Anti-aging composition for improving facial wrinkles and preparation method thereof

Abstract

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

Description

Technical Field

[0001] The invention relates to the technical field of anti-aging composition preparation, and in particular to an anti-aging composition for improving facial wrinkles and a preparation method thereof. Background Art

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

[0003] From a physiological perspective, as we age, collagen and elastic fibers in the skin gradually disappear, cell metabolism slows down, and the skin loses its original firmness and elasticity, which leads to wrinkles. At the same time, external environmental factors such as long-term exposure to ultraviolet rays can damage the skin's barrier function, induce the production of a large number of free radicals, accelerate the aging process of the skin, and cause wrinkles to appear prematurely.

[0004] In the fast-paced life of modern society, problems such as staying up late, high stress, and bad eating habits occur frequently, further aggravating skin aging. Consumers are in urgent need of products that can effectively improve facial wrinkles and achieve skin rejuvenation. However, existing anti-aging products have many limitations. Most traditional skin care products can only act on the surface of the skin and it is difficult to penetrate into the underlying skin to repair damaged tissue from the root. Although some cosmetic methods, such as medical cosmetic surgery, have certain effects, they are costly, risky, and the postoperative recovery process is cumbersome.

[0005] Health drinks are gradually becoming a popular anti-aging method as a relatively mild and convenient way. They can be taken orally, and the active ingredients are absorbed by the digestive system and act on the skin all over the body, nourishing the skin from the inside out, regulating body functions, and helping to reduce wrinkles.

[0006] Currently, there are many shortcomings in the health drinks on the market that are specifically 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 active ingredients added is insufficient or the ingredients are not reasonably matched, and they cannot accurately target the key factors that cause facial wrinkles, such as being unable to effectively stimulate collagen regeneration and inhibit free radical damage to the skin. It is difficult to effectively improve skin sagging, reduce the depth and number of wrinkles, and cannot meet the skin rejuvenation effect expected by consumers.

[0007] From the perspective of ingredient safety, some health drinks add some potentially risky chemical ingredients in pursuit of short-term effects. These ingredients may burden the liver and kidney functions of the human body after long-term use, and even cause adverse reactions such as allergies, making consumers face health risks while pursuing beauty.

[0008] Therefore, we propose an anti-aging composition for improving facial wrinkles and a preparation method thereof, which can be applied to prepare health drinks to solve the above problems. Summary of the invention

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

[0010] A method for preparing an anti-aging composition for improving facial wrinkles comprises the following steps: S1: Preparation of peanut peptide-selenium chelate Peanut protein is extracted from peanut meal, and the peanut protein is enzymatically hydrolyzed with 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; S2: Preparation of Grifola frondosa fermentation broth The yam powder and the noni fruit powder are extracted with ethanol to obtain the yam alcohol extract and the noni pomace; the maitake mushroom block is inoculated and activated, and then added into the fermentation medium containing the noni fruit alcohol extract and the yam alcohol extract for fermentation to obtain the maitake mushroom fermentation liquid; S3: Noni pomace fermented with Eurotium cristatum A mixture of noni pomace and deionized water is inoculated with Eurotium cristatum to prepare a starter, and then the remaining mixed system is used as a fermentation matrix, and the starter is added to perform fermentation to prepare a noni pomace fermentation product; S4: Preparation of anti-aging composition The anti-aging composition is prepared by compounding sodium hyaluronate, collagen peptide, peanut peptide-selenium chelate, Grifola frondosa fermentation liquid and Noni pomace fermentation product.

[0011] Furthermore, the preparation of the peanut peptide-selenium chelate in step S1 specifically comprises the following steps: S1.1: Add peanut meal to deionized water and adjust the pH to 9-9.2, extract with stirring in a 50-60°C water bath for 2-3 hours, then centrifuge at 4000-5000 r / min for 15-20 minutes, adjust the pH of the supernatant after centrifugation to 4.5-4.8, then centrifuge again at 4000-5000 r / min for 15-20 minutes to obtain a precipitate, and freeze-dry the precipitate to obtain peanut protein; S1.2: 20-30 parts by weight of peanut protein are added to 1000-1200 parts by weight of deionized water, and then ultrasonicated at 240-250W power and 55-60°C for 30-40min. After the ultrasonication is completed, the pH is adjusted to 8-9, and then alkaline protease is added. The amount of alkaline protease added is 4-5% of the substrate mass in the system. After reacting for 50-60min, the enzyme is inactivated at 95-98°C for 10-12min to obtain the first enzymatic solution; S1.3: Heat the first enzymatic hydrolysis solution to 50-60°C, then adjust the pH to 5-6, and then add flavor protease, the amount of flavor protease added is 4-5% of the substrate mass in the system, react for 50-60 minutes, inactivate the enzyme at 95-98°C for 10-12 minutes, and then centrifuge at 5000-5500r / min for 15-20 minutes, 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 form a polypeptide solution with a concentration of 15-20%, then add NaOH solution to adjust the pH value to 7.5-8, then add 6-8 parts by weight of selenium yeast and mix, react at 38-40°C for 20-30 minutes, cool after the reaction is completed, and then centrifuge at 5000-5200r / min for 15-20 minutes to obtain a supernatant, add 3-5 times the volume of anhydrous ethanol to the supernatant, centrifuge again to obtain the precipitate, and then freeze-dry to obtain peanut peptide-selenium chelate.

[0012] Furthermore, in step S1.1, the peanut meal and deionized water are mixed in a mass ratio of 1:10-11.

[0013] Furthermore, the peanut meal in step S1.1 is peanut meal having a protein content of 48-50%.

[0014] Furthermore, the preparation of Grifola frondosa fermented liquid in step S2 specifically comprises the following steps: S2.1: 20-30 parts by weight of yam powder are added to 100-200 parts by weight of ethanol with a concentration of 80-90%, and the mixture is shaken and extracted at 25-30°C for 24-25h, the supernatant is filtered out, and after reduced pressure rotary evaporation, distilled water is added to prepare 0.1g / mL of yam alcohol extract, 10-20 parts by weight of noni fruit powder are added to 100-200 parts by weight of ethanol with a concentration of 80-90%, and the mixture is shaken and extracted at 25-30°C for 24-25h, the supernatant is filtered out, and after reduced pressure rotary evaporation, distilled water is added to prepare 0.1g / mL of noni fruit alcohol extract and noni pomace; S2.2: Prepare culture medium: PDA culture medium is 200.0g / L peeled potato, 20.0g / L glucose, 2.0g / L peptone, 2.0g / L KH2PO4, 1.0g / L MgSO4·7H2O, 20.0g / L agar, seed culture medium is 30.0g / L glucose, 0.5g / L KH2PO4, 2.0g / L peptone, 0.5g / L MgSO4·7H2O; 6.0g / L yeast extract, fermentation medium is 50.0g / L glucose, 0.5g / L KH2PO4, 5.0g / L peptone, 5.0g / L noni alcohol extract, 8.0g / L yam alcohol extract, 2.0g / L MgSO4·7H2O, 10.0g / L yeast extract; S2.3: Take 1cm 2 The mycelium block of Grifola frondosa was inoculated into PDA culture medium and cultured in a constant temperature biochemical incubator at 25-27°C for 8-10 days. Then, the mycelium on the slope was gently scraped with an inoculation spoon and inoculated into the seed culture medium. The culture was carried out in a shaker at 25-27°C and 150-200 r / min for 4-5 days. Then, the seed liquid was aspirated from the seed liquid with a pipette at a 10% inoculation rate and inoculated into the fermentation medium. The culture was carried out in a shaker at 25-27°C and 150-200 r / min for 7-13 days to obtain Grifola frondosa fermentation liquid.

[0015] Further, step S3 of fermenting Noni pomace with Eurotium cristatum specifically comprises the following steps: S3.1: inoculating the cristatum cristatum into a PDA medium, and culturing it in a constant temperature incubator at 27-28°C for 5-6 days to obtain activated cristatum cristatum, and drying the noni pomace in step S2.1 at 50-60°C to a constant weight to obtain dried noni pomace, and setting it aside; S3.2: Noni pomace and deionized water are stirred and mixed in a ratio of 1:1 to obtain a mixed system, 2-3 parts by weight of the mixed system are sterilized at 120-121°C for 25-30 minutes, then cooled to room temperature, activated Eurotium cristatum is inoculated into the mixed system, and then fermented at 28-30°C for 7-8 days to obtain a starter; S3.3: 15-20 parts by weight of the mixed system in step S3.2 are used as fermentation substrates and sterilized at 120-121°C for 25-30 minutes. After cooling to room temperature, the fermentation agent is added, and the mixture is shaken and mixed, and the mixture is fermented at 28-30°C for 4-5 days. After the fermentation is completed, the mixture is freeze-dried and crushed through 60-70 mesh to obtain the fermented noni pomace.

[0016] Furthermore, in step S3.2, the inoculation amount of Eurotium cristatum is 2-3% of the mixed system.

[0017] Furthermore, in step S3.3, the amount of the fermentation agent added is 4-5% of the fermentation substrate.

[0018] Furthermore, step S4 of preparing the anti-aging composition specifically comprises the following steps: S4.1: 3-5 parts by weight of sodium hyaluronate and 5-8 parts by weight of collagen peptide are placed in a high-speed stirrer, and stirred at a speed of 500-800 r / min for 10-15 min to preliminarily mix them evenly to obtain a mixture; S4.2: Then, 1-2 parts by weight of peanut peptide-selenium chelate, 3-5 parts by weight of Grifola frondosa fermentation liquid and 2-3 parts by weight of Noni pomace fermentation product are added to the mixture and stirred at 300-500 r / min for 20-30 min to obtain an anti-aging composition.

[0019] An anti-aging composition for improving facial wrinkles is prepared by any of the above-mentioned methods for preparing an anti-aging composition for improving facial wrinkles.

[0020] Compared with the prior art, the present invention has at least the following beneficial effects: 1. The present invention adopts sodium hyaluronate, collagen peptide, peanut peptide-selenium chelate, maitake mushroom fermentation liquid and noni pomace fermentation product to prepare an anti-aging composition. Collagen peptide can directly penetrate into the dermis, transmit synthesis signals to fibroblasts, start the collagen regeneration program, awaken cell vitality, and stimulate collagen synthesis. Peanut peptide-selenium chelate can promote selenium absorption and enhance the ability to scavenge free radicals. Maitake mushroom fermentation liquid and noni pomace fermentation product improve the ability to scavenge free radicals in the body from multiple angles. Through the synergistic effect of each component, the anti-aging composition can effectively stimulate collagen regeneration, inhibit free radical damage to the skin, and delay aging. In addition, the anti-aging composition does not contain chemical ingredients and will not burden the liver and kidney functions of the human body.

[0021] 2. The present invention adds peanut peptide-selenium chelate to the composition, and the selenium atom is coordinated and combined with the functional groups on the peanut peptide to form a relatively stable chelate, which effectively protects selenium from the harsh environment of the gastrointestinal tract, allowing it to reach the absorption site in a more complete form, thereby increasing the possibility of selenium being absorbed by the intestine. Peanut peptide has a hydrophilic group. After chelating with selenium, it significantly improves the solubility of selenium in aqueous media, allowing it to be better dispersed in the intestinal digestive juice, increasing the surface area in contact with intestinal epithelial cells, promoting selenium transmembrane transport into cells, and improving absorption efficiency. Peanut peptide can be used as a carrier to bind to specific receptors on the cell surface , selenium is brought into cells through receptor-mediated endocytosis, which further ensures that selenium can enter cells smoothly and lays the foundation for its subsequent function. In addition, the active groups carried by peanut peptides themselves (such as amino groups, carboxyl groups, etc.) will expose more sites that can react with free radicals after chelating with selenium. These sites can synergize with the antioxidant sites of selenium, enabling it to capture free radicals from multiple angles, enhance the scavenging ability of active oxygen free radicals such as hydrogen peroxide, superoxide anion free radicals, and hydroxyl free radicals, and protect cells from oxidative damage in all directions, thereby effectively inhibiting the damage of free radicals to the skin, slowing down the aging process from the root, and reducing the growth of wrinkles.

[0022] 3. The present invention adopts Eurotium guanidinum to ferment Noni pomace. After Eurotium guanidinum ferments Noni pomace, multiple antioxidant substances can be produced in the fermentation process, such as functional ingredients such as polysaccharide compounds, thereby improving the concentration of functional ingredients in Noni pomace fermentation products. High concentrations of antioxidant substances can effectively remove free radicals in the body, thereby reducing oxidative damage to cells, delaying the process of cell aging to a certain extent, and Noni pomace can also be recycled. At the same time, Eurotium guanidinum itself can regulate the balance of intestinal microorganisms in the intestine as a probiotic. When this bacterium enters the intestine along with the fermented Noni pomace, it can inhibit the growth of harmful bacteria, promote the proliferation of beneficial bacteria, enhance the body's immunity, reduce the body function decline caused by factors such as diseases, thereby indirectly playing an anti-aging effect.

[0023] 4. The present invention assists the fermentation of Grifola frondosa by using yam alcohol extract and noni fruit alcohol extract. The yam alcohol extract contains components such as p-hydroxybenzyl alcohol, p-hydroxybenzaldehyde and allantoin that promote the growth of Grifola frondosa. Grifola frondosa itself is rich in polysaccharides. With the assistance of yam alcohol extract and noni fruit alcohol extract, additional carbon sources and other nutrients can be provided to Grifola frondosa during the fermentation process, thereby promoting its growth and metabolism and further increasing the yield of polysaccharides. At the same time, after the yam alcohol extract and noni fruit alcohol extract participate in the fermentation of Grifola frondosa, the in vitro antioxidant activity of Grifola frondosa polysaccharide can be improved, and the ability to remove free radicals in the body can be improved, thereby reducing the damage of free radicals to collagen and elastic fibers in skin cells and delaying aging. Moreover, the fermented Grifola frondosa polysaccharide can regulate the signal pathway in the cell, induce the expression of antioxidant enzymes, increase the activity of antioxidant enzymes in the body, improve the ability to resist oxidative stress, reduce the damage of oxidative stress to skin cells, delay the process of skin aging, and thus reduce the generation of wrinkles. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] The accompanying drawings, which are incorporated herein and constitute a part of the specification, illustrate embodiments of the invention and, together with the description, further serve to explain the principles of the invention and to enable those skilled in the relevant art to make and use the invention.

[0025] Figure 1 This is a flow chart of a method for preparing an anti-aging composition for improving facial wrinkles adopted in an embodiment of the present invention.

[0026] Figure 2 It is the glucose standard curve in the embodiment of the present invention. DETAILED DESCRIPTION

[0027] The following is a detailed description of a method for preparing an anti-aging composition for improving facial wrinkles provided by the present invention in conjunction with the accompanying drawings and specific embodiments. It is also noted that, in order to make the embodiments more detailed, the following embodiments are the best and preferred embodiments, and those skilled in the art may also adopt other alternatives to implement some known technologies; and the accompanying drawings are only for a more specific description of the embodiments, and are not intended to specifically limit the present invention.

[0028] Grifola frondosa: strain number: 5.404, China General Microbiological Culture Collection Management Center.

[0029] Eurotium cristatum: strain number: CICC2099, China Industrial Microbiology Culture Collection Management Center.

[0030] Example 1 A method for preparing an anti-aging composition for improving facial wrinkles, such as Figure 1 As shown, the following steps are included: S1: Preparation of peanut peptide-selenium chelate 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. The mixture was stirred and extracted for 2 hours in a 50°C water bath, and then centrifuged at 4000 r / min for 15 minutes. The pH of the supernatant after centrifugation was adjusted to 4.5, and then centrifuged again at 4000 r / min for 15 minutes to obtain a precipitate. The precipitate was freeze-dried to obtain peanut protein; S1.2: 20 parts by weight of peanut protein were added to 1000 parts by weight of deionized water, and then ultrasonicated at 240 W power and 55° C. for 30 min. After the ultrasonication, the pH was adjusted to 8, and then alkaline protease was added. The amount of alkaline protease added was 4% of the substrate mass in the system. After reacting for 50 min, the enzyme was inactivated at 95° C. for 10 min to obtain the first enzymatic solution; S1.3: The first enzymatic hydrolysis solution was heated to 50°C, and then the pH was adjusted to 5, and then flavor protease was added. The amount of flavor protease added was 4% of the substrate mass in the system. After reacting for 50 minutes, the enzyme was inactivated at 95°C for 10 minutes, and then centrifuged at 5000r / min for 15 minutes. The supernatant was collected and freeze-dried to obtain peanut peptide powder; S1.4: 20 parts by weight of peanut peptide powder were added to deionized water, stirred and mixed to form a 15% peptide solution, then NaOH solution was added to adjust the pH value to 7.5, and then 6 parts by weight of selenium yeast were added and mixed, and the mixture was reacted at 38°C for 20 minutes. After the reaction was completed, the mixture was cooled, and then centrifuged at 5000 r / min for 15 minutes to obtain a supernatant. 3 times the volume of anhydrous ethanol was added to the supernatant, and the mixture was centrifuged again to obtain a precipitate, which was then freeze-dried to obtain a peanut peptide-selenium chelate; S2: Preparation of Grifola frondosa fermentation broth S2.1: 20 parts by weight of yam powder are added to 100 parts by weight of ethanol with a concentration of 80%, and the mixture is shaken and extracted at 25°C for 24 hours. The supernatant is filtered out by suction, and after rotary evaporation under reduced pressure, distilled water is added to prepare 0.1 g / mL of yam alcohol extract. 10 parts by weight of noni fruit powder are added to 100 parts by weight of ethanol with a concentration of 80%, and the mixture is shaken and extracted at 25°C for 24 hours. The supernatant is filtered out by suction, and after rotary evaporation under reduced pressure, distilled water is added to prepare 0.1 g / mL of noni fruit alcohol extract and noni pomace; S2.2: Prepare culture medium: PDA culture medium is 200.0g / L peeled potato, 20.0g / L glucose, 2.0g / L peptone, 2.0g / L KH2PO4, 1.0g / L MgSO4·7H2O, 20.0g / L agar, seed culture medium is 30.0g / L glucose, 0.5g / L KH2PO4, 2.0g / L peptone, 0.5g / L MgSO4·7H2O; 6.0g / L yeast extract, fermentation medium is 50.0g / L glucose, 0.5g / L KH2PO4, 5.0g / L peptone, 5.0g / L noni alcohol extract, 8.0g / L yam alcohol extract, 2.0g / L MgSO4·7H2O, 10.0g / L yeast extract; S2.3: Take 1cm 2 The mycelium block of Grifola frondosa was inoculated into PDA medium, placed in a constant temperature biochemical incubator at 25°C for 8 days, then the mycelium on the slope was gently scraped with an inoculation spoon and inoculated into the seed medium, and incubated in a shaker at 25°C and 150 r / min for 4 days, then the seed liquid was drawn from the seed liquid with a pipette at a 10% inoculation amount and inoculated into the fermentation medium, and incubated in a shaker at 25°C and 150 r / min for 7 days to obtain Grifola frondosa fermentation liquid; S3: Noni pomace fermented with Eurotium cristatum S3.1: inoculating the PDA medium with Eurotium cristatum, and culturing it in a constant temperature incubator at 27°C for 5 days to obtain activated Eurotium cristatum, which is set aside; drying the noni pomace in step S2.1 at 50°C to constant weight to obtain dried noni pomace, which is set aside; S3.2: Noni pomace and deionized water are stirred and mixed in a ratio of 1:1 to obtain a mixed system, 2 parts by weight of the mixed system are sterilized at 120°C for 25 minutes, and then cooled to room temperature, and activated Eurotium cristatum is inoculated into the mixed system, the inoculation amount of Eurotium cristatum is 2% of the mixed system, and then fermented at 28°C for 7 days to obtain a starter; S3.3: sterilize 15 parts by weight of the mixed system in step S3.2 as a fermentation substrate at 120°C for 25 minutes, cool to room temperature, add a fermentation agent, and add an amount of the fermentation agent of 4% of the fermentation substrate. Shake and mix well, and ferment at 28°C for 4 days. After the fermentation is completed, freeze-dry and crush through 60 mesh to obtain a fermented noni pomace. S4: Preparation of anti-aging composition S4.1: 3 parts by weight of sodium hyaluronate and 5 parts by weight of collagen peptide were placed in a high-speed stirrer, and stirred at a speed of 500 r / min for 10 min to preliminarily mix them evenly to obtain a mixture; S4.2: Then, 1 part by weight of peanut peptide-selenium chelate, 3 parts by weight of Grifola frondosa fermentation liquid and 2 parts by weight of Noni pomace fermentation product were added to the mixture and stirred at 300 r / min for 20 min to obtain an anti-aging composition.

[0031] Example 2 A method for preparing an anti-aging composition for improving facial wrinkles, such as Figure 1 As shown, the following steps are included: S1: Preparation of peanut peptide-selenium chelate 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. The mixture was stirred and extracted for 3 hours in a 60°C water bath, and then centrifuged at 5000 r / min for 20 minutes. The pH of the supernatant after centrifugation was adjusted to 4.5, and then centrifuged again at 5000 r / min for 20 minutes to obtain a precipitate. The precipitate was freeze-dried to obtain peanut protein; S1.2: 20 parts by weight of peanut protein were added to 1000 parts by weight of deionized water, and then ultrasonicated at 250 W power and 60° C. for 40 min. After the ultrasonication, the pH was adjusted to 8, and then alkaline protease was added. The amount of alkaline protease added was 4% of the substrate mass in the system. After reacting for 60 min, the enzyme was inactivated at 98° C. for 12 min to obtain the first enzymatic solution; S1.3: The first enzymatic hydrolysis solution was heated to 60°C, and then the pH was adjusted to 5, and then flavor protease was added. The amount of flavor protease added was 4% of the substrate mass in the system. After reacting for 60 minutes, the enzyme was inactivated at 98°C for 12 minutes, and then centrifuged at 5500r / min for 20 minutes. The supernatant was collected and freeze-dried to obtain peanut peptide powder; S1.4: 20 parts by weight of peanut peptide powder were added to deionized water, stirred and mixed to form a 15% peptide solution, then NaOH solution was added to adjust the pH value to 7.5, and then 6 parts by weight of selenium yeast were added and mixed, and the mixture was reacted at 40°C for 30 minutes. After the reaction was completed, the mixture was cooled, and then centrifuged at 5200 r / min for 20 minutes to obtain a supernatant. 3 times the volume of anhydrous ethanol was added to the supernatant, and the precipitate was obtained after centrifugation again, and then freeze-dried to obtain a peanut peptide-selenium chelate; S2: Preparation of Grifola frondosa fermentation broth S2.1: 20 parts by weight of yam powder are added to 100 parts by weight of ethanol with a concentration of 80%, and the mixture is shaken and extracted at 30°C for 25 hours. The supernatant is filtered out by suction, and after rotary evaporation under reduced pressure, distilled water is added to prepare 0.1 g / mL of yam alcohol extract. 10 parts by weight of noni fruit powder are added to 100 parts by weight of ethanol with a concentration of 80%, and the mixture is shaken and extracted at 30°C for 25 hours. The supernatant is filtered out by suction, and after rotary evaporation under reduced pressure, distilled water is added to prepare 0.1 g / mL of noni fruit alcohol extract and noni pomace; S2.2: Prepare culture medium: PDA culture medium is 200.0g / L peeled potato, 20.0g / L glucose, 2.0g / L peptone, 2.0g / L KH2PO4, 1.0g / L MgSO4·7H2O, 20.0g / L agar, seed culture medium is 30.0g / L glucose, 0.5g / L KH2PO4, 2.0g / L peptone, 0.5g / L MgSO4·7H2O; 6.0g / L yeast extract, fermentation medium is 50.0g / L glucose, 0.5g / L KH2PO4, 5.0g / L peptone, 5.0g / L noni alcohol extract, 8.0g / L yam alcohol extract, 2.0g / L MgSO4·7H2O, 10.0g / L yeast extract; S2.3: Take 1cm 2 The mycelium block of Grifola frondosa was inoculated into PDA medium and placed in a constant temperature biochemical incubator at 27°C for 8-10 days. Then, the mycelium on the slope was gently scraped with an inoculation spoon and inoculated into the seed medium. The culture was carried out in a shaking incubator at 27°C and 200 r / min for 5 days. Then, the seed liquid was drawn from the seed liquid with a pipette according to 10% inoculation amount and inoculated into the fermentation medium. The culture was carried out in a shaking incubator at 27°C and 200 r / min for 13 days to obtain Grifola frondosa fermentation liquid. S3: Noni pomace fermented with Eurotium cristatum S3.1: inoculating the PDA medium with Eurotium cristatum, and culturing it in a constant temperature incubator at 28°C for 6 days to obtain activated Eurotium cristatum, which is set aside; drying the noni pomace in step S2.1 at 60°C to constant weight to obtain dried noni pomace, which is set aside; S3.2: Noni pomace and deionized water are stirred and mixed in a ratio of 1:1 to obtain a mixed system, 2 parts by weight of the mixed system are sterilized at 121°C for 30 minutes, and then cooled to room temperature, and activated Eurotium cristatum is inoculated into the mixed system, the inoculation amount of Eurotium cristatum is 2% of the mixed system, and then fermented at 30°C for 8 days to obtain a starter; S3.3: sterilize 15 parts by weight of the mixed system in step S3.2 as a fermentation substrate at 121°C for 30 minutes, cool to room temperature, add a fermentation agent, and add an amount of the fermentation agent of 4% of the fermentation substrate. Shake and mix well, and ferment at 30°C for 5 days. After the fermentation is completed, freeze-dry and crush through 70 mesh to obtain a fermented noni pomace. S4: Preparation of anti-aging composition S4.1: 3 parts by weight of sodium hyaluronate and 5 parts by weight of collagen peptide were placed in a high-speed stirrer, and stirred at a speed of 800 r / min for 15 min to preliminarily mix them evenly to obtain a mixture; S4.2: Then, 1 part by weight of peanut peptide-selenium chelate, 3 parts by weight of Grifola frondosa fermentation liquid and 2 parts by weight of Noni pomace fermentation product were added to the mixture and stirred at 500 r / min for 30 minutes to obtain an anti-aging composition.

[0032] Example 3 A method for preparing an anti-aging composition for improving facial wrinkles, such as Figure 1 As shown, the following steps are included: S1: Preparation of peanut peptide-selenium chelate 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. The mixture was stirred and extracted for 2 hours in a 50°C water bath, and then centrifuged at 4000 r / min for 15 minutes. The pH of the supernatant after centrifugation was adjusted to 4.8, and then centrifuged again at 4000 r / min for 15 minutes to obtain a precipitate. The precipitate was freeze-dried to obtain peanut protein; S1.2: 30 parts by weight of peanut protein were added to 1200 parts by weight of deionized water, and then ultrasonicated at 240 W power and 55° C. for 30 min. After the ultrasonication, the pH was adjusted to 9, and then alkaline protease was added. The amount of alkaline protease added was 5% of the substrate mass in the system. After reacting for 50 min, the enzyme was inactivated at 95° C. for 10 min to obtain the first enzymatic solution; S1.3: The first enzymatic hydrolysis solution was heated to 50°C, and then the pH was adjusted to 6, and then flavor protease was added. The amount of flavor protease added was 5% of the substrate mass in the system. After reacting for 50 minutes, the enzyme was inactivated at 95°C for 10 minutes, and then centrifuged at 5000r / min for 15 minutes. The supernatant was collected and freeze-dried to obtain peanut peptide powder; S1.4: 30 parts by weight of peanut peptide powder were added to deionized water, stirred and mixed to form a 20% peptide solution, then NaOH solution was added to adjust the pH value to 8, and then 8 parts by weight of selenium yeast were added and mixed, and the mixture was reacted at 38°C for 20 minutes. After the reaction was completed, the mixture was cooled, and then centrifuged at 5000 r / min for 15 minutes to obtain a supernatant. 5 times the volume of anhydrous ethanol was added to the supernatant, and the mixture was centrifuged again to obtain a precipitate, which was then freeze-dried to obtain a peanut peptide-selenium chelate; S2: Preparation of Grifola frondosa fermentation broth S2.1: 30 parts by weight of yam powder are added to 200 parts by weight of 90% ethanol, and the mixture is shaken and extracted at 25°C for 24 hours. The supernatant is filtered out by suction, and after rotary evaporation under reduced pressure, distilled water is added to prepare 0.1 g / mL yam alcohol extract. 20 parts by weight of noni fruit powder are added to 200 parts by weight of 90% ethanol, and the mixture is shaken and extracted at 25°C for 24 hours. The supernatant is filtered out by suction, and after rotary evaporation under reduced pressure, distilled water is added to prepare 0.1 g / mL noni fruit alcohol extract and noni pomace; S2.2: Prepare culture medium: PDA culture medium is 200.0g / L peeled potato, 20.0g / L glucose, 2.0g / L peptone, 2.0g / L KH2PO4, 1.0g / L MgSO4·7H2O, 20.0g / L agar, seed culture medium is 30.0g / L glucose, 0.5g / L KH2PO4, 2.0g / L peptone, 0.5g / L MgSO4·7H2O; 6.0g / L yeast extract, fermentation medium is 50.0g / L glucose, 0.5g / L KH2PO4, 5.0g / L peptone, 5.0g / L noni alcohol extract, 8.0g / L yam alcohol extract, 2.0g / L MgSO4·7H2O, 10.0g / L yeast extract; S2.3: Take 1cm 2 The mycelium block of Grifola frondosa was inoculated into PDA medium, placed in a constant temperature biochemical incubator at 25°C for 8 days, then the mycelium on the slope was gently scraped with an inoculation spoon and inoculated into the seed medium, and incubated in a shaker at 25°C and 150 r / min for 4 days, then the seed liquid was drawn from the seed liquid with a pipette at a 10% inoculation amount and inoculated into the fermentation medium, and incubated in a shaker at 25°C and 150 r / min for 7 days to obtain Grifola frondosa fermentation liquid; S3: Noni pomace fermented with Eurotium cristatum S3.1: inoculating the PDA medium with Eurotium cristatum, and culturing it in a constant temperature incubator at 27°C for 5 days to obtain activated Eurotium cristatum, which is set aside; drying the noni pomace in step S2.1 at 50°C to constant weight to obtain dried noni pomace, which is set aside; S3.2: Noni pomace and deionized water are stirred and mixed in a ratio of 1:1 to obtain a mixed system, 3 parts by weight of the mixed system are sterilized at 120°C for 25 minutes, and then cooled to room temperature, and activated Eurotium cristatum is inoculated into the mixed system, the inoculation amount of Eurotium cristatum is 3% of the mixed system, and then fermented at 28°C for 7 days to obtain a starter; S3.3: sterilize 20 parts by weight of the mixed system in step S3.2 at 120°C for 25 minutes as a fermentation substrate, cool to room temperature, add a fermentation agent, and add 5% of the fermentation substrate. Shake and mix well, and ferment at 28°C for 4 days. After the fermentation is completed, freeze-dry and crush through 60 mesh to obtain a fermented noni pomace. S4: Preparation of anti-aging composition S4.1: 5 parts by weight of sodium hyaluronate and 8 parts by weight of collagen peptide were placed in a high-speed stirrer, and stirred at a speed of 500 r / min for 10 min to preliminarily mix them evenly to obtain a mixture; S4.2: Then, 2 parts by weight of peanut peptide-selenium chelate, 5 parts by weight of Grifola frondosa fermentation liquid and 3 parts by weight of Noni pomace fermentation product were added to the mixture and stirred at 300 r / min for 20 minutes to obtain an anti-aging composition.

[0033] Comparative Example 1 Compared with Example 1, the difference of Comparative Example 1 is that Comparative Example 1 removes step S1, replaces the peanut peptide-selenium chelate in step S4.2 with peanut peptide and selenium yeast, and prepares the anti-aging composition without changing the other steps, which is recorded as Comparative Example 1.

[0034] Comparative Example 2 Compared with Example 1, Comparative Example 2 is different in that the Grifola frondosa fermented liquid in Steps S2.2-2.3 and Step S4.2 is removed in Comparative Example 2, and the other steps remain unchanged to prepare the anti-aging composition, which is recorded as Comparative Example 2.

[0035] Comparative Example 3 Compared with Example 1, the difference of Comparative Example 3 is that, in Comparative Example 3, the noni fruit alcohol extract in step S2.2 is removed, and the other steps remain unchanged to prepare the anti-aging composition, which is recorded as Comparative Example 3.

[0036] Comparative Example 4 Compared with Example 1, the difference of Comparative Example 4 is that, in Comparative Example 4, the yam alcohol extract in step S2.2 is removed, and the other steps remain unchanged to prepare the anti-aging composition, which is recorded as Comparative Example 4.

[0037] Comparative Example 5 Compared with Example 1, the difference of Comparative Example 5 is that the fermented noni pomace in step S3 and step S4.2 is removed, and the other steps remain unchanged to prepare the anti-aging composition, which is recorded as Comparative Example 5.

[0038] Free radical scavenging rate: Take 20 mg of the oral solution respectively, dissolve it with anhydrous ethanol, and make up to 250 mL to obtain the pre-prepared test sample; accurately weigh 20 mg of 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH), dissolve it with anhydrous ethanol and make up to 250 mL; put 2 mL of the pre-prepared test sample and 2 mL of DPPH solution in a test tube, place it in a constant temperature box at 37°C for 1 hour, use anhydrous ethanol as a blank control, measure the absorbance of each test sample at 517 nm, and calculate the free radical scavenging rate, measure three groups of parallel samples for each test sample, and calculate the average value.

[0039] The free radical scavenging rate is calculated as follows: free radical scavenging rate (%) = [(1-(A1-A2)) / A3]×100%.

[0040] Among them, A3 is the absorbance of 2 mL of anhydrous ethanol plus 2 mL of DPPH solution; A1 is the absorbance of 2 mL of the test agent plus 2 mL of DPPH solution; A2 is the absorbance of 2 mL of the test agent plus 2 M anhydrous ethanol solution.

[0041] Rate of change of skin wrinkles: The skin fast three-dimensional imaging system PRIMOS was used to detect the skin wrinkles of the volunteers before using the oral liquid and 30 days after using the oral liquid (wherein, the smaller the detection value, the fewer the skin wrinkles), and the change rate (%) of skin wrinkles was calculated and recorded. The calculation method of the change rate of skin wrinkles is: change rate (%) = [(skin wrinkles after using the oral liquid-skin wrinkles before using the oral liquid) / skin wrinkles before using the oral liquid] × 100%.

[0042] The anti-aging compositions prepared in Examples 1-3 and Comparative Examples 1-5 were added to the PQQ Caviar Collagen Tripeptide Drink Oral Liquid of Xinchen Biology (Guangdong) Co., Ltd. at a ratio of 5%. Thirty healthy female volunteers aged 30 to 40 were randomly selected in each group, and the volunteers took 30 mL of the oral liquid once in the morning and evening every day for 30 days. The volunteers were tested for the change rate of skin wrinkles, and the average value of the test results of each group of volunteers was recorded. The test results refer to Table 1.

[0043] Table 1. Measurement results of Examples 1-3 and Comparative Examples 1-5

[0044] It can be seen from the data in Table 1 that the anti-aging composition prepared by the present invention effectively stimulates collagen regeneration, inhibits the damage of free radicals to the skin, and achieves the effect of reducing wrinkles and anti-aging. It can be seen from the data in Comparative Example 1 that the addition of peanut peptide-selenium chelate to the composition has better scavenging ability for active oxygen free radicals than directly adding peanut peptide and selenium yeast, and 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.

[0045] From the data of Comparative Examples 2-4, it can be seen that the ability to scavenge free radicals by assisting the fermentation of Grifola frondosa with yam alcohol extract and noni fruit alcohol extract can be improved, and the yam alcohol extract and noni fruit alcohol 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 effects.

[0046] From the data of Comparative Example 5, it can be seen that fermenting noni pomace with Eurotium cristatum can effectively remove free radicals in the body, thereby reducing oxidative damage to cells and delaying the process of cell aging to a certain extent.

[0047] The phenol-sulfuric acid method was used to determine the polysaccharide content in the fermentation broth of Grifola frondosa: Drawing of the standard curve: First, prepare 1mg / mL glucose standard solution, use a pipette to draw 0mL, 0.1mL, 0.2mL, 0.3mL, 0.4mL, 0.5mL, 0.6mL, 0.7mL, 0.8mL, 0.9mL, and 1mL of glucose standard solution into a 25mL stoppered test tube, add water to make up to 2.0mL, then add 1mL of 6% phenol solution, and then add 5mL of concentrated sulfuric acid. After shaking and letting it stand for 20min, measure the absorbance at 490nm. Make 3 parallels for each group, take the average value, and draw the standard curve with glucose content as the X-axis and absorbance as the Y-axis, such as Figure 2 shown.

[0048] Determination of samples: Take 5mL of fermentation liquid in a 50mL centrifuge tube, add 4 times the volume of 95% ethanol, place it in a 4℃ refrigerator for alcohol precipitation for 24h, centrifuge at 4000r / min for 15min after balancing, discard the supernatant, rinse the precipitate with 95% ethanol 3 times, dry it at 60℃, then add distilled water to redissolve and make up to 50mL.

[0049] The polysaccharide content of the Grifola frondosa fermentation broth of Example 1 and Comparative Examples 3-4 was determined by referring to the above method, and the determination results are shown in Table 2.

[0050] Table 2. Determination results of polysaccharide content in Grifola frondosa fermentation broth of Example 1 and Comparative Examples 3-4

[0051] From the data in Table 2, it can be seen that the use of yam alcohol extract and noni fruit alcohol extract to assist the fermentation of Grifola frondosa can increase the content of polysaccharides, thereby improving the antioxidant capacity.

[0052] The fermented Noni pomace prepared in Example 1 and the unfermented Noni pomace in step S2.1 were used as samples, and the polysaccharide content was determined by the phenol-sulfuric acid method. The determination results are shown in Table 3.

[0053] Sample determination: Take 5g of sample in a 50mL centrifuge tube, add 4 times the volume of 95% ethanol, place in a 4℃ refrigerator for alcohol precipitation for 24h, centrifuge at 4000r / min for 15min after balancing, discard the supernatant, rinse the precipitate with 95% ethanol 3 times, dry at 60℃, then add distilled water to redissolve and make up to 50mL.

[0054] Table 3. Determination results of polysaccharide content in fermented Noni pomace and unfermented Noni pomace in Example 1

[0055] From the data in Table 3, it can be seen that fermenting noni pomace with Eurotium cristatum can promote the synthesis of functional components of polysaccharide compounds, thereby increasing the concentration of functional components in the fermented noni pomace.

[0056] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A method for preparing an anti-aging composition for improving facial wrinkles, characterized in that: The steps include: S1: Preparation of peanut peptide-selenium chelate Peanut protein is extracted from peanut meal, and the peanut protein is enzymatically hydrolyzed with 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; S2: Preparation of Grifola frondosa fermentation broth The yam powder and the noni fruit powder are extracted with ethanol to obtain the yam alcohol extract and the noni pomace; the maitake mushroom block is inoculated and activated, and then added into the fermentation medium containing the noni fruit alcohol extract and the yam alcohol extract for fermentation to obtain the maitake mushroom fermentation liquid; S3: Noni pomace fermented with Eurotium cristatum A mixture of noni pomace and deionized water is inoculated with Eurotium cristatum to prepare a starter, and then the remaining mixed system is used as a fermentation matrix, and the starter is added to perform fermentation to prepare a noni pomace fermentation product; S4: Preparation of anti-aging composition The anti-aging composition is prepared by compounding sodium hyaluronate, collagen peptide, peanut peptide-selenium chelate, Grifola frondosa fermentation liquid 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: Step S1: Preparation of peanut peptide-selenium chelate, specifically comprising the following steps: S1.1: Add peanut meal to deionized water and adjust the pH to 9-9.2, extract with stirring in a 50-60°C water bath for 2-3 hours, then centrifuge at 4000-5000 r / min for 15-20 minutes, adjust the pH of the supernatant after centrifugation to 4.5-4.8, then centrifuge again at 4000-5000 r / min for 15-20 minutes to obtain a precipitate, and freeze-dry the precipitate to obtain peanut protein; S1.2: 20-30 parts by weight of peanut protein are added to 1000-1200 parts by weight of deionized water, and then ultrasonicated at 240-250W power and 55-60°C for 30-40min. After the ultrasonication is completed, the pH is adjusted to 8-9, and then alkaline protease is added. The amount of alkaline protease added is 4-5% of the substrate mass in the system. After reacting for 50-60min, the enzyme is inactivated at 95-98°C for 10-12min to obtain the first enzymatic solution; S1.3: Heat the first enzymatic hydrolysis solution to 50-60°C, then adjust the pH to 5-6, and then add flavor protease, the amount of flavor protease added is 4-5% of the substrate mass in the system, react for 50-60 minutes, inactivate the enzyme at 95-98°C for 10-12 minutes, and then centrifuge at 5000-5500r / min for 15-20 minutes, 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 form a polypeptide solution with a concentration of 15-20%, then add NaOH solution to adjust the pH value to 7.5-8, then add 6-8 parts by weight of selenium yeast and mix, react at 38-40°C for 20-30 minutes, cool after the reaction is completed, and then centrifuge at 5000-5200r / min for 15-20 minutes to obtain a supernatant, add 3-5 times the volume of anhydrous ethanol to the supernatant, centrifuge again to obtain the precipitate, and then freeze-dry to obtain peanut peptide-selenium chelate.

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

4. The method for preparing an anti-aging composition for improving facial wrinkles according to claim 2, characterized in that: The peanut meal in step S1.1 is peanut meal with a protein content of 48-50%.

5. The method for preparing an anti-aging composition for improving facial wrinkles according to claim 2, characterized in that: Step S2: Preparation of Grifola frondosa fermented liquid, specifically comprising the following steps: S2.1: 20-30 parts by weight of yam powder are added to 100-200 parts by weight of ethanol with a concentration of 80-90%, and the mixture is shaken and extracted at 25-30°C for 24-25h, the supernatant is filtered out, and after reduced pressure rotary evaporation, distilled water is added to prepare 0.1g / mL of yam alcohol extract, 10-20 parts by weight of noni fruit powder are added to 100-200 parts by weight of ethanol with a concentration of 80-90%, and the mixture is shaken and extracted at 25-30°C for 24-25h, the supernatant is filtered out, and after reduced pressure rotary evaporation, distilled water is added to prepare 0.1g / mL of noni fruit alcohol extract and noni pomace; S2.2: Prepare culture medium: PDA culture medium is 200.0g / L peeled potato, 20.0g / L glucose, 2.0g / L peptone, 2.0g / L KH2PO4, 1.0g / L MgSO4·7H2O, 20.0g / L agar, seed culture medium is 30.0g / L glucose, 0.5g / L KH2PO4, 2.0g / L peptone, 0.5g / L MgSO4·7H2O; 6.0g / L yeast extract, fermentation medium is 50.0g / L glucose, 0.5g / L KH2PO4, 5.0g / L peptone, 5.0g / L noni alcohol extract, 8.0g / L yam alcohol extract, 2.0g / L MgSO4·7H2O, 10.0g / L yeast extract; S2.3: Take 1cm 2 The mycelium block of Grifola frondosa was inoculated into PDA culture medium and cultured in a constant temperature biochemical incubator at 25-27°C for 8-10 days. Then, the mycelium on the slope was gently scraped with an inoculation spoon and inoculated into the seed culture medium. The culture was carried out in a shaker at 25-27°C and 150-200 r / min for 4-5 days. Then, the seed liquid was aspirated from the seed liquid with a pipette at a 10% inoculation rate and inoculated into the fermentation medium. The culture was carried out in a shaker at 25-27°C and 150-200 r / min for 7-13 days to obtain Grifola frondosa fermentation liquid.

6. The method for preparing an anti-aging composition for improving facial wrinkles according to claim 5, characterized in that: Step S3 ferments the noni pomace with Eurotium cristatum, specifically comprising the following steps: S3.1: inoculating the cristatum cristatum into a PDA medium, and culturing it in a constant temperature incubator at 27-28°C for 5-6 days to obtain activated cristatum cristatum, and drying the noni pomace in step S2.1 at 50-60°C to a constant weight to obtain dried noni pomace, and setting it aside; S3.2: Noni pomace and deionized water are stirred and mixed in a ratio of 1:1 to obtain a mixed system, 2-3 parts by weight of the mixed system are sterilized at 120-121°C for 25-30 minutes, then cooled to room temperature, activated Eurotium cristatum is inoculated into the mixed system, and then fermented at 28-30°C for 7-8 days to obtain a starter; S3.3: 15-20 parts by weight of the mixed system in step S3.2 are used as fermentation substrates and sterilized at 120-121°C for 25-30 minutes. After cooling to room temperature, the fermentation agent is added, and the mixture is shaken and mixed, and the mixture is fermented at 28-30°C for 4-5 days. After the fermentation is completed, the mixture is freeze-dried and crushed through 60-70 mesh to obtain the fermented noni pomace.

7. The method for preparing an anti-aging composition for improving facial wrinkles according to claim 6, characterized in that: In step S3.2, the inoculation amount of Eurotium cristatum is 2-3% of the mixed system.

8. The method for preparing an anti-aging composition for improving facial wrinkles according to claim 6, characterized in that: In step S3.3, the amount of fermentation agent added is 4-5% of the fermentation substrate.

9. The method for preparing an anti-aging composition for improving facial wrinkles according to claim 8, characterized in that: Step S4 is the preparation of the anti-aging composition, which specifically comprises the following steps: S4.1: 3-5 parts by weight of sodium hyaluronate and 5-8 parts by weight of collagen peptide are placed in a high-speed stirrer, and stirred at a speed of 500-800 r / min for 10-15 min to preliminarily mix them evenly to obtain a mixture; S4.2: Then, 1-2 parts by weight of peanut peptide-selenium chelate, 3-5 parts by weight of Grifola frondosa fermentation liquid and 2-3 parts by weight of Noni pomace fermentation product are added to the mixture and stirred at 300-500 r / min for 20-30 min to obtain an anti-aging composition.

10. An anti-aging composition for improving facial wrinkles, characterized in that: The composition is prepared by the method for preparing an anti-aging composition for improving facial wrinkles as described in any one of claims 1 to 9.

Images