Nutritional food for improving night vision function and preparation method thereof

Abstract

The invention discloses a nutritional food capable of improving a night vision function and a preparation method of the nutritional food. The composition is prepared from anthocyanin-sterculia nobilis acid nano-particles, fucoxanthin-vitamin A-resveratrol nano-particles and zinc taurate. The functions of retinal rod cells can be remarkably improved, the blood microcirculation of eyes can be promoted, and the vision sensitivity and the asthenopia resistance in dark environments such as night, weak light and low visibility can be remarkably improved; the product can be prepared into capsules, tablets, electuary, oral liquid or beverage.

Description

Technical Field

[0001] The invention relates to the technical field of food processing, and in particular to a nutritious food for improving night vision function and a preparation method thereof. Background Art

[0002] With the advancement of science and technology and the needs of rapid social development, people are becoming more and more active in activities without light or in dim light environments (dark environments). These activities not only include night driving, which is closely related to people's daily lives, but also involve important areas such as scientific exploration in dark environments (such as deep sea, underground scientific research, etc.), military activities (such as night military operations, submarine operations, etc.), and production and construction (such as tunnel construction, mineral mining, etc.). However, humans have poor visual sensitivity and weak ability to capture environmental signals in dark environments, which leads to slow brain reaction sensitivity and limb movement coordination. This greatly affects people's work / work efficiency in dark environments and greatly increases the risk of safety accidents. Therefore, nutritional or functional foods that significantly improve visual sensitivity in dark environments have become products that are in urgent need.

[0003] Studies have shown that some special dietary nutrients can regulate retinal cell function, thereby improving night vision or improving night blindness. These dietary nutrients include anthocyanins, fucoxanthin, etc. However, anthocyanins and fucoxanthin have poor digestion stability and are difficult to cross the intestinal mucus barrier and intestinal epithelial barrier, resulting in low oral bioavailability and difficulty in fully exerting their biological and nutritional functions. Although the current conventional micro-nano encapsulation technology has effectively improved the stability of nutrients such as anthocyanins and carotenoids, it is still difficult to take into account the nutritional needs of efficient intestinal absorption. Summary of the invention

[0004] In view of the deficiencies in the prior art, the purpose of the present invention is to provide a nutritious food with synergistic product ingredients, efficient absorption of core functional ingredients, and significant effect of improving night vision through product formula design and production process innovation.

[0005] To achieve the above purpose, a nutritious food for improving night vision function is provided, characterized in that it is prepared from anthocyanin-sterculic acid nanoparticles, fucoxanthin-vitamin A-resveratrol nanoparticles, and zinc taurine; the weight ratio of the anthocyanin-sterculic acid nanoparticles, fucoxanthin-vitamin A-resveratrol nanoparticles, and zinc taurine is 40-60:5-10:0.1-1.

[0006] Furthermore, the anthocyanin-sterculic acid nanoparticles are spherical particles formed by self-assembly of an aqueous solution of anthocyanin-anionic polysaccharide self-assembled complex and an aqueous solution of sterculic acid-mannose grafted protein ligand complex, preferably in a mass ratio of 1:1 to 1:4, and the sterculic acid-mannose grafted protein ligand complex wraps the anthocyanin-anionic polysaccharide self-assembled complex in the inner core, and the particle size is 300 to 500 nm;

[0007] Preferably, the preparation method of the anthocyanin-sterculic acid nanoparticles is:

[0008] The pH of the aqueous solution of the sterculia acid-mannose grafted protein ligand complex is adjusted to acidic, preferably, the acidic pH is 3.0±0.5; then, it is added to the aqueous solution of the anthocyanin-anionic polysaccharide self-assembled complex, and the reaction is preferably carried out in the dark for 30±5 minutes, so that the sterculia acid-mannose grafted protein ligand complex is tightly adsorbed on the outer layer of the anthocyanin-anionic polysaccharide self-assembled complex, and anthocyanin-sterculia acid nanoparticles are obtained after freeze-drying; preferably, the mass ratio of the anthocyanin-anionic polysaccharide self-assembled complex to the sterculia acid-mannose grafted protein ligand complex is 1:1 to 1:4;

[0009] Preparation of the aqueous solution of the sterculia acid-mannose grafted protein ligand complex: 1) Mixing protein and mannose and dissolving them in deionized water, preferably, the mass ratio of protein to mannose is 1:5; after freeze-drying, placing the mixed powder in a sealed container for reaction, preferably, the reaction conditions in the sealed container are a sealed container with a relative humidity of 79±1%, and reacting at 65±5°C for 48±3h; obtaining a mannose grafted protein with a grafting degree of not less than 20%; 2) Dissolving the mannose grafted protein in deionized water and adjusting the pH to alkaline, preferably, the alkaline pH is 11.0±1; then adding dissolved sterculia acid, and then adjusting the pH to neutral; allowing sterculia acid to enter the hydrophobic cavity of the mannose grafted protein in the form of a ligand to obtain a sterculia acid-mannose grafted protein ligand complex aqueous solution; the mass ratio of the mannose grafted protein to sterculia acid is 1:0.02 to 1:0.08;

[0010] Preparation of the anthocyanin-anionic polysaccharide self-assembly complex: dissolving anthocyanin and anionic polysaccharide in deionized water in proportion, and adjusting the pH to acidic, preferably, the weight ratio of anthocyanin to anionic polysaccharide is 1:40-86; the acidic pH is 3.0±0.5; then performing ultrasonic treatment to obtain an aqueous solution of anthocyanin-anionic polysaccharide self-assembly complex;

[0011] Preferably, the anthocyanin is one of anthocyanin extracts of bilberry, grape, raspberry, and black wolfberry, and the total anthocyanin content is ≥ 90% by mass; and / or

[0012] The anionic polysaccharide is one of sulfated polysaccharide, hyaluronic acid, pectin and sodium alginate; and / or

[0013] The protein is one of bovine serum albumin, whey protein, ovalbumin and soy protein.

[0014] Furthermore, the fucoxanthin-vitamin A-resveratrol nanoparticles are spherical particles formed by self-assembly of an aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles and mannose-grafted chitosan in a mass ratio of 15:1 to 15:4, and the mannose-grafted chitosan wraps the fucoxanthin-vitamin A-resveratrol-protein particles in the inner core, and the particle size is 400 to 600 nm;

[0015] Preferably, the preparation method of the fucoxanthin-vitamin A-resveratrol nanoparticles is:

[0016] Adding mannose-grafted chitosan to an aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles, wherein the concentration of mannose-grafted chitosan is 0.1 mass % to 0.4 mass %, and reacting at room temperature in the dark for preferably 30±5 min, so that the mannose-grafted chitosan is adsorbed on the outer layer of the protein particles, and then freeze-dried to obtain fucoxanthin-vitamin A-resveratrol nanoparticles;

[0017] The preparation of the mannose-grafted chitosan comprises: dissolving mannose in a chitosan solution so that the mass ratio of chitosan to mannose is 1:3; adjusting the pH of the chitosan-mannose mixed solution to an acidic reaction, preferably, pH 5.5±1, and reacting at 65±5° C. for 60±12 h; cooling after the reaction, and freeze-drying to obtain a water-soluble mannose-grafted chitosan;

[0018] The aqueous solution of the fucoxanthin-vitamin A-resveratrol-protein particles is prepared by mixing and dissolving fucoxanthin, vitamin A, resveratrol and oleic acid, adding them to alkaline deionized water, and ultrasonicating them to homogenize and aggregate the above-mentioned fat-soluble components to obtain mixed micelles of fucoxanthin, vitamin A, resveratrol and oleic acid; mixing the mixed micelles with an alkaline protein solution, reacting them at room temperature in the dark, and then adjusting the pH back to neutral to obtain an aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles with a protein shell; preferably, the mass ratio of fucoxanthin, vitamin A, resveratrol and oleic acid is 2:1:1:1; the ultrasonication time is 2±0.5min; the concentration of the protein solution is 2% by mass, and the pH of the alkaline deionized water and the alkaline protein solution is independently 11.0±1; the reaction time in the dark is 30±5min;

[0019] Preferably, the protein is one of bovine serum albumin, whey protein, ovalbumin and soy protein.

[0020] Furthermore, the nutritional food is a capsule, tablet, granule, oral liquid or beverage.

[0021] The present invention also provides a method for preparing the nutritious food for improving night vision function, which is characterized by comprising the following steps: The method for preparing the anthocyanin-sterculic acid nanoparticles is:

[0022] S1. Preparation of anthocyanin-sterculic acid nanoparticles:

[0023] S1.1 Preparation of aqueous solution of sterculic acid-mannose grafted protein ligand complex: protein and mannose are mixed and dissolved in deionized water, and the mixed powder is placed in a sealed container for reaction after freeze-drying to obtain mannose-grafted protein; preferably, the mass ratio of protein to mannose is 1:5; the mass concentration of protein is 2%; the reaction conditions in the sealed container are a relative humidity of 79±1% in a sealed container, and a reaction temperature of 65±5°C for 48±3h; the grafting degree of mannose-grafted protein is 20±4%;

[0024] The mannose-grafted protein is dissolved in deionized water, and the pH is adjusted to alkaline, and then dissolved sterculia acid is added, and then the pH is adjusted to neutral, so that sterculia acid enters the hydrophobic cavity of the mannose-grafted protein in the form of a ligand, and a sterculia acid-mannose-grafted protein ligand complex aqueous solution is obtained; preferably, the alkaline pH is 11.0±1; preferably, the mass ratio of the mannose-grafted protein to sterculia acid is 1:0.02 to 1:0.08;

[0025] S1.2 Preparation of anthocyanin-anionic polysaccharide self-assembled complex aqueous solution: dissolve anthocyanin and anionic polysaccharide in deionized water, and adjust the pH to acidic; then perform ultrasonic treatment to obtain an aqueous solution of anthocyanin-anionic polysaccharide self-assembled complex; preferably, the weight ratio of anthocyanin to anionic polysaccharide is 1:40-86; the acidic pH is 3.0±1;

[0026] The above steps S1.1 and S1.2 can be performed in different orders or simultaneously;

[0027] S1.3 The pH of the aqueous solution of the sterculic acid-mannose grafted protein ligand complex is adjusted to acidic, and then added to the aqueous solution of the anthocyanin-anionic polysaccharide self-assembled complex, and the reaction is carried out in the dark, so that the sterculic acid-mannose grafted protein ligand complex is tightly adsorbed on the outer layer of the anthocyanin-anionic polysaccharide self-assembled complex, and anthocyanin-sterculic acid nanoparticles are obtained after freeze-drying; preferably, the acidic pH is 3.0±1; the mass ratio of the anthocyanin-anionic polysaccharide self-assembled complex to the sterculic acid-mannose grafted protein ligand complex is 1:1 to 1:4; the reaction time in the dark is 30±5min;

[0028] S2. The preparation method of fucoxanthin-vitamin A-resveratrol nanoparticles is:

[0029] S2.1 Preparation of mannose-grafted chitosan: dissolving mannose in a chitosan solution, preferably, making the mass ratio of chitosan to mannose be 1:3; adjusting the pH of the chitosan-mannose mixed solution to acidic and reacting; cooling after the reaction, and freeze-drying to obtain water-soluble mannose-grafted chitosan; preferably, the final concentration of the chitosan solution is 5±1 g / L, and the solvent is 1% by volume acetic acid aqueous solution; the pH of the acidic reaction is 5.5±1, and the reaction conditions are 65±5°C for 60±12h;

[0030] S2.2 Preparation of aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles: fucoxanthin, vitamin A, resveratrol and oleic acid are mixed and dissolved, and then added into alkaline deionized water, and ultrasonicated to homogenize and aggregate the above fat-soluble components to obtain mixed micelles of fucoxanthin, vitamin A, resveratrol and oleic acid; the mixed micelles are mixed with alkaline protein solution, reacted at room temperature in the dark, and then the pH is adjusted back to neutral to obtain fucoxanthin-vitamin A-resveratrol-protein particles with a protein shell; preferably, the weight ratio of fucoxanthin, vitamin A, resveratrol and oleic acid is 2:1:1:1; the ultrasonication time is 2±0.5min; the concentration of the protein solution is 2% by mass, and the pH of the alkaline deionized water and the alkaline protein solution is independently 11.0±1; the reaction time in the dark is 30±5min;

[0031] The above steps S2.1 and S2.2 can be performed in different orders or simultaneously.

[0032] S2.3 adding mannose-grafted chitosan to the aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles, wherein the concentration of mannose-grafted chitosan is 0.1 mass % to 0.4 mass %, and reacting at room temperature in the dark for preferably 30±5 min, so that the mannose-grafted chitosan is adsorbed on the outer layer of the protein particles, and lyophilizing to obtain fucoxanthin-vitamin A-resveratrol nanoparticles;

[0033] The above steps S1 and S2 can be exchanged in order or performed simultaneously;

[0034] S3. Mix anthocyanin-sterculic acid nanoparticles, fucoxanthin-vitamin A-resveratrol nanoparticles, zinc taurine, water or solid filler excipients according to proportion.

[0035] Further, the method further comprises: S4. homogenizing and filtering the aqueous solution mixture, or sieving the solid mixture.

[0036] Furthermore, it also includes: S5. filling after sterilization, or packaging after tableting or granulation.

[0037] Furthermore, in the preparation of the aqueous solution of anthocyanin-anionic polysaccharide self-assembled complex in step S1.2, the ultrasonic power is 100 to 500 W, and the ultrasonic time is 1 to 5 min; and / or

[0038] The mass ratio of anthocyanin to anionic polysaccharide is 1:40 to 1:80.

[0039] Furthermore, in the preparation of the mannose grafted chitosan in step S2.1, the viscosity of the chitosan is 150 mPa·s, and the grafting degree of the mannose grafted chitosan is 35±3%.

[0040] The present invention also protects the nutritional food for improving night vision function or the nutritional food for improving night vision function prepared by the preparation method, which has the effect of improving night vision function.

[0041] The ultrasound in step 2.2 can promote the interaction of these fat-soluble components and form smaller aggregates with uniform particles. Without ultrasound, the aggregates formed are larger and uneven, which is not conducive to the subsequent protein encapsulation.

[0042] In order to develop nutritious foods with significant effects, the present invention has made the following innovations in key technologies for product development: (1) With the help of the formula design theory of "synergism" in traditional Chinese medicine, dozens of potential dietary active substances and anthocyanins were studied and screened for compatibility, and it was found that fucoxanthin and taurine chelated zinc had a significant synergistic gain effect on the night vision improvement function of anthocyanins; (2) Taking full advantage of the physiological characteristics of the intestinal epithelium expressing mannose receptors, mannose was modified on the surface of anthocyanin and fucoxanthin nanoparticles, thereby promoting the aggregation of anthocyanins and fucoxanthin on the outside of intestinal epithelial cells; at the same time, sterculic acid was introduced into the anthocyanin carrier to promote the intestinal absorption of anthocyanins by upregulating intestinal epithelial glucose transporter; (3) The introduction of vitamin A into the carrier can promote the cellular uptake of fucoxanthin, and the introduction of resveratrol can inhibit the re-excretion of fucoxanthin into the intestinal cavity, thereby effectively improving the oral bioavailability of fucoxanthin.

[0043] Compared with the prior art, the present invention has the following advantages:

[0044] (1) Innovative use of intestinal epithelial affinity adsorption-transporter regulation coupling technology to prepare highly intestinal absorbable anthocyanin nanoparticles. Compared with the existing preparation technology, mannose is modified on the surface of the delivery carrier, thereby giving the delivery system the ability to affinity adsorb to the intestinal epithelial mannose receptor, allowing anthocyanins to accumulate on the outside of the intestinal epithelial cells, increasing the chance of being absorbed and transported by the epithelial cells; at the same time, sterculia acid, which can upregulate the expression of the intestinal epithelial glucose transporter GLUT2, is integrated into the delivery system, thereby promoting the transmembrane transport of anthocyanins and greatly improving the oral bioavailability of anthocyanins.

[0045] (2) Innovative use of intestinal epithelial affinity adsorption-lipid transport protein regulation coupling technology to prepare highly intestinal absorbable fucoxanthin nanoparticles. Compared with the existing preparation technology, mannose modification was performed on the surface of the delivery carrier, thereby giving the delivery system the ability to affinity adsorb to the intestinal epithelial mannose receptor, allowing fucoxanthin to accumulate on the outside of the intestinal epithelial cells, increasing the chance of being absorbed and transported by the epithelial cells; at the same time, vitamin A that can promote SR-B1 expression and resveratrol that can inhibit P-gp expression are integrated into the delivery system, which can not only promote the uptake of fucoxanthin by intestinal epithelial cells, but also effectively inhibit the excretion of the ingested fucoxanthin into the intestinal cavity, greatly improving the oral bioavailability of fucoxanthin.

[0046] (3) The product is a composition of anthocyanin-sterculic acid nanoparticles, fucoxanthin-vitamin A-resveratrol nanoparticles, and zinc taurine. The present invention adopts the "synergistic" formula design technology and screens out a nutritional formula from dozens of dietary active ingredients through animal experiments, confirming that anthocyanin, fucoxanthin, and zinc taurine have a synergistic nutritional effect in improving night vision function. DETAILED DESCRIPTION

[0047] Embodiments of the present invention are described in detail below, and the examples of the embodiments are intended to be used to explain the present invention, but should not be construed as limiting the present invention. In the embodiments, if specific techniques or conditions are not specified, they are carried out according to the techniques or conditions described in the literature in this area or according to the product specification. If the manufacturer is not specified for reagents or instruments used, they are all conventional products that can be obtained commercially.

[0048] A method for preparing a nutritious food for improving night vision function, characterized in that it comprises the following steps: The method for preparing anthocyanin-sterculic acid nanoparticles is:

[0049] S1. Preparation of anthocyanin-sterculic acid nanoparticles:

[0050] S1.1 Preparation of aqueous solution of sterculic acid-mannose grafted protein ligand complex: protein and mannose are mixed and dissolved in deionized water, and the mixed powder is placed in a sealed container for reaction after freeze-drying to obtain mannose-grafted protein; preferably, the mass ratio of protein to mannose is 1:5; the mass concentration of protein is 2%; the reaction conditions in the sealed container are a relative humidity of 79±1% in a sealed container, and a reaction temperature of 65±5°C for 48±3h; the grafting degree of mannose-grafted protein is 20±4%;

[0051] The mannose-grafted protein is dissolved in deionized water, and the pH is adjusted to alkaline, and then dissolved sterculia acid is added, and then the pH is adjusted to neutral, so that sterculia acid enters the hydrophobic cavity of the mannose-grafted protein in the form of a ligand, and a sterculia acid-mannose-grafted protein ligand complex aqueous solution is obtained; preferably, the alkaline pH is 11.0±1; preferably, the mass ratio of the mannose-grafted protein to sterculia acid is 1:0.02 to 1:0.08;

[0052] S1.2 Preparation of anthocyanin-anionic polysaccharide self-assembled complex aqueous solution: dissolve anthocyanin and anionic polysaccharide in deionized water, and adjust the pH to acidic; then perform ultrasonic treatment to obtain an aqueous solution of anthocyanin-anionic polysaccharide self-assembled complex; preferably, the weight ratio of anthocyanin to anionic polysaccharide is 1:40-86; the acidic pH is 3.0±1;

[0053] The above steps S1.1 and S1.2 can be performed in different orders or simultaneously;

[0054] S1.3 The pH of the aqueous solution of the sterculic acid-mannose grafted protein ligand complex is adjusted to acidic, and then added to the aqueous solution of the anthocyanin-anionic polysaccharide self-assembled complex, and the reaction is carried out in the dark, so that the sterculic acid-mannose grafted protein ligand complex is tightly adsorbed on the outer layer of the anthocyanin-anionic polysaccharide self-assembled complex, and anthocyanin-sterculic acid nanoparticles are obtained after freeze-drying; preferably, the acidic pH is 3.0±1; the mass ratio of the anthocyanin-anionic polysaccharide self-assembled complex to the sterculic acid-mannose grafted protein ligand complex is 1:1 to 1:4; the reaction time in the dark is 30±5min;

[0055] S2. The preparation method of fucoxanthin-vitamin A-resveratrol nanoparticles is:

[0056] S2.1 Preparation of mannose-grafted chitosan: dissolving mannose in a chitosan solution, preferably, making the mass ratio of chitosan to mannose be 1:3; adjusting the pH of the chitosan-mannose mixed solution to acidic and reacting; cooling after the reaction, and freeze-drying to obtain water-soluble mannose-grafted chitosan; preferably, the final concentration of the chitosan solution is 5±1 g / L, and the solvent is 1% by volume acetic acid aqueous solution; the pH of the acidic reaction is 5.5±1, and the reaction conditions are 65±5°C for 60±12h;

[0057] S2.2 Preparation of aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles: fucoxanthin, vitamin A, resveratrol and oleic acid are mixed and dissolved, and then added into alkaline deionized water, and ultrasonicated to homogenize and aggregate the above fat-soluble components to obtain mixed micelles of fucoxanthin, vitamin A, resveratrol and oleic acid; the mixed micelles are mixed with alkaline protein solution, reacted at room temperature in the dark, and then the pH is adjusted back to neutral to obtain fucoxanthin-vitamin A-resveratrol-protein particles with a protein shell; preferably, the weight ratio of fucoxanthin, vitamin A, resveratrol and oleic acid is 2:1:1:1; the ultrasonication time is 2±0.5min; the concentration of the protein solution is 2% by mass, and the pH of the alkaline deionized water and the alkaline protein solution is independently 11.0±1; the reaction time in the dark is 30±5min;

[0058] The above steps S2.1 and S2.2 can be performed in different orders or simultaneously.

[0059] S2.3 adding mannose-grafted chitosan to the aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles, wherein the concentration of mannose-grafted chitosan is 0.1 mass % to 0.4 mass %, and reacting at room temperature in the dark for preferably 30±5 min, so that the mannose-grafted chitosan is adsorbed on the outer layer of the protein particles, and lyophilizing to obtain fucoxanthin-vitamin A-resveratrol nanoparticles;

[0060] The above steps S1 and S2 can be exchanged in order or performed simultaneously;

[0061] S3. Mix anthocyanin-sterculic acid nanoparticles, fucoxanthin-vitamin A-resveratrol nanoparticles, zinc taurine, water or solid filler excipients according to proportion.

[0062] Further, the method further comprises: S4. homogenizing and filtering the aqueous solution mixture, or sieving the solid mixture.

[0063] Furthermore, it also includes: S5. filling after sterilization, or packaging after tableting or granulation.

[0064] Furthermore, in the preparation of the aqueous solution of anthocyanin-anionic polysaccharide self-assembled complex in step S1.2, the ultrasonic power is 100 to 500 W, and the ultrasonic time is 1 to 5 min; and / or

[0065] The mass ratio of anthocyanin to anionic polysaccharide is 1:40 to 1:80.

[0066] Furthermore, in the preparation of the mannose grafted chitosan in step S2.1, the viscosity of the chitosan is 150 mPa·s, and the grafting degree of the mannose grafted chitosan is 35±3%.

[0067] Example 1

[0068] S1. Preparation of anthocyanin-sterculic acid nanoparticles:

[0069] S1.1 Mix bovine serum albumin and mannose in a mass ratio of 1:5 and dissolve in deionized water, wherein the mass concentration of the protein is 2%; place the mixed powder of bovine serum albumin and mannose in a sealed container with a relative humidity of 78% after freeze-drying, and react at 60°C for 51 hours to obtain mannose-grafted protein; dissolve the mannose-grafted protein in deionized water, and adjust the pH to 10.0, add dissolved sterculia acid (a cosolvent can be used to assist dissolution, such as anhydrous ethanol or a menthol-thymol mixture (molar ratio of 1:1), the same applies below), so that the mass ratio of mannose-grafted protein to sterculia acid is 1:0.02, and then adjust the pH to 7.0 to obtain a sterculia acid-mannose grafted protein ligand complex; adjust the pH of the sterculia acid-mannose grafted protein ligand complex aqueous solution to 3.0;

[0070] S1.2. The blueberry anthocyanin extract (total anthocyanin content of 90% by mass) and sulfated polysaccharide were dissolved in deionized water at a mass ratio of 1:40, and the pH was adjusted to 3.0; then, the solution was treated under ultrasonic conditions at a power of 100 W for 5 min to obtain an aqueous solution of anthocyanin-sulfated polysaccharide self-assembled complex;

[0071] The above steps S1.1 and S1.2 can be performed in different orders or simultaneously;

[0072] S1.3 The pH of the aqueous solution of the sterculic acid-mannose grafted protein ligand complex was adjusted to 2.0, and then added to the aqueous solution of the anthocyanin-sulfated polysaccharide self-assembly complex. The mass ratio of the anthocyanin-anionic polysaccharide self-assembly complex and the sterculic acid-mannose grafted protein ligand complex was 1:1. The reaction was carried out at room temperature in the dark for 25 minutes, so that the sterculic acid-mannose grafted protein ligand complex was tightly adsorbed on the outer layer of the anthocyanin-sulfated polysaccharide self-assembly complex. After freeze-drying, anthocyanin-sterculic acid nanoparticles with a particle size of 300 nm were obtained.

[0073] S2. Preparation of Fucoxanthin-Vitamin A-Resveratrol Nanoparticles:

[0074] S2.1 Dissolve chitosan in 1% by volume acetic acid aqueous solution, and the final concentration of chitosan is 5 g / L; dissolve mannose in the chitosan solution, and make the mass ratio of chitosan to mannose be 1:3; adjust the pH of the chitosan and mannose mixed solution to 4.5, and react at 60°C for 72 hours; cool down after the reaction, and obtain water-soluble mannose-grafted chitosan after freeze-drying;

[0075] S2.2 Fucoxanthin, vitamin A, resveratrol and oleic acid were mixed in a mass ratio of 2:1:1:1, dissolved in ethanol, added into deionized water of pH 10.0, and sonicated for 2.5 min to obtain mixed micelles of fucoxanthin, vitamin A, resveratrol and oleic acid; a bovine serum albumin aqueous solution with a concentration of 2% by mass was prepared, and the pH was adjusted to 10.0; the mixed micelles were then mixed with the protein solution, reacted at room temperature in the dark for 25 min, and the pH was adjusted back to 7.0 to obtain fucoxanthin-vitamin A-resveratrol-protein particles with a protein shell;

[0076] The above steps S2.1 and S2.2 can be performed in different orders or simultaneously.

[0077] S2.3 Add mannose-grafted chitosan to the aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles, the concentration of mannose-grafted chitosan is 0.1% by mass, the mass ratio of fucoxanthin-vitamin A-resveratrol-protein particles to mannose-grafted chitosan is 15:1, react at room temperature in the dark for 25 minutes, so that the mannose-grafted chitosan covers the outer layer of the protein particles, and after freeze-drying, obtain fucoxanthin-vitamin A-resveratrol nanoparticles with a particle size of 400 nm;

[0078] The above steps S1 and S2 can be exchanged in order or performed simultaneously;

[0079] S3 Weigh the above components in a ratio of 40 parts by weight of anthocyanin-sterculic acid nanoparticles, 5 parts by weight of fucoxanthin-vitamin A-resveratrol nanoparticles, and 0.1 parts by weight of taurine zinc, add appropriate amounts of magnesium stearate and starch to the formula, mix well,

[0080] After S4 is screened, it is filled into hard capsules using a fully automatic capsule filling machine, with each capsule containing 0.5g, to obtain capsule-type nutritious food.

[0081] Example 2

[0082] S1. Preparation of anthocyanin-sterculic acid nanoparticles:

[0083] S1.1 Mix whey protein and mannose in a mass ratio of 1:5 and dissolve in deionized water, wherein the mass concentration of protein is 2%; place the mixed powder of whey protein and mannose in a closed container with a relative humidity of 79% after freeze-drying, and react at 65°C for 48 hours to obtain mannose-grafted protein; dissolve the mannose-grafted protein in deionized water, and adjust the pH to 11.0, add sterculia acid with ethanol as the solubilizer, so that the mass ratio of mannose-grafted protein to sterculia acid is 1:0.04, and then adjust the pH to 7.0 to obtain a sterculia acid-mannose grafted protein ligand complex;

[0084] S1.2 dissolving grape anthocyanin extract (total anthocyanin content of 92.5% by mass) and hyaluronic acid in deionized water at a mass ratio of 1:60, and adjusting the pH to 3.0; then treating under ultrasonic conditions at a power of 300 W for 3 minutes to obtain an aqueous solution of anthocyanin-hyaluronic acid self-assembled complex;

[0085] The above steps S1.1 and S1.2 can be performed in different orders or simultaneously;

[0086] S1.3 The pH of the aqueous solution of the sterculic acid-mannose grafted protein ligand complex was adjusted to 3.0, and then added to the aqueous solution of the anthocyanin-hyaluronic acid self-assembled complex, the mass ratio of the anthocyanin-hyaluronic acid self-assembled complex to the sterculic acid-mannose grafted protein ligand complex was 1:2, and the reaction was carried out at room temperature in the dark for 30 minutes, so that the sterculic acid-mannose grafted protein ligand complex was tightly adsorbed on the outer layer of the anthocyanin-hyaluronic acid self-assembled complex, and anthocyanin-sterculic acid nanoparticles with a particle size of 380 nm were obtained after freeze-drying;

[0087] Preparation of S2 Fucoxanthin-Vitamin A-Resveratrol Nanoparticles:

[0088] S2.1 Dissolve chitosan in 1% by volume acetic acid aqueous solution, and the final concentration of chitosan is 5 g / L; dissolve mannose in the chitosan solution, and make the mass ratio of chitosan to mannose be 1:3; adjust the pH of the chitosan and mannose mixed solution to 5.5, and react at 65°C for 60 hours; cool down after the reaction, and obtain water-soluble mannose-grafted chitosan after freeze-drying;

[0089] S2.2 Fucoxanthin, vitamin A, resveratrol and oleic acid were mixed in a mass ratio of 2:1:1:1, dissolved with ethanol, added into deionized water of pH 11.0, and sonicated for 2 min to obtain mixed micelles of fucoxanthin, vitamin A, resveratrol and oleic acid; a whey protein aqueous solution with a concentration of 2% by mass was prepared, and the pH was adjusted to 11.0; the mixed micelles were then mixed with the protein solution, reacted at room temperature in the dark for 30 min, and the pH was adjusted back to 7.0 to obtain fucoxanthin-vitamin A-resveratrol-protein particles with a protein shell;

[0090] The above steps S2.1 and S2.2 can be performed in different orders or simultaneously.

[0091] S2.3 Add mannose-grafted chitosan to the aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles, the concentration of mannose-grafted chitosan is 0.2% by mass, the mass ratio of fucoxanthin-vitamin A-resveratrol-protein particles to mannose-grafted chitosan is 15:2, react at room temperature in the dark for 30 minutes, so that the mannose-grafted chitosan covers the outer layer of the protein particles, and after freeze-drying, obtain fucoxanthin-vitamin A-resveratrol nanoparticles with a particle size of 490 nm.

[0092] The above steps S1 and S2 can be exchanged in order or performed simultaneously;

[0093] S3: weigh the above components in a ratio of 50 parts by weight of anthocyanin-sterculic acid nanoparticles, 7 parts by weight of fucoxanthin-vitamin A-resveratrol nanoparticles, and 0.5 parts by weight of zinc taurine, then add appropriate amounts of one or more auxiliary materials selected from dextrin, starch, microcrystalline cellulose, and lactose, mix well, and moisten with appropriate amounts of 70% (v / v) ethanol to prepare a soft material;

[0094] S4 is granulated through a 30-mesh sieve, dried at 70-80°C, sieved through a 60-mesh sieve, and compressed into tablets, each tablet weighing 0.2 g, to obtain a tablet-type nutritional food.

[0095] Example 3

[0096] S1. Preparation of anthocyanin-sterculic acid nanoparticles:

[0097] S1.1 Mix ovalbumin and mannose in a mass ratio of 1:5 and dissolve in deionized water, wherein the mass concentration of the protein is 2%; place the mixed powder of ovalbumin and mannose in a closed container with a relative humidity of 80% after freeze-drying, and react at 70°C for 45 hours to obtain mannose-grafted protein; dissolve the mannose-grafted protein in deionized water, and adjust the pH to 12.0, add sterculia acid with the aid of ethanol solubilization, so that the mass ratio of mannose-grafted protein to sterculia acid is 1:0.06, and then adjust the pH to 7.0 to obtain a sterculia acid-mannose grafted protein ligand complex;

[0098] S1.2 Dissolve raspberry anthocyanin extract (total anthocyanin content of 91.8% by mass) and pectin in deionized water at a mass ratio of 1:70, and adjust the pH to 3.0; then treat under ultrasonic conditions at a power of 400 W for 2 minutes to obtain an aqueous solution of anthocyanin-pectin self-assembled complex;

[0099] The above steps S1.1 and S1.2 can be performed in different orders or simultaneously;

[0100] S1.3 The pH of the aqueous solution of the sterculic acid-mannose grafted protein ligand complex is adjusted to 4.0, and then added to the aqueous solution of the anthocyanin-pectin self-assembled complex, the mass ratio of the anthocyanin-pectin self-assembled complex to the sterculic acid-mannose grafted protein ligand complex is 1:3, and the reaction is carried out at room temperature in the dark for 35 minutes, so that the sterculic acid-mannose grafted protein ligand complex is tightly adsorbed on the outer layer of the anthocyanin-pectin self-assembled complex, and anthocyanin-sterculic acid nanoparticles with a particle size of 440 nm are obtained after freeze-drying;

[0101] S2. Preparation of Fucoxanthin-Vitamin A-Resveratrol Nanoparticles:

[0102] S2.1 Dissolve chitosan in 1% by volume acetic acid aqueous solution, and the final concentration of chitosan is 5 g / L; dissolve mannose in the chitosan solution, and make the mass ratio of chitosan to mannose be 1:3; adjust the pH of the chitosan and mannose mixed solution to 6.5, and react at 70°C for 48 hours; cool down after the reaction, and obtain water-soluble mannose-grafted chitosan after freeze-drying;

[0103] S2.2 Fucoxanthin, vitamin A, resveratrol and oleic acid were mixed in a mass ratio of 2:1:1:1, dissolved in ethanol, added into deionized water of pH 12.0, and sonicated for 1.5 min to obtain mixed micelles of fucoxanthin, vitamin A, resveratrol and oleic acid; an ovalbumin aqueous solution with a concentration of 2% by mass was prepared, and the pH was adjusted to 12.0; the mixed micelles were then mixed with the protein solution, reacted at room temperature in the dark for 35 min, and the pH was adjusted back to 7.0 to obtain fucoxanthin-vitamin A-resveratrol-protein particles with a protein shell;

[0104] The above steps S2.1 and S2.2 can be performed in different orders or simultaneously.

[0105] S2.3 Add mannose-grafted chitosan to the aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles, the concentration of mannose-grafted chitosan is 0.3% by mass, the mass ratio of fucoxanthin-vitamin A-resveratrol-protein particles to mannose-grafted chitosan is 15:3, react at room temperature in the dark for 35 minutes, so that the mannose-grafted chitosan covers the outer layer of the protein particles, and after freeze-drying, obtain fucoxanthin-vitamin A-resveratrol nanoparticles with a particle size of 530 nm.

[0106] The above steps S1 and S2 can be exchanged in order or performed simultaneously;

[0107] S3: Weigh the above components in a ratio of 55 parts by weight of anthocyanin-sterculic acid nanoparticles, 9 parts by weight of fucoxanthin-vitamin A-resveratrol nanoparticles, and 0.7 parts by weight of taurine zinc, then add an appropriate amount of one or more of soluble starch, dextrin, sucrose, lactose, and mannitol, and moisten with an appropriate amount of 70% (v / v) ethanol to prepare a soft material.

[0108] S4 is granulated through a 20-mesh sieve, dried at 70-80°C, and granulated into 60-mesh granules to obtain a granule-type nutritional food.

[0109] Example 4

[0110] S1. Preparation of anthocyanin-sterculic acid nanoparticles:

[0111] S1.1 Soy protein and mannose are mixed in a mass ratio of 1:5 and dissolved in deionized water, wherein the mass concentration of protein is 2%; after freeze-drying, the mixed powder of soy protein and mannose is placed in a sealed container with a relative humidity of 79%, and reacted at 65°C for 48 hours to obtain mannose-grafted protein; the mannose-grafted protein is dissolved in deionized water, and the pH is adjusted to 11.0, and sterculia acid with ethanol as the solubilizer is added to make the mass ratio of mannose-grafted protein to sterculia acid 1:0.08, and then the pH is adjusted to 7.0 to obtain a sterculia acid-mannose grafted protein ligand complex;

[0112] S1.2 The anthocyanin extract of Lycium ruthenicum (total anthocyanin content of 93.2% by mass) and sodium alginate were dissolved in deionized water at a mass ratio of 1:80, and the pH was adjusted to 3.0; then, the mixture was treated under ultrasonic conditions at a power of 500 W for 1 min to obtain an aqueous solution of anthocyanin-sodium alginate self-assembled complex;

[0113] The above steps S1.1 and S1.2 can be performed in different orders or simultaneously;

[0114] S1.3 The pH of the aqueous solution of the sterculia acid-mannose grafted protein ligand complex was adjusted to 3.0, and then added to the aqueous solution of the anthocyanin-sodium alginate self-assembled complex. The mass ratio of the anthocyanin-sodium alginate self-assembled complex and the sterculia acid-mannose grafted protein ligand complex was 1:4. The reaction was carried out at room temperature in the dark for 30 minutes, so that the sterculia acid-mannose grafted protein ligand complex was tightly adsorbed on the outer layer of the anthocyanin-sodium alginate self-assembled complex. After freeze-drying, anthocyanin-sterculia acid nanoparticles with a particle size of 500 nm were obtained.

[0115] S2. Preparation of Fucoxanthin-Vitamin A-Resveratrol Nanoparticles:

[0116] S2.1 Dissolve chitosan in 1% by volume acetic acid aqueous solution, and the final concentration of chitosan is 5 g / L; dissolve mannose in the chitosan solution, and make the mass ratio of chitosan to mannose be 1:3; adjust the pH of the chitosan and mannose mixed solution to 5.5, and react at 65°C for 72 hours; cool down after the reaction, and obtain water-soluble mannose-grafted chitosan after freeze-drying;

[0117] S2.2 Fucoxanthin, vitamin A, resveratrol and oleic acid were mixed in a mass ratio of 2:1:1:1, dissolved in ethanol, added into deionized water of pH 11.0, and sonicated for 2 minutes to obtain mixed micelles of fucoxanthin, vitamin A, resveratrol and oleic acid; a 2% by mass soy protein aqueous solution was prepared, and the pH was adjusted to 11.0; the mixed micelles were then mixed with the protein solution, reacted at room temperature in the dark for 30 minutes, and the pH was adjusted back to 7.0 to obtain fucoxanthin-vitamin A-resveratrol-protein particles with a protein shell;

[0118] The above steps S2.1 and S2.2 can be performed in different orders or simultaneously.

[0119] S2.3 Add mannose-grafted chitosan to the aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles, the concentration of mannose-grafted chitosan is 0.4% by mass, the mass ratio of fucoxanthin-vitamin A-resveratrol-protein particles to mannose-grafted chitosan is 15:4, react at room temperature in the dark for 30 minutes, so that the mannose-grafted chitosan covers the outer layer of the protein particles, and after freeze-drying, obtain fucoxanthin-vitamin A-resveratrol nanoparticles with a particle size of 600 nm.

[0120] The above steps S1 and S2 can be exchanged in order or performed simultaneously;

[0121] S3: Weigh the above components in a ratio of 60 parts by weight of anthocyanin-sterculic acid nanoparticles, 10 parts by weight of fucoxanthin-vitamin A-resveratrol nanoparticles, and 1 part by weight of taurine zinc, then dissolve them in water, add appropriate amounts of sweeteners and stabilizers, mix well, and homogenize.

[0122] S4 filtering, filling and sterilizing to obtain beverage-like functional food.

[0123] Example 5 (Functional evaluation test)

[0124] (1) Effect of intestinal epithelial affinity adsorption-transporter regulation coupling technology on the absorption-enhancing effect of anthocyanins

[0125] Taking Example 1 as an example, the test sample and its preparation method are as follows:

[0126] Table 1 Functional evaluation test samples

[0127]

[0128] Sample treatment: Dissolve the above-mentioned sample in deionized water at a mass-to-volume ratio of 1:10 to obtain the test solution.

[0129] Experimental animals: SPF grade male ICR mice, 15-20 g, purchased from Shanghai Slake Experimental Animal Co., Ltd. Basic feed ingredients for mice: carbohydrate 64%, protein 21%, fat 4%, fiber 5%, water 6%. The above feeds were purchased from Shanghai Slake Experimental Animal Co., Ltd. The temperature of the feeding room was 23±2℃, and the relative humidity was 55±5%.

[0130] Animal grouping and treatment method: After 1 week of adaptive feeding, the mice were randomly divided into 6 groups according to their body weight, with 7 mice in each group, namely, the blank group (administered with deionized water), the 5 groups in Table 1 (specifically, the bilberry anthocyanin extract group, the bilberry anthocyanin-sulfated polysaccharide nanoparticle group, the bilberry anthocyanin-sulfated polysaccharide-protein nanoparticle group, the bilberry anthocyanin-sulfated polysaccharide-mannose modified protein nanoparticle group, and the bilberry anthocyanin-sulfated polysaccharide-sterculic acid-mannose modified protein nanoparticle group). Each mouse was gavaged with 200 μL of the test solution each time (5 groups: the gavage dose of anthocyanin in each group was 50.0 mg / kg) or deionized water (blank group). After 4 hours of gavage, the mouse eyeballs were removed to collect blood, and the changes in the content of anthocyanin in plasma were analyzed by high performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS); the expression of glucose transporter GLUT2 in the small intestine was analyzed by Elisa kit method to evaluate the action pathway of sterculic acid in promoting the intestinal absorption of anthocyanin.

[0131] Experimental results:

[0132] Table 2. Effect of intestinal epithelial affinity adsorption-transporter regulation coupling technology on the absorption-promoting effect of anthocyanins

[0133]

[0134] HPLC analysis of the total response concentration of anthocyanins in the serum of mice in each treatment group is shown in Table 2. The results show that the total response concentration of unencapsulated bilberry anthocyanins in serum was only 7.54 nmol / mL after oral gavage. After being encapsulated by sulfated polysaccharides, the absorption effect was improved to a certain extent, and its total response concentration in serum reached 11.01 nmol / mL, an increase of 46.02%; further, the protein shell was combined with the outer layer of bilberry anthocyanin-sulfated polysaccharide nanoparticles, and the serum response concentration of anthocyanins reached 16.57 nmol / mL. It is worth noting that when its protein shell was modified by mannose, the serum response concentration of anthocyanins was further increased to 34.49 nmol / mL, which was 3.57 times higher than that of the bilberry anthocyanin extract group. It shows that mannose-modified particles can bind to the intestinal mannose receptor, promote the accumulation of anthocyanins on the outside of the small intestinal epithelial cells, and increase the chance of being absorbed by the small intestinal cells. When sterculia acid was incorporated into the delivery system, the absorption effect was greatly improved, and the total serum response concentration of anthocyanins climbed to 82.64nmol / mL. Compared with the bilberry anthocyanin extract group, the intestinal absorption effect was increased by 9.96 times. At the same time, the expression of GLUT2 in the mouse intestine also increased by 3.64 times. It shows that the use of the intestinal epithelial affinity adsorption-transporter regulation coupling technology of the present invention to prepare anthocyanin nanoparticles can effectively improve the oral bioavailability of anthocyanins, thereby laying a foundation for its function of improving vision.

[0135] (2) Effect of intestinal epithelial affinity adsorption-lipid transport protein regulation coupling technology on the absorption-enhancing effect of fucoxanthin

[0136] Taking Example 1 as an example, the test sample and its preparation method are as follows:

[0137] Table 3 Table of the test samples for the effect of intestinal epithelial affinity adsorption-lipid transport protein regulation coupling technology on the absorption promotion of fucoxanthin

[0138]

[0139] Sample treatment: Take the above test sample and dissolve it in deionized water at a mass volume ratio of 1:10 (if necessary, a co-solvent can be used to assist dissolution, such as anhydrous ethanol or menthol-thymol mixture (molar ratio 1:1)), which is the test solution.

[0140] Experimental animals: SPF grade male ICR mice, 15-20 g, purchased from Shanghai Slake Experimental Animal Co., Ltd. Basic feed ingredients for mice: carbohydrate 64%, protein 21%, fat 4%, fiber 5%, water 6%. The above feeds were purchased from Shanghai Slake Experimental Animal Co., Ltd. The temperature of the feeding room was 23±2℃, and the relative humidity was 55±5%.

[0141] Animal grouping and treatment methods: After 1 week of adaptive feeding, the mice were randomly divided into 7 groups according to their body weight, with 7 mice in each group, namely, the blank group (administered with deionized water), the 6 groups in Table 3 (i.e., fucoxanthin group, fucoxanthin-protein nanoparticle group, fucoxanthin-protein-chitosan nanoparticle group, fucoxanthin-protein-mannose modified chitosan nanoparticle group, fucoxanthin-vitamin A-protein-mannose modified chitosan nanoparticle group, and fucoxanthin-vitamin A-resveratrol-protein-mannose modified chitosan nanoparticle group). Each mouse was gavaged with 200 μL of the test solution each time (6 groups: the gavage dose of fucoxanthin in each group was 10.0 mg / kg) or deionized water (blank group). After 4 hours of gavage, the eyeballs of the mice were removed to collect blood, and the changes in the content of fucoxanthin and its metabolites in plasma were analyzed by high performance liquid chromatography (HPLC); the expression levels of intestinal SR-B1 and P-gp were analyzed by Elisa kit method.

[0142] Experimental results:

[0143] Table 4. Effect of intestinal epithelial affinity adsorption-lipid transport protein regulation coupling technology on the absorption-enhancing effect of fucoxanthin

[0144]

[0145] HPLC analysis of the total response concentration of fucoxanthin in the serum of mice in each treatment group is shown in Table 4. The results show that after oral gavage, the total response concentration of unencapsulated fucoxanthin in serum was only 38.01 pmol / mL. After being encapsulated by bovine serum albumin, the absorption effect was improved to a certain extent, and its total response concentration in serum reached 57.50 pmol / mL, which was only increased by 51.27%. When the protein shell of fucoxanthin-protein nanoparticles was covered with chitosan, the absorption effect of fucoxanthin was further improved, especially after chitosan was modified with mannose, the corresponding serum concentration of fucoxanthin was increased to 163.51 pmol / mL. It shows that the mannose in the delivery system can bind to the mannose receptors of intestinal epithelial cells, promote the accumulation of fucoxanthin on the outside of intestinal epithelial cells, and increase the chance of fucoxanthin being absorbed by intestinal epithelial cells. When vitamin A was incorporated into the delivery system, the expression of SR-B1 protein responsible for the transmembrane transport of fucoxanthin increased significantly, which was 2.57 times that of the blank group, and the serum response concentration of fucoxanthin also increased to 275.64pmol / mL, an increase of 6.25 times. It is worth noting that at this time, the expression of P-gp also increased significantly, which was 2.15 times that of the blank group. When resveratrol, which inhibits the expression of P-gp, was further incorporated into the delivery system, SR-B1 did not change significantly, but the expression of P-gp was significantly inhibited, falling to 1.25 times the level of the blank group. At this time, the total serum response concentration of fucoxanthin climbed to 406.35pmol / mL, an increase of 9.69 times. It is explained that the preparation of fucoxanthin nanoparticles using the intestinal epithelial affinity adsorption-lipid transporter protein regulation coupling technology of the present invention can achieve efficient oral absorption of fucoxanthin, thereby laying a foundation for its function of improving vision.

[0146] (3) Evaluation of dark vision function

[0147] Test samples (12 groups): bilberry anthocyanin extract, fucoxanthin, zinc taurine, bilberry anthocyanin extract + fucoxanthin (mass ratio of 40:5 mixed evenly), bilberry anthocyanin extract + fucoxanthin + zinc taurine (mass ratio of 40:5:0.1 mixed evenly), bilberry anthocyanin extract + fucoxanthin + zinc taurine (mass ratio of 50:7:0.5 mixed evenly), bilberry anthocyanin extract + fucoxanthin + zinc taurine (mass ratio of 55:9:0.7 mixed evenly), bilberry anthocyanin extract + fucoxanthin + zinc taurine (mass ratio of 60:10:1 mixed evenly), and the formula products obtained in Examples 1 to 4 of the present invention.

[0148] Sample treatment: Dissolve the test sample directly in deionized water at a mass-to-volume ratio of 1:10 to obtain the test solution.

[0149] Experimental animals and feeding environment: SPF grade male ICR mice, 15-20 g, were purchased from Shanghai Slake Experimental Animal Co., Ltd. The basic feed ingredients for mice: carbohydrate 64%, protein 21%, fat 4%, fiber 5%, water 6%. The above feeds were purchased from Shanghai Slake Experimental Animal Co., Ltd. The feeding room temperature was 23±2℃, and the relative humidity was 55±5%.

[0150] Animal grouping and treatment method: The mice were adaptively raised for one week and fed freely. After one week, they were randomly divided into 13 groups according to body weight, namely, a blank group and 12 test groups, with 7 test animals in each group. Each mouse was gavaged with 200 μL of the test solution (12 groups) or deionized water (blank group) each time, and the gavage dose was 50 mg / kg. After 4 weeks, the dark-adapted 0.01ERG b-wave amplitude of the retina of each group of animals was detected by an electro-visual physiology instrument to evaluate the function of the rod cells in the retina, thereby reflecting the changes in night vision function.

[0151] Experimental results:

[0152] Table 5 Functional evaluation of product formulations on rod cells (dark vision)

[0153]

[0154]

[0155] Rod cells are mainly responsible for visual sensitivity in dark environments. The larger the b-wave amplitude of the retinal dark adaptation 0.01ERG, the better the rod cell function and the stronger the night vision. The results are shown in Table 5. Although the use of bilberry anthocyanin extract, fucoxanthin, and zinc taurine alone can improve the function of rod cells, the effect is very limited. When these three substances are combined in a specific ratio, the function of rod cells is greatly improved, and the b-wave amplitude is increased from 51.25±3.28μV in the blank group to 90.19±4.25~101.68±4.77μV, indicating that anthocyanin, fucoxanthin, and zinc taurine have a synergistic effect of improving night vision function within the mass ratio range set by the present invention. When the products of Examples 1 to 4 were used for intervention, the night vision function of mice was further improved. The products of Examples 1 to 4 can further increase the b-wave amplitude of 0.01ERG to 163.35±8.42, 178.24±7.84, 169.65±7.39, and 182.68±8.13 μV. This effect should be attributed to the innovative molecular encapsulation technology adopted in the present invention, which promotes the intestinal absorption efficiency of anthocyanins and fucoxanthin, thereby further enhancing the nutritional effect of the formula in improving night vision function.

[0156] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention without departing from the principles and intent of the present invention.

Claims

1. A nutritious food for improving night vision function, characterized in that: The invention is prepared from anthocyanin-sterculic acid nanoparticles, fucoxanthin-vitamin A-resveratrol nanoparticles and zinc taurine; the weight ratio of the anthocyanin-sterculic acid nanoparticles, fucoxanthin-vitamin A-resveratrol nanoparticles and zinc taurine is 40-60:5-10:0.1-1.

2. The nutritious food for improving night vision function according to claim 1, characterized in that: The anthocyanin-sterculin acid nanoparticles are spherical particles formed by self-assembly of an aqueous solution of anthocyanin-anionic polysaccharide self-assembled complex and an aqueous solution of sterculin acid-mannose grafted protein ligand complex, preferably in a mass ratio of 1:1 to 1:4, and the sterculin acid-mannose grafted protein ligand complex wraps the anthocyanin-anionic polysaccharide self-assembled complex in the inner core, and the particle size is 300 to 500 nm; Preferably, the preparation method of the anthocyanin-sterculic acid nanoparticles is: The pH of the aqueous solution of the sterculia acid-mannose grafted protein ligand complex is adjusted to acidic, preferably, the acidic pH is 3.0±0.5; then, it is added to the aqueous solution of the anthocyanin-anionic polysaccharide self-assembled complex, and the reaction is preferably carried out in the dark for 30±5 minutes, so that the sterculia acid-mannose grafted protein ligand complex is tightly adsorbed on the outer layer of the anthocyanin-anionic polysaccharide self-assembled complex, and anthocyanin-sterculia acid nanoparticles are obtained after freeze-drying; preferably, the mass ratio of the anthocyanin-anionic polysaccharide self-assembled complex to the sterculia acid-mannose grafted protein ligand complex is 1:1 to 1:4; Preparation of the aqueous solution of the sterculia acid-mannose grafted protein ligand complex: 1) Mixing protein and mannose and dissolving them in deionized water, preferably, the mass ratio of protein to mannose is 1:5; after freeze-drying, placing the mixed powder in a sealed container for reaction, preferably, the reaction conditions in the sealed container are a sealed container with a relative humidity of 79±1%, and reacting at 65±5°C for 48±3h; obtaining a mannose grafted protein with a grafting degree of not less than 20%; 2) Dissolving the mannose grafted protein in deionized water and adjusting the pH to alkaline, preferably, the alkaline pH is 11.0±1; then adding dissolved sterculia acid, and then adjusting the pH to neutral; allowing sterculia acid to enter the hydrophobic cavity of the mannose grafted protein in the form of a ligand to obtain a sterculia acid-mannose grafted protein ligand complex aqueous solution; the mass ratio of the mannose grafted protein to sterculia acid is 1:0.02 to 1:0.08; Preparation of the anthocyanin-anionic polysaccharide self-assembly complex: dissolving anthocyanin and anionic polysaccharide in deionized water in proportion, and adjusting the pH to acidic, preferably, the weight ratio of anthocyanin to anionic polysaccharide is 1:40-86; the acidic pH is 3.0±0.5; then performing ultrasonic treatment to obtain an aqueous solution of anthocyanin-anionic polysaccharide self-assembly complex; Preferably, the anthocyanin is one of anthocyanin extracts of bilberry, grape, raspberry, and black wolfberry, and the total anthocyanin content is ≥ 90% by mass; and / or The anionic polysaccharide is one of sulfated polysaccharide, hyaluronic acid, pectin and sodium alginate; and / or The protein is one of bovine serum albumin, whey protein, ovalbumin and soy protein.

3. The nutritious food for improving night vision function according to claim 1, characterized in that: The fucoxanthin-vitamin A-resveratrol nanoparticles are spherical particles formed by self-assembly of an aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles and mannose-grafted chitosan in a mass ratio of 15:1 to 15:4, and the mannose-grafted chitosan wraps the fucoxanthin-vitamin A-resveratrol-protein particles in the inner core, and the particle size is 400 to 600 nm; Preferably, the preparation method of the fucoxanthin-vitamin A-resveratrol nanoparticles is: Adding mannose-grafted chitosan to an aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles, wherein the concentration of mannose-grafted chitosan is 0.1 mass % to 0.4 mass %, and reacting at room temperature in the dark for preferably 30±5 min, so that the mannose-grafted chitosan is adsorbed on the outer layer of the protein particles, and then freeze-dried to obtain fucoxanthin-vitamin A-resveratrol nanoparticles; The preparation of the mannose-grafted chitosan comprises: dissolving mannose in a chitosan solution so that the mass ratio of chitosan to mannose is 1:3; adjusting the pH of the chitosan-mannose mixed solution to an acidic reaction, preferably, pH 5.5±1, and reacting at 65±5° C. for 60±12 h; cooling after the reaction, and freeze-drying to obtain a water-soluble mannose-grafted chitosan; The aqueous solution of the fucoxanthin-vitamin A-resveratrol-protein particles is prepared by mixing and dissolving fucoxanthin, vitamin A, resveratrol and oleic acid, adding them to alkaline deionized water, and ultrasonicating them to homogenize and aggregate the above-mentioned fat-soluble components to obtain mixed micelles of fucoxanthin, vitamin A, resveratrol and oleic acid; mixing the mixed micelles with an alkaline protein solution, reacting them at room temperature in the dark, and then adjusting the pH back to neutral to obtain an aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles with a protein shell; preferably, the mass ratio of fucoxanthin, vitamin A, resveratrol and oleic acid is 2:1:1:1; the ultrasonication time is 2±0.5min; the concentration of the protein solution is 2% by mass, and the pH of the alkaline deionized water and the alkaline protein solution is independently 11.0±1; the reaction time in the dark is 30±5min; Preferably, the protein is one of bovine serum albumin, whey protein, ovalbumin and soy protein.

4. The nutritious food for improving night vision function according to claim 1, characterized in that: The nutritional food is a capsule, a tablet, an granule, an oral liquid or a beverage.

5. The method for preparing the nutritious food for improving night vision function according to claim 1, characterized in that: The method comprises the following steps: The preparation method of anthocyanin-sterculic acid nanoparticles is: S1. Preparation of anthocyanin-sterculic acid nanoparticles: S1.1 Preparation of aqueous solution of sterculic acid-mannose grafted protein ligand complex: protein and mannose are mixed and dissolved in deionized water, and the mixed powder is placed in a sealed container for reaction after freeze-drying to obtain mannose-grafted protein; preferably, the mass ratio of protein to mannose is 1:5; the mass concentration of protein is 2%; the reaction conditions in the sealed container are a relative humidity of 79±1% in a sealed container, and a reaction temperature of 65±5°C for 48±3h; the grafting degree of mannose-grafted protein is 20±4%; The mannose-grafted protein is dissolved in deionized water, and the pH is adjusted to alkaline, and then dissolved sterculia acid is added, and then the pH is adjusted to neutral, so that sterculia acid enters the hydrophobic cavity of the mannose-grafted protein in the form of a ligand, and a sterculia acid-mannose-grafted protein ligand complex aqueous solution is obtained; preferably, the alkaline pH is 11.0±1; preferably, the mass ratio of the mannose-grafted protein to sterculia acid is 1:0.02 to 1:0.08; S1.2 Preparation of anthocyanin-anionic polysaccharide self-assembled complex aqueous solution: dissolve anthocyanin and anionic polysaccharide in deionized water, and adjust the pH to acidic; then perform ultrasonic treatment to obtain an aqueous solution of anthocyanin-anionic polysaccharide self-assembled complex; preferably, the weight ratio of anthocyanin to anionic polysaccharide is 1:40-86; the acidic pH is 3.0±1; The above steps S1.1 and S1.2 can be performed in different orders or simultaneously; S1.3 The pH of the aqueous solution of the sterculic acid-mannose grafted protein ligand complex is adjusted to acidic, and then added to the aqueous solution of the anthocyanin-anionic polysaccharide self-assembled complex, and the reaction is carried out in the dark, so that the sterculic acid-mannose grafted protein ligand complex is tightly adsorbed on the outer layer of the anthocyanin-anionic polysaccharide self-assembled complex, and anthocyanin-sterculic acid nanoparticles are obtained after freeze-drying; preferably, the acidic pH is 3.0±1; the mass ratio of the anthocyanin-anionic polysaccharide self-assembled complex to the sterculic acid-mannose grafted protein ligand complex is 1:1 to 1:4; the reaction time in the dark is 30±5min; S2. The preparation method of fucoxanthin-vitamin A-resveratrol nanoparticles is: S2.1 Preparation of mannose-grafted chitosan: dissolving mannose in a chitosan solution, preferably, making the mass ratio of chitosan to mannose be 1:3; adjusting the pH of the chitosan-mannose mixed solution to acidic and reacting; cooling after the reaction, and freeze-drying to obtain water-soluble mannose-grafted chitosan; preferably, the final concentration of the chitosan solution is 5±1 g / L, and the solvent is 1% by volume acetic acid aqueous solution; the pH of the acidic reaction is 5.5±1, and the reaction conditions are 65±5°C for 60±12h; S2.2 Preparation of aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles: fucoxanthin, vitamin A, resveratrol and oleic acid are mixed and dissolved, and then added into alkaline deionized water, and ultrasonicated to homogenize and aggregate the above fat-soluble components to obtain mixed micelles of fucoxanthin, vitamin A, resveratrol and oleic acid; the mixed micelles are mixed with alkaline protein solution, reacted at room temperature in the dark, and then the pH is adjusted back to neutral to obtain fucoxanthin-vitamin A-resveratrol-protein particles with a protein shell; preferably, the weight ratio of fucoxanthin, vitamin A, resveratrol and oleic acid is 2:1:1:1; the ultrasonication time is 2±0.5min; the concentration of the protein solution is 2% by mass, and the pH of the alkaline deionized water and the alkaline protein solution is independently 11.0±1; the reaction time in the dark is 30±5min; The above steps S2.1 and S2.2 can be performed in different orders or simultaneously. S2.3 adding mannose-grafted chitosan to the aqueous solution of fucoxanthin-vitamin A-resveratrol-protein particles, wherein the concentration of mannose-grafted chitosan is 0.1 mass % to 0.4 mass %, and reacting at room temperature in the dark for preferably 30±5 min, so that the mannose-grafted chitosan is adsorbed on the outer layer of the protein particles, and lyophilizing to obtain fucoxanthin-vitamin A-resveratrol nanoparticles; The above steps S1 and S2 can be exchanged in order or performed simultaneously; S3. Mix anthocyanin-sterculic acid nanoparticles, fucoxanthin-vitamin A-resveratrol nanoparticles, zinc taurine, water or solid filler excipients according to proportion.

6. The method for preparing the nutritious food for improving night vision function as claimed in claim 5, characterized in that: Also includes: S4. Homogenize and filter the aqueous mixture, or sieve the solid mixture.

7. The method for preparing the nutritious food for improving night vision function as claimed in claim 6, characterized in that: Also includes: S5. Filling after sterilization, or packaging after tableting or granulation.

8. The method for preparing the nutritious food for improving night vision function according to any one of claims 5 to 7, characterized in that: In the preparation of the aqueous solution of anthocyanin-anionic polysaccharide self-assembled complex in step S1.2, the ultrasonic power is 100 to 500 W and the ultrasonic time is 1 to 5 min; and / or The mass ratio of anthocyanin to anionic polysaccharide is 1:40 to 1:

80.

9. The method for preparing the nutritious food for improving night vision function according to any one of claims 5 to 7, characterized in that: In the preparation of the mannose grafted chitosan in step S2.1, the viscosity of the chitosan is 150 mPa·s, and the grafting degree of the mannose grafted chitosan is 35±3%.

10. The nutritious food for improving night vision function according to any one of claims 1 to 4 or the nutritious food for improving night vision function prepared by any one of the preparation methods according to claims 5 to 9 has the effect of improving night vision function.

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