An anti-aging composition for improving facial wrinkles and a preparation method thereof

Abstract

The application provides an anti-aging composition for improving facial wrinkles and a preparation method thereof, and belongs to the technical field of anti-aging composition preparation; the preparation process comprises the following steps: preparation of a chelate, preparation of a grifola frondosa fermentation liquor, fermentation of morinda citrifolia pomace by eurotium cristatum, and preparation of the anti-aging composition. The anti-aging composition prepared by compounding sodium hyaluronate, collagen peptide, the chelate, the grifola frondosa fermentation liquor and the morinda citrifolia pomace fermentation liquor can effectively stimulate collagen regeneration and inhibit the damage of free radicals to the skin, and meets the skin rejuvenation effect expected by consumers.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of anti-aging composition preparation, and particularly relates to an anti-aging composition for improving facial wrinkles and a preparation method thereof. BACKGROUND

[0002] With the improvement of living standards and the increasing attention of people to their own image, facial anti-aging has become a hot topic in the field of beauty and skin care. Facial wrinkles, as one of the most intuitive external manifestations of aging, have a relatively complex formation mechanism.

[0003] From a physiological point of view, with the increase of age, the collagen and elastic fibers in the skin gradually lose, and the cell metabolism slows down, so that the skin loses its original tightness and elasticity, and then wrinkles are produced. At the same time, long-term exposure to ultraviolet radiation will damage the skin barrier function, induce a large number of free radicals, accelerate the skin aging process, and cause wrinkles to appear prematurely.

[0004] Under the fast-paced life in modern society, problems such as staying up late, high stress, and unhealthy eating habits are frequent, which further aggravates skin aging. Consumers are extremely urgent for products that can effectively improve facial wrinkles and achieve skin rejuvenation. However, existing anti-aging products have many limitations. Traditional skin care products can mostly only act on the surface of the skin and are difficult to penetrate the bottom layer of the skin to repair damaged tissues from the root.

[0005] Health drinks, as a relatively mild and convenient anti-aging approach, have gradually entered the public's field of vision. They can be absorbed through the digestive system after oral administration, and then act on the skin of the whole body, nourishing the skin from the inside out, regulating body functions, and assisting in reducing wrinkle formation.

[0006] At present, there are many shortcomings in the health drinks on the market that are specially designed to improve facial wrinkles. On the one hand, in terms of efficacy, the anti-wrinkle effect of most products is not significant. The concentration of the effective components they add is insufficient or the component combination is unreasonable, which cannot accurately target the key factors that cause facial wrinkles, such as being unable to effectively stimulate collagen regeneration, inhibit the damage of free radicals to the skin, and effectively improve skin relaxation, reduce wrinkle depth and number, and cannot meet the skin rejuvenation effect expected by consumers.

[0007] From the safety of ingredients, some health drinks add some chemical ingredients with potential risks in pursuit of short-term efficacy. These ingredients may cause burden on the liver and kidney functions of the human body after long-term consumption, and even cause allergic reactions and other adverse reactions, so that consumers face health risks while pursuing beauty.

[0008] Therefore, the anti-aging composition for improving facial wrinkles and a preparation method thereof are provided, which can be applied to the preparation of health drinks to solve the above problems. SUMMARY

[0009] In view of the deficiencies in the prior art, the purpose of the present application is to provide an anti-aging composition for improving facial wrinkles and a preparation method thereof.

[0010] A preparation method of an anti-aging composition for improving facial wrinkles, comprising the following steps:

[0011] S1: Preparation of peanut peptide-selenium chelate

[0012] Peanut protein is extracted from peanut meal, and peanut protein is subjected to enzymolysis of alkaline protease and flavor protease to obtain peanut peptide powder, and then the peanut peptide powder is reacted with selenium yeast to prepare peanut peptide-selenium chelate;

[0013] S2: Preparation of grifola ferroxidans fermentation broth

[0014] Yam powder and noni fruit powder are subjected to alcohol extraction to obtain yam alcohol extract and noni fruit residue, and grifola ferroxidans mycelial blocks are inoculated and activated, and then added to a fermentation medium containing noni fruit alcohol extract and yam alcohol extract for fermentation to obtain grifola ferroxidans fermentation broth;

[0015] S3: Fermentation of noni fruit residue by eurotium cristatum

[0016] Eurotium cristatum is used to inoculate a mixed system of noni fruit residue and deionized water to prepare a fermentation agent, and then the remaining mixed system is used as a fermentation substrate to add the fermentation agent for fermentation to prepare noni fruit residue fermentation product;

[0017] S4: Preparation of anti-aging composition

[0018] The anti-aging composition is prepared by compounding sodium hyaluronate, collagen peptide, peanut peptide-selenium chelate, grifola ferroxidans fermentation broth and noni fruit residue fermentation product.

[0019] Further, the preparation of peanut peptide-selenium chelate in step S1 specifically comprises the following steps:

[0020] S1.1: The peanut meal is added to deionized water, and the pH is adjusted to 9-9.2, and stirred in a 50-60℃ water bath for 2-3h, and then centrifuged at 4000-5000r / min for 15-20min, and the supernatant after centrifugation is adjusted to pH 4.5-4.8, and then centrifuged again at 4000-5000r / min for 15-20min to obtain the precipitate, which is freeze-dried to obtain peanut protein;

[0021] S1.2: 20-30 parts by weight of peanut protein is added to 1000-1200 parts by weight of deionized water, then ultrasonic is performed at 240-250 W power and 55-60 DEG C for 30-40 min, after ultrasonic is completed, the pH is adjusted to 8-9, then alkaline protease is added, the alkaline protease is added in an amount of 4-5% of the mass of the substrate in the system, after reaction for 50-60 min, the enzyme is inactivated at 95-98 DEG C for 10-12 min, to obtain a first enzymolysis solution;

[0022] S1.3: the first enzymolysis solution is heated to 50-60 DEG C, then the pH is adjusted to 5-6, then flavor protease is added, the flavor protease is added in an amount of 4-5% of the mass of the substrate in the system, after reaction for 50-60 min, the enzyme is inactivated at 95-98 DEG C for 10-12 min, then centrifugation is performed at 5000-5500 r / min for 15-20 min, the supernatant is collected and freeze-dried to obtain peanut peptide powder;

[0023] S1.4: 20-30 parts by weight of peanut peptide powder is added to deionized water, stirring and mixing to prepare a polypeptide solution with a concentration of 15-20%, then NaOH solution is added to adjust the pH value to 7.5-8, then 6-8 parts by weight of selenium yeast is added and mixed, and then reaction is performed at 38-40 DEG C for 20-30 min, after reaction is completed, cooling is performed, then centrifugation is performed at 5000-5200 r / min for 15-20 min, to obtain a supernatant, 3-5 times the volume of anhydrous ethanol is added to the supernatant, centrifugation is performed again, and then the precipitate is obtained, and then freeze-drying is performed to obtain peanut peptide-selenium chelate.

[0024] Further, in step S1.1, the peanut meal is mixed with deionized water at a mass ratio of 1:10-11.

[0025] Further, in step S1.1, the peanut meal is peanut meal with a protein content of 48-50%.

[0026] Further, the preparation of the grifola ferroxida fermentation liquor in step S2 specifically includes the following steps:

[0027] S2.1: 20-30 parts by weight of yam powder is added to 100-200 parts by weight of ethanol with a concentration of 80-90%, and oscillation extraction is performed at 25-30 DEG C for 24-25 h, the supernatant is obtained by suction filtration, and after rotary evaporation under reduced pressure, distilled water is added to prepare yam alcohol extract with a concentration of 0.1 g / mL; 10-20 parts by weight of Morinda citrifolia powder is added to 100-200 parts by weight of ethanol with a concentration of 80-90%, and oscillation extraction is performed at 25-30 DEG C for 24-25 h, the supernatant is obtained by suction filtration, and after rotary evaporation under reduced pressure, distilled water is added to prepare Morinda citrifolia alcohol extract with a concentration of 0.1 g / mL and Morinda citrifolia residue;

[0028] S2.2: Prepare the medium: PDA medium is peeled potato 200.0 g / L, glucose 20.0 g / L, peptone 2.0 g / L, KH2PO4 2.0 g / L, MgSO4·7H2O 1.0 g / L, agar 20.0 g / L, seed medium is glucose 30.0 g / L, KH2PO4 0.5 g / L, peptone 2.0 g / L, MgSO4·7H2O 0.5 g / L; yeast extract 6.0 g / L, fermentation medium is glucose 50.0 g / L, KH2PO4 0.5 g / L, peptone 5.0 g / L, noli fruit alcohol extract 5.0 g / L, yam alcohol extract 8.0 g / L, MgSO4·7H2O 2.0 g / L, yeast extract 10.0 g / L;

[0029] S2.3: Take 1 cm 2 of the Grifola frondosa mycelium block is inoculated on the PDA medium and placed in a constant temperature biochemical incubator at 25-27℃ for 8-10d, then the mycelium on the slope is scraped with an inoculation spoon and inoculated on the seed medium, cultured at 25-27℃, 150-200r / min in a shaker for 4-5d, then the seed liquid is taken from the seed liquid with a pipette gun at a 10% inoculation amount and inoculated into the fermentation medium, cultured at 25-27℃, 150-200r / min in a shaker for 7-13d to obtain Grifola frondosa fermentation liquor.

[0030] Further, step S3 fermenting noli fruit residue with Eurotium cristatum, specifically comprising the following steps:

[0031] S3.1: inoculate Eurotium cristatum on PDA medium and culture in a constant temperature incubator at 27-28℃ for 5-6d to obtain activated Eurotium cristatum, which is ready for use, and dry the noli fruit residue in step S2.1 at 50-60℃ to constant weight to obtain dried noli fruit residue, which is ready for use;

[0032] S3.2: mix the noli fruit residue with deionized water at a ratio of 1:1 to obtain a mixed system, sterilize 2-3 parts by weight of the mixed system at 120-121℃ for 25-30min, then cool to room temperature, inoculate the activated Eurotium cristatum into the mixed system, then ferment at 28-30℃ for 7-8d to obtain a fermentation agent;

[0033] S3.3: take 15-20 parts by weight of the mixed system in step S3.2 as a fermentation substrate and sterilize at 120-121℃ for 25-30min, then cool to room temperature, add the fermentation agent, shake and mix uniformly, then ferment at 28-30℃ for 4-5d, and then freeze-dry, crush to 60-70 mesh to obtain noli fruit residue fermentation product.

[0034] Further, the inoculation amount of Eurotium cristatum in step S3.2 is 2-3% of the mixed system.

[0035] Further, the amount of the ferment added in step S3.3 is 4-5% of the fermentation substrate.

[0036] Further, the preparation of the anti-aging composition in step S4 specifically comprises the following steps:

[0037] S4.1: 3-5 parts by weight of sodium hyaluronate and 5-8 parts by weight of collagen peptide are put into a high-speed stirrer, and stirred at a speed of 500-800 r / min for 10-15 min to make them preliminarily mixed uniformly to obtain a mixture;

[0038] S4.2: Then 1-2 parts by weight of peanut peptide-selenium chelate, 3-5 parts by weight of grifola ferment and 2-3 parts by weight of Morinda citrifolia residue ferment are added to the mixture, and stirred at 300-500 r / min for 20-30 min to obtain the anti-aging composition.

[0039] An anti-aging composition for improving facial wrinkles, which is prepared by the preparation method of the anti-aging composition for improving facial wrinkles according to any one of the above.

[0040] Compared with the prior art, the present application has at least the following beneficial effects:

[0041] 1: The anti-aging composition is prepared by compounding sodium hyaluronate, collagen peptide, peanut peptide-selenium chelate, grifola ferment and Morinda citrifolia residue ferment, the collagen peptide can directly penetrate into the dermis layer, transmit synthesis signals to fibroblasts, start the collagen regeneration program, awaken cell vitality, stimulate collagen synthesis, the peanut peptide-selenium chelate can promote the absorption of selenium and enhance the free radical scavenging ability, the grifola ferment and Morinda citrifolia residue ferment improve the ability to scavenge free radicals in the body from multiple angles, through the synergistic effect of the components, the effect of effectively stimulating collagen regeneration, inhibiting the damage of free radicals to the skin and delaying aging is achieved, and the anti-aging composition is free of chemical components and does not cause burden to the liver and kidney functions of the human body.

[0042] 2. This invention incorporates a peanut peptide-selenium chelate into the composition. Selenium atoms coordinate with functional groups on the peanut peptide, forming a relatively stable chelate. This effectively protects selenium from the harsh environment of the gastrointestinal tract, allowing it to reach the absorption site in a more intact form, thereby increasing the likelihood of selenium absorption by the intestines. The peanut peptide, with its hydrophilic groups, significantly improves the solubility of selenium in aqueous media after chelation with selenium, enabling better dispersion in intestinal digestive fluids. This increases the surface area in contact with intestinal epithelial cells, promoting transmembrane transport of selenium into cells and improving absorption efficiency. Furthermore, the peanut peptide can act as a carrier, binding to specific receptors on the cell surface. Through receptor-mediated endocytosis, selenium is carried into cells, further ensuring its smooth entry and laying the foundation for its subsequent effects. Furthermore, the active groups (such as amino and carboxyl groups) carried by peanut peptides themselves, after chelating with selenium, expose more sites that can react with free radicals. These sites can synergize with the antioxidant sites of selenium, enabling it to capture free radicals from multiple angles and enhance its ability to scavenge reactive oxygen species such as hydrogen peroxide, superoxide anion radicals, and hydroxyl radicals. This provides comprehensive protection for cells from oxidative damage, effectively inhibiting free radical damage to the skin, slowing down the aging process from the root, and reducing the formation of wrinkles.

[0043] 3. This invention utilizes *Aspergillus cristatus* to ferment noni pomace. During fermentation, *Aspergillus cristatus* produces various antioxidants, such as polysaccharides and other functional components, thereby increasing the concentration of these functional components in the fermented noni pomace. This high concentration of antioxidants effectively eliminates free radicals in the body, reducing oxidative damage to cells and, to some extent, slowing down the aging process. Furthermore, the noni pomace can be recycled. Simultaneously, *Aspergillus cristatus* itself, as a probiotic, can regulate the balance of intestinal microbiota. When this bacterium enters the intestines along with the fermented noni pomace, it inhibits the growth of harmful bacteria, promotes the proliferation of beneficial bacteria, enhances the body's immunity, and reduces the decline in bodily functions caused by diseases and other factors, thus indirectly playing an anti-aging role.

[0044] 4、The present application promotes the growth of Grifola by fermentation of the yam alcohol extract and the Morinda citrifolia alcohol extract, the yam alcohol extract contains p-hydroxybenzyl alcohol, p-hydroxybenzaldehyde and allantoin and other components that promote the growth of Grifola, and Grifola itself contains rich polysaccharides, under the assistance of the yam alcohol extract and the Morinda citrifolia alcohol extract, the fermentation process can provide additional carbon sources and other nutrients for Grifola, promote its growth and metabolism, further improve the yield of polysaccharides, and the yam alcohol extract and the Morinda citrifolia alcohol extract can improve the in-vitro antioxidant activity of Grifola polysaccharides after participating in the fermentation of Grifola, improve the ability to scavenge free radicals in the body, thereby reducing the damage of free radicals to collagen and elastic fibers in skin cells, delaying aging, and the fermented Grifola polysaccharides can regulate the signal pathway in cells, induce the expression of antioxidant enzymes, increase the activity of antioxidant enzymes in the body, improve the antioxidant stress resistance, reduce the damage of oxidative stress to skin cells, delay the process of skin aging, and thereby reduce the generation of wrinkles. BRIEF DESCRIPTION OF DRAWINGS

[0045] The accompanying drawings, which are incorporated herein and constitute part of the specification, illustrate embodiments of the application and, together with the description, further serve to explain the principles of the application and to enable a person skilled in the relevant art to implement and use the application.

[0046] Figure 1 A preparation method of an anti-aging composition for improving facial wrinkles is adopted in the embodiments of the present application.

[0047] Figure 2 A glucose standard curve in the embodiments of the present application. DETAILED DESCRIPTION

[0048] The preparation method of an anti-aging composition for improving facial wrinkles provided by the present application will be described in detail below in combination with the drawings and specific embodiments. It should be noted that, in order to make the embodiments more detailed, the following embodiments are the best and preferred embodiments, and other alternative ways can also be adopted by those skilled in the art to implement them; and the drawings are only used to more specifically describe the embodiments, and are not intended to specifically limit the present application.

[0049] Grifola: strain number: 5.404, China General Microbial Culture Collection Center.

[0050] Eurotium cristatum: strain number: CICC2099, China Industrial Microbial Culture Collection Center.

[0051] Example 1

[0052] A preparation method of an anti-aging composition for improving facial wrinkles, as shown in Figure 1 , includes the following steps:

[0053] S1: Preparation of peanut peptide-selenium chelate

[0054] S1.1: Peanut meal with a protein content of 48% was added to deionized water at a mass ratio of 1:10, and the pH was adjusted to 9. It was extracted in a water bath at 50°C for 2h, and then centrifuged at 4000r / min for 15min. The supernatant was adjusted to pH 4.5, and then centrifuged again at 4000r / min for 15min to obtain a precipitate. The precipitate was freeze-dried to obtain peanut protein;

[0055] S1.2: 20 parts by weight of peanut protein were added to 1000 parts by weight of deionized water, and then ultrasonicated at 240W power and 55°C for 30min. After ultrasonication, the pH was adjusted to 8, and then alkaline protease was added in an amount of 4% of the mass of the substrate in the system. After 50min of reaction, the enzyme was inactivated at 95°C for 10min to obtain a first enzymatic hydrolysate;

[0056] S1.3: The first enzymatic hydrolysate was heated to 50°C, and then the pH was adjusted to 5. Flavor protease was then added in an amount of 4% of the mass of the substrate in the system. After 50min of reaction, the enzyme was inactivated at 95°C for 10min, and then centrifuged at 5000r / min for 15min. The supernatant was collected and freeze-dried to obtain peanut peptide powder;

[0057] S1.4: 20 parts by weight of peanut peptide powder were added to deionized water, and stirred to prepare a polypeptide solution with a concentration of 15%. NaOH solution was then added to adjust the pH to 7.5. 6 parts by weight of selenium yeast were added and mixed, and then reacted at 38°C for 20min. After cooling, the mixture was centrifuged at 5000r / min for 15min to obtain a supernatant. 3 volumes of anhydrous ethanol were added to the supernatant, which was centrifuged again. The precipitate was collected and freeze-dried to obtain peanut peptide-selenium chelate;

[0058] S2: Preparation of Grifola frondosa fermentation broth

[0059] S2.1: 20 parts by weight of yam powder were added to 100 parts by weight of 80% ethanol, and oscillation extraction was performed at 25°C for 24h. The supernatant was obtained by suction filtration, and then distilled water was added to prepare a yam alcohol extract with a concentration of 0.1g / mL. 10 parts by weight of Morinda citrifolia powder were added to 100 parts by weight of 80% ethanol, and oscillation extraction was performed at 25°C for 24h. The supernatant was obtained by suction filtration, and then distilled water was added to prepare a Morinda citrifolia alcohol extract with a concentration of 0.1g / mL and Morinda citrifolia residue;

[0060] S2.2: Preparation of culture medium: PDA medium is 200.0 g / L of peeled potato, 20.0 g / L of glucose, 2.0 g / L of peptone, 2.0 g / L of KH2PO4, 1.0 g / L of MgSO4·7H2O, 20.0 g / L of agar; seed culture medium is 30.0 g / L of glucose, 0.5 g / L of KH2PO4, 2.0 g / L of peptone, 0.5 g / L of MgSO4·7H2O; yeast extract 6.0 g / L; fermentation medium is 50.0 g / L of glucose, 0.5 g / L of KH2PO4, 5.0 g / L of peptone, 5.0 g / L of noni fruit alcohol extract, 8.0 g / L of yam alcohol extract, 2.0 g / L of MgSO4·7H2O, 10.0 g / L of yeast extract;

[0061] S2.3: 1 cm 2 of the Grifola frondosa mycelium was inoculated on the PDA medium and placed in a 25°C constant temperature biochemical incubator for culture for 8 days, after which the mycelium on the slope was scraped with an inoculation spoon and inoculated on the seed culture medium, and cultured at 25°C in a 150 r / min shaker for 4 days, after which the seed liquid was inoculated into the fermentation medium at a 10% inoculation amount using a pipette gun, and cultured at 25°C in a 150 r / min shaker for 7 days to obtain the Grifola frondosa fermentation liquor;

[0062] S3: Fermentation of noni fruit residue by Eurotium cristatum

[0063] S3.1: The Eurotium cristatum was inoculated on the PDA medium and cultured in a 27°C constant temperature incubator for 5 days to obtain activated Eurotium cristatum, which was reserved for use; the noni fruit residue in step S2.1 was dried at 50°C to constant weight to obtain dried noni fruit residue, which was reserved for use;

[0064] S3.2: The noni fruit residue was mixed with deionized water at a ratio of 1:1 to obtain a mixed system, 2 parts by weight of the mixed system were sterilized at 120°C for 25 min, and then cooled to room temperature, the activated Eurotium cristatum was inoculated into the mixed system, the inoculation amount of Eurotium cristatum was 2% of the mixed system, and then fermented at 28°C for 7 days to obtain a fermentation agent;

[0065] S3.3: 15 parts by weight of the mixed system in step S3.2 were used as a fermentation substrate and sterilized at 120°C for 25 min, and then cooled to room temperature, the fermentation agent was added in an amount of 4% of the fermentation substrate, and then shaken and mixed, and then fermented at 28°C for 4 days, after which freeze-drying was performed, and then pulverized through a 60-mesh sieve to obtain noni fruit residue fermentation product;

[0066] S4: Preparation of anti-aging composition

[0067] S4.1: 3 parts by weight of sodium hyaluronate and 5 parts by weight of collagen peptide were put into a high-speed stirrer, stirred at a speed of 500 r / min for 10 min to preliminarily mix and uniformly, and a mixture was obtained;

[0068] S4.2: Then 1 part by weight of peanut peptide-selenium chelate, 3 parts by weight of grifola ferments and 2 parts by weight of Morinda citrifolia residue ferment were added to the mixture and stirred at 300 r / min for 20 min to obtain an anti-aging composition.

[0069] Example 2

[0070] A preparation method of an anti-aging composition for improving facial wrinkles, as shown in Figure 1 , comprises the following steps:

[0071] S1: Preparation of peanut peptide-selenium chelate

[0072] S1.1: Peanut meal with a protein content of 48% was added to deionized water at a mass ratio of 1:10, and the pH was adjusted to 9. It was stirred in a water bath at 60°C for 3 h, then centrifuged at 5000 r / min for 20 min. The supernatant after centrifugation was adjusted to pH 4.5, then centrifuged again at 5000 r / min for 20 min to obtain a precipitate. The precipitate was freeze-dried to obtain peanut protein;

[0073] S1.2: 20 parts by weight of peanut protein was added to 1000 parts by weight of deionized water, then ultrasonic treatment was carried out at 250 W power and 60°C for 40 min. After ultrasonic treatment, the pH was adjusted to 8, then alkaline protease was added. The amount of alkaline protease added was 4% of the mass of the substrate in the system. After 60 min of reaction, the enzyme was inactivated at 98°C for 12 min to obtain a first enzymolysis solution;

[0074] S1.3: The first enzymolysis solution was heated to 60°C, then the pH was adjusted to 5, then flavor protease was added. The amount of flavor protease added was 4% of the mass of the substrate in the system. After 60 min of reaction, the enzyme was inactivated at 98°C for 12 min, then centrifuged at 5500 r / min for 20 min. The supernatant was collected and freeze-dried to obtain peanut peptide powder;

[0075] S1.4: 20 parts by weight of peanut peptide powder was added to deionized water, stirred and mixed to prepare a polypeptide solution with a concentration of 15%, then NaOH solution was added to adjust the pH to 7.5. Then 6 parts by weight of selenium yeast was added and mixed, and then reacted at 40°C for 30 min. After the reaction was completed, it was cooled, then centrifuged at 5200 r / min for 20 min to obtain a supernatant. 3 times the volume of anhydrous ethanol was added to the supernatant, then centrifuged again to collect the precipitate, then freeze-dried to obtain peanut peptide-selenium chelate;

[0076] S2: Preparation of Grifola frondosa fermentation broth

[0077] S2.1: 20 parts by weight of yam powder was added to 100 parts by weight of ethanol with a concentration of 80%, and oscillation extraction was carried out at 30°C for 25h. The supernatant was obtained by suction filtration, and after rotary evaporation under reduced pressure, distilled water was added to prepare yam alcohol extract with a concentration of 0.1g / mL. 10 parts by weight of Morinda citrifolia powder was added to 100 parts by weight of ethanol with a concentration of 80%, and oscillation extraction was carried out at 30°C for 25h. The supernatant was obtained by suction filtration, and after rotary evaporation under reduced pressure, distilled water was added to prepare Morinda citrifolia alcohol extract with a concentration of 0.1g / mL and Morinda citrifolia residue;

[0078] S2.2: Preparation of culture medium: PDA medium was prepared with peeled potatoes 200.0g / L, glucose 20.0g / L, peptone 2.0g / L, KH2PO42.0g / L, MgSO4·7H2O 1.0g / L, and agar 20.0g / L. Seed culture medium was prepared with glucose 30.0g / L, KH2PO40.5g / L, peptone 2.0g / L, MgSO4·7H2O 0.5g / L, and yeast extract 6.0g / L. Fermentation medium was prepared with glucose 50.0g / L, KH2PO40.5g / L, peptone 5.0g / L, Morinda citrifolia alcohol extract 5.0g / L, yam alcohol extract 8.0g / L, MgSO4·7H2O 2.0g / L, and yeast extract 10.0g / L;

[0079] S2.3: 1cm 2 of Grifola frondosa mycelium was inoculated on PDA medium and placed in a 27°C constant temperature biochemical incubator for 8-10d. Then, the mycelium on the slant was scraped with an inoculation spoon and inoculated on seed culture medium. The seed culture medium was cultured at 27°C and 200r / min in a shaker for 5d. Then, the seed liquid was inoculated into the fermentation medium at a 10% inoculation amount by using a pipette gun. The fermentation medium was cultured at 27°C and 200r / min in a shaker for 13d to obtain Grifola frondosa fermentation broth;

[0080] S3: Fermentation of Morinda citrifolia residue by Eurotium cristatum

[0081] S3.1: Eurotium cristatum was inoculated on PDA medium and cultured in a 28°C constant temperature incubator for 6d to obtain activated Eurotium cristatum, which was prepared for use. The Morinda citrifolia residue in step S2.1 was dried at 60°C to constant weight to obtain dried Morinda citrifolia residue, which was prepared for use;

[0082] S3.2: The Morinda citrifolia residue was mixed with deionized water at a ratio of 1:1 to obtain a mixed system. 2 parts by weight of the mixed system were sterilized at 121°C for 30min, and then cooled to room temperature. The activated Eurotium cristatum was inoculated into the mixed system at a ratio of 2% of the mixed system. Then, the mixture was fermented at 30°C for 8d to obtain a fermentation agent.

[0083] S3.3: 15 parts by weight of the mixture in step S3.2 was sterilized at 121°C for 30 min as a fermentation substrate, after cooling to room temperature, a fermenting agent was added, the amount of the fermenting agent added was 4% of the fermentation substrate, after shaking and mixing, fermentation was carried out at 30°C for 5 days, after the fermentation was completed, freeze-drying was carried out, crushing was carried out to 70 meshes, and a morinda citrifolia residue ferment was obtained;

[0084] S4: Preparation of an anti-aging composition

[0085] S4.1: 3 parts by weight of sodium hyaluronate and 5 parts by weight of collagen peptide were put into a high-speed stirrer, stirred at a speed of 800 r / min for 15 min, and preliminarily mixed uniformly to obtain a mixture;

[0086] S4.2: Then 1 part by weight of peanut peptide-selenium chelate, 3 parts by weight of grifola ferment liquor and 2 parts by weight of morinda citrifolia residue ferment were added to the mixture and stirred at 500 r / min for 30 min to obtain an anti-aging composition.

[0087] Example 3

[0088] A preparation method of an anti-aging composition for improving facial wrinkles, as shown in Figure 1 , comprises the following steps:

[0089] S1: Preparation of peanut peptide-selenium chelate

[0090] S1.1: Peanut meal with a protein content of 50% was added to deionized water at a mass ratio of 1:11, and the pH was adjusted to 9.2, and stirred in a water bath at 50°C for 2h, then centrifuged at 4000r / min for 15min, the supernatant after centrifugation was adjusted to pH 4.8, then centrifuged again at 4000r / min for 15min, the precipitate was obtained, and the precipitate was freeze-dried to obtain peanut protein;

[0091] S1.2: 30 parts by weight of peanut protein was added to 1200 parts by weight of deionized water, then ultrasonic treatment was carried out at 240W power and 55°C for 30min, after the ultrasonic treatment was completed, the pH was adjusted to 9, then alkaline protease was added, the amount of alkaline protease added was 5% of the mass of the substrate in the system, after reaction for 50min, the enzyme was inactivated at 95°C for 10min, to obtain a first enzymolysis solution;

[0092] S1.3: The first enzymolysis solution was heated to 50°C, then the pH was adjusted to 6, then flavor protease was added, the amount of flavor protease added was 5% of the mass of the substrate in the system, after reaction for 50min, the enzyme was inactivated at 95°C for 10min, then centrifuged at 5000r / min for 15min, the supernatant was collected and freeze-dried to obtain peanut peptide powder;

[0093] S1.4: 30 parts by weight of peanut peptide powder was added to deionized water, and stirred to mix to prepare a polypeptide solution with a concentration of 20%, then NaOH solution was added to adjust the pH value to 8, and then 8 parts by weight of selenium yeast was added and mixed, and then reacted at 38°C for 20 min, after the reaction was completed, it was cooled, then centrifuged at 5000 r / min for 15 min to obtain the supernatant, 5 times the volume of anhydrous ethanol was added to the supernatant, and then centrifuged again to obtain the precipitate, which was then freeze-dried to obtain peanut peptide-selenium chelate;

[0094] S2: Preparation of Grifola frondosa fermentation broth

[0095] S2.1: 30 parts by weight of yam powder was added to 200 parts by weight of 90% concentration ethanol, and oscillation extraction was carried out at 25°C for 24 h, the supernatant was obtained by suction filtration, and after rotary evaporation under reduced pressure, distilled water was added to prepare 0.1 g / mL yam alcohol extract. 20 parts by weight of Morinda citrifolia powder was added to 200 parts by weight of 90% concentration ethanol, and oscillation extraction was carried out at 25°C for 24 h, the supernatant was obtained by suction filtration, and after rotary evaporation under reduced pressure, distilled water was added to prepare 0.1 g / mL Morinda citrifolia alcohol extract and Morinda citrifolia residue;

[0096] S2.2: Preparation of culture medium: PDA culture medium is 200.0 g / L peeled potato, 20.0 g / L glucose, 2.0 g / L peptone, 2.0 g / L KH2PO4, 1.0 g / L MgSO4·7H2O, and 20.0 g / L agar. The seed culture medium is 30.0 g / L glucose, 0.5 g / L KH2PO4, 2.0 g / L peptone, 0.5 g / L MgSO4·7H2O, and 6.0 g / L yeast extract. The fermentation medium is 50.0 g / L glucose, 0.5 g / L KH2PO4, 5.0 g / L peptone, 5.0 g / L Morinda citrifolia alcohol extract, 8.0 g / L yam alcohol extract, 2.0 g / L MgSO4·7H2O, and 10.0 g / L yeast extract.

[0097] S2.3: 1 cm 2 of Grifola frondosa mycelium was inoculated on PDA culture medium and placed in a 25°C constant temperature biochemical incubator for 8 d, then the mycelium on the slant was scraped with an inoculation spoon and inoculated on the seed culture medium, and cultured at 25°C, 150 r / min in a shaking bed for 4 d, then the seed liquid was inoculated into the fermentation medium at a 10% inoculation amount using a pipette gun, and cultured at 25°C, 150 r / min in a shaking bed for 7 d to obtain Grifola frondosa fermentation broth;

[0098] S3: Fermentation of Morinda citrifolia residue by Eurotium cristatum

[0099] S3.1: Emericella coronata was inoculated in PDA culture medium and cultured in a constant temperature incubator at 27°C for 5 days to obtain activated Emericella coronata, which was prepared for use. The Morinda citrifolia pomace in step S2.1 was dried at 50°C to a constant weight to obtain dried Morinda citrifolia pomace, which was prepared for use;

[0100] S3.2: The Morinda citrifolia pomace was mixed with deionized water at a ratio of 1:1 to obtain a mixed system. 3 parts by weight of the mixed system were sterilized at 120°C for 25 min, and then cooled to room temperature. The activated Emericella coronata was inoculated into the mixed system, and the inoculation amount of Emericella coronata was 3% of the mixed system. Then, fermentation was carried out at 28°C for 7 days to obtain a fermentation agent;

[0101] S3.3: 20 parts by weight of the mixed system in step S3.2 were sterilized at 120°C for 25 min as a fermentation substrate. After cooling to room temperature, the fermentation agent was added, and the addition amount of the fermentation agent was 5% of the fermentation substrate. After shaking and mixing uniformly, fermentation was carried out at 28°C for 4 days. After fermentation was completed, freeze-drying was carried out, and the product was pulverized to 60 mesh to obtain Morinda citrifolia pomace fermentation product;

[0102] S4: Preparation of anti-aging composition

[0103] S4.1: 5 parts by weight of sodium hyaluronate and 8 parts by weight of collagen peptide were put into a high-speed stirrer and stirred at a speed of 500 r / min for 10 min to preliminarily mix them uniformly to obtain a mixture;

[0104] S4.2: Then, 2 parts by weight of peanut peptide-selenium chelate, 5 parts by weight of Grifola frondosa fermentation broth, and 3 parts by weight of Morinda citrifolia pomace fermentation product were added to the mixture and stirred at 300 r / min for 20 min to obtain an anti-aging composition.

[0105] Comparative Example 1

[0106] Comparative Example 1 is different from Example 1 in that the peanut peptide-selenium chelate in step S4.2 of Comparative Example 1 is replaced by peanut peptide and selenium yeast, and the remaining steps are unchanged to prepare the anti-aging composition, which is denoted as Comparative Example 1.

[0107] Comparative Example 2

[0108] Comparative Example 2 is different from Example 1 in that the Grifola frondosa fermentation broth in steps S2.2-2.3 and step S4.2 of Comparative Example 2 is removed, and the remaining steps are unchanged to prepare the anti-aging composition, which is denoted as Comparative Example 2.

[0109] Comparative Example 3

[0110] Comparative Example 3 is different from Example 1 in that the Morinda citrifolia pomace extract in step S2.2 of Comparative Example 3 is removed, and the remaining steps are unchanged to prepare the anti-aging composition, which is denoted as Comparative Example 3.

[0111] Comparative Example 4

[0112] Comparative Example 4 is different from Example 1 in that Comparative Example 4 is prepared by removing the yam alcohol extract in step S2.2, and the rest of the steps remain unchanged to prepare the anti-aging composition, which is denoted as Comparative Example 4.

[0113] Comparative Example 5

[0114] Comparative Example 5 is different from Example 1 in that Comparative Example 5 is prepared by removing the fermentation of noni fruit residue in steps S3 and S4.2, and the rest of the steps remain unchanged to prepare the anti-aging composition, which is denoted as Comparative Example 5.

[0115] Radical scavenging rate:

[0116] Take 20 mg of the oral solution, respectively, dissolve it in anhydrous ethanol, and make up to 250 mL to obtain a pre-prepared sample to be tested; accurately weigh 20 mg of 1,1-diphenyl-2-trinitrobenzene hydrazine (DPPH), dissolve it in anhydrous ethanol, and make up to 250 mL; place 2 mL of the pre-prepared sample to be tested and 2 mL of the DPPH solution in a test tube, and place it in a thermostat at 37°C for 1 h. Take anhydrous ethanol as a blank control, measure the absorbance of each sample to be tested at 517 nm, and calculate the radical scavenging rate. Each sample to be tested is measured in triplicate, and the average value is obtained.

[0117] The calculation method of the radical scavenging rate is: radical scavenging rate (%) = [(1-(A1-A2)) / A3]x100%.

[0118] Wherein, A3 is the absorbance of 2 mL of anhydrous ethanol plus 2 mL of DPPH solution;

[0119] A1 is the absorbance of 2 mL of the sample to be tested plus 2 mL of DPPH solution;

[0120] A2 is the absorbance of 2 mL of the sample to be tested plus 2 mL of anhydrous ethanol solution.

[0121] Change rate of skin wrinkles:

[0122] The skin rapid three-dimensional imaging system PRIMOS is used to detect the skin wrinkles of the volunteers before using the oral solution and after using the oral solution for 30 days (wherein the smaller the detection value, the fewer the skin wrinkles). The change rate of skin wrinkles (%) is calculated and recorded. The calculation method of the change rate of skin wrinkles is: change rate (%) = [(skin wrinkles after using the oral solution-skin wrinkles before using the oral solution) / skin wrinkles before using the oral solution]x100%.

[0123] The anti-aging compositions prepared in Examples 1-3 and Comparative Examples 1-5 were added to PQQ fish caviar collagen tripeptide oral liquid produced by Xinchensheng (Guangdong) Co., Ltd. at a proportion of 5%, and 30 healthy female volunteers were randomly selected from each group, the volunteers were aged 30-40 years old, and the volunteers orally took 30 mL of the oral liquid once a day in the morning and evening, and persisted in using for 30 days. The change rate of skin wrinkles of the volunteers was detected, and the average value of the detection results of the volunteers in each group was recorded. The determination results are shown in Table 1.

[0124] Table 1. Determination results of Examples 1-3 and Comparative Examples 1-5

[0125]

[0126] From the data in Table 1, it can be seen that the anti-aging composition prepared in the present application can effectively stimulate collagen regeneration, inhibit the damage of free radicals to the skin, and achieve the effect of reducing wrinkles and anti-aging. From the data of Comparative Example 1, it can be seen that the addition of peanut peptide-selenium chelate in the composition is better than the direct addition of peanut peptide and selenium yeast in the ability of removing active oxygen free radicals, which can effectively inhibit the damage of free radicals to the skin, slow down the aging process from the root, and reduce the growth of wrinkles.

[0127] From the data of Comparative Examples 2-4, it can be seen that the fermentation of Grifola frondosa assisted by yam ethanol extract and noni fruit ethanol extract can improve the ability to scavenge free radicals, and the yam ethanol extract and noni fruit ethanol extract can produce a synergistic effect to promote the fermentation of Grifola frondosa, thereby improving the ability to scavenge free radicals and improving the antioxidant and anti-wrinkle effect.

[0128] From the data of Comparative Example 5, it can be seen that the fermentation of noni fruit residue by Eurotium cristatum can effectively scavenge free radicals in the body, thereby reducing the oxidative damage of cells and delaying the aging process of cells to a certain extent.

[0129] The content of polysaccharides in the Grifola frondosa fermentation broth was determined by the phenol-sulfuric acid method:

[0130] Preparation of standard curve: first, prepare a 1 mg / mL glucose standard solution, and use a pipette to take 0 mL, 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, 0.5 mL, 0.6 mL, 0.7 mL, 0.8 mL, 0.9 mL, and 1 mL of the glucose standard solution into 25 mL test tubes with stoppers, add water to 2.0 mL, then add 1 mL of 6% phenol solution, and then add 5 mL of concentrated sulfuric acid, shake well, and then stand for 20 min, then measure the absorbance at 490 nm, and take the average value of three parallel groups. The standard curve is drawn with the glucose content as the X-axis and the absorbance as the y-axis, as shown in Figure 2 .

[0131] Sample determination: 5 mL of fermentation broth was taken in a 50 mL centrifuge tube, 4 times the volume of 95% ethanol was added, and it was placed in a 4°C refrigerator for alcohol precipitation for 24 h. After balancing, it was centrifuged at 4000 r / min for 15 min, the supernatant was discarded, the precipitate was washed with 95% ethanol for 3 times, dried at 60°C, then redissolved with distilled water and constant volume to 50 mL.

[0132] The determination of the polysaccharide content of the fermentation broth of Morchella esculenta in Example 1 and Comparative Examples 3-4 was carried out according to the above method, and the determination results are shown in Table 2.

[0133] Table 2. Determination results of the polysaccharide content of the fermentation broth of Morchella esculenta in Example 1 and Comparative Examples 3-4

[0134]

[0135] As can be seen from the data in Table 2, the fermentation of Morchella esculenta assisted by the yam alcohol extract and the Morinda citrifolia alcohol extract can increase the content of polysaccharide, thereby improving the antioxidant capacity.

[0136] The Morinda citrifolia residue fermentate prepared in Example 1 and the Morinda citrifolia residue without fermentation in step S2.1 were used as samples, and the determination of the polysaccharide content was carried out by the phenol-sulfuric acid method, and the determination results are shown in Table 3.

[0137] Sample determination: 5 g of sample was taken in a 50 mL centrifuge tube, 4 times the volume of 95% ethanol was added, and it was placed in a 4°C refrigerator for alcohol precipitation for 24 h. After balancing, it was centrifuged at 4000 r / min for 15 min, the supernatant was discarded, the precipitate was washed with 95% ethanol for 3 times, dried at 60°C, then redissolved with distilled water and constant volume to 50 mL.

[0138] Table 3. Determination results of the polysaccharide content of the Morinda citrifolia residue fermentate and the Morinda citrifolia residue without fermentation in Example 1

[0139]

[0140] As can be seen from the data in Table 3, the fermentation of Morinda citrifolia residue by Eurotium cristatum can promote the synthesis of functional components of polysaccharide compounds, thereby increasing the concentration of functional components in the Morinda citrifolia residue fermentate.

[0141] The above examples are only illustrative of the principles and effects of the present application, and are not intended to limit the present application. Any person skilled in the art can modify or change the above examples without departing from the spirit and scope of the present application. Therefore, all equivalent modifications or changes made by those skilled in the art without departing from the spirit and technical thought disclosed by the present application should be covered by the claims of the present application.

Claims

1. A method for preparing an anti-aging composition for improving facial wrinkles, characterized in that, Includes the following steps: S1: Preparation of peanut peptide-selenium chelate S1.1: Add peanut meal to deionized water and adjust the pH to 9-9.

2. In a water bath at 50-60℃, stir and extract for 2-3 hours. Then centrifuge at 4000-5000 r / min for 15-20 min. Adjust the pH of the supernatant after centrifugation to 4.5-4.8, and then centrifuge again at 4000-5000 r / min for 15-20 min to obtain the precipitate. Freeze-dry the precipitate to obtain peanut protein. S1.2: Add 20-30 parts by weight of peanut protein to 1000-1200 parts by weight of deionized water, then sonicate at 240-250W power and 55-60℃ for 30-40 minutes. After sonication, adjust the pH to 8-9, then add alkaline protease at 4-5% of the substrate mass in the system. After reacting for 50-60 minutes, inactivate the enzyme at 95-98℃ for 10-12 minutes to obtain the first enzymatic hydrolysis solution. S1.3: Heat the first enzymatic hydrolysis solution to 50-60℃, then adjust the pH to 5-6, and then add flavor protease. The amount of flavor protease added is 4-5% of the substrate in the system. After reacting for 50-60 min, inactivate the enzyme at 95-98℃ for 10-12 min, then centrifuge at 5000-5500 r / min for 15-20 min, collect the supernatant and freeze-dry to obtain peanut peptide powder. S1.4: Add 20-30 parts by weight of peanut peptide powder to deionized water, stir and mix to prepare a peptide solution with a concentration of 15-20%, then add NaOH solution to adjust the pH to 7.5-8, then add 6-8 parts by weight of selenium yeast and mix. React at 38-40℃ for 20-30 min. After the reaction is complete, cool and then centrifuge at 5000-5200 r / min for 15-20 min to obtain the supernatant. Add 3-5 times the volume of anhydrous ethanol to the supernatant, centrifuge again and take the precipitate. Then freeze-dry to obtain peanut peptide-selenium chelate. S2: Preparation of Grifola frondosa fermentation broth S2.1: Add 20-30 parts by weight of yam powder to 100-200 parts by weight of 80-90% ethanol, and extract by shaking at 25-30℃ for 24-25 hours. Filter the supernatant, evaporate under reduced pressure, and add distilled water to prepare a 0.1 g / mL yam ethanol extract. Add 10-20 parts by weight of noni fruit powder to 100-200 parts by weight of 80-90% ethanol, and extract by shaking at 25-30℃ for 24-25 hours. Filter the supernatant, evaporate under reduced pressure, and add distilled water to prepare a 0.1 g / mL noni fruit ethanol extract and noni fruit pomace. S2.2: Preparation of culture media: PDA medium consists of peeled potato 200.0 g / L, glucose 20.0 g / L, peptone 2.0 g / L, KH2PO4 2.0 g / L, MgSO4·7H2O 1.0 g / L, and agar 20.0 g / L; seed culture medium consists of glucose 30.0 g / L, KH2PO4 0.5 g / L, peptone 2.0 g / L, and MgSO4·7H2O 0.5 g / L; yeast extract 6.0 g / L; fermentation medium consists of glucose 50.0 g / L, KH2PO4 0.5 g / L, peptone 5.0 g / L, noni fruit alcohol extract 5.0 g / L, yam alcohol extract 8.0 g / L, MgSO4·7H2O 2.0 g / L, and yeast extract 10.0 g / L. S2.3: Take 1cm 2 Grifola frondosa mycelial blocks were inoculated into PDA medium and cultured in a constant temperature biochemical incubator at 25-27℃ for 8-10 days. Then, the mycelium on the slant was gently scraped with an inoculation spoon and inoculated into seed medium. The culture was carried out in a shaker at 25-27℃ and 150-200 r / min for 4-5 days. Then, the seed liquid was pipetted into the fermentation medium at a 10% inoculation rate and cultured in a shaker at 25-27℃ and 150-200 r / min for 7-13 days to obtain Grifola frondosa fermentation broth. S3: Noni pomace fermented with *Eurotium cristatum* S3.1: Inoculate *Aspergillus cristatus* into PDA medium and culture in a constant temperature incubator at 27-28℃ for 5-6 days to obtain activated *Aspergillus cristatus* for later use. Dry the noni pomace from step S2.1 at 50-60℃ to constant weight to obtain dried noni pomace for later use. S3.2: Mix noni fruit pomace and deionized water in a 1:1 ratio to obtain a mixed system. Sterilize 2-3 parts by weight of the mixed system at 120-121℃ for 25-30 minutes, then cool to room temperature. Inoculate the activated Aspergillus cristatus into the mixed system and ferment at 28-30℃ for 7-8 days to obtain the starter culture. S3.3: Sterilize the 15-20 parts by weight of the mixture in step S3.2 at 120-121℃ for 25-30 minutes as the fermentation substrate, cool to room temperature, add the fermentation agent, shake and mix well, and ferment at 28-30℃ for 4-5 days. After fermentation, freeze dry and pulverize through 60-70 mesh to obtain noni pomace fermentation product. S4: Preparation of Anti-aging Composition An anti-aging composition was prepared by compounding sodium hyaluronate, collagen peptides, peanut peptide-selenium chelate, Grifola frondosa fermentation broth, and Noni pomace fermentation product.

2. The method for preparing an anti-aging composition for improving facial wrinkles according to claim 1, characterized in that, In step S1.1, peanut meal and deionized water are mixed at a mass ratio of 1:10-11.

3. The method for preparing an anti-aging composition for improving facial wrinkles according to claim 1, characterized in that, In step S1.1, the peanut meal is peanut meal with a protein content of 48-50%.

4. The method for preparing an anti-aging composition for improving facial wrinkles according to claim 1, characterized in that, In step S3.2, the inoculum amount of *Eurotium cristatum* is 2-3% of the mixed system.

5. A method for preparing an anti-aging composition for improving facial wrinkles according to claim 1, characterized in that, In step S3.3, the amount of fermentation agent added is 4-5% of the fermentation substrate.

6. A method for preparing an anti-aging composition for improving facial wrinkles according to claim 1, characterized in that, The preparation of the anti-aging composition in step S4 specifically includes the following steps: S4.1: Add 3-5 parts by weight of sodium hyaluronate and 5-8 parts by weight of collagen peptides to a high-speed stirrer and stir at 500-800 r / min for 10-15 min to make them initially mixed evenly to obtain a mixture; S4.2: Then add 1-2 parts by weight of peanut peptide-selenium chelate, 3-5 parts by weight of Grifola frondosa fermentation broth and 2-3 parts by weight of Noni pomace fermentation broth to the mixture and stir at 300-500 r / min for 20-30 min to obtain the anti-aging composition.

7. An anti-aging composition for improving facial wrinkles, characterized in that, It is prepared by the method of preparing an anti-aging composition for improving facial wrinkles as described in any one of claims 1-6.

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