Liver-protecting food as well as preparation method and application thereof

Abstract

The invention discloses a liver protection food which comprises an astaxanthin compound preparation, a fucoxanthin compound preparation, taurine, protein peptide and cuttlefish juice powder in a mass ratio of (20-40): (10-30): (1-5): (1-5): (1-5). According to the liver-protecting food provided by the invention, a multi-component'synergistic 'formula design concept of a traditional Chinese medicine theory is used for reference, and on the basis of promoting efficient intestinal absorption of the liver-protecting components such as astaxanthin and fucoxanthin, the components are subjected to formula design, so that the synergistic effect of liver protection is achieved, the liver-protecting effect of the liver-protecting food is greatly improved, and the liver-protecting food is suitable for being eaten by people. Therefore, the traditional Chinese medicine composition has a good liver-protecting effect.

Description

Technical Field

[0001] The present invention relates to the technical field of health food, and in particular to a liver protection food and a preparation method and application thereof. Background Art

[0002] Wine culture plays an important role in people's daily social activities, life, leisure, entertainment and other activities. However, alcohol intake will increase the burden of liver metabolism and detoxification. Excessive and long-term drinking may even cause health problems such as alcoholic fatty liver, hepatitis, and cirrhosis. However, existing liver protection drugs have problems such as large side effects and high prices. Therefore, functional foods that are green, safe, and have significant liver protection effects have become products that are urgently needed.

[0003] Existing studies have shown that some special dietary nutrients, such as astaxanthin and fucoxanthin, can effectively regulate alcohol metabolism and improve the liver's detoxification function, thereby achieving a liver-protecting effect. However, most of these nutrients are fat-soluble and have low bioavailability when taken orally, which greatly reduces their biological and nutritional functions and prevents them from fully exerting their due effects. At the same time, astaxanthin and fucoxanthin are prone to decomposition under acid-base reagents and heating conditions, and decompose during the preparation of edible products, reducing the effective ingredients of astaxanthin and fucoxanthin in the product, greatly reducing the efficacy of the product. Summary of the invention

[0004] The purpose of the present invention is to provide a liver-protecting food which has significant liver-protecting effect, high absorption efficiency of core components, and takes into account the green, safe and nutritious nature of the product.

[0005] To achieve the above-mentioned purpose, the first aspect of the present invention proposes a liver protection food, comprising an astaxanthin compound preparation, a fucoxanthin compound preparation, taurine, protein peptides, and cuttlefish juice powder, wherein the mass ratio thereof is 20-40:10-30:1-5:1-5:1-5.

[0006] Furthermore, the astaxanthin composite preparation includes astaxanthin, soybean saponin and triterpenic acid, and the mass ratio of astaxanthin, soybean saponin and triterpenic acid is 1:25-40:0.5-2.

[0007] Furthermore, the fucoxanthin compound preparation includes fucoxanthin, glycyrrhizic acid and isoquercetin, and the mass ratio of fucoxanthin, glycyrrhizic acid and isoquercetin is 1:15-30:0.5-2.

[0008] Furthermore, the astaxanthin composite preparation is prepared by mixing and dissolving astaxanthin, soybean saponin and triterpenic acid to obtain a first fused micelle, adding the first fused micelle to a denatured protein solution, mixing and homogenizing to obtain a first complex aqueous solution with protein as an outer shell and astaxanthin, soybean saponin and triterpenic acid as an inner core, adding antifreeze peptides and polysaccharides to mix, adjusting the pH to acidic at low temperature to obtain a first complex suspension with polysaccharides adsorbed on the surface of the protein shell, and centrifuging and drying the first complex suspension to obtain the astaxanthin composite preparation.

[0009] Furthermore, the fucoxanthin composite preparation is prepared by mixing and dissolving fucoxanthin, glycyrrhizic acid and isoquercetin to obtain a second fused micelle, adding the second fused micelle to a denatured protein solution, mixing and homogenizing to obtain a second complex aqueous solution with protein as the shell and fucoxanthin, glycyrrhizic acid and isoquercetin as the core, adding antifreeze peptide and mannose-grafted chitosan to mix, adjusting the pH to acidic at low temperature to obtain a second complex suspension with mannose-grafted chitosan adsorbed on the protein shell, and centrifuging and drying the second complex suspension to obtain the fucoxanthin composite preparation;

[0010] Preferably, mannose is dissolved in an acid solution of chitosan to obtain a mixed solution, the pH of the mixed solution is adjusted and then heated for reaction, and after the reaction is completed, ice bathing and freeze drying are performed to obtain water-soluble mannose-grafted chitosan;

[0011] Preferably, the concentration of the chitosan acid solution is 5.0 g / L.

[0012] The second aspect of the present invention provides a method for preparing a liver-protecting food, which is used to prepare the liver-protecting food provided in the first aspect of the present invention, comprising:

[0013] S1: astaxanthin, soybean saponin, and triterpenic acid are mixed and dissolved to obtain a first fusion micelle, and the first fusion micelle is added to a denatured protein solution, and mixed and homogenized to obtain a first complex aqueous solution with protein as the shell and astaxanthin, soybean saponin, and triterpenic acid as the core, and antifreeze peptides and polysaccharides are added and mixed, and the pH is adjusted to acidic at low temperature so that the protein shell has an opposite charge to the polysaccharide, and a first complex suspension with polysaccharides adsorbed on the surface of the protein shell is obtained, and the first complex suspension is centrifuged and dried to obtain an astaxanthin composite preparation;

[0014] S2: fucoxanthin, glycyrrhizic acid and isoquercetin are mixed and dissolved to obtain a second fused micelle, the second fused micelle is added to the denatured protein solution, mixed and homogenized to obtain a second complex aqueous solution with protein as the shell and fucoxanthin, glycyrrhizic acid and isoquercetin as the core, antifreeze peptide and mannose-grafted chitosan are added and mixed, the pH is adjusted to acidic at low temperature to obtain a second complex suspension with mannose-grafted chitosan adsorbed on the protein shell, and the second complex suspension is centrifuged, frozen and dried to obtain a fucoxanthin composite preparation;

[0015] The order of S1 and S2 is not limited and they can be switched or performed simultaneously.

[0016] S3: the prepared astaxanthin composite preparation and fucoxanthin composite preparation are mixed with taurine, protein peptide, cuttlefish juice powder, water or filler excipients, and then sterilized to prepare the finished product;

[0017] Preferably, in step S1, the pH is adjusted to 3.0-4.0 at low temperature to obtain a first complex suspension in which polysaccharides are adsorbed on the surface of the protein shell;

[0018] Preferably, in step S2, the pH is adjusted to 5.0-6.0 at low temperature to obtain a second complex suspension in which mannose-grafted chitosan is adsorbed on the surface of the protein shell;

[0019] Preferably, the polysaccharide in step S1 is any one of pectin, hyaluronic acid, fucoidan, and galactan sulfate;

[0020] Preferably, the denatured protein solution in step S1 and step S2 is obtained by treating the protein solution by any one of electric field, ultrasound, microwave and high static pressure; more preferably, the electric field is a pulsed electric field with an intensity of 30-40 kV / cm for 6-8 min;

[0021] Preferably, the homogenization is high pressure homogenization, more preferably homogenization under high pressure of 60 MPa-120 MPa for 1-5 min;

[0022] Preferably, the mass ratio of astaxanthin to protein in the first complex aqueous solution in step S1 is 1:15-30, and the mass ratio of fucoxanthin to protein in the second complex aqueous solution in step S2 is 1:15-30; more preferably, the protein is any one of bovine serum albumin, ovalbumin, whey protein, peanut protein, and soy protein;

[0023] Preferably, the molecular weight of the antifreeze peptide is 500-2000Da.

[0024] Further, mannose is dissolved in an acid solution of chitosan to obtain a mixed solution, the pH of the mixed solution is adjusted and then heated for reaction, and after the reaction is completed, ice bathing and freeze drying are performed to obtain water-soluble mannose-grafted chitosan;

[0025] Preferably, the concentration of the chitosan acid solution is 5.0 g / L;

[0026] Preferably, the mass ratio of chitosan to mannose in the mixed solution is 1:3;

[0027] Preferably, the pH of the mixed solution is adjusted to 5-6;

[0028] Preferably, the heating reaction is carried out at 60-70° C. for 70-80 h.

[0029] Furthermore, the mass concentration of the polysaccharide in the first complex suspension is 0.2%-0.5%; and / or

[0030] The mass concentration of mannose-grafted chitosan in the second complex suspension is 0.1%-0.5%.

[0031] The liver-protecting food provided in the first aspect of the present invention and the liver-protecting food obtained by the preparation method provided in the second aspect have the purpose of protecting the liver.

[0032] The liver-protecting food proposed in the first aspect of the present invention and the liver-protecting food obtained by the preparation method proposed in the second aspect are one or more of capsules, tablets, granules, oral liquids or beverages.

[0033] The liver-protecting food proposed in the present invention draws on the multi-component "synergistic" formula design concept of traditional Chinese medicine theory. On the basis of promoting the efficient intestinal absorption of liver-protecting ingredients such as astaxanthin and fucoxanthin, the above ingredients are formulated to play a synergistic and synergistic role in liver protection, thereby greatly improving the liver-protecting efficacy of the liver-protecting food and making it have a good liver-protecting effect.

[0034] This liver protection food has a good oral absorption effect. In view of the fact that astaxanthin and fucoxanthin are closely regulated by lipid transport proteins during transmembrane transport, the present invention innovatively uses mannose-grafted chitosan to modify the fucoxanthin composite preparation, so that the liver protection food of the present invention can specifically bind to the mannose receptors of intestinal epithelial cells. By regulating the expression of lipid transport proteins responsible for the absorption of astaxanthin / fucoxanthin across the intestinal epithelium, the intestinal absorption efficiency of astaxanthin and fucoxanthin is effectively improved. At the same time, bioenhancers that can promote intestinal absorption are introduced into the delivery system: soybean saponin / triterpenic acid, glycyrrhizic acid / isoquercetin. The oral absorption efficiency of astaxanthin is increased by 29.3-30.2 times, and the oral absorption efficiency of fucoxanthin is increased by 19.5-20.4 times.

[0035] The present invention also proposes a method for preparing liver-protecting food. In view of the characteristics that astaxanthin and fucoxanthin are sensitive to acid-base reagents and easily decomposed by heat, in the process of preparing astaxanthin and fucoxanthin as a composite preparation, astaxanthin and fucoxanthin are first prepared as the first fusion micelle and the second fusion micelle, respectively, and then a denatured protein aqueous solution is added to prepare the first complex aqueous solution and the second complex aqueous solution. The hydrophobic amino acids inside the denatured protein are exposed, so that they can self-assemble with the aforementioned micelles, and the protein is adsorbed outside the micelle to form a core-shell structure with a micelle group at the center and a protein shell. The first complex aqueous solution is a first fusion micelle and a second fusion micelle encapsulated with protein as the shell, so that astaxanthin and fucoxanthin are encapsulated inside the protein to form a shell-core structure.

[0036] The process can use any physical means and homogenization technology that can denature proteins, without using any heating conditions or acid-base reagents, so that the processing environment of protein-encapsulated astaxanthin and fucoxanthin is always at a low temperature and neutral pH. Pulsed electric field is a low-cost and stable physical processing method with stable product quality, and does not use any chemical reagents, so food safety is guaranteed.

[0037] At the same time, low-temperature ion-driven self-assembly technology is used to further modify the polysaccharide surface molecules of the shell-core structure encapsulating astaxanthin or fucoxanthin. Low temperature conditions, such as -2°C, are used during the processing process, which effectively reduces the damage of the processing conditions to astaxanthin and fucoxanthin. The encapsulation rates of astaxanthin and fucoxanthin reached 98% and 95% respectively.

[0038] The particle size of the astaxanthin composite preparation prepared by the method is 600-700nm, the particle size of the fucoxanthin composite preparation is 550nm-680nm, and the retention rates of astaxanthin and fucoxanthin composite preparations are both ≥90% after being heated at 121°C for 15min. The retention rate of common astaxanthin-containing products on the market under the same conditions is less than 20%, and the loss rate is more than 80%, while the loss rate of the astaxanthin composite preparation and fucoxanthin composite preparation prepared by the present invention can be controlled within 10%. DETAILED DESCRIPTION

[0039] 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, and should not be construed as limiting the present invention. In the embodiments, if specific techniques or conditions are not indicated, the techniques or conditions described in the literature in this area or the product specifications are carried out. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products that can be obtained commercially. In the following embodiments, if not clearly stated, "%" refers to weight percentage.

[0040] 1. The preparation method of the liver-protecting food of the present invention is as follows:

[0041] S1: astaxanthin, soybean saponin, and triterpenic acid are mixed and dissolved to obtain a first fusion micelle, and the first fusion micelle is added to a denatured protein solution, and mixed and homogenized to obtain a first complex aqueous solution with protein as the shell and astaxanthin, soybean saponin, and triterpenic acid as the core, and antifreeze peptides and polysaccharides are added and mixed, and the pH is adjusted to acidic at low temperature so that the protein shell has an opposite charge to the polysaccharide, and a first complex suspension with polysaccharides adsorbed on the surface of the protein shell is obtained, and the first complex suspension is centrifuged and dried to obtain an astaxanthin composite preparation;

[0042] S2: fucoxanthin, glycyrrhizic acid and isoquercetin are mixed and dissolved to obtain a second fused micelle, the second fused micelle is added to the denatured protein solution, mixed and homogenized to obtain a second complex aqueous solution with protein as the shell and fucoxanthin, glycyrrhizic acid and isoquercetin as the core, antifreeze peptide and mannose-grafted chitosan are added and mixed, the pH is adjusted to acidic at low temperature to obtain a second complex suspension with mannose-grafted chitosan adsorbed on the protein shell, and the second complex suspension is centrifuged, frozen and dried to obtain a fucoxanthin composite preparation;

[0043] The order of S1 and S2 is not limited and they can be switched or performed simultaneously.

[0044] S3: the prepared astaxanthin composite preparation and fucoxanthin composite preparation are mixed with taurine, protein peptide, cuttlefish juice powder, water or filler excipients, and then sterilized to prepare the finished product;

[0045] Preferably, in step S1, the pH is adjusted to 3.0-4.0 at low temperature to obtain a first complex suspension in which polysaccharides are adsorbed on the surface of the protein shell;

[0046] Preferably, in step S2, the pH is adjusted to 5.0-6.0 at low temperature to obtain a second complex suspension in which mannose-grafted chitosan is adsorbed on the surface of the protein shell;

[0047] Preferably, the polysaccharide in step S1 is any one of pectin, hyaluronic acid, fucoidan, and galactan sulfate;

[0048] Preferably, the denatured protein solution in step S1 and step S2 is obtained by treating the protein solution by any one of electric field, ultrasound, microwave and high static pressure; more preferably, the electric field is a pulsed electric field with an intensity of 30-40 kV / cm for 6-8 min;

[0049] Preferably, the homogenization is high pressure homogenization, more preferably homogenization under high pressure of 60 MPa-120 MPa for 1-5 min;

[0050] Preferably, the mass ratio of astaxanthin to protein in the first complex aqueous solution in step S1 is 1:15-30, and the mass ratio of fucoxanthin to protein in the second complex aqueous solution in step S2 is 1:15-30; more preferably, the protein is any one of bovine serum albumin, ovalbumin, whey protein, peanut protein, and soy protein;

[0051] Preferably, the molecular weight of the antifreeze peptide is 500-2000Da.

[0052] 2. The preparation method of mannose grafted chitosan in the present invention is as follows:

[0053] Dissolving mannose in an acid solution of chitosan to obtain a mixed solution, adjusting the pH of the mixed solution and then heating the mixed solution for reaction, and after the reaction is completed, placing the mixed solution in an ice bath and freeze-drying to obtain a water-soluble mannose-grafted chitosan;

[0054] Preferably, the concentration of the chitosan acid solution is 5.0 g / L;

[0055] Preferably, the mass ratio of chitosan to mannose in the mixed solution is 1:3;

[0056] Preferably, the pH of the mixed solution is adjusted to 5-6;

[0057] Preferably, the heating reaction is carried out at 60-70° C. for 70-80 h.

[0058] In the present invention, a denatured protein solution is used to embed the fusion micelles. Any physical means that can denature proteins can be used to denature the protein solution in the present invention. For ease of comparison, a pulsed electric field is used to denature the protein solution in the following examples.

[0059] Example 1

[0060] S1.1 Preparation of astaxanthin composite preparation: Astaxanthin, soybean saponin, and triterpene acid are weighed in a mass ratio of 1:30:1, and after dissolving with a small amount of anhydrous ethanol, stirred evenly, and ultrasonicated for 2 minutes to obtain the first fusion micelle. Bovine serum albumin is dissolved in deionized water to obtain a protein solution with a mass concentration of 2%. The first fusion micelle is added to the protein solution treated with a pulsed electric field of 30kV / cm for 6 minutes and stirred to make the mass ratio of astaxanthin to bovine serum albumin 1:20. A high-pressure homogenizer is used for homogenization at a pressure of 60MPa for 5 minutes to obtain a first complex aqueous solution. The first fusion micelle containing astaxanthin is encapsulated by a protein shell. Antifreeze peptide with an average molecular weight of 500Da is added to the first complex aqueous solution to make the mass concentration of antifreeze peptide in the first complex aqueous solution 1%, and then pectin is added to make the mass concentration of pectin in the first complex aqueous solution 0.2%. After mixing, it is placed in a cold storage at -2°C. The solution temperature was reduced to -2°C, and the pH of the system was adjusted to 4.0 to obtain the first complex suspension. Finally, a core-shell structure was obtained in which astaxanthin was encapsulated inside the protein shell and polysaccharides were adsorbed on the outside. The encapsulation efficiency of astaxanthin in the first complex suspension was 98%, and the particle size of the core-shell structure encapsulating astaxanthin was 600.0nm. The first complex suspension was centrifuged and dried to obtain an astaxanthin composite preparation dry powder. The obtained astaxanthin composite preparation dry powder was redissolved in deionized water and heated at 121°C for 15 minutes, and the retention rate of astaxanthin was 90%.

[0061] S1.2 Preparation of fucoxanthin composite preparation: Weigh fucoxanthin, glycyrrhizic acid, and isoquercetin in a mass ratio of 1:20:1, use a small amount of anhydrous ethanol to dissolve, stir evenly, and ultrasonicate for 2 minutes to obtain the second fusion micelle. Dissolve bovine serum albumin in deionized water to obtain a protein solution with a mass concentration of 2%. Add the second fusion micelle to the protein solution treated with a pulsed electric field of 30kV / cm for 6 minutes and stir to mix. The mass ratio of fucoxanthin to protein is 1:20. Use a high-pressure homogenizer to homogenize at a pressure of 60MPa for 5 minutes to obtain a second complex aqueous solution. The second fusion micelle containing fucoxanthin is encapsulated by a protein shell. Add an antifreeze peptide with an average molecular weight of 500Da into the second complex aqueous solution. The mass concentration of the antifreeze peptide in the solution is 1%. Add mannose-grafted chitosan to make its mass concentration in the solution 0.1%. After mixing, place it in a cold storage at -2°C. When the solution temperature dropped to -2°C, the pH of the system was adjusted to 5.0 to obtain a second complex suspension. Finally, a core-shell structure was obtained in which fucoxanthin was encapsulated inside the protein shell and mannose-grafted chitosan was adsorbed on the outside. The encapsulation efficiency of fucoxanthin in the second complex suspension was 95%, and the particle size of the core-shell structure encapsulated with fucoxanthin was 550.0nm. The second complex suspension was centrifuged and dried to obtain a fucoxanthin composite preparation dry powder. The fucoxanthin composite preparation dry powder was redissolved in deionized water and heated at 121°C for 15 minutes, and the retention rate of fucoxanthin was 90%.

[0062] The preparation method of mannose grafted chitosan is as follows: chitosan is dissolved in an acetic acid aqueous solution with a volume concentration of 1%, and the final concentration of chitosan in the system is 5g / L. Mannose is added to the system so that the mass ratio of chitosan to mannose is 1:3. After mixing, the pH of the solution is adjusted to 5.5, and the reaction is carried out at 65°C for 72h. After the reaction is completed, the solution is placed in an ice bath for 10min, and lyophilized to obtain mannose grafted chitosan.

[0063] S1.3 Preparation of liver protection food: Weigh the above components in the proportion of 20 parts by weight of astaxanthin compound preparation, 10 parts by weight of fucoxanthin compound preparation, 5 parts by weight of taurine, 5 parts by weight of bird's nest peptide, and 5 parts by weight of cuttlefish juice powder, add appropriate amount of magnesium stearate and starch to the formula, mix well, sieve and fill into hard capsules with a fully automatic capsule filling machine, with a filling amount of 0.5g per capsule, to obtain capsule-type functional food.

[0064] Example 2

[0065] S2.1 Preparation of astaxanthin composite preparation: Astaxanthin, soybean saponin, and triterpene acid were weighed in a mass ratio of 1:30:1, and after dissolving with a small amount of anhydrous ethanol, they were stirred evenly and ultrasonicated for 2 minutes to obtain the first fusion micelle. Ovalbumin was dissolved in deionized water to obtain a protein solution with a mass concentration of 2%. The first fusion micelle was added to the protein solution treated with a pulsed electric field of 30 kV / cm for 6 minutes and stirred to make the mass ratio of astaxanthin to protein 1:20. A high-pressure homogenizer was used for homogenization at a pressure of 80 MPa for 4 minutes to obtain the first complex aqueous solution. The first fusion micelle containing astaxanthin was encapsulated by a protein shell. Antifreeze peptide with an average molecular mass of 1000 Da was added to the first complex aqueous solution to make the mass concentration of antifreeze peptide in the solution 1%, and then fucoidan was added to make the mass concentration of fucoidan in the first complex aqueous solution 0.3%. After mixing, it was placed in a cold storage at -2°C. The solution temperature was reduced to -2°C, and the pH of the system was adjusted to 3.0 to obtain the first complex suspension. Finally, a core-shell structure was obtained in which astaxanthin was encapsulated inside the protein shell and polysaccharides were adsorbed on the outside. The encapsulation efficiency of astaxanthin in the first complex suspension was 98.4%, and the particle size of the core-shell structure encapsulating astaxanthin was 640.0nm. The first complex suspension was centrifuged and dried to obtain an astaxanthin composite preparation dry powder. The obtained astaxanthin composite preparation dry powder was redissolved in deionized water and heated at 121°C for 15 minutes, and the retention rate of astaxanthin was 91.3%.

[0066] S2.2 Preparation of fucoxanthin composite preparation: Weigh fucoxanthin, glycyrrhizic acid, and isoquercetin in a mass ratio of 1:20:1, use a small amount of anhydrous ethanol to dissolve, stir evenly, and ultrasonicate for 2 minutes to obtain the second fusion micelle. Dissolve ovalbumin in deionized water to obtain a protein solution with a mass concentration of 2%. Add the second fusion micelle to the protein solution treated with a pulsed electric field of 30kV / cm for 6 minutes and stir to mix. The mass ratio of fucoxanthin to ovalbumin is 1:20. Use a high-pressure homogenizer to homogenize at a pressure of 80MPa for 4 minutes to obtain a second complex aqueous solution. The second fusion micelle containing fucoxanthin is encapsulated by a protein shell. Add an antifreeze peptide with an average molecular mass of 1000Da to the second complex aqueous solution. The mass concentration of the antifreeze peptide in the solution is 1%. Add mannose-grafted chitosan to make its mass concentration in the solution 0.2%. After mixing, place it in a cold storage at -2°C. When the solution temperature dropped to -2°C, the pH of the system was adjusted to 5.5 to obtain a second complex suspension. Finally, a core-shell structure was obtained in which fucoxanthin was encapsulated inside the protein shell and mannose-grafted chitosan was adsorbed on the outside. The encapsulation efficiency of fucoxanthin in the second complex suspension was 96.2%, and the particle size of the core-shell structure encapsulated with fucoxanthin was 600.0nm. The second complex suspension was centrifuged and dried to obtain a fucoxanthin composite preparation dry powder. The fucoxanthin composite preparation dry powder was redissolved in deionized water and heated at 121°C for 15 minutes, and the retention rate of fucoxanthin was 91.8%.

[0067] The preparation method of mannose grafted chitosan is as follows: chitosan is dissolved in an acetic acid aqueous solution with a volume concentration of 1%, and the final concentration of chitosan in the system is 5g / L. Mannose is added to the system so that the mass ratio of chitosan to mannose is 1:3. After mixing, the pH of the solution is adjusted to 5.0, and the reaction is carried out at 60°C for 70h. After the reaction is completed, the solution is placed in an ice bath for 10min, and the solution is freeze-dried to obtain mannose grafted chitosan.

[0068] S2.3 Preparation of liver-protecting functional food: weigh the above components in the proportion of 25 parts by weight of astaxanthin compound preparation, 15 parts by weight of fucoxanthin compound preparation, 4 parts by weight of taurine, 4 parts by weight of oyster peptide, and 4 parts by weight of cuttlefish juice powder, and then add appropriate amount of dextrin, starch, microcrystalline cellulose and lactose or more, mix evenly, moisten with appropriate amount of 70% (v / v) ethanol, make a soft material, granulate through a 30-mesh sieve, dry at 70-80°C, sieve the whole granules through a 60-mesh sieve, and press into tablets to obtain tablets, each tablet is 0.2g, and a tablet-type functional food is obtained.

[0069] Example 3

[0070] S3.1 Preparation of astaxanthin composite preparation: Astaxanthin, soybean saponin, and triterpene acid are weighed in a mass ratio of 1:30:1, and after dissolving with a small amount of anhydrous ethanol, stirred evenly, and ultrasonicated for 2 minutes to obtain the first fusion micelle. Whey protein is dissolved in deionized water to obtain a protein solution with a mass concentration of 2%. The first fusion micelle is added to the protein solution treated with a pulsed electric field of 30kV / cm for 6 minutes and stirred to make the mass ratio of astaxanthin to protein 1:20. A high-pressure homogenizer is used for homogenization at a pressure of 100MPa for 2 minutes to obtain a first complex aqueous solution. The first fusion micelle containing astaxanthin is encapsulated by a protein shell. Antifreeze peptide with an average molecular weight of 1500Da is added to the first complex aqueous solution to make the mass concentration of antifreeze peptide in the solution 1%, and then sulfated galactan is added to make the mass concentration of sulfated galactan in the first complex aqueous solution 0.4%. After mixing, it is placed in a cold storage at -2°C. The solution temperature was reduced to -2°C, and the pH of the system was adjusted to 3.5 to obtain the first complex suspension. Finally, a core-shell structure was obtained in which astaxanthin was encapsulated inside the protein shell and polysaccharides were adsorbed on the outside. The encapsulation efficiency of astaxanthin in the first complex suspension was 98.6%, and the particle size of the core-shell structure encapsulating astaxanthin was 660.0nm. The first complex suspension was centrifuged and dried to obtain an astaxanthin composite preparation dry powder. The obtained astaxanthin composite preparation dry powder was redissolved in deionized water and heated at 121°C for 15 minutes, and the retention rate of astaxanthin was 92.4%.

[0071] S3.2 Preparation of fucoxanthin composite preparation: Weigh fucoxanthin, glycyrrhizic acid, and isoquercetin at a mass ratio of 1:20:1, use a small amount of anhydrous ethanol to assist dissolution, stir evenly, and ultrasonicate for 2 minutes to obtain the second fusion micelle. Dissolve whey protein in deionized water to obtain a protein solution with a mass concentration of 2%. Add the second fusion micelle to the protein solution treated with a pulsed electric field of 30kV / cm for 6 minutes and stir and mix. The mass ratio of fucoxanthin to protein is 1:20. Use a high-pressure homogenizer to homogenize at a pressure of 100MPa for 2 minutes to obtain a second complex aqueous solution. The second fusion micelle containing fucoxanthin is encapsulated by a protein shell. Add an antifreeze peptide with an average molecular weight of 1500Da to the second complex aqueous solution. The mass concentration of the antifreeze peptide in the solution is 1%. Add mannose-grafted chitosan to make its mass concentration in the solution 0.3%. After mixing, place it in a cold storage at -2°C. When the solution temperature dropped to -2°C, the pH of the system was adjusted to 6.0 to obtain a second complex suspension. Finally, a core-shell structure was obtained in which fucoxanthin was encapsulated inside the protein shell and mannose-grafted chitosan was adsorbed on the outside. The encapsulation efficiency of fucoxanthin in the second complex suspension was 96.8%, and the particle size of the core-shell structure encapsulated with fucoxanthin was 645.0 nm. The second complex suspension was centrifuged and dried to obtain a fucoxanthin composite preparation dry powder. The fucoxanthin composite preparation dry powder was redissolved in deionized water and heated at 121°C for 15 minutes, and the retention rate of fucoxanthin was 92.1%.

[0072] The preparation method of mannose grafted chitosan is as follows: chitosan is dissolved in an acetic acid aqueous solution with a volume concentration of 1%, and the final concentration of chitosan in the system is 5g / L. Mannose is added to the system so that the mass ratio of chitosan to mannose is 1:3. After mixing, the pH of the solution is adjusted to 6.0, and the reaction is carried out at 60°C for 80h. After the reaction is completed, the solution is placed in an ice bath for 10min, and lyophilized to obtain mannose grafted chitosan.

[0073] S3.3 Preparation of liver-protecting functional food: weigh the above components in the proportion of 30 parts by weight of astaxanthin compound preparation, 20 parts by weight of fucoxanthin compound preparation, 3 parts by weight of taurine, 3 parts by weight of corn peptide, and 3 parts by weight of cuttlefish juice powder, and then add appropriate amount of soluble starch, dextrin, sucrose, lactose and mannitol, moisten with appropriate amount of 70% (v / v) ethanol, make a soft material, granulate through a 20-mesh sieve, dry at 70-80℃, and granulate into 60-mesh granules to obtain functional food of the granule type.

[0074] Example 4

[0075] S4.1 Preparation of astaxanthin composite preparation: Astaxanthin, soybean saponin, and triterpene acid are weighed in a mass ratio of 1:30:1, and after dissolving with a small amount of anhydrous ethanol, they are stirred evenly and ultrasonicated for 2 minutes to obtain the first fusion micelle. Peanut protein is dissolved in deionized water to obtain a protein solution with a mass concentration of 2%. The first fusion micelle is added to the protein solution treated with a pulsed electric field with an intensity of 30kV / cm for 6 minutes and stirred to make the mass ratio of astaxanthin to protein 1:20. A high-pressure homogenizer is used for homogenization at a pressure of 120MPa for 1 minute to obtain a first complex aqueous solution, and the first fusion micelle containing astaxanthin is encapsulated by a protein shell. Antifreeze peptide with an average molecular mass of 2000Da is added to the first complex aqueous solution to make the mass concentration of antifreeze peptide in the solution 1%, and then hyaluronic acid is added to make the mass concentration of hyaluronic acid in the first complex aqueous solution 0.5%, and after mixing, it is placed in a cold storage at -2°C. The solution temperature is reduced to -2°C, and the pH of the system is adjusted to 4.0 to obtain the first complex suspension. Finally, a core-shell structure with astaxanthin encapsulated inside the protein shell and polysaccharides adsorbed outside was obtained. The encapsulation efficiency of astaxanthin in the first complex suspension was 98.8%, and the particle size of the core-shell structure encapsulating astaxanthin was 700.0 nm. The first complex suspension was centrifuged and dried to obtain an astaxanthin composite preparation dry powder. The obtained astaxanthin composite preparation dry powder was redissolved in deionized water and heated at 100°C for 15 minutes, and the retention rate of astaxanthin was 93.1%.

[0076] S4.2 Preparation of fucoxanthin composite preparation: Weigh fucoxanthin, glycyrrhizic acid, and isoquercetin in a mass ratio of 1:20:1, use a small amount of anhydrous ethanol to assist dissolution, stir evenly, and ultrasonicate for 2 minutes to obtain the second fusion micelle. Dissolve peanut protein in deionized water to obtain a protein solution with a mass concentration of 2%. Add the second fusion micelle to the protein solution treated with a pulsed electric field of 30kV / cm for 6 minutes and stir to mix. The mass ratio of fucoxanthin to protein is 1:20. Use a high-pressure homogenizer to homogenize at a pressure of 120MPa for 1 minute to obtain a second complex aqueous solution. The second fusion micelle containing fucoxanthin is encapsulated by a protein shell. Add an antifreeze peptide with an average molecular weight of 2000Da to the second complex aqueous solution. The mass concentration of the antifreeze peptide in the solution is 1%. Add mannose-grafted chitosan to make its mass concentration in the solution 0.5%. After mixing, place it in a cold storage at -2°C. When the solution temperature dropped to -2°C, the pH of the system was adjusted to 6.0 to obtain a second complex suspension. Finally, a core-shell structure was obtained in which fucoxanthin was encapsulated inside the protein shell and mannose-grafted chitosan was adsorbed on the outside. The encapsulation efficiency of fucoxanthin in the second complex suspension was 97.1%, and the particle size of the core-shell structure encapsulated with fucoxanthin was 680.0nm. The second complex suspension was centrifuged and dried to obtain a fucoxanthin composite preparation dry powder. The fucoxanthin composite preparation dry powder was redissolved in deionized water and heated at 121°C for 15 minutes, and the retention rate of fucoxanthin was 92.9%.

[0077] The preparation method of mannose grafted chitosan is as follows: chitosan is dissolved in an acetic acid aqueous solution with a volume concentration of 1%, and the final concentration of chitosan in the system is 5g / L. Mannose is added to the system so that the mass ratio of chitosan to mannose is 1:3. After mixing, the pH of the solution is adjusted to 5.5, and the reaction is carried out at 65°C for 72h. After the reaction is completed, the solution is placed in an ice bath for 10min, and lyophilized to obtain mannose grafted chitosan.

[0078] S4.3 Preparation of liver-protecting functional food: Weigh the above components in the proportion of 40 parts by weight of astaxanthin compound preparation, 30 parts by weight of fucoxanthin compound preparation, 1 part by weight of taurine, 1 part by weight of collagen peptide, and 1 part by weight of cuttlefish juice powder, then dissolve them in water, add appropriate amounts of sweeteners and stabilizers, mix well, homogenize, filter, fill, and sterilize to obtain a beverage-type functional food.

[0079] Embodiment 5:

[0080] (1) Evaluation of oral absorption of astaxanthin compound preparations

[0081] Table 1. Astaxanthin test samples and their preparation methods

[0082]

[0083]

[0084] 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 solubilization if necessary), and convert the dose to ensure that the astaxanthin dose for each group is 10.0 mg / kg, which is the test solution.

[0085] Experimental animals: SPF male ICR mice, 15-20 g, were purchased from Shanghai Slake Laboratory Animal Co., Ltd.

[0086] 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.

[0087] The temperature of the breeding room was 23±2℃ and the relative humidity was 55±5%.

[0088] Animal grouping and treatment method: After 1 week of adaptive feeding, the mice were randomly divided into 18 groups according to their body weight, with 7 mice in each group, including a blank group (gavage with deionized water) and 17 groups of astaxanthin test samples as described in Table 1.

[0089] Each mouse was given a dose of 10.0 mg / kg. After 4 hours, the mouse eyeballs were removed and blood was collected. The astaxanthin content in the serum was analyzed by high-performance liquid chromatography (HPLC). The expression levels of SR-B1, NPC1L1, 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-delivery system on promoting intestinal absorption of astaxanthin.

[0090] Experimental results:

[0091] Table 2. Oral absorption effect of astaxanthin and expression of key transport proteins

[0092]

[0093]

[0094] HPLC was used to analyze the astaxanthin concentration in the serum of mice in each treatment group, and the results are shown in Table 2. The results showed that the serum concentration of unencapsulated astaxanthin was only 30.48 ng / mL after oral gavage. When astaxanthin and soybean saponin were combined at a mass ratio of 1:30, a good absorption-promoting effect was shown, and the serum concentration of astaxanthin increased to 223.46 ng / mL; when the ratio of soybean saponin was reduced to 1:25, the serum concentration of astaxanthin decreased (P<0.01), but it still had a good effect compared with the astaxanthin group, which was 173.22 ng / mL; when the ratio of soybean saponin was increased to 1:35, the serum concentration of astaxanthin did not change significantly compared with the effect of the mass ratio of 1:30 (P<0.05), and both had a high absorption effect. When the mass ratio of astaxanthin to soybean saponin is 1:10, although the serum concentration of astaxanthin decreased compared with other test groups (P<0.01), it decreased to 182.43ng / mL, but it still had a good absorption effect compared with the astaxanthin group. Correspondingly, the expression levels of SR-B1 and NPC1L1, the transporters responsible for astaxanthin across the intestinal epithelium, changed accordingly, indicating that soybean saponin can promote the intestinal absorption of astaxanthin by upregulating the expression of transporters. It is worth noting that the higher the expression level of the efflux transporter P-gp, the lower the serum concentration of astaxanthin. It shows that P-gp is involved in the efflux of astaxanthin and inhibits the accumulation of astaxanthin in cells. Considering the factors of comprehensive cost and the absorption effect of astaxanthin, the optimal mass ratio of astaxanthin to soybean saponin is 1:30, but there is a good intestinal absorption effect in the ratio range of 1:25-40 proposed in this embodiment.

[0095] Further introduction of triterpenic acid, when the mass ratio of astaxanthin + soybean saponin + triterpenic acid was 1:30:1, the serum concentration of astaxanthin reached 698.39pmol / mL, which was 21.91 times higher than that of the astaxanthin group (P<0.01). By observing the expression of transporters, the expression levels of SR-B1 and NPC1L1, which promote the intestinal epithelial uptake of astaxanthin, were further increased, while the expression of the efflux transporter P-gp was significantly inhibited. If the ratio of triterpenic acid is reduced, so that the mass ratio of astaxanthin + soybean saponin + triterpenic acid is 1:30:0.5, the serum concentration of astaxanthin is 574.25pmol / mL, which still has a significant improvement in intestinal absorption compared to the serum concentration of astaxanthin (223.46pmol / mL) when only astaxanthin and soybean saponin are used. If the proportion of triterpenic acid in the combination is further increased, when the mass ratio of astaxanthin + soybean saponin + triterpenic acid is 1:30:1.5, compared with the ratio of 1:30:1, the serum concentration of astaxanthin has no significant change (P<0.05), and the serum of the test group has a higher concentration of astaxanthin; but when the mass ratio of astaxanthin + soybean saponin + triterpenic acid is 1:30:2, the serum concentration of astaxanthin drops to 556.79pmol / mL, and at the same time, the transporters SR-B1 and NPC1L1 responsible for the cellular uptake of astaxanthin are significantly inhibited, and further increasing the concentration of triterpenic acid is not conducive to the cell's uptake of astaxanthin. Therefore, the mass ratio of astaxanthin, soybean saponin and triterpenic acid is in the range of 1:25-40:0.5-2. Compared with the test group using only astaxanthin and soybean saponin, the test group using astaxanthin, soybean saponin and triterpenic acid has a higher astaxanthin concentration in the blood. Considering the comprehensive cost and serum concentration of astaxanthin, astaxanthin, soybean saponin and triterpene acid have a strong synergistic absorption-promoting effect when the mass ratio is 1:30:1.

[0096] On this basis, astaxanthin, soybean saponin and triterpenic acid were co-loaded in the form of fused micelles according to the method of the present invention, and the absorption effect of the protein-encapsulated astaxanthin complex and the astaxanthin composite preparations of Examples 1-4 were evaluated. The results showed that the intestinal absorption effect of astaxanthin can be further improved by encapsulating the first fused micelle formed by astaxanthin-soybean saponin-triterpenic acid with protein alone. At the same time, the protein addition ratio also has a certain effect on the absorption of astaxanthin. When the mass ratio of astaxanthin to protein is 1:15, the serum concentration of astaxanthin is 721.34pmol / mL; when the protein ratio is further increased to make the mass ratio of astaxanthin to protein reach 1:20, the serum concentration of astaxanthin reaches the maximum, which is 784.52pmol / mL. When the protein ratio was further increased to 1:25 or 1:30, the serum concentration of astaxanthin decreased, especially at 1:30, it decreased significantly (P<0.05), down to 718.62pmol / mL. In the range of 1:15-30 of the mass ratio of astaxanthin concentration to protein, the concentration of astaxanthin in the blood of the test group was higher than that of the astaxanthin-soy saponin-triterpenic acid fusion micelle test group that was not embedded by protein. Taking all factors into consideration, the optimal mass ratio of astaxanthin, soybean saponin, triterpenic acid and protein is 1:30:1:20.

[0097] When the protein particles are further modified with polysaccharide surface molecules, the absorption effect is further improved: after the astaxanthin composite preparations of Examples 1-4 were gavaged into mice, the serum concentrations reached 923.54pmol / mL, 928.75pmol / mL, 940.12pmol / mL, and 950.98pmol / mL, respectively. Compared with the unencapsulated astaxanthin, the absorption effect was increased by 29.3 times, 29.47 times, 29.84 times, and 30.2 times, respectively, with a very significant improvement effect (P<0.01). However, the expression of the transport proteins SR-B1 and NPC1L1 responsible for the absorption of astaxanthin across the intestinal epithelium did not change significantly.

[0098] This shows that the beneficial effect of the present invention on efficient intestinal absorption of astaxanthin is achieved through the absorption-promoting nutrient combination technology, pulsed electric field coupled high-voltage homogenization technology, and low-temperature ion-driven self-assembly technology. It can not only regulate the expression of intestinal epithelial transport proteins, but also improve the digestive stability of astaxanthin, thereby achieving efficient oral absorption of astaxanthin, and the absorption efficiency is increased by 29.3-30.2 times.

[0099] (2) Evaluation of oral absorption of fucoxanthin compound preparations

[0100] Table 3. Fucoxanthin test samples and their preparation methods

[0101]

[0102] 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 solubilization if necessary), and convert the dose to ensure that the dose of fucoxanthin in each group is 10.0 mg / kg, which is the test solution.

[0103] Experimental animals: SPF male ICR mice, 15-20 g, were purchased from Shanghai Slake Laboratory Animal Co., Ltd.

[0104] 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.

[0105] The temperature of the breeding room was 23±2℃ and the relative humidity was 55±5%.

[0106] Animal grouping and treatment method: After 1 week of adaptive feeding, the mice were randomly divided into 18 groups according to body weight, with 7 mice in each group, including a blank group (administered with deionized water) and 17 groups of fucoxanthin test samples described in Table 3. Each mouse was given a dose of 10.0 mg / kg. After 4 hours, the eyeballs of the mice were removed to collect blood, and the content of fucoxanthin and its metabolites in plasma was analyzed by high performance liquid chromatography (HPLC); the expression levels of intestinal SR-B1, CD36 and P-gp 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 intestinal absorption of fucoxanthin.

[0107] Table 4. Oral absorption effect of fucoxanthin and expression of key transport proteins

[0108]

[0109]

[0110] 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 after oral gavage was only 42.31pmol / mL. When fucoxanthin and glycyrrhizic acid were combined at a mass ratio of 1:20, a better absorption-promoting effect was shown, and the serum corresponding concentration of fucoxanthin increased to 247.35pmol / mL, an increase of 4.85 times (P<0.01). Correspondingly, the expression levels of SR-B1 and CD36, the transporters responsible for the absorption of fucoxanthin across the intestinal epithelium, increased accordingly, but the expression level of the efflux transporter P-gp also increased at the same time. If the ratio of glycyrrhizic acid is reduced to a mass ratio of fucoxanthin to glycyrrhizic acid of 1:15, the serum corresponding concentration of fucoxanthin decreased significantly to 179.24pmol / mL, but it still had a better absorption effect than the fucoxanthin group (i.e., the content of fucoxanthin in serum was significantly increased). When glycyrrhizic acid is added to make the mass ratio of fucoxanthin to glycyrrhizic acid reach 1:25 or 1:30, the corresponding serum concentration of fucoxanthin also decreases, especially at 1:30, the corresponding serum concentration of fucoxanthin decreases to 209.63pmol / mL, showing a significant decrease (P<0.01). At this time, we can see that the expression of the efflux transporter P-gp gradually increases. It shows that higher glycyrrhizic acid will lead to an increase in the expression of P-gp, and the fucoxanthin that has been taken up by intestinal cells will be excreted into the intestinal cavity through exocytosis, resulting in a deterioration of the overall absorption function. Therefore, in the range of 1:15-30 of the mass ratio of fucoxanthin to glycyrrhizic acid described in the present invention, the concentration of fucoxanthin in serum is higher, and has a better absorption effect. Considering the comprehensive cost and absorption effect, the optimal mass ratio of fucoxanthin to glycyrrhizic acid is 1:20.

[0111] Further introduction of P-gp inhibitor isoquercetin, making the mass ratio of fucoxanthin to isoquercetin 1:0.5, the serum response concentration of fucoxanthin increased significantly (P<0.01), reaching 452.25pmol / mL; when the mass ratio of fucoxanthin to isoquercetin was further increased to 1:1, the serum response concentration of fucoxanthin reached 573.28pmol / mL, which was 12.55 times higher than that of the fucoxanthin group. By observing the expression of transporters, compared with the fucoxanthin + glycyrrhizic acid (mass ratio 1:20) group, the expression levels of transporters SR-B1 and CD36 that promote fucoxanthin absorption did not change significantly (P<0.05), but the expression of efflux transporter P-gp was significantly inhibited (P<0.01). When the ratio of isoquercetin was further increased, and the mass ratio of fucoxanthin to isoquercetin was increased to 1:1.5 or 1:2, the serum response concentration of fucoxanthin showed a downward trend. At the same time, it was found that the expression levels of SR-B1 and CD36 decreased. This shows that high concentrations of isoquercetin will inhibit the expression of transporters that promote the uptake of fucoxanthin, resulting in a poorer overall absorption effect. Therefore, fucoxanthin, glycyrrhizic acid, and isoquercetin have a strong synergistic absorption-promoting effect within the mass ratio range of 1:15-30:0.5-2.

[0112] On this basis, according to the method of the present invention, fucoxanthin, glycyrrhizic acid and isoquercetin were co-loaded in the form of fused micelles, and the absorption effect of the protein-encapsulated fucoxanthin complex and the fucoxanthin composite preparations of Examples 1-4 were evaluated. The results showed that the encapsulation of the second fused micelle formed by fucoxanthin-glycyrrhizic acid-isoquercetin with protein alone can improve the intestinal absorption effect of fucoxanthin to a certain extent. However, the protein ratio also has a certain effect. For example, when the mass ratio of fucoxanthin to protein increases from 1:15 to 1:20, the serum response concentration of fucoxanthin increases from 584.37pmol / mL to 615.44pmol / mL. When the mass ratio of fucoxanthin to protein is further increased to 1:25 or 1:30, the absorption effect of fucoxanthin is somewhat reduced. This may be due to the increase in the protein ratio, which changes the structure and digestion characteristics of the complex. Therefore, the mass ratio of fucoxanthin, glycyrrhizic acid, isoquercetin and protein in the range of 1:15-30:0.5-2:15-30 has a better absorption-promoting effect.

[0113] When the surface molecular modification of the protein particles is further performed using a ligand polysaccharide (mannose grafted chitosan) with intestinal epithelial mannose receptors, the absorption effect is greatly improved. After the fucoxanthin composite preparations of Examples 1-4 were gavaged into mice, the serum response concentrations reached 867.36pmol / mL, 876.51pmol / mL, 889.27pmol / mL, and 905.43pmol / mL, respectively. Compared with the unencapsulated astaxanthin, the absorption effect was increased by 19.5 times, 19.7 times, 20.0 times, and 20.4 times, respectively, with a very significant absorption-promoting effect (P<0.01). It is worth noting that the expression levels of the transporters SR-B1 and NPC1L1 responsible for the absorption of fucoxanthin across the intestinal epithelium, as well as the efflux transporter P-pg, did not change significantly.

[0114] This shows that the beneficial effect of the present invention on achieving efficient intestinal absorption of fucoxanthin is due to the use of absorption-promoting nutrient combination technology on the one hand, and the use of pulsed electric field coupled high-voltage homogenization technology and low-temperature ion-driven self-assembly technology to modify the surface molecules of the delivery carrier with intestinal affinity adsorption effect on the other hand, thereby achieving efficient oral absorption of fucoxanthin, and the absorption efficiency is increased by 19.5-20.4 times.

[0115] (3) Evaluation of product liver protection function

[0116] Test samples: astaxanthin, fucoxanthin, astaxanthin + fucoxanthin physical mixture (mass ratio 20:10), astaxanthin + fucoxanthin + taurine physical mixture (mass ratio 20:10:5), astaxanthin + fucoxanthin + taurine + bird's nest peptide physical mixture (mass ratio 20:10:5:5), astaxanthin + fucoxanthin + taurine + bird's nest peptide + cuttlefish juice powder physical mixture (mass ratio 20:10:5:5:5), and the formula products obtained in Examples 1-4 of the present invention.

[0117] Sample treatment: Dissolve the test sample directly in deionized water at a mass-to-volume ratio of 1:10 (use appropriate amount of ethanol to assist dissolution if necessary), and convert the dose. The dosage of all test samples is 50.0 mg / kg, which is the test solution.

[0118] Experimental animals and breeding environment: SPF-grade male C57BL / 6J mice, 15-20 g, were purchased from Shanghai Slake Laboratory Animal Co., Ltd.

[0119] 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.

[0120] The temperature of the breeding room was 23±2℃ and the relative humidity was 55±5%.

[0121] Animal grouping and treatment methods: Mice were adaptively fed for one week and fed freely. After one week, they were randomly divided into 12 groups according to body weight, namely the normal group, the model group and the above 10 test groups, with 7 experimental animals in each group. The acute alcoholic liver injury model was established by inducing mice with ethanol. Except for the normal group, all other groups were gavaged with 56% ethanol (10 mg / kg) once a day for 3 consecutive days. After modeling, the drug-treated group was gavaged with different test samples once a day, and the normal group and the model group were gavaged with equal amounts of water for 7 consecutive days. After fasting for 12 hours, blood was collected and liver function was evaluated by analyzing the levels of transaminases AST and ALT. The changes in liver inflammation were evaluated by detecting the levels of IL-1β, IL-6 and TNF-α in liver tissue.

[0122] Experimental results:

[0123] Table 5. Evaluation table of liver protection function of each component and product

[0124]

[0125] An acute alcoholic hepatitis model was established using a mouse model. Astaxanthin and fucoxanthin were selected from a number of dietary ingredients as the core functional ingredients for combined liver protection. The liver protection effect of the formula combination of astaxanthin, fucoxanthin, taurine, protein peptide, and cuttlefish juice powder was investigated. Further relying on the strategy of improving the oral bioavailability of the core ingredients, the liver protection effect of the delivery system composition (Examples 1-4) constructed by special encapsulation technology was evaluated, and the results are shown in Table 5.

[0126] Although astaxanthin and fucoxanthin alone can play a certain role in protecting the liver, the effect is very limited. When astaxanthin and fucoxanthin were combined in a mass ratio of 20:10, the serum AST and ALT of mice were significantly reduced from 50.8U / L and 48.4U / L in the model group to 40.2U / L and 37.4U / L (P<0.01), and the level of liver inflammatory factors also decreased significantly (P<0.01). When taurine was further introduced, the mass ratio of astaxanthin, fucoxanthin and taurine was 20:10:5, respectively, and serum AST and ALT were further significantly reduced (P<0.01); the same trend was observed when protein peptides and cuttlefish juice powder were further introduced in sequence.

[0127] When the mass ratio of astaxanthin + fucoxanthin + taurine + protein peptide + cuttlefish juice powder was 20:10:5:5:5, serum AST and ALT were reduced to 27.6U / L (45.7% decrease) (P<0.01) and 20.1U / L (58.5% decrease) (P<0.01), respectively, and the levels of liver inflammatory factors IL-1β, IL-6 and TNF-α were also reduced to 56.4ng / L, 24.5mg / L and 547.6ng / mL, respectively, down 38.8% (P<0.01), 47.3% (P<0.01) and 30.4% (P<0.01), respectively. This indicates that astaxanthin, fucoxanthin, taurine, protein peptide and cuttlefish juice powder have synergistic liver protection effects under this ratio.

[0128] When the products of Examples 1-4 were used for intervention, the liver protection effect was more significantly improved. The products of Examples 1-4 were able to further reduce the serum AST and ALT of mice to 20.9-22.3U / L (a decrease of 56.1-58.9%) (P<0.01) and 16.3-17.6U / L (a decrease of 63.6-66.3%) (P<0.01), respectively, and the levels of liver inflammatory factors IL-1β, IL-6 and TNF-α were further reduced to 45.8-46.7ng / L (P<0.01), 18.6-19.1mg / L (P<0.01) and 483.4-492.6ng / mL (P<0.01), respectively, which were very close to the liver function indicators of mice in the normal group.

[0129] It can be seen that the formula product disclosed in the present invention has a good liver protection function, and the beneficial effects of the present invention are attributed to the nutritional synergistic formula design technology adopted on the one hand, and the innovative molecular encapsulation technology adopted by the present invention on the other hand, which promotes the intestinal absorption efficiency of astaxanthin and fucoxanthin, thereby further improving the efficacy of the formula product in liver protection function.

[0130] 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 liver-protecting food, characterized in that: The invention comprises an astaxanthin compound preparation, a fucoxanthin compound preparation, taurine, protein peptide and cuttlefish juice powder, and the mass ratio thereof is 20-40:10-30:1-5:1-5:1-5:1-5.

2. The liver-protecting food according to claim 1, characterized in that: The astaxanthin composite preparation comprises astaxanthin, soybean saponin and triterpenic acid, and the mass ratio of the astaxanthin, soybean saponin and triterpenic acid is 1:25-40:0.5-2.

3. The liver-protecting food according to claim 1, characterized in that: The fucoxanthin composite preparation comprises fucoxanthin, glycyrrhizic acid and isoquercetin, and the mass ratio of the fucoxanthin, glycyrrhizic acid and isoquercetin is 1:15-30:0.5-2.

4. The liver-protecting food according to claim 1, characterized in that: The astaxanthin composite preparation is prepared by mixing and dissolving astaxanthin, soybean saponin and triterpenic acid to obtain a first fused micelle, adding the first fused micelle to a denatured protein solution, mixing and homogenizing to obtain a first complex aqueous solution with protein as an outer shell and astaxanthin, soybean saponin and triterpenic acid as an inner core, adding antifreeze peptides and polysaccharides to mix, adjusting the pH to acidic at low temperature to obtain a first complex suspension with polysaccharides adsorbed on the surface of the protein shell, and centrifuging and drying the first complex suspension to obtain the astaxanthin composite preparation.

5. The liver-protecting food according to claim 1, characterized in that: The fucoxanthin composite preparation is prepared by mixing and dissolving fucoxanthin, glycyrrhizic acid and isoquercetin to obtain a second fused micelle, adding the second fused micelle to a denatured protein solution, mixing and homogenizing to obtain a second complex aqueous solution with protein as the shell and fucoxanthin, glycyrrhizic acid and isoquercetin as the core, adding antifreeze peptide and mannose-grafted chitosan to mix, adjusting the pH to acidic at low temperature to obtain a second complex suspension with mannose-grafted chitosan adsorbed on the protein shell, and centrifuging and drying the second complex suspension to obtain the fucoxanthin composite preparation; Preferably, mannose is dissolved in an acid solution of chitosan to obtain a mixed solution, the pH of the mixed solution is adjusted and then heated for reaction, and after the reaction is completed, ice bathing and freeze drying are performed to obtain water-soluble mannose-grafted chitosan; Preferably, the concentration of the chitosan acid solution is 5.0 g / L.

6. A method for preparing a liver-protecting food, characterized in that: Used for preparing the liver protection food according to any one of claims 1 to 5, comprising: S1: astaxanthin, soybean saponin, and triterpenic acid are mixed and dissolved to obtain a first fused micelle, and the first fused micelle is added to a denatured protein solution, mixed and homogenized to obtain a first complex aqueous solution with protein as the shell and astaxanthin, soybean saponin, and triterpenic acid as the core, and antifreeze peptides and polysaccharides are added and mixed, and the pH is adjusted to acidic at low temperature so that the protein shell has an opposite charge to the polysaccharide to obtain a first complex suspension with polysaccharide adsorbed on the surface of the protein shell, and the first complex suspension is centrifuged and dried to obtain an astaxanthin composite preparation; S2: fucoxanthin, glycyrrhizic acid and isoquercetin are mixed and dissolved to obtain a second fused micelle, the second fused micelle is added to the denatured protein solution, mixed and homogenized to obtain a second complex aqueous solution with protein as the shell and fucoxanthin, glycyrrhizic acid and isoquercetin as the core, antifreeze peptide and mannose-grafted chitosan are added and mixed, the pH is adjusted to acidic at low temperature to obtain a second complex suspension with mannose-grafted chitosan adsorbed on the protein shell, and the second complex suspension is centrifuged, frozen and dried to obtain a fucoxanthin composite preparation; The order of S1 and S2 is not limited and they can be switched or performed simultaneously. S3: the prepared astaxanthin composite preparation and fucoxanthin composite preparation are mixed with taurine, protein peptide, cuttlefish juice powder, water or filler excipients, and then sterilized to prepare the finished product; Preferably, in step S1, the pH is adjusted to 3.0-4.0 at low temperature to obtain a first complex suspension in which polysaccharides are adsorbed on the surface of the protein shell; Preferably, in step S2, the pH is adjusted to 5.0-6.0 at low temperature to obtain a second complex suspension in which mannose-grafted chitosan is adsorbed on the surface of the protein shell; Preferably, the polysaccharide in step S1 is any one of pectin, hyaluronic acid, fucoidan, and galactan sulfate; Preferably, the denatured protein solution in step S1 and step S2 is obtained by treating the protein solution by any one of electric field, ultrasound, microwave and high static pressure; more preferably, the electric field is a pulse electric field with an intensity of 30-40 kV / cm for 6-8 min; Preferably, the homogenization is high-pressure homogenization, more preferably homogenization under a high pressure of 60 MPa-120 MPa for 1-5 min; Preferably, the mass ratio of astaxanthin to protein in the first complex aqueous solution in step S1 is 1:15-30, and the mass ratio of fucoxanthin to protein in the second complex aqueous solution in step S2 is 1:15-30; more preferably, the protein is any one of bovine serum albumin, ovalbumin, whey protein, peanut protein, and soy protein; Preferably, the molecular weight of the antifreeze peptide is 500-2000Da.

7. The method for preparing the liver-protecting food according to claim 6, characterized in that: Dissolving mannose in an acid solution of chitosan to obtain a mixed solution, adjusting the pH of the mixed solution and then heating the mixed solution for reaction, and after the reaction is completed, ice bathing and freeze drying to obtain water-soluble mannose-grafted chitosan; Preferably, the concentration of the chitosan acid solution is 5.0 g / L; Preferably, the mass ratio of chitosan to mannose in the mixed solution is 1:3; Preferably, the pH of the mixed solution is adjusted to 5-6; Preferably, the heating reaction is carried out at 60-70° C. for 70-80 hours.

8. The method for preparing the liver-protecting food according to claim 6, characterized in that: The mass concentration of the polysaccharide in the first complex suspension is 0.2%-0.5%; and / or The mass concentration of the mannose-grafted chitosan in the second complex suspension is 0.1%-0.5%.

9. The liver-protecting food according to any one of claims 1 to 5 and the liver-protecting food obtained by the preparation method according to any one of claims 6 to 8 are used for protecting the liver.

10. The liver-protecting food according to any one of claims 1 to 5 and the liver-protecting food obtained by the preparation method according to any one of claims 6 to 8 are in the form of one or more of capsules, tablets, granules, oral liquids or beverages.

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