emulsion. Therefore, the stability of the OVA-FA-KC stabilized emulsion is better than that of the OVA-FA-SA stabilized emulsion in microscopic view.
Example 6
(1) Accurately weighing 2g of Ovalbumin (OVA) powder, dispersing the powder in 200g of deionized water, continuously stirring at room temperature for 2 hours, adjusting the pH to 7.0+/-0.1 by using acid-base solution, and then placing the powder at a low temperature of 4 ℃ for 24 hours to fully hydrate the protein so as to obtain the ovalbumin storage solution with the mass concentration of 1%.
(2) 1g of Ferulic Acid (FA) powder is weighed and added into 200g of deionized water, and is continuously stirred for 2 hours at room temperature, the pH is adjusted to 7.0+/-0.1 by alkali liquor, and the ferulic acid is fully and uniformly mixed to obtain the ferulic acid solution with the mass concentration of 0.5 percent.
(3) Mixing the ovalbumin solution in the step (1) and the ferulic acid solution in the step (2) according to the volume ratio of 4:1, dropwise adding the ferulic acid solution into the ovalbumin solution under magnetic stirring, wherein the magnetic stirring speed is 800-1400 rpm, the stirring time is 60-120 min, and the pH value of the ovalbumin-ferulic acid mixed solution is regulated to 6.0+/-0.1 to obtain the ovalbumin-ferulic acid nanoparticle solution.
(4) 1g of Sodium Alginate (SA), carrageenan (KC), hyaluronic Acid (HA) and Agar (Agar) powder are respectively weighed, then are respectively added into 200g of deionized water, are continuously stirred for 2 hours at room temperature, and are adjusted to have pH value of 6.0+/-0.1 by alkali liquor, so that polysaccharide is fully and uniformly mixed, and a polysaccharide solution with mass concentration of 0.5% is obtained.
(5) Mixing the ovalbumin-ferulic acid nanoparticle solution obtained in the step (3) with the polysaccharide solution obtained in the step (4) according to the volume ratio of 1:1, namely, dropwise adding the polysaccharide solution into the ovalbumin-ferulic acid nanoparticle solution under magnetic stirring, wherein the magnetic stirring speed is 400-1200 rpm, and the stirring time is 30-120 min, so as to obtain the ovalbumin-ferulic acid-polysaccharide composite solution.
(6) Mixing the ovalbumin-ferulic acid-polysaccharide composite solution obtained in the step (5) with medium-chain triglyceride according to the volume ratio of 5:7 and 5:9, namely respectively dropwise adding the medium-chain triglyceride into the ovalbumin-ferulic acid-polysaccharide composite solution under magnetic stirring, wherein the magnetic stirring speed is 200-1500 rpm, and the stirring time is 30min.
(7) Homogenizing the mixed solution obtained in the step (6) for a certain time (2 min) under the condition of 12000rpm, thus obtaining the stable emulsion of the ovalbumin-ferulic acid-polysaccharide ternary system.
(8) The emulsion in step (7) was left at 37℃for two weeks, and the particle size potential (determined by Dynamic Light Scattering (DLS)) of the emulsion was observed and measured.
This example examined the storage stability of an ovalbumin-ferulic acid-polysaccharide complex stable emulsion. FIG. 7 shows the change in the stable emulsions of OVA-FA-SA and OVA-FA-KC at volume ratios of 5:7 and 5:9 during 14 days of storage at 37 ℃. For both the OVA-FA-SA and OVA-FA-KC stable emulsions, the average particle size increased slightly with increasing storage time and the emulsion remained stable, no significant separation was observed for storage 7d (FIG. 7A), indicating that the emulsion exhibited good stability over this storage time, mainly due to increased electrostatic repulsion and steric hindrance between droplets. After 7d storage, the average particle size of the emulsion increased slightly due to electrostatic repulsion. However, as is clear from fig. 7B, the particle size of the OVA-FA-KC stable emulsion is significantly increased, and fig. 7A shows that no significant layering phenomenon is found after 14 days of storage, probably because the specific surface area of the oil droplets is gradually reduced with the increase of the volume of the oil droplets, and the oil droplets are covered with the OVA-FA-KC complex, so that the specific surface area of the oil droplets remains stable and no layering phenomenon occurs. From fig. 7C, it can be seen that the absolute value of the potential of the emulsion gradually decreases with the increase of the storage time. Thus, it was found that the storage stability of the OVA-FA-KC emulsion was better than that of OVA-FA-SA.
Example 7
(1) Accurately weighing 2g of Ovalbumin (OVA) powder, dispersing the powder in 200g of deionized water, continuously stirring at room temperature for 2 hours, adjusting the pH to 7.0+/-0.1 by using acid-base solution, and then placing the powder at a low temperature of 4 ℃ for 24 hours to fully hydrate the protein so as to obtain the ovalbumin storage solution with the mass concentration of 1%.
(2) 1g of Ferulic Acid (FA) powder is weighed and added into 200g of deionized water, and is continuously stirred for 2 hours at room temperature, the pH is adjusted to 7.0+/-0.1 by alkali liquor, and the ferulic acid is fully and uniformly mixed to obtain the ferulic acid solution with the mass concentration of 0.5 percent.
(3) Mixing the ovalbumin solution in the step (1) and the ferulic acid solution in the step (2) according to the volume ratio of 4:1, dropwise adding the ferulic acid solution into the ovalbumin solution under magnetic stirring, wherein the magnetic stirring speed is 800-1400 rpm, the stirring time is 60-120 min, and the pH value of the ovalbumin-ferulic acid mixed solution is regulated to 6.0+/-0.1 to obtain the ovalbumin-ferulic acid nanoparticle solution.
(4) 1g of Sodium Alginate (SA), carrageenan (KC) and Hyaluronic Acid (HA) are respectively weighed, then added into 200g of deionized water respectively, continuously stirred for 2 hours at room temperature, and the pH value is adjusted to 6.0+/-0.1 by alkali liquor, so that polysaccharide is fully and uniformly mixed, and a polysaccharide solution with the mass concentration of 0.5% is obtained.
(5) Mixing the ovalbumin-ferulic acid nanoparticle solution obtained in the step (3) with the polysaccharide solution obtained in the step (4) according to the volume ratio of 1:1, namely, dropwise adding the polysaccharide solution into the ovalbumin-ferulic acid nanoparticle solution under magnetic stirring, wherein the magnetic stirring speed is 400-1200 rpm, and the stirring time is 30-120 min, so as to obtain the ovalbumin-ferulic acid-polysaccharide composite solution.
(6) Weighing 4g of rutin (Ru) powder, adding the rutin powder into 350mL of Medium Chain Triglyceride (MCT), continuously stirring for 30-90 min at room temperature, centrifuging for 2-10 min at 5000rpm, and obtaining supernatant as rutin-loaded Medium Chain Triglyceride (MCT).
(7) Mixing the ovalbumin-ferulic acid-polysaccharide composite solution obtained in the step (5) with the rutin-loaded medium-chain triglyceride obtained in the step (6) according to the volume ratio of 5:9, namely respectively dropwise adding the rutin-loaded medium-chain triglyceride into the ovalbumin-ferulic acid-polysaccharide composite solution under magnetic stirring, wherein the magnetic stirring speed is 200-1500 rpm, and the stirring time is 30min.
(8) Homogenizing the mixed solution obtained in the step (6) for 2min at 12000rpm to obtain emulsion (OVA-FA-SA-Ru, OVA-FA-KC-Ru, OVA-FA-HA-Ru) with stable ternary system of ovalbumin-ferulic acid-polysaccharide loaded with rutin.
This example examined the effect of the loading of fat-soluble polyphenols on the rheological properties of the emulsion (with the emulsion OVA-FA-SA, OVA-FA-KC, OVA-FA-HA prepared under the same conditions and stabilized with a ternary system without rutin). The results are shown in FIG. 8: as can be seen from the graph, the storage modulus and the loss modulus of the OVA-FA-SA-Ru and the OVA-FA-KC-Ru are obviously higher than those of the OVA-FA-SA and the OVA-FA-KC. The results indicate that the loading of the fat-soluble polyphenols can increase the storage modulus and loss modulus of the ternary emulsion. The storage modulus and the loss modulus of the emulsion of the OVA-FA-HA loaded fat-soluble polyphenol are obviously lower than those of the OVA-FA-SA and the OVA-FA-KC loaded fat-soluble polyphenol emulsion although the storage modulus and the loss modulus of the emulsion are increased relative to the single OVA-FA-HA emulsion. After the OVA-FA-SA and the OVA-FA-KC are loaded by the fat-soluble polyphenol, the storage modulus and the loss modulus of the OVA-FA-SA and the OVA-FA-KC are obviously increased relative to the single OVA-FA-SA and the single OVA-FA-KC, and meanwhile, the storage modulus and the loss modulus of the OVA-FA-SA and the single OVA-FA-KC do not show great difference, and the storage modulus and the loss modulus of the OVA-FA-SA and the loss modulus of the OVA-FA-KC are greatly increased relative to the single OVA-FA-SA and the single OVA-FA-KC. Furthermore, during the increase in test frequency, G' of each emulsion was significantly higher than G ", representing the solid nature of the gel.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.

Claims (7)

1. A method for preparing an ovalbumin-ferulic acid-polysaccharide composite emulsion, which is characterized by comprising the following steps:
(1) Adding egg albumin into water, stirring uniformly, regulating the pH value to 7.0+/-0.1, and standing to fully hydrate the protein to obtain an egg albumin dispersion liquid;
(2) Adding ferulic acid into water, stirring uniformly, and regulating pH value to 7.0+ -0.1 to obtain ferulic acid solution;
(3) Adding the ferulic acid solution obtained in the step (2) into the ovalbumin dispersion liquid obtained in the step (1), stirring and mixing uniformly, and regulating the pH value to 6.0+/-0.1 to obtain ovalbumin-ferulic acid solution;
(4) Adding polysaccharide into water, stirring uniformly, and regulating pH value to 6.0+ -0.1 to obtain polysaccharide solution; wherein the polysaccharide is carrageenan;
(5) Adding the ovalbumin-ferulic acid solution obtained in the step (3) into the polysaccharide solution obtained in the step (4), and uniformly stirring and mixing to obtain an ovalbumin-ferulic acid-polysaccharide compound solution;
(6) Dropwise adding medium-chain triglyceride into the ovalbumin-ferulic acid-polysaccharide compound solution obtained in the step (5), stirring and uniformly mixing, and homogenizing to obtain an ovalbumin-ferulic acid-polysaccharide compound emulsion;
the mass concentration of the ovalbumin dispersion liquid in the step (1) is 1%;
the mass concentration of the ferulic acid solution in the step (2) is 0.5%;
the volume ratio of the ovalbumin dispersion liquid to the ferulic acid solution in the step (3) is 4:1;
the mass concentration of the ovalbumin-ferulic acid solution in the step (3) is 1%;
the mass concentration of the polysaccharide solution in the step (4) is 0.5%;
the volume ratio of the ovalbumin-ferulic acid solution to the polysaccharide solution in the step (5) is 1:1;
the volume ratio of the ovalbumin-ferulic acid-polysaccharide complex solution to the medium chain triglyceride in the step (6) is 5: 7-9.
2. The method for preparing an ovalbumin-ferulic acid-polysaccharide complex emulsion according to claim 1, wherein the method comprises the following steps:
the conditions for sufficient hydration of the protein by standing as described in step (1) are: standing at a low temperature of 4 ℃ for 12-24 hours;
the stirring conditions in the steps (1), (2) and (4) are as follows: stirring continuously at room temperature for more than 2 hours;
the regulator used for regulating the pH value in the steps (1), (2), (3) and (4) is an HCl solution with the concentration of 0.1-1 mol/L and an NaOH solution with the concentration of 0.1-1 mol/L;
the stirring conditions in the step (3) are as follows: stirring at 500-1400 rpm for 30-120 min;
the stirring conditions in the step (5) are as follows: stirring at 400-1200 rpm for 10-120 min;
the stirring conditions in the step (6) are as follows: stirring at 200-1500 rpm for 10-100 min;
the homogenization conditions described in step (6) are: homogenizing at 2000-20000 rpm for 1-20 min.
3. An ovalbumin-ferulic acid-polysaccharide complex emulsion, characterized in that: prepared by the method of any one of claims 1-2.
4. Use of an ovalbumin-ferulic acid-polysaccharide complex emulsion according to claim 3 in the field of food, pharmaceutical, health care or cosmetics.
5. Use of an ovalbumin-ferulic acid-polysaccharide complex emulsion according to claim 3 for the preparation of a drug delivery system.
6. A preparation method of an ovalbumin-ferulic acid-polysaccharide complex emulsion loaded with a drug is characterized by comprising the following steps: steps (1) to (5) of the method for preparing an ovalbumin-ferulic acid-polysaccharide complex emulsion according to any one of claims 1 to 2, and the steps of:
(6) Adding the medicine into the medium chain triglyceride, and uniformly stirring and mixing to obtain the medium chain triglyceride loaded with the medicine;
(7) Dropwise adding the medium chain triglyceride loaded with the drug obtained in the step (6) into the ovalbumin-ferulic acid-polysaccharide compound solution obtained in the step (5), uniformly stirring and mixing, and homogenizing to obtain the ovalbumin-ferulic acid-polysaccharide compound emulsion loaded with the drug.
7. The method for preparing a drug-loaded ovalbumin-ferulic acid-polysaccharide complex emulsion of claim 6, wherein the method comprises the steps of:
the medicine in the step (6) is a fat-soluble medicine;
the fat-soluble medicine is at least one of fat-soluble polyphenol compounds and fat-soluble vitamin substances;
the dosage of the medicine in the step (6) is calculated according to the medium chain triglyceride of 85-90 mL in each gram of medicine;
the volume ratio of the ovalbumin-ferulic acid-polysaccharide complex solution and the medium chain triglyceride loaded with the drug in the step (7) is 5: 5-9.
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Citations (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7005158B1 (en) * 2003-06-30 2006-02-28 University Of Florida Research Foundation, Inc. Methods of improving the properties of egg proteins
JP2016037461A (en) * 2014-08-06 2016-03-22 ユーハ味覚糖株式会社 Production method of nanoparticle which uses naturally occurring component as substrate
CN105601701A (en) * 2016-01-21 2016-05-25 中国农业大学 Protein-polyphenol covalent compound and preparation method and application thereof
CN108719999A (en) * 2018-05-07 2018-11-02 中国农业大学 A kind of preparation method and applications of protein-polyphenol-polysaccharide covalent compound
CN108783149A (en) * 2018-05-17 2018-11-13 南京农业大学 A kind of production method of curcumin-ovalbumin compound anti-oxidation protein beverage
CN108976819A (en) * 2018-07-16 2018-12-11 广州大学 A kind of preparation method of soybean protein isolate-chitosan composite particle
CN109247425A (en) * 2018-08-22 2019-01-22 湖北工业大学 A kind of preparation method of the ovalbumin lotion with duplex shell structure
CN109453138A (en) * 2018-11-28 2019-03-12 江苏大学 A kind of load medicine albumin microparticle or nanoparticle and preparation method thereof
CN110710577A (en) * 2018-07-12 2020-01-21 华中农业大学 Preparation method of high-oil-loading-capacity emulsion rich in fibrous polysaccharide
CN111227161A (en) * 2020-01-09 2020-06-05 扬州大学 Ovalbumin-carvacrol nanoparticle and preparation method and antibacterial application thereof
CN111357971A (en) * 2020-04-23 2020-07-03 广州大学 Double-crosslinked pickering emulsion and preparation method and application thereof
CN111393671A (en) * 2020-03-27 2020-07-10 广州大学 Glycinin-chitosan composite gel and preparation method thereof

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10117839B2 (en) * 2014-08-05 2018-11-06 Intervet Inc. Encapsulation of hydrophobic biologically active compounds
CN107923896A (en) * 2015-04-24 2018-04-17 高露洁-棕榄公司 Porous protein body as active agent carrier

Patent Citations (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7005158B1 (en) * 2003-06-30 2006-02-28 University Of Florida Research Foundation, Inc. Methods of improving the properties of egg proteins
JP2016037461A (en) * 2014-08-06 2016-03-22 ユーハ味覚糖株式会社 Production method of nanoparticle which uses naturally occurring component as substrate
CN105601701A (en) * 2016-01-21 2016-05-25 中国农业大学 Protein-polyphenol covalent compound and preparation method and application thereof
CN108719999A (en) * 2018-05-07 2018-11-02 中国农业大学 A kind of preparation method and applications of protein-polyphenol-polysaccharide covalent compound
CN108783149A (en) * 2018-05-17 2018-11-13 南京农业大学 A kind of production method of curcumin-ovalbumin compound anti-oxidation protein beverage
CN110710577A (en) * 2018-07-12 2020-01-21 华中农业大学 Preparation method of high-oil-loading-capacity emulsion rich in fibrous polysaccharide
CN108976819A (en) * 2018-07-16 2018-12-11 广州大学 A kind of preparation method of soybean protein isolate-chitosan composite particle
CN109247425A (en) * 2018-08-22 2019-01-22 湖北工业大学 A kind of preparation method of the ovalbumin lotion with duplex shell structure
CN109453138A (en) * 2018-11-28 2019-03-12 江苏大学 A kind of load medicine albumin microparticle or nanoparticle and preparation method thereof
CN111227161A (en) * 2020-01-09 2020-06-05 扬州大学 Ovalbumin-carvacrol nanoparticle and preparation method and antibacterial application thereof
CN111393671A (en) * 2020-03-27 2020-07-10 广州大学 Glycinin-chitosan composite gel and preparation method thereof
CN111357971A (en) * 2020-04-23 2020-07-03 广州大学 Double-crosslinked pickering emulsion and preparation method and application thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
Ferulic acid-ovalbumin protein nanoparticles: Structure and foaming behavior;Chang Keifei等;Food Research International;第136卷;页码109311 *
卵清蛋白基复合载体的构建及其负载山奈酚/单宁酸的研究;史航;中国优秀硕士学位论文全文数据库 工程科技Ⅰ辑(第12期);第10-11页"1.5研究目的及意义"和"1.6主要研究内容"，第29-30页"3.3方法"，第45-46页"3.5本章小结"，第64-65页"5总结、创新点与展望"部分 *

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