Functional food for reducing fat and losing weight and preparation method thereof

Abstract

The invention discloses a functional food for reducing fat and losing weight and a preparation method thereof. Comprising the following components in parts by weight: 30 to 50 parts of limonin-rutin-isoquercitrin-folic acid nanoparticles, 20 to 40 parts of fucoxanthin-glycyrrhizic acid-isoquercitrin-mannose nanoparticles, 0.1 to 0.5 part of vitamin D3-linoleic acid-isoquercitrin-hyaluronic acid nanoparticles, and the balance of water or solid filler auxiliary materials. The surfaces of limonin, fucoxanthin and vitamin D3 nanoparticles are respectively modified by folic acid, mannose and hyaluronic acid, and a biological enhancer and isoquercitrin are introduced, so that efficient intestinal absorption of weight-losing components such as limonin, fucoxanthin and vitamin D3 is promoted, meanwhile, the components are subjected to formula design, and the weight-losing effect is improved. The fat-reducing and weight-losing effects of the formula product are greatly improved, the fat accumulation is effectively reduced, and the body weight is reduced; the product can be prepared into capsules, tablets, electuary, oral liquid or beverage.

Description

Technical Field

[0001] The invention relates to the field of functional foods, and in particular to a functional food for reducing fat and weight and a preparation method thereof. Background Art

[0002] With the rapid development of society, unhealthy diet, irregular work and rest schedule, high stress, lack of aerobic exercise and other factors have led to more and more people becoming overweight or even obese. Obesity can lead to disorders of sugar and lipid metabolism, increasing the risk of cardiovascular disease, diabetes, fatty liver and other diseases; at the same time, it can also affect personal image and life, causing great psychological burden. However, existing weight loss drugs have problems such as large side effects, poor weight loss effects, and easy rebound. Therefore, functional foods that are green, safe, and have significant weight loss effects have become products that are urgently needed.

[0003] Studies have shown that some special dietary nutrients can effectively regulate sugar and lipid metabolism, thereby achieving the effect of reducing fat and losing weight. These dietary nutrients include limonin, fucoxanthin, vitamin D 3 Etc. Most of these nutrients are fat-soluble and 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 can effectively improve the water solubility and stability of the above nutrients, it is still difficult to take into account the nutritional needs of efficient intestinal absorption. This is because conventional micro-nano encapsulation particles are difficult to achieve the aggregation of nutrients on the outside of intestinal epithelial cells, and lack the driving effect to promote the interaction between nutrients and intestinal epithelial cells. At the same time, these nutrients are strictly regulated by lipid transport proteins when transported across the membrane. In addition, there are multidrug resistance proteins in intestinal epithelial cells, which mediate the cells to excrete the ingested nutrients into the intestinal cavity. Therefore, in order to enable these nutrients to fully exert their biological functions, it is necessary to break through these key technologies: (1) fully enrich or adsorb on the outside of intestinal epithelial cells to increase the chance of absorption by intestinal cells; (2) promote the expression of transport proteins responsible for transmembrane transport of nutrients to promote cellular uptake and absorption; (3) inhibit the expression of P-gp multidrug resistance protein to reduce the nutrients taken up by intestinal epithelial cells from being re-excreted into the intestinal cavity.

[0004] Products that address these key technologies are urgently needed to enable nutrients to fully exert their biological functions. Summary of the invention

[0005] The purpose of the present invention is to provide a functional food with synergistic product ingredients, efficient absorption of core functional ingredients, and significant lipid-lowering and weight-loss effects through product formula design and production process innovation.

[0006] To achieve the above-mentioned object, the present invention provides a functional food for reducing fat and weight, characterized in that it comprises limonin-rutin-isoquercetin-folic acid nanoparticles, fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles, vitamin D3-linoleic acid-isoquercetin-hyaluronic acid nanoparticles, and water or solid filler auxiliary materials; wherein the limonin-rutin-isoquercetin-folic acid nanoparticles, fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles, vitamin D3-linoleic acid-isoquercetin-hyaluronic acid nanoparticles, and water or solid filler auxiliary materials; 3 -The weight ratio of linoleic acid-isoquercetin-hyaluronic acid nanoparticles is (30-50):(20-40):(0.1-0.5).

[0007] Furthermore, in the limonin-rutin-isoquercetin-folic acid nanoparticles, the mass ratio of limonin, rutin, and isoquercetin is 5:(0.5-2):(0.5-2);

[0008] Preferably, the preparation method of limonin-rutin-isoquercetin-folic acid nanoparticles is:

[0009] S1.1 Preparation of folic acid-sodium alginate complex: folic acid and sodium alginate are mixed and dissolved in deionized water, and the mixed powder of folic acid and sodium alginate is placed in a container for reaction after freeze-drying to obtain a folic acid-sodium alginate complex; preferably, the mass ratio of folic acid to sodium alginate is 1:(10-30); the reaction in the container is placed in a closed container with a relative humidity of 80% and reacted at 60°C for 24 hours;

[0010] S1.2 Preparation of limonin-rutin-isoquercetin self-assembly complex: After dissolving limonin, rutin and isoquercetin, add them into the protein aqueous solution and mix well, and denature the protein to obtain an aqueous solution of limonin-rutin-isoquercetin self-assembly complex; preferably, the mass ratio of limonin, rutin and isoquercetin is 5:(0.5-2):(0.5-2), the concentration of the protein aqueous solution is 2% to 4% by mass, the mass ratio of limonin to protein is 1:(20-40), and the protein is denatured by ultrasound, pulsed electric field, high static pressure, and microwave treatment; more preferably, the ultrasound condition is 350±50W for 3±1min;

[0011] The above S1.1 and S1.2 can be switched in order or performed simultaneously;

[0012] S1.3 adding folic acid-sodium alginate complex to the aqueous solution of limonin-rutin-isoquercetin self-assembled complex, preferably, the concentration of folic acid-sodium alginate complex in the aqueous solution is 0.1 mass % to 0.5 mass %; adjusting the pH to acidic so that the protein shell has the opposite electrical properties to folic acid-sodium alginate, reacting at room temperature in the dark so that the folic acid-sodium alginate complex is tightly adsorbed on the protein shell of the limonin-rutin-isoquercetin self-assembled complex, centrifuging, and freeze-drying to obtain limonin-rutin-isoquercetin-folic acid nanoparticles;

[0013] Preferably, the acidity is pH 3.0-4.0; and the reaction time at room temperature and in the dark is 30±10 min.

[0014] Furthermore, in the fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles, the mass ratio of fucoxanthin, glycyrrhizic acid, and isoquercetin is 5:(5-30):(1-3);

[0015] Preferably, the preparation method of fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles is:

[0016] S2.1 Preparation of mannose-grafted chitosan: dissolve mannose in an acetic acid aqueous solution of chitosan, adjust the pH of the chitosan-mannose mixed solution to weak acidity, react; ice bath, freeze-dry to obtain water-soluble mannose-grafted chitosan; preferably, the concentration of the acetic acid aqueous solution is 1% by volume, and the final concentration of chitosan in the acetic acid aqueous solution is 5g / L; the mass ratio of chitosan to mannose is 1:(2-5); the weakly acidic pH is 4.0-5.5, and the reaction conditions are 72h at 65°C; the ice bath time is 10min;

[0017] S2.2 Preparation of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles: Dissolve fucoxanthin, glycyrrhizic acid and isoquercetin and add them to a protein aqueous solution, adjust the pH to alkaline, partially denature the protein and expose the hydrophobic amino acids, react at room temperature in the dark, and adjust the pH back to neutral after the reaction to obtain an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles with a protein shell; preferably, the mass ratio of fucoxanthin, glycyrrhizic acid and isoquercetin is 5:(5-30):(1-3), the concentration of the protein aqueous solution is 2% to 5% by mass, the mass ratio of fucoxanthin to protein is 1:(20-50), the alkaline pH is 9.0-11.0, and the reaction time at room temperature in the dark is 30±10 min;

[0018] The above S2.1 and S2.2 can be performed in reverse order or simultaneously;

[0019] S2.3 adding mannose-grafted chitosan to an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles, preferably, the concentration of mannose-grafted chitosan in the aqueous solution is 0.1 mass % to 0.5 mass %; reacting at room temperature in the dark so that the mannose-grafted chitosan covers the outer layer of the protein particles, concentrating under reduced pressure, and freeze-drying to obtain fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles;

[0020] Preferably, the reaction time at room temperature and in the dark is 30±10 min.

[0021] Furthermore, the vitamin D 3 -Linoleic acid-isoquercetin-hyaluronic acid nanoparticles, vitamin D 3 The dosage ratio of linoleic acid and isoquercetin is 6:(10-25):(0.5-2);

[0022] Preferably, vitamin D 3 -The preparation method of linoleic acid-isoquercetin-hyaluronic acid nanoparticles is:

[0023] S3.1 Vitamin D 3 - Linoleic acid-isoquercetin-protein particle preparation: Vitamin D 3 After the linoleic acid and isoquercetin are dissolved, they are added to the protein aqueous solution, and the pH is adjusted to alkaline to partially denature the protein and expose the hydrophobic amino acids. After the reaction is carried out at room temperature and away from light, the pH is adjusted back to neutral to obtain vitamin D with a protein shell. 3 - an aqueous solution of linoleic acid-isoquercetin-protein particles; preferably, vitamin D 3 The mass ratio of linoleic acid and isoquercetin is 6:(10-25):(0.5-2), the concentration of the protein aqueous solution is 1% to 3% by mass, and vitamin D 3 The mass ratio of protein is 1:(10-30), the alkaline pH is 9.0-11.0, and the reaction time is 30±10min at room temperature and in the dark;

[0024] S3.2 Vitamin D 3 - Obtaining linoleic acid-isoquercetin-hyaluronic acid nanoparticles: Adding hyaluronic acid to the vitamin D 3 -linoleic acid-isoquercetin-protein particles in an aqueous solution, preferably, the concentration of hyaluronic acid in the aqueous solution is 0.05 mass % to 0.2 mass %; and the pH is adjusted to acidic so that the protein shell has an opposite electrical property to that of the hyaluronic acid, react at room temperature in the dark so that the hyaluronic acid is fully adsorbed on the outer layer of the protein particles, centrifuged, and freeze-dried to obtain vitamin D 3 -linoleic acid-isoquercetin-hyaluronic acid nanoparticles; preferably, the acidic pH is 3.0-4.0, and the reaction time at room temperature and away from light is 30±10 min.

[0025] Furthermore, the functional food is a capsule, a tablet, an granule, an oral liquid or a beverage.

[0026] The present invention also provides a method for preparing the functional food for reducing fat and weight, which comprises the following steps:

[0027] S1. Preparation of limonin-rutin-isoquercetin-folic acid nanoparticles:

[0028] S1.1 Preparation of folic acid-sodium alginate complex: folic acid and sodium alginate are mixed and dissolved in deionized water, and the mixed powder of folic acid and sodium alginate is placed in a container for reaction after freeze-drying to obtain a folic acid-sodium alginate complex; preferably, the mass ratio of folic acid to sodium alginate is 1:(10-30); the reaction in the container is placed in a closed container with a relative humidity of 80% and reacted at 60°C for 24 hours;

[0029] S1.2 Preparation of limonin-rutin-isoquercetin self-assembly complex: After dissolving limonin, rutin and isoquercetin, add them into the protein aqueous solution and mix well, and denature the protein to obtain an aqueous solution of limonin-rutin-isoquercetin self-assembly complex; preferably, the mass ratio of limonin, rutin and isoquercetin is 5:(0.5-2):(0.5-2), the concentration of the protein aqueous solution is 2% to 4% by mass, the mass ratio of limonin to protein is 1:(20-40), and the protein is denatured by ultrasound, pulsed electric field, high static pressure, and microwave treatment; more preferably, the ultrasound condition is 350±50W for 3±1min;

[0030] The above S1.1 and S1.2 can be switched in order or performed simultaneously;

[0031] S1.3. Adding folic acid-sodium alginate complex to the aqueous solution of limonin-rutin-isoquercetin self-assembled complex, preferably, the concentration of folic acid-sodium alginate complex in the aqueous solution is 0.1 mass % to 0.5 mass %; adjusting the pH to acidic so that the protein shell has the opposite electrical properties to the dietary polysaccharide, reacting at room temperature in the dark so that the folic acid-sodium alginate complex is tightly adsorbed on the protein shell of the limonin-rutin-isoquercetin self-assembled complex, centrifuging, and freeze-drying to obtain limonin-rutin-isoquercetin-folic acid nanoparticles;

[0032] Preferably, the acidity is pH 3.0-4.0; the reaction time at room temperature and in the dark is 30±10 min;

[0033] S2. Preparation of fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles

[0034] S2.1 Preparation of mannose-grafted chitosan: dissolve mannose in an acetic acid aqueous solution of chitosan, adjust the pH of the chitosan-mannose mixed solution to weak acidity, react; ice bath, freeze-dry to obtain water-soluble mannose-grafted chitosan; preferably, the concentration of the acetic acid aqueous solution is 1% by volume, and the final concentration of chitosan in the acetic acid aqueous solution is 5g / L; the mass ratio of chitosan to mannose is 1:(2-5); the weakly acidic pH is 4.0-5.5, and the reaction conditions are 72h at 65°C; the ice bath time is 10min;

[0035] S2.2 Preparation of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles: Dissolve fucoxanthin, glycyrrhizic acid and isoquercetin and add them to a protein aqueous solution, adjust the pH to alkaline, partially denature the protein and expose the hydrophobic amino acids, react at room temperature in the dark, and adjust the pH back to neutral after the reaction to obtain an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles with a protein shell; preferably, the mass ratio of fucoxanthin, glycyrrhizic acid and isoquercetin is 5:(5-30):(1-3), the concentration of the protein aqueous solution is 2% to 5% by mass, the mass ratio of fucoxanthin to protein is 1:(20-50), the alkaline pH is 9.0-11.0, and the reaction time at room temperature in the dark is 30±10 min;

[0036] The above S2.1 and S2.2 can be performed in reverse order or simultaneously;

[0037] S2.3 adding mannose-grafted chitosan to an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles, preferably, the concentration of mannose-grafted chitosan in the aqueous solution is 0.1 mass % to 0.5 mass %; reacting at room temperature in the dark so that the mannose-grafted chitosan covers the outer layer of the protein particles, concentrating under reduced pressure, and freeze-drying to obtain fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles;

[0038] Preferably, the reaction time at room temperature and away from light is 30±10 min;

[0039] S3. Vitamin D 3 -Preparation of linoleic acid-isoquercetin-hyaluronic acid nanoparticles:

[0040] S3.1 Vitamin D 3 - Linoleic acid-isoquercetin-protein particle preparation: Vitamin D 3 After the linoleic acid and isoquercetin are dissolved, they are added to the protein aqueous solution and the pH is adjusted to alkaline to partially denature the protein and expose the hydrophobic amino acids. After the reaction is carried out at room temperature and away from light, the pH is adjusted back to neutral to obtain vitamin D with a protein shell. 3 - an aqueous solution of linoleic acid-isoquercetin-protein particles; preferably, vitamin D3 The mass ratio of linoleic acid and isoquercetin is 6:(10-25):(0.5-2), the concentration of the protein aqueous solution is 1% to 3% by mass, and vitamin D 3 The mass ratio of protein is 1:(10-30), the alkaline pH is 9.0-11.0, and the reaction time is 30±10min at room temperature and in the dark;

[0041] S3.2 Vitamin D 3 - Obtaining linoleic acid-isoquercetin-hyaluronic acid nanoparticles: Adding hyaluronic acid to vitamin D 3 -linoleic acid-isoquercetin-protein particles in an aqueous solution, and adjust the pH to acidic, react at room temperature in the dark, so that hyaluronic acid is fully adsorbed on the outer layer of the protein particles, centrifuged, and freeze-dried to obtain vitamin D 3 - linoleic acid-isoquercetin-hyaluronic acid nanoparticles; preferably, the acidic pH is 3.0-4.0, and the reaction time at room temperature and away from light is 30±10 min;

[0042] The above S1, S2, and S3 can be interchanged in order or performed simultaneously;

[0043] S4. The required limonin-rutin-isoquercetin-folic acid nanoparticles, fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles, vitamin D 3 -linoleic acid-isoquercetin-hyaluronic acid nanoparticles, water or solid filler excipients are mixed evenly.

[0044] Furthermore, the method further comprises the following step: S5. if the obtained mixture is an aqueous solution, homogenizing and filtering the aqueous solution mixture; if the obtained mixture is a solid mixture, sieving the solid mixture.

[0045] Furthermore, the method further comprises the following steps: S6. sterilizing after filling, or packaging after tableting or granulation.

[0046] Furthermore, the protein is one of bovine serum albumin, whey protein, ovalbumin, and soy protein.

[0047] Further, in step S1, the folic acid-sodium alginate complex is added to the aqueous solution of the limonin-rutin-isoquercetin self-assembled complex, and the concentration of the folic acid-sodium alginate complex in the aqueous solution is 0.1% to 0.5% by mass; and / or

[0048] In the step S2, mannose-grafted chitosan is added to the aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles, and the concentration of mannose-grafted chitosan in the aqueous solution is 0.1% to 0.5% by mass; and / or

[0049] In the step S3.2, hyaluronic acid is added to the aqueous solution of vitamin D3-linoleic acid-isoquercetin-protein particles, and the concentration of hyaluronic acid in the aqueous solution is 0.05 mass % to 0.2 mass %.

[0050] In the preparation of the limonin-rutin-isoquercetin self-assembly complex, limonin, rutin, and isoquercetin are dissolved, added to the protein aqueous solution and mixed, and the protein is denatured to obtain an aqueous solution of the limonin-rutin-isoquercetin self-assembly complex. The method for denaturing the protein is not limited to ultrasound, and can also be a pulsed electric field, high static pressure, or microwave treatment. The ultrasound condition is 350±50W for 3±1min.

[0051] According to the process of the present invention, the particle size of the prepared limonin-rutin-isoquercetin-folic acid nanoparticles is 320nm-570nm. The particle size of the fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles is 400nm-650nm. Vitamin D 3 -The particle size of linoleic acid-isoquercetin-hyaluronic acid nanoparticles is 250nm-340nm. Among them, limonin nanoparticles, fucoxanthin nanoparticles and vitamin D 3 After heating at 121°C for 15 min, the retention rate of the active ingredients was ≥90%.

[0052] The present invention has the following advantages:

[0053] (1) Taking full advantage of the physiological characteristics of intestinal epithelium, such as its ability to express folate receptors, mannose receptors, and hyaluronic acid receptors, the 3 The surface of the nanoparticles was modified with folic acid, mannose and hyaluronic acid, respectively, which can promote the absorption of limonin, fucoxanthin and vitamin D. 3 Adsorption and aggregation on the outside of intestinal epithelial cells;

[0054] (2) Given that CD36, SR-B1, and VDR are responsible for limonin, fucoxanthin, and vitamin D, respectively 3 The main transporters transported across intestinal epithelial cells were introduced into their corresponding delivery systems with bioenhancers that can promote the expression of the corresponding transporters: rutin, glycyrrhizic acid and linoleic acid, thereby improving the intestinal epithelial cells' uptake efficiency of the above three nutrients;

[0055] (3) Given that P-gp is responsible for the transport of limonin, fucoxanthin, and vitamin D into cells 3 The common multidrug resistance protein excreted into the intestinal cavity, isoquercetin that can inhibit P-gp expression was incorporated into the three vectors, so that the nutrients taken in by the intestinal epithelial cells were not excreted into the intestinal cavity, thereby effectively increasing the levels of limonin, fucoxanthin, and vitamin D. 3oral bioavailability;

[0056] (4) It fully draws on the multi-component "synergistic" formula design concept of traditional Chinese medicine theory to promote the production of limonin, fucoxanthin, and vitamin D. 3 On the basis of efficient intestinal absorption of weight loss ingredients such as chlorinated parabens, the above ingredients are formulated to give full play to the synergistic effect of lowering fat and losing weight, thus greatly improving the fat-lowering and weight loss efficacy of the formula products.

[0057] The present invention not only emphasizes the efficient oral absorption of nutrients in terms of preparation technology, but also the product has undergone strict formula design, fully ensuring the green safety and nutritional functionality of the product. DETAILED DESCRIPTION

[0058] 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.

[0059] The preparation method of the functional food for reducing fat and weight comprises the following steps:

[0060] S1. Preparation of limonin-rutin-isoquercetin-folic acid nanoparticles:

[0061] S1.1 Preparation of folic acid-sodium alginate complex: folic acid and sodium alginate are mixed and dissolved in deionized water, and the mixed powder of folic acid and sodium alginate is placed in a container for reaction after freeze-drying to obtain a folic acid-sodium alginate complex; preferably, the mass ratio of folic acid to sodium alginate is 1:(10-30); the reaction in the container is placed in a closed container with a relative humidity of 80% and reacted at 60°C for 24 hours;

[0062] S1.2 Preparation of limonin-rutin-isoquercetin self-assembly complex: After dissolving limonin, rutin and isoquercetin, add them into the protein aqueous solution and mix well, and denature the protein to obtain an aqueous solution of limonin-rutin-isoquercetin self-assembly complex; preferably, the mass ratio of limonin, rutin and isoquercetin is 5:(0.5-2):(0.5-2), the concentration of the protein aqueous solution is 2% to 4% by mass, the mass ratio of limonin to protein is 1:(20-40), and the protein is denatured by ultrasound, pulsed electric field, high static pressure, and microwave treatment; more preferably, the ultrasound condition is 350±50W for 3±1min;

[0063] The above S1.1 and S1.2 can be switched in order or performed simultaneously;

[0064] S1.3. Adding folic acid-sodium alginate complex to the aqueous solution of limonin-rutin-isoquercetin self-assembled complex, preferably, the concentration of folic acid-sodium alginate complex in the aqueous solution is 0.1 mass % to 0.5 mass %; adjusting the pH to acidic so that the protein shell has the opposite electrical properties to the dietary polysaccharide, reacting at room temperature in the dark so that the folic acid-sodium alginate complex is tightly adsorbed on the protein shell of the limonin-rutin-isoquercetin self-assembled complex, centrifuging, and freeze-drying to obtain limonin-rutin-isoquercetin-folic acid nanoparticles;

[0065] Preferably, the acidity is pH 3.0-4.0; the reaction time at room temperature and in the dark is 30±10 min;

[0066] S2. Preparation of fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles

[0067] S2.1 Preparation of mannose-grafted chitosan: dissolve mannose in an acetic acid aqueous solution of chitosan, adjust the pH of the chitosan-mannose mixed solution to weak acidity, react; ice bath, freeze-dry to obtain water-soluble mannose-grafted chitosan; preferably, the concentration of the acetic acid aqueous solution is 1% by volume, and the final concentration of chitosan in the acetic acid aqueous solution is 5g / L; the mass ratio of chitosan to mannose is 1:(2-5); the weakly acidic pH is 4.0-5.5, and the reaction conditions are 72h at 65°C; the ice bath time is 10min;

[0068] S2.2 Preparation of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles: Dissolve fucoxanthin, glycyrrhizic acid and isoquercetin and add them to a protein aqueous solution, adjust the pH to alkaline, partially denature the protein and expose the hydrophobic amino acids, react at room temperature in the dark, and adjust the pH back to neutral after the reaction to obtain an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles with a protein shell; preferably, the mass ratio of fucoxanthin, glycyrrhizic acid and isoquercetin is 5:(5-30):(1-3), the concentration of the protein aqueous solution is 2% to 5% by mass, the mass ratio of fucoxanthin to protein is 1:(20-50), the alkaline pH is 9.0-11.0, and the reaction time at room temperature in the dark is 30±10 min;

[0069] The above S2.1 and S2.2 can be performed in reverse order or simultaneously;

[0070] S2.3 adding mannose-grafted chitosan to an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles, preferably, the concentration of mannose-grafted chitosan in the aqueous solution is 0.1 mass % to 0.5 mass %; reacting at room temperature in the dark so that the mannose-grafted chitosan covers the outer layer of the protein particles, concentrating under reduced pressure, and freeze-drying to obtain fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles;

[0071] Preferably, the reaction time at room temperature and away from light is 30±10 min;

[0072] S3. Vitamin D 3 -Preparation of linoleic acid-isoquercetin-hyaluronic acid nanoparticles:

[0073] S3.1 Vitamin D 3 - Linoleic acid-isoquercetin-protein particle preparation: Vitamin D 3 After the linoleic acid and isoquercetin are dissolved, they are added to the protein aqueous solution and the pH is adjusted to alkaline to partially denature the protein and expose the hydrophobic amino acids. After the reaction is carried out at room temperature and away from light, the pH is adjusted back to neutral to obtain vitamin D with a protein shell. 3 - an aqueous solution of linoleic acid-isoquercetin-protein particles; preferably, vitamin D 3 The mass ratio of linoleic acid and isoquercetin is 6:(10-25):(0.5-2), the concentration of the protein aqueous solution is 1% to 3% by mass, and vitamin D 3 The mass ratio of protein is 1:(10-30), the alkaline pH is 9.0-11.0, and the reaction time is 30±10min at room temperature and in the dark;

[0074] S3.2 Vitamin D 3 - Obtaining linoleic acid-isoquercetin-hyaluronic acid nanoparticles: Adding hyaluronic acid to vitamin D 3 -linoleic acid-isoquercetin-protein particles in an aqueous solution, and adjust the pH to acidic, react at room temperature in the dark, so that hyaluronic acid is fully adsorbed on the outer layer of the protein particles, centrifuged, and freeze-dried to obtain vitamin D 3 - linoleic acid-isoquercetin-hyaluronic acid nanoparticles; preferably, the acidic pH is 3.0-4.0, and the reaction time at room temperature and away from light is 30±10 min;

[0075] The above S1, S2, and S3 can be interchanged in order or performed simultaneously;

[0076] S4. The required limonin-rutin-isoquercetin-folic acid nanoparticles, fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles, vitamin D 3 -linoleic acid-isoquercetin-hyaluronic acid nanoparticles, water or solid filler excipients are mixed evenly.

[0077] Furthermore, the method further comprises the following step: S5. if the obtained mixture is an aqueous solution, homogenizing and filtering the aqueous solution mixture; if the obtained mixture is a solid mixture, sieving the solid mixture.

[0078] Furthermore, the method further comprises the following steps: S6. sterilizing after filling, or packaging after tableting or granulation.

[0079] Furthermore, the protein is one of bovine serum albumin, whey protein, ovalbumin, and soy protein.

[0080] Further, in step S1, the folic acid-sodium alginate complex is added to the aqueous solution of the limonin-rutin-isoquercetin self-assembled complex, and the concentration of the folic acid-sodium alginate complex in the aqueous solution is 0.1% to 0.5% by mass; and / or

[0081] In the step S2, mannose-grafted chitosan is added to the aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles, and the concentration of mannose-grafted chitosan in the aqueous solution is 0.1% to 0.5% by mass; and / or

[0082] In the step S3.2, hyaluronic acid is added to the aqueous solution of vitamin D3-linoleic acid-isoquercetin-protein particles, and the concentration of hyaluronic acid in the aqueous solution is 0.05 mass % to 0.2 mass %.

[0083] Example 1

[0084] S1. Preparation of limonin-rutin-isoquercetin-folic acid nanoparticles:

[0085] S1.1 Limonin, rutin, and isoquercetin were weighed and dissolved in a mass ratio of 5:0.5:0.5 (ethanol can be used as a solubilizing agent, and the same applies to other embodiments), and then added to a 2% protein aqueous solution, the mass ratio of limonin to bovine serum albumin was 1:20, and ultrasonicated for 4 minutes at a power of 300 W to obtain an aqueous solution of a limonin-rutin-isoquercetin self-assembled complex;

[0086] S1.2 Mix folic acid and sodium alginate in a mass ratio of 1:10 and dissolve in deionized water. After freeze-drying, place the mixed powder of folic acid and sodium alginate in a sealed container with a relative humidity of 80%, and react at 60° C. for 24 hours to obtain a folic acid-sodium alginate complex;

[0087] The above S1.1 and S1.2 can be performed in reverse order or simultaneously; the same applies to each of the following embodiments;

[0088] S1.3 Add folic acid-sodium alginate complex to the aqueous solution of limonin-rutin-isoquercetin self-assembled complex, the concentration of folic acid-sodium alginate complex in the aqueous solution is 0.1% by mass, then adjust the pH to 3.0, react at room temperature in the dark for 20 minutes, centrifuge, and freeze-dry to obtain limonin-rutin-isoquercetin-folic acid nanoparticles with a particle size of 320 nm.

[0089] S2. Preparation of fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles

[0090] S2.1 Fucoxanthin, glycyrrhizic acid and isoquercetin were mixed in a mass ratio of 5:5:1 and dissolved with ethanol, and then added to a 2% protein aqueous solution, the mass ratio of fucoxanthin to bovine serum albumin was 1:20, and the pH was adjusted to 9.0, reacted at room temperature in the dark for 20 minutes, and the pH was adjusted back to 7.0 to obtain an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles with a protein shell;

[0091] S2.2 Dissolve chitosan in 1% by volume acetic acid aqueous solution to a final concentration of 5 g / L; dissolve mannose in the chitosan solution to make the mass ratio of chitosan to mannose 1:2; adjust the pH of the chitosan-mannose mixed solution to 4.0, and react at 65°C for 72 hours; after the reaction is completed, place in an ice bath for 10 minutes, and freeze-dry to obtain water-soluble mannose-grafted chitosan;

[0092] The above S2.1 and S2.2 can be performed in reverse order or simultaneously; the same applies to each of the following embodiments;

[0093] S2.3 Add mannose-grafted chitosan to the aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles, the concentration of mannose-grafted chitosan in the aqueous solution is 0.1% by mass, react at room temperature in the dark for 20 minutes, concentrate under reduced pressure, and freeze-dry to obtain fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles with a particle size of 400 nm.

[0094] S3. Vitamin D 3 Preparation of linoleic acid-isoquercetin-hyaluronic acid nanoparticles

[0095] S3.1 Vitamin D 3 , linoleic acid, and isoquercetin were mixed in a mass ratio of 6:10:0.5, dissolved in ethanol, and added to a 1% protein aqueous solution. 3 The mass ratio of the mixture to bovine serum albumin was 1:10, and the pH was adjusted to 9.0. After reacting at room temperature and away from light for 20 minutes, the pH was adjusted back to 7.0 to obtain vitamin D with a protein shell. 3 - an aqueous solution of linoleic acid-isoquercetin-protein particles;

[0096] S3.2 Adding hyaluronic acid to vitamin D 3 -In the aqueous solution of linoleic acid-isoquercetin-protein particles, the concentration of hyaluronic acid in the aqueous solution is 0.05% by mass, and the pH is adjusted to 3.0, and the reaction is carried out at room temperature and in the dark for 20 minutes to allow the hyaluronic acid to be fully adsorbed on the outer layer of the protein particles, and then centrifuged and freeze-dried to obtain vitamin D 3-Linoleic acid-isoquercetin-hyaluronic acid nanoparticles, particle size is 250nm.

[0097] The above S1, S2, and S3 can be interchanged in order or performed simultaneously; the same applies to each of the following embodiments;

[0098] S4. 30 parts by weight of limonin-rutin-isoquercetin-folic acid nanoparticles, 20 parts by weight of fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles, and vitamin D 3 - Linoleic acid-isoquercetin-hyaluronic acid nanoparticles 0.1 parts by weight Weigh the above components, add appropriate amounts of magnesium stearate and starch to the formula, and mix well;

[0099] S5.Sieving;

[0100] S6. After sterilization, use a fully automatic capsule filling machine to fill it into hard capsules, with each capsule filling 0.5g, to obtain capsule functional food.

[0101] Example 2

[0102] S1. Preparation of limonin-rutin-isoquercetin-folic acid nanoparticles:

[0103] S1.1 Weigh limonin, rutin and isoquercetin in a mass ratio of 5:1:1 and dissolve them in ethanol. Then add them to a 3% protein aqueous solution, the mass ratio of limonin to whey protein is 1:30, and ultrasonicate for 2 minutes at a power of 350 W to obtain an aqueous solution of a limonin-rutin-isoquercetin self-assembled complex;

[0104] S1.2 Mix folic acid and sodium alginate in a mass ratio of 1:20 and dissolve in deionized water. After freeze-drying, place the mixed powder of folic acid and sodium alginate in a sealed container with a relative humidity of 80%, and react at 60° C. for 24 hours to obtain a folic acid-sodium alginate complex;

[0105] S1.3 Add folic acid-sodium alginate complex to the aqueous solution of limonin-rutin-isoquercetin self-assembled complex, the concentration of folic acid-sodium alginate complex in the aqueous solution is 0.3% by mass, then adjust the pH to 3.5, react at room temperature in the dark for 25 minutes, centrifuge, and freeze-dry to obtain limonin-rutin-isoquercetin-folic acid nanoparticles with a particle size of 425 nm.

[0106] S2. Preparation of fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles

[0107] S2.1 Fucoxanthin, glycyrrhizic acid and isoquercetin were mixed in a mass ratio of 5:10:2 and dissolved with ethanol. The mixture was then added to a 3% protein aqueous solution. The mass ratio of fucoxanthin to whey protein was 1:30. The pH was adjusted to 9.5. The mixture was reacted at room temperature in the dark for 25 minutes. The pH was adjusted back to 7.0 to obtain an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles with a protein shell;

[0108] S2.2 Dissolve chitosan in 1% by volume acetic acid aqueous solution to a final concentration of 5 g / L; dissolve mannose in the chitosan solution to make the mass ratio of chitosan to mannose 1:3; adjust the pH of the chitosan-mannose mixed solution to 4.5, and react at 65°C for 72 hours; after the reaction, ice bath for 10 minutes, and freeze-dry to obtain water-soluble mannose-grafted chitosan;

[0109] S2.3 Add mannose-grafted chitosan to the aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles, the concentration of mannose-grafted chitosan in the aqueous solution is 0.3% by mass, react at room temperature in the dark for 25 minutes, concentrate under reduced pressure, and freeze-dry to obtain fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles with a particle size of 509 nm.

[0110] S3. Vitamin D 3 Preparation of linoleic acid-isoquercetin-hyaluronic acid nanoparticles

[0111] S3.1 Vitamin D 3 , linoleic acid, and isoquercetin were mixed in a mass ratio of 6:15:1, dissolved in ethanol, and added to a 2% protein aqueous solution. 3 The mass ratio of the mixture to whey protein was 1:20, and the pH was adjusted to 10.0. After reacting at room temperature and away from light for 25 minutes, the pH was adjusted back to 7.0 to obtain vitamin D with a protein shell. 3 - an aqueous solution of linoleic acid-isoquercetin-protein particles;

[0112] S3.2 Adding hyaluronic acid to vitamin D 3 -In the aqueous solution of linoleic acid-isoquercetin-protein particles, the concentration of hyaluronic acid in the aqueous solution is 0.1% by mass, and the pH is adjusted to 3.5, and the reaction is carried out at room temperature and in the dark for 25 minutes to allow the hyaluronic acid to be fully adsorbed on the outer layer of the protein particles, and then centrifuged and freeze-dried to obtain vitamin D 3 -Linoleic acid-isoquercetin-hyaluronic acid nanoparticles, particle size is 278nm.

[0113] S4 is composed of 40 parts by weight of limonin-rutin-isoquercetin-folic acid nanoparticles, 30 parts by weight of fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles, and vitamin D 3-linoleic acid-isoquercetin-hyaluronic acid nanoparticles 0.3 parts by weight Weigh the above components, then add appropriate amounts of dextrin, starch, microcrystalline cellulose and lactose, one or more auxiliary materials, mix well, moisten with appropriate amount of 70% (v / v) ethanol to make a soft material,

[0114] S5 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 functional food.

[0115] Example 3

[0116] S1. Preparation of limonin-rutin-isoquercetin-folic acid nanoparticles:

[0117] S1.1 Weigh limonin, rutin, and isoquercetin in a mass ratio of 5:1.5:1.5, dissolve them in ethanol, and then add them to a 3.5% protein aqueous solution. The mass ratio of limonin to ovalbumin is 1:35. Ultrasound is performed at a power of 400 W for 1 min to obtain an aqueous solution of a limonin-rutin-isoquercetin self-assembled complex.

[0118] S1.2 Mix folic acid and sodium alginate in a mass ratio of 1:25 and dissolve in deionized water. After freeze-drying, place the mixed powder of folic acid and sodium alginate in a sealed container with a relative humidity of 80%, and react at 60° C. for 24 hours to obtain a folic acid-sodium alginate complex.

[0119] S1.3 Add folic acid-sodium alginate complex to the aqueous solution of limonin-rutin-isoquercetin self-assembled complex, the concentration of folic acid-sodium alginate complex in the aqueous solution is 0.4% by mass, then adjust the pH to 4.0, react at room temperature in the dark for 30 minutes, centrifuge, and freeze-dry to obtain limonin-rutin-isoquercetin-folic acid nanoparticles with a particle size of 500 nm.

[0120] S2. Preparation of fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles

[0121] S2.1 Fucoxanthin, glycyrrhizic acid and isoquercetin were mixed in a mass ratio of 5:20:2 and dissolved in ethanol. The mixture was then added to a 4% protein aqueous solution. The mass ratio of fucoxanthin to ovalbumin was 1:40. The pH was adjusted to 10.0. The mixture was reacted at room temperature in the dark for 30 minutes. The pH was adjusted back to 7.0 to obtain an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles with a protein shell;

[0122] S2.2 Dissolve chitosan in 1% by volume acetic acid aqueous solution to a final concentration of 5 g / L; dissolve mannose in the chitosan solution to make the mass ratio of chitosan to mannose 1:4; adjust the pH of the chitosan-mannose mixed solution to 5.0, and react at 65°C for 72 hours; after the reaction is completed, place in an ice bath for 10 minutes, and freeze-dry to obtain water-soluble mannose-grafted chitosan;

[0123] S2.3 Add mannose-grafted chitosan to the aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles, the concentration of mannose-grafted chitosan in the aqueous solution is 0.4% by mass, react at room temperature in the dark for 30 minutes, concentrate under reduced pressure, and freeze-dry to obtain fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles with a particle size of 590 nm.

[0124] S3. Vitamin D 3 Preparation of linoleic acid-isoquercetin-hyaluronic acid nanoparticles

[0125] S3.1 Vitamin D 3 , linoleic acid, and isoquercetin were mixed in a mass ratio of 6:20:1.5, dissolved in ethanol, and added to a 2.5 mass% protein aqueous solution. 3 The mass ratio of the mixture to egg white protein was 1:25, and the pH was adjusted to 10.5. After reacting at room temperature and away from light for 30 minutes, the pH was adjusted back to 7.0 to obtain vitamin D with a protein shell. 3 - an aqueous solution of linoleic acid-isoquercetin-protein particles;

[0126] S3.2 Adding hyaluronic acid to vitamin D 3 -In the aqueous solution of linoleic acid-isoquercetin-protein particles, the concentration of hyaluronic acid in the aqueous solution is 0.15% by mass, and the pH is adjusted to 4.0, and the reaction is carried out at room temperature and in the dark for 30 minutes to allow the hyaluronic acid to be fully adsorbed on the outer layer of the protein particles, and then centrifuged and freeze-dried to obtain vitamin D 3 -Linoleic acid-isoquercetin-hyaluronic acid nanoparticles, particle size is 310nm.

[0127] 45 parts by weight of limonin-rutin-isoquercetin-folic acid nanoparticles, 35 parts by weight of fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles, and vitamin D 3 - 0.4 parts by weight of linoleic acid-isoquercetin-hyaluronic acid nanoparticles Weigh the above components, then add appropriate amounts of one or more of soluble starch, dextrin, sucrose, lactose and mannitol, and moisten with appropriate amounts of 70% (v / v) ethanol to prepare a soft material;

[0128] S4 is granulated through a 20-mesh sieve, dried at 70-80°C, and granulated into 60-mesh granules to obtain a functional food of the granule type.

[0129] Example 4

[0130] S1. Preparation of limonin-rutin-isoquercetin-folic acid nanoparticles:

[0131] S1.1 Limonin, rutin and isoquercetin were weighed in a mass ratio of 5:2:2 and dissolved in ethanol. Then, they were added to a 4% protein aqueous solution. The mass ratio of limonin to soy protein was 1:40. Ultrasound was performed at a power of 350 W for 3 min to obtain an aqueous solution of a limonin-rutin-isoquercetin self-assembled complex.

[0132] S1.2 Mix folic acid and sodium alginate in a mass ratio of 1:30 and dissolve in deionized water. After freeze-drying, place the mixed powder of folic acid and sodium alginate in a sealed container with a relative humidity of 80%, and react at 60° C. for 24 hours to obtain a folic acid-sodium alginate complex.

[0133] S1.3 Add folic acid-sodium alginate complex to the aqueous solution of limonin-rutin-isoquercetin self-assembled complex, the concentration of folic acid-sodium alginate complex in the aqueous solution is 0.5% by mass, then adjust the pH to 4.0, react at room temperature in the dark for 40 minutes, centrifuge, and freeze-dry to obtain limonin-rutin-isoquercetin-folic acid nanoparticles with a particle size of 570 nm.

[0134] S2. Preparation of fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles

[0135] S2.1 Fucoxanthin, glycyrrhizic acid and isoquercetin were mixed in a mass ratio of 5:30:3 and dissolved in ethanol. The mixture was then added to a 5% protein aqueous solution. The mass ratio of fucoxanthin to soy protein was 1:50. The pH was adjusted to 11.0. The mixture was reacted at room temperature in the dark for 30 minutes. The pH was adjusted back to 7.0 to obtain an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles with a protein shell;

[0136] S2.2 Dissolve chitosan in 1% by volume acetic acid aqueous solution to a final concentration of 5 g / L; dissolve mannose in the chitosan solution to make the mass ratio of chitosan to mannose 1:5; adjust the pH of the chitosan-mannose mixed solution to 5.5, and react at 65°C for 72 hours; after the reaction, ice bath for 10 minutes, and freeze-dry to obtain water-soluble mannose-grafted chitosan;

[0137] S2.3 Add mannose-grafted chitosan to the aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles, the concentration of mannose-grafted chitosan in the aqueous solution is 0.5% by mass, react at room temperature in the dark for 40 minutes, concentrate under reduced pressure, and freeze-dry to obtain fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles with a particle size of 650 nm.

[0138] S3. Vitamin D 3 Preparation of linoleic acid-isoquercetin-hyaluronic acid nanoparticles

[0139] S3.1 Vitamin D 3 , linoleic acid, and isoquercetin were mixed in a mass ratio of 6:25:2, dissolved in ethanol, and added to a 3% protein aqueous solution. 3 The mass ratio of the mixture to soybean protein was 1:30, and the pH was adjusted to 11.0. After reacting at room temperature and away from light for 40 minutes, the pH was adjusted back to 7.0 to obtain vitamin D with a protein shell. 3 - an aqueous solution of linoleic acid-isoquercetin-protein particles;

[0140] S3.2 Adding hyaluronic acid to vitamin D 3 -In the aqueous solution of linoleic acid-isoquercetin-protein particles, the concentration of hyaluronic acid in the aqueous solution is 0.2% by mass, and the pH is adjusted to 4.0, and the reaction is carried out at room temperature and in the dark for 40 minutes to allow the hyaluronic acid to be fully adsorbed on the outer layer of the protein particles, and then centrifuged and freeze-dried to obtain vitamin D 3 -Linoleic acid-isoquercetin-hyaluronic acid nanoparticles, particle size is 340nm.

[0141] S4 is composed of 50 parts by weight of limonin-rutin-isoquercetin-folic acid nanoparticles, 40 parts by weight of fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles, and vitamin D 3 - 0.5 parts by weight of linoleic acid-isoquercetin-hyaluronic acid nanoparticles Weigh the above components, dissolve them in water, add appropriate amounts of sweeteners and stabilizers, and mix well;

[0142] S5 homogenization, filtration,

[0143] S6 filling, sterilization, and you get functional food like beverage.

[0144] Functional evaluation test:

[0145] (1) Evaluation of oral absorption of limonin

[0146] The tested samples and their preparation methods are shown in Table 1:

[0147] Table 1 Oral absorption effect of limonin Test samples and their preparation methods

[0148]

[0149] Sample treatment: Dissolve the test sample in deionized water at a mass-to-volume ratio of 1:10 (add a small amount of ethanol to aid dissolution if necessary), and convert the dose. The dose of limonin is 10.0 mg / kg, which is the test solution.

[0150] Experimental animals: SPF male ICR mice, 15-20 g, were purchased from Shanghai Slake Laboratory Animal Co., Ltd.

[0151] The basic feed composition of 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%.

[0152] Animal grouping and treatment methods: After 1 week of adaptive feeding, the mice were randomly divided into 6 groups according to body weight, with 9 mice in each group, namely, blank group (administered with deionized water), limonin group, limonin-sodium alginate nanoparticle group, limonin-folate nanoparticle group, limonin-rutin-folate nanoparticle group, and limonin-rutin-isoquercetin-folate nanoparticle group (the preparation methods are shown in Table 1). Each mouse was given a dose of 10.0 mg / kg. After 4 hours, the mouse eyeballs were removed and the changes in the content of limonin in plasma were analyzed by high performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS); the expression levels of CD36 and P-pg in the small intestine were analyzed by Elisa kit method to verify the beneficial effects and action pathways of the design of the nutrient co-carrying delivery system on promoting the intestinal absorption of limonin.

[0153] The experimental results are shown in Table 2:

[0154] Table 2. Oral absorption of limonin and expression of key transport proteins

[0155]

[0156]

[0157] HPLC and LC-MS were used to analyze the total response concentration of limonin in the serum of mice in each treatment group, and the results are shown in Table 2. The results show that after oral gavage, the total response concentration of unencapsulated limonin in serum was only 80.24 ng / mL. When protein and sodium alginate were used as wall materials and the preparation process of Example 1 was used for encapsulation, the absorption effect was improved to a certain extent, and its total response concentration in serum was raised to 145.32 ng / mL, an increase of 81.11%; under the same conditions, when sodium alginate was modified with folic acid, the serum response concentration of limonin in the obtained limonin-folic acid nanoparticle group was improved, and the serum response concentration reached 365.88 ng / mL, which was 3.56 times higher than that of the limonin group, indicating that folic acid modification promoted the aggregation of limonin on the outside of intestinal epithelial cells and increased the chance of being taken up by intestinal epithelial cells. When rutin, which promotes CD36 expression, is fused to the delivery system, the expression of CD36 in the mouse intestine increases to 3.43 times that of the blank group, and the serum response value of limonin is further increased to 885.69 ng / mL, which is 10.04 times higher than that of the limonin group; when isoquercetin, which inhibits P-gp, is further fused to the delivery system, the serum response value of limonin climbs to 1895.65 ng / mL, which is 22.62 times higher than that of the limonin group. This shows that the limonin-rutin-isoquercetin-folic acid nanoparticle delivery system designed by the present invention can effectively improve the oral bioavailability of limonin, thereby laying a foundation for its lipid-lowering and weight-loss functions.

[0158] (2) Evaluation of oral absorption of fucoxanthin

[0159] The tested samples and their preparation methods are shown in Table 3:

[0160] Table 3 Oral absorption effect of fucoxanthin Test samples and their preparation methods

[0161]

[0162]

[0163] Sample processing: Dissolve the test sample in deionized water at a mass-to-volume ratio of 1:10 (add a small amount of ethanol to aid dissolution if necessary), and convert the dosage. The dosage of fucoxanthin is 10.0 mg / kg, which is the test solution.

[0164] 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%.

[0165] Animal grouping and treatment methods: After 1 week of adaptive feeding, the mice were randomly divided into 6 groups according to their body weight, with 9 mice in each group, namely, blank group (administered with deionized water), fucoxanthin group, fucoxanthin-chitosan nanoparticle group, fucoxanthin-mannose nanoparticle group, fucoxanthin-glycyrrhizic acid-mannose nanoparticle group, and fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticle group. Each mouse was given a dose of 10.0 mg / kg. After 4 hours, the mouse eyeballs were removed and the changes in the content of fucoxanthin and its metabolites in plasma were analyzed by high performance liquid chromatography (HPLC); the expression of intestinal SR-B1 and P-gp was analyzed by Elisa kit method to verify the beneficial effects and action pathways of the design of nutrient co-carrying delivery system on promoting intestinal absorption of limonin.

[0166] The experimental results are shown in Table 4:

[0167] Table 4. Oral absorption effect of fucoxanthin and expression of key transport proteins

[0168]

[0169]

[0170] 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 the total response concentration of unencapsulated fucoxanthin in serum was only 41.28 pmol / mL after oral gavage. When protein and chitosan were used as wall materials and the preparation process of Example 1 was used for encapsulation, the absorption effect was improved to a certain extent, and the total response concentration of serum was raised to 60.67 pmol / mL, an increase of 46.97%; under the same conditions, when chitosan was modified with mannose, the fucoxanthin serum response concentration of the obtained fucoxanthin-mannose nanoparticle group was improved, and the serum response concentration reached 135.24 pmol / mL, which was 2.28 times higher than that of the fucoxanthin group, indicating that mannose modification promoted the aggregation of fucoxanthin on the outside of intestinal epithelial cells and increased the chance of being taken up by intestinal epithelial cells. When glycyrrhizic acid, which promotes SR-B1 expression, is incorporated into the delivery system, the expression of SR-B1 in the mouse intestine increases to 3.28 times that of the blank group, and the serum response value of fucoxanthin is further increased to 387.29pmol / mL, which is 8.38 times higher than that of the fucoxanthin group; when isoquercetin, which inhibits P-gp, is further incorporated into the delivery system, the serum response value of fucoxanthin climbs to 591.43pmol / mL, which is 13.33 times higher than that of the fucoxanthin group. This indicates that the fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticle delivery system designed by the present invention can effectively improve the oral bioavailability of fucoxanthin, thereby laying a foundation for its lipid-lowering and weight-loss functions.

[0171] (3) Vitamin D 3 Evaluation of oral absorption effect

[0172] The tested samples and their preparation methods are shown in Table 5:

[0173] Table 5 Vitamin D 3 Table of oral absorption effects of tested samples and their preparation methods

[0174]

[0175]

[0176] Sample processing: Dissolve the test sample in deionized water at a mass-to-volume ratio of 1:10 (add a small amount of ethanol to aid dissolution if necessary), and convert the dose. The dose of vitamin D3 is 0.10 mg / kg, which is the test solution.

[0177] 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%.

[0178] Animal grouping and treatment methods: After 1 week of adaptive feeding, the mice were randomly divided into 6 groups according to their body weight, with 9 mice in each group, namely, blank group (oral administration of deionized water), vitamin D3 group, vitamin D3-bovine serum albumin nanoparticle group, vitamin D3-hyaluronic acid nanoparticle group, vitamin D3-linoleic acid-hyaluronic acid nanoparticle group, and vitamin D3-linoleic acid-isoquercetin-hyaluronic acid nanoparticle group. Each mouse was given a dose of 0.1 mg / kg. After 4 hours, the mouse eyeballs were removed and the changes in the content of hydrogenated vitamin D3 (1,25(OH)2D3) in plasma were analyzed by high performance liquid chromatography (HPLC); the expression of intestinal VDR and P-gp was analyzed by Elisa kit method to verify the beneficial effects and action pathways of the design of nutrient co-carrying delivery system on promoting intestinal absorption of vitamin D3.

[0179] The experimental results are shown in Table 6:

[0180] Table 6. Vitamin D 3 Table of oral absorption effect and key transporter protein expression

[0181]

[0182] HPLC analysis of 1,25(OH) in the serum of mice in each treatment group 2 D 3 The results are shown in Table 6. The results show that the unencapsulated vitamin D 3 After oral gavage, the serum response concentration was only 1182.27 pg / mL. When bovine serum albumin was used as the wall material and the preparation process of Example 1 was adopted for embedding, the absorption effect was improved to a certain extent, and the serum response concentration was raised to 1329.75 pg / mL, which was higher than that of vitamin D. 3 The group increased by 12.47%; under the same conditions, when the protein shell was modified with hyaluronic acid, the vitamin D 3 - Vitamin D in hyaluronic acid nanoparticles 3 The serum response concentration was improved and reached 1537.66 pg / mL, which is similar to vitamin D 3 Compared with the control group, it increased by 30.06%, indicating that hyaluronic acid modification promoted the 3The accumulation on the outside of intestinal epithelial cells increases the chance of being taken up by intestinal epithelial cells. When linoleic acid, which promotes VDR expression, was incorporated into the delivery system, the expression of VDR in the mouse intestine increased to 1.98 times that of the blank group. 3 The serum response value of vitamin D also increased further to 2069.93 pg / mL, which is consistent with 3 Compared with the control group, it increased by 75.08%; when isoquercetin, which inhibits P-gp, was further integrated into the delivery system, vitamin D 3 The serum response value climbed to 2819.04 pg / mL, which was consistent with the vitamin D 3 Compared with the control group, it increased by 1.38 times. 3 -Linoleic acid-isoquercetin-hyaluronic acid nanoparticle delivery system can effectively increase vitamin D 3 The oral bioavailability of this medicine is good, thus laying a foundation for its lipid-lowering and weight-loss function.

[0183] (4) Evaluation of product lipid-lowering and weight-loss functions

[0184] Test samples: limonin, fucoxanthin, vitamin D 3 , physical mixture of limonin + fucoxanthin (mass ratio 30:20), limonin + fucoxanthin + vitamin D 3 Physical mixture (mass ratio 30:20:0.1), limonin + fucoxanthin + vitamin D 3 Physical mixture (mass ratio 40:30:0.3), limonin + fucoxanthin + vitamin D 3 Physical mixture (mass ratio 45:35:0.4), limonin + fucoxanthin + vitamin D 3 A physical mixture (mass ratio 50:40:0.5), and the formulation products obtained in Examples 1 to 4 of the present invention.

[0185] Sample processing: The test samples were directly dissolved in deionized water at a mass-to-volume ratio of 1:10 (using appropriate amount of ethanol to assist dissolution when necessary), and the dose conversion was performed. The dosage of all test samples was 50.0 mg / kg (based on the body weight of mice), which was the test solution.

[0186] Experimental animals and feeding environment: SPF male ICR mice, 15-20g, purchased from Shanghai Slake Experimental Animal Co., Ltd. Basic feed ingredients for mice: 64% carbohydrates, 21% protein, 4% fat, 5% fiber, 6% water. High-fat feed ingredients for mice: 78.8% basic feed, 10% egg yolk powder, 10% lard, 1% edible grade cholesterol, 0.2% pig bile salt (cholate). The above feeds were purchased from Shanghai Slake Experimental Animal Co., Ltd. Feeding room temperature: 23±2℃, relative humidity 55±5%.

[0187] Animal grouping and treatment method: The mice were adaptively fed for one week and fed freely. After one week, they were randomly divided into 14 groups according to body weight, namely the normal group, the model group and the above 12 test samples, with 7 experimental animals in each group. The mice in the normal group were fed with normal standard feed (i.e., fed with feed according to the basic feed ingredients of mice), and the mice in the model group were fed with high-fat feed. After about 12 weeks of modeling, when the average weight of the mice in the model group exceeded 20% or more of the normal group, it indicated that the modeling of obese mice was successful. After the modeling was successful, the normal group continued to be fed with normal standard feed, the model group continued to be fed with high-fat feed, and the intervention group (i.e., the above 12 groups) was fed with high-fat feed + test samples, and continued to be fed for 6 weeks. After the experiment, the mice in each group were weighed and the epididymal fat index (epididymal fat mass / mouse body weight × 100%) was calculated to evaluate the lipid-lowering and weight-loss effects of each test sample.

[0188] The experimental results are shown in Table 7:

[0189] Table 7 Evaluation table of lipid-lowering and weight-loss functions of each component and product formula

[0190]

[0191] In view of the vitamin D3 deficiency in obese people and the function of potential dietary ingredients in regulating lipid metabolism, limonin and fucoxanthin were selected from a large number of dietary ingredients as the core functional ingredients for combined weight loss. The lipid-lowering effect of the formula combination of limonin, fucoxanthin and vitamin D3 within a specific ratio range was investigated, and further relying on the strategy of improving the oral bioavailability of the core ingredients, the weight loss efficacy of the delivery system composition constructed by special encapsulation technology was evaluated. The results are shown in Table 7. Although limonin, fucoxanthin and vitamin D3 can reduce weight and fat when used alone, the effect is very limited. When limonin and fucoxanthin were combined in a mass ratio of 30:20, the body weight and epididymal fat index of mice dropped from 50.75g and 6.95% in the model group to 38.26g and 4.82%, respectively; when vitamin D was further introduced 3, the mass ratios of limonin, fucoxanthin, and vitamin D3 were 30:20:0.1, 40:30:0.3, 45:35:0.4, and 50:40:0.5, respectively. The body weight and epididymal fat index of mice were further significantly reduced, indicating that limonin, fucoxanthin, and vitamin D 3 The three ingredients have a synergistic lipid-lowering and weight-loss effect within this ratio range. When the products of Examples 1 to 4 were used for intervention, the weight loss effect of mice was more significantly improved. The products of Examples 1 to 4 can further reduce the body weight and epididymal fat index of mice to 25.69g to 27.13g and 3.04% to 3.54%, respectively, which are very close to the body weight of 23.62g and epididymal index of 2.52% of the normal group of mice. It can be seen that this effect is due to the innovative molecular encapsulation technology used in the present invention, which promotes the intestinal absorption efficiency of limonin, fucoxanthin and vitamin D3, so that the nutritional effect of the formula in terms of lipid-lowering and weight-loss function is further improved.

[0192] 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 functional food for reducing fat and weight, characterized in that: It includes limonin-rutin-isoquercetin-folic acid nanoparticles, fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles, vitamin D3-linoleic acid-isoquercetin-hyaluronic acid nanoparticles, and water or solid filler excipients; wherein the weight ratio of limonin-rutin-isoquercetin-folic acid nanoparticles, fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles, and vitamin D3-linoleic acid-isoquercetin-hyaluronic acid nanoparticles is (30-50):(20-40):(0.1-0.5).

2. The lipid-lowering and weight-loss functional food according to claim 1, characterized in that: In the limonin-rutin-isoquercetin-folic acid nanoparticles, the mass ratio of limonin, rutin and isoquercetin is 5:(0.5-2):(0.5-2); Preferably, the preparation method of limonin-rutin-isoquercetin-folic acid nanoparticles is: S1.1 Preparation of folic acid-sodium alginate complex: folic acid and sodium alginate are mixed and dissolved in deionized water, and the mixed powder of folic acid and sodium alginate is placed in a container for reaction after freeze-drying to obtain a folic acid-sodium alginate complex; preferably, the mass ratio of folic acid to sodium alginate is 1:(10-30); the reaction in the container is placed in a closed container with a relative humidity of 80% and reacted at 60°C for 24 hours; S1.2 Preparation of limonin-rutin-isoquercetin self-assembly complex: After dissolving limonin, rutin and isoquercetin, add them into the protein aqueous solution and mix well, and denature the protein to obtain an aqueous solution of limonin-rutin-isoquercetin self-assembly complex; preferably, the mass ratio of limonin, rutin and isoquercetin is 5:(0.5-2):(0.5-2), the concentration of the protein aqueous solution is 2% to 4% by mass, the mass ratio of limonin to protein is 1:(20-40), and the protein is denatured by ultrasound, pulsed electric field, high static pressure, and microwave treatment; more preferably, the ultrasound condition is 350±50W for 3±1min; The above S1.1 and S1.2 can be switched in order or performed simultaneously; S1.3 adding folic acid-sodium alginate complex to the aqueous solution of limonin-rutin-isoquercetin self-assembled complex, preferably, the concentration of folic acid-sodium alginate complex in the aqueous solution is 0.1 mass % to 0.5 mass %; adjusting the pH to acidic so that the protein shell has the opposite electrical properties to folic acid-sodium alginate, reacting at room temperature in the dark so that the folic acid-sodium alginate complex is tightly adsorbed on the protein shell of the limonin-rutin-isoquercetin self-assembled complex, centrifuging, and freeze-drying to obtain limonin-rutin-isoquercetin-folic acid nanoparticles; Preferably, the acidity is pH 3.0-4.0; and the reaction time at room temperature and in the dark is 30±10 min.

3. The lipid-lowering and weight-loss functional food according to claim 1, characterized in that: In the fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles, the mass ratio of fucoxanthin, glycyrrhizic acid, and isoquercetin is 5:(5-30):(1-3); Preferably, the preparation method of fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles is: S2.1 Preparation of mannose-grafted chitosan: dissolve mannose in an acetic acid aqueous solution of chitosan, adjust the pH of the chitosan-mannose mixed solution to weak acidity, react; ice bath, freeze-dry to obtain water-soluble mannose-grafted chitosan; preferably, the concentration of the acetic acid aqueous solution is 1% by volume, and the final concentration of chitosan in the acetic acid aqueous solution is 5g / L; the mass ratio of chitosan to mannose is 1:(2-5); the weakly acidic pH is 4.0-5.5, and the reaction conditions are 72h at 65°C; the ice bath time is 10min; S2.2 Preparation of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles: Dissolve fucoxanthin, glycyrrhizic acid and isoquercetin and add them to a protein aqueous solution, adjust the pH to alkaline, partially denature the protein and expose the hydrophobic amino acids, react at room temperature in the dark, and adjust the pH back to neutral after the reaction to obtain an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles with a protein shell; preferably, the mass ratio of fucoxanthin, glycyrrhizic acid and isoquercetin is 5:(5-30):(1-3), the concentration of the protein aqueous solution is 2% to 5% by mass, the mass ratio of fucoxanthin to protein is 1:(20-50), the alkaline pH is 9.0-11.0, and the reaction time at room temperature in the dark is 30±10 min; The above S2.1 and S2.2 can be performed in reverse order or simultaneously; S2.3 adding mannose-grafted chitosan to an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles, preferably, the concentration of mannose-grafted chitosan in the aqueous solution is 0.1 mass % to 0.5 mass %; reacting at room temperature in the dark so that the mannose-grafted chitosan covers the outer layer of the protein particles, concentrating under reduced pressure, and freeze-drying to obtain fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles; Preferably, the reaction time at room temperature and in the dark is 30±10 min.

4. The lipid-lowering and weight-loss functional food according to claim 1, characterized in that: In the vitamin D3-linoleic acid-isoquercetin-hyaluronic acid nanoparticles, the dosage ratio of vitamin D3, linoleic acid, and isoquercetin is 6:(10-25):(0.5-2); Preferably, the preparation method of vitamin D3-linoleic acid-isoquercetin-hyaluronic acid nanoparticles is: S3.1 Preparation of vitamin D3-linoleic acid-isoquercetin-protein particles: Vitamin D3, linoleic acid and isoquercetin are dissolved and added to a protein aqueous solution, and the pH is adjusted to alkaline to partially denature the protein and expose the hydrophobic amino acids. After reacting at room temperature in the dark, the pH is adjusted back to neutral to obtain an aqueous solution of vitamin D3-linoleic acid-isoquercetin-protein particles with a protein shell; preferably, the mass ratio of vitamin D3, linoleic acid and isoquercetin is 6:(10-25):(0.5-2), the concentration of the protein aqueous solution is 1% to 3% by mass, the mass ratio of vitamin D3 to protein is 1:(10-30), the alkaline pH is 9.0-11.0, and the reaction time at room temperature in the dark is 30±10 min; S3.2 Obtaining vitamin D3-linoleic acid-isoquercetin-hyaluronic acid nanoparticles: adding hyaluronic acid to the aqueous solution of the vitamin D3-linoleic acid-isoquercetin-protein particles, preferably, the concentration of hyaluronic acid in the aqueous solution is 0.05% by mass to 0.2% by mass; and adjusting the pH to acidic so that the protein shell has an opposite electrical property to that of the hyaluronic acid, reacting at room temperature in the dark so that the hyaluronic acid is fully adsorbed on the outer layer of the protein particles, centrifuging, and freeze-drying to obtain vitamin D3-linoleic acid-isoquercetin-hyaluronic acid nanoparticles; preferably, the acidic pH is 3.0-4.0, and the reaction time at room temperature in the dark is 30±10min.

5. The functional food for reducing fat and weight as claimed in any one of claims 1 to 4, characterized in that: The functional food is a capsule, a tablet, an granule, an oral liquid or a beverage.

6. A method for preparing the functional food for reducing fat and weight according to claim 1, comprising the following steps: S1. Preparation of limonin-rutin-isoquercetin-folic acid nanoparticles: S1.1 Preparation of folic acid-sodium alginate complex: folic acid and sodium alginate are mixed and dissolved in deionized water, and the mixed powder of folic acid and sodium alginate is placed in a container for reaction after freeze-drying to obtain a folic acid-sodium alginate complex; preferably, the mass ratio of folic acid to sodium alginate is 1:(10-30); the reaction in the container is placed in a closed container with a relative humidity of 80% and reacted at 60°C for 24 hours; S1.2 Preparation of limonin-rutin-isoquercetin self-assembly complex: After dissolving limonin, rutin and isoquercetin, add them into the protein aqueous solution and mix well, and denature the protein to obtain an aqueous solution of limonin-rutin-isoquercetin self-assembly complex; preferably, the mass ratio of limonin, rutin and isoquercetin is 5:(0.5-2):(0.5-2), the concentration of the protein aqueous solution is 2% to 4% by mass, the mass ratio of limonin to protein is 1:(20-40), and the protein is denatured by ultrasound, pulsed electric field, high static pressure, and microwave treatment; more preferably, the ultrasound condition is 350±50W for 3±1min; The above S1.1 and S1.2 can be switched in order or performed simultaneously; S1.

3. Adding folic acid-sodium alginate complex to the aqueous solution of limonin-rutin-isoquercetin self-assembled complex, preferably, the concentration of folic acid-sodium alginate complex in the aqueous solution is 0.1 mass % to 0.5 mass %; adjusting the pH to acidic so that the protein shell has the opposite electrical properties to the dietary polysaccharide, reacting at room temperature in the dark so that the folic acid-sodium alginate complex is tightly adsorbed on the protein shell of the limonin-rutin-isoquercetin self-assembled complex, centrifuging, and freeze-drying to obtain limonin-rutin-isoquercetin-folic acid nanoparticles; Preferably, the acidity is pH 3.0-4.0; the reaction time at room temperature and in the dark is 30±10 min; S2. Preparation of fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles S2.1 Preparation of mannose-grafted chitosan: dissolve mannose in an acetic acid aqueous solution of chitosan, adjust the pH of the chitosan-mannose mixed solution to weak acidity, react; ice bath, freeze-dry to obtain water-soluble mannose-grafted chitosan; preferably, the concentration of the acetic acid aqueous solution is 1% by volume, and the final concentration of chitosan in the acetic acid aqueous solution is 5g / L; the mass ratio of chitosan to mannose is 1:(2-5); the weakly acidic pH is 4.0-5.5, and the reaction conditions are 72h at 65°C; the ice bath time is 10min; S2.2 Preparation of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles: Dissolve fucoxanthin, glycyrrhizic acid and isoquercetin and add them to a protein aqueous solution, adjust the pH to alkaline, partially denature the protein and expose the hydrophobic amino acids, react at room temperature in the dark, and adjust the pH back to neutral after the reaction to obtain an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles with a protein shell; preferably, the mass ratio of fucoxanthin, glycyrrhizic acid and isoquercetin is 5:(5-30):(1-3), the concentration of the protein aqueous solution is 2% to 5% by mass, the mass ratio of fucoxanthin to protein is 1:(20-50), the alkaline pH is 9.0-11.0, and the reaction time at room temperature in the dark is 30±10 min; The above S2.1 and S2.2 can be performed in reverse order or simultaneously; S2.3 adding mannose-grafted chitosan to an aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles, preferably, the concentration of mannose-grafted chitosan in the aqueous solution is 0.1 mass % to 0.5 mass %; reacting at room temperature in the dark so that the mannose-grafted chitosan covers the outer layer of the protein particles, concentrating under reduced pressure, and freeze-drying to obtain fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles; Preferably, the reaction time at room temperature and away from light is 30±10 min; S3. Preparation of Vitamin D3-linoleic acid-isoquercetin-hyaluronic acid nanoparticles: S3.1 Preparation of vitamin D3-linoleic acid-isoquercetin-protein particles: Vitamin D3, linoleic acid and isoquercetin are dissolved and added to a protein aqueous solution, and the pH is adjusted to alkaline to partially denature the protein and expose the hydrophobic amino acids. After reacting at room temperature in the dark, the pH is adjusted back to neutral to obtain an aqueous solution of vitamin D3-linoleic acid-isoquercetin-protein particles with a protein shell; preferably, the mass ratio of vitamin D3, linoleic acid and isoquercetin is 6:(10-25):(0.5-2), the concentration of the protein aqueous solution is 1% to 3% by mass, the mass ratio of vitamin D3 to protein is 1:(10-30), the alkaline pH is 9.0-11.0, and the reaction time at room temperature in the dark is 30±10 min; S3.2 Obtaining vitamin D3-linoleic acid-isoquercetin-hyaluronic acid nanoparticles: adding hyaluronic acid to an aqueous solution of vitamin D3-linoleic acid-isoquercetin-protein particles, and adjusting the pH to acidic, reacting at room temperature in the dark, allowing the hyaluronic acid to be fully adsorbed on the outer layer of the protein particles, centrifuging, and freeze-drying to obtain vitamin D3-linoleic acid-isoquercetin-hyaluronic acid nanoparticles; preferably, the acidic pH is 3.0-4.0, and the reaction time at room temperature in the dark is 30±10 min; The above S1, S2, and S3 can be interchanged in order or performed simultaneously; S4. Mix the required limonin-rutin-isoquercetin-folic acid nanoparticles, fucoxanthin-glycyrrhizic acid-isoquercetin-mannose nanoparticles, vitamin D3-linoleic acid-isoquercetin-hyaluronic acid nanoparticles, water or solid filler excipients.

7. The preparation method according to claim 6, characterized in that: The method further comprises the following step: S5. if the obtained mixture is an aqueous solution, homogenizing and filtering the aqueous solution mixture; if the obtained mixture is a solid mixture, sieving the solid mixture.

8. The preparation method according to claim 7, characterized in that: The method further comprises the following steps: S6. sterilizing after filling, or packaging after tableting or granulation.

9. The preparation method according to any one of claims 6 to 8, characterized in that: The protein is one of bovine serum albumin, whey protein, ovalbumin and soy protein.

10. The preparation method according to any one of claims 6 to 8, characterized in that: In the step S1, the folic acid-sodium alginate complex is added to the aqueous solution of the limonin-rutin-isoquercetin self-assembled complex, and the concentration of the folic acid-sodium alginate complex in the aqueous solution is 0.1% to 0.5% by mass; and / or In the step S2, mannose-grafted chitosan is added to the aqueous solution of fucoxanthin-glycyrrhizic acid-isoquercetin-protein particles, and the concentration of mannose-grafted chitosan in the aqueous solution is 0.1% to 0.5% by mass; and / or In the step S3.2, hyaluronic acid is added to the aqueous solution of vitamin D3-linoleic acid-isoquercetin-protein particles, and the concentration of hyaluronic acid in the aqueous solution is 0.05 mass % to 0.2 mass %.

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