An anti-inflammatory repair composition and methods of making and using the same

Abstract

The present application relates to the technical field of daily chemical industry, and in particular to an anti-inflammatory repair composition, a preparation method and application thereof. The composition comprises a rhizoma anemarrhenae extract obtained by fermenting rhizoma anemarrhenae by a chrysosporium fungus. The present application provides a composition that enhances the moisture retention capacity of the skin, helps form a moisturizing barrier, and prevents moisture loss from the skin. The composition contains flavonoids and polysaccharide compounds, which can scavenge free radicals and play an antioxidant role, thereby slowing down skin aging and protecting the skin from damage caused by environmental stress. The formula of the present application inhibits the formation of melanin, helps improve uneven skin tone, and enhances the brightness of the skin.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of daily chemical technology, and in particular to an anti-inflammatory repair composition, a preparation method thereof and application thereof. BACKGROUND

[0002] Anemarrhena asphodeloides, commonly known as Zhi-mu, is a common medicinal herb used in traditional Chinese medicine. It belongs to the Liliaceae family and is typically found growing in dry or semi-dry mountainous regions and grasslands in China, Korea, and Mongolia. The rhizome of Zhi-mu contains various bioactive components, particularly saponins, which have significant pharmacological effects and are widely used in traditional medicine.

[0003] Zhi-mu root is rich in various bioactive components, with one of the most important being Anemarrhenasaponins, a class of compounds with strong pharmacological activity. Zhi-mu also contains beneficial polysaccharides, flavonoids, volatile oils, amino acids, and minerals.

[0004] Anemarrhenasaponins are one of the most important active components in Zhi-mu, with multiple pharmacological effects such as anti-inflammatory, anti-tumor, and immune regulation. Zhi-mu polysaccharides are another active component with immune regulation and antioxidant effects. Flavonoids have anti-inflammatory, antioxidant, and antibacterial biological activities.

[0005] Zhi-mu has a significant yin-nourishing and dryness-moisturizing effect, effectively improving dryness and dehydration of the skin. It helps to replenish skin moisture and enhance skin hydration, keeping the skin soft and smooth. For dry skin, dehydrated skin, or skin affected by climate dryness, Zhi-mu extract is often used for moisturizing and repair. This effect can be achieved through the polysaccharide components and saponins in Zhi-mu, which have good hydration and can enhance the skin barrier function, lock in moisture, and reduce water loss.

[0006] The flavonoids and polysaccharides in Zhi-mu have antioxidant effects, effectively resisting free radical damage and slowing down the aging process. Antioxidant components can help neutralize oxidative damage on the skin surface caused by environmental factors such as UV rays, pollution, and stress, reducing the rate of cell aging.

[0007] Studies have shown that Zhi-mu extract can promote skin cell repair and regeneration, improve skin elasticity and firmness, and thus combat signs of aging. It can also stimulate collagen synthesis, improving fine lines and laxity.

[0008] However, the water solubility of timosaponin is poor. Saponins generally have certain amphiphilicity, i.e. both hydrophilic and hydrophobic structural parts. Timosaponin contains a glycosyl part, which is hydrophilic, but because it also contains a large amount of hydrophobic steroidal skeleton in the molecule, the overall water solubility is poor. At the same time, the extract of Anemarrhena asphodeloides is sensitive to acid and alkali values, and requires a relatively high applicable environment and storage environment.

[0009] To this end, the present application provides an anti-inflammatory repair composition and a preparation method and application thereof. SUMMARY

[0010] To solve the above technical problems, the present application provides an anti-inflammatory repair composition, which comprises an extract of Anemarrhena asphodeloides obtained by fermentation of Anemarrhena asphodeloides by Chrysosporium merdarium.

[0011] The second aspect of the present application provides an antioxidant repair composition, which comprises an extract of Anemarrhena asphodeloides obtained by fermentation of Anemarrhena asphodeloides by Chrysosporium merdarium.

[0012] As an embodiment of the present application, the composition further comprises tea extract, licorice extract, and Oldenlandia diffusa extract.

[0013] As an embodiment of the present application, the mass ratio of the extract of Anemarrhena asphodeloides, tea extract, licorice extract, and Oldenlandia diffusa extract is 1-3:5-15:2-8:1-3.

[0014] As an embodiment of the present application, the mass ratio of the extract of Anemarrhena asphodeloides, tea extract, licorice extract, and Oldenlandia diffusa extract is 3:10:6:2.

[0015] As an embodiment of the present application, the preparation method of the extract of Anemarrhena asphodeloides is as follows:

[0016] S01 strain activation: Chrysosporium merdarium is activated on PDA plate solid culture medium, and cultured at 28℃ for 7 days until obvious colonies are grown;

[0017] S02 activation of symbiotic bacteria: yeast is cultured in YPD medium for 24 hours for standby;

[0018] S03 preparation of liquid seed solution: the activated strain is inoculated into a liquid culture medium containing glucose and yeast extract, and cultured in a constant temperature shaker at 28℃ and 150rpm for 48 hours to prepare a seed solution.

[0019] S04 preliminary liquid fermentation: Anemarrhena asphodeloides powder is added to the liquid culture medium, the pH is adjusted to 5.5, and 5% Chrysosporium merdarium seed solution is inoculated; the fermentation conditions are set as aerobic fermentation at 28℃ and 150rpm for 3 days;

[0020] S05: adding symbiotic bacteria in the middle stage: when the liquid fermentation is carried out to the third day, the activated yeast is added, and the culture is continued for 3-4 days;

[0021] S06: transferring into solid fermentation: the fermentation liquid is adsorbed on wheat bran, and is placed in a constant humidity incubator for static fermentation at 25℃ for 7 days;

[0022] S07: obtaining final fermentation ingredients: after the whole fermentation product of step S06 is mixed with the same volume of deionized water and stirred and soaked, the upper clear liquid is obtained by centrifugal separation, and then low-temperature reduced-pressure concentration and freeze-drying are performed, so that the Zhi-mu extract is obtained.

[0023] As an embodiment of the present application, the components of the S04 liquid culture medium comprise:

[0024] glucose 20-30 g / L, peptone 5-10 g / L, yeast extract 2-5 g / L, ammonium nitrate 1-2 g / L, magnesium sulfate heptahydrate 0.5 g / L, vitamin B1 0.5 g / L, gum arabic 0.05 g / L, and water 1 L.

[0025] The third aspect of the present application provides a skin care product comprising the repair antioxidant composition.

[0026] As an embodiment of the present application, the raw materials for preparing the skin care product comprise:

[0027] glycerin: 4 parts

[0028] p-hydroxyacetophenone: 0.3 parts

[0029] carbomer: 0.6 parts

[0030] arginine: 0.6 parts

[0031] the repair antioxidant composition: 10-15 parts

[0032] 1,2-hexanediol: 0.5 parts

[0033] water is added to 100 parts.

[0034] The fourth aspect of the present application provides the composition for use in the field of daily chemicals.

[0035] By adopting the above technical solution, the present application has the following beneficial effects:

[0036] The present application provides a composition that enhances the skin's ability to retain moisture, helps form a moisture barrier, and prevents moisture loss from the skin; the composition contains flavonoids and polysaccharide compounds, which can scavenge free radicals and have antioxidant effects, thereby slowing down skin aging and protecting the skin from environmental stress damage.

[0037] The formula of the present application inhibits the formation of melanin, helps to improve uneven skin tone, and enhances the brightness of the skin. DETAILED DESCRIPTION

[0038] The technical solutions of the present application will be described clearly and completely below. Obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0039] The present application provides a repairing antioxidant composition, which comprises Anemarrhena asphodeloides extract, tea leaf extract, liquorice extract, and Speranskia extract, and the mass ratio is 3:10:6:2.

[0040] As an embodiment of the present application, the preparation method of the Anemarrhena asphodeloides extract is as follows:

[0041] S01 strain activation: activate the golden flower fungus on PDA plate solid culture medium, cultivate at 28℃ for 7 days, until obvious colonies grow;

[0042] S02 activation of symbiotic bacteria: cultivate the yeast in YPD medium for 24 hours, and reserve;

[0043] S03 preparation of liquid seed solution: inoculate the activated strain in the liquid culture medium containing glucose and yeast extract, cultivate in a constant temperature shaker at 28℃ and 150rpm for 48 hours, to prepare the seed solution.

[0044] S04 preliminary liquid fermentation: add Anemarrhena asphodeloides powder to the liquid culture medium, adjust the pH to 5.5, inoculate 5% golden flower fungus seed solution; the fermentation conditions are set as aerobic fermentation at 28℃ and 150rpm for 3 days;

[0045] S05 intermediate addition of symbiotic bacteria: when the liquid fermentation is carried out for 3 days, add green tea powder and inoculate the activated yeast (inoculate 3%), continue to cultivate for 3-4 days;

[0046] S06 transfer to solid fermentation: adsorb the fermentation liquid on wheat bran, and place in a constant humidity incubator for static fermentation at 25℃ for 7 days;

[0047] S07 obtain the final fermentation ingredient: mix and stir the whole fermentation product of step S06 and the same volume of deionized water after soaking, then centrifuge to take the supernatant, then low-temperature reduce-pressure concentration, freeze-drying, to obtain the Anemarrhena asphodeloides extract.

[0048] As an embodiment of the present application, the components of the liquid culture medium in S04 comprise:

[0049] Glucose 20-30 g / L, peptone 5-10 g / L, yeast extract 2-5 g / L, ammonium nitrate 1-2 g / L, magnesium sulfate heptahydrate 0.5 g / L, vitamin B1 0.5 g / L, gum arabic 0.05 g / L, water 1 L.

[0050] Among them, the golden flower fungus is Eurotium cristatum, China Industrial Microbial Strain Preservation and Management Center.

[0051] The yeast is Rhodosporidium toruloides, China Industrial Microbial Strain Preservation and Management Center.

[0052] The mass ratio of the rhizoma anemarrhenae powder and the green tea powder is 3:1.

[0053] As an embodiment of the present application, the tea leaves are black tea and white tea, and the mass ratio is 2:3, and the tea leaf extraction method is as follows:

[0054] Crushing: crushing the black tea leaves / white tea leaves, and passing through 100 meshes;

[0055] Suspending the crushed tea leaf powder in a buffer solution at 50 DEG C (0.1M citric acid-phosphate buffer, pH 4.5-5.5), adding cellulase and pectinase (total mass 8%, mass ratio 3:5), and carrying out enzymatic hydrolysis treatment for 30 minutes;

[0056] Ultrasonic extraction: placing the tea leaf suspension after enzymatic hydrolysis in an ultrasonic extraction instrument, and treating at 40 kHz frequency and 50 DEG C for 30 minutes.

[0057] Microwave extraction: immediately transferring the solution to a microwave extraction device, heating at 300 W power for 5 minutes; then using CO2 as a supercritical fluid, setting the temperature at 35 DEG C and the pressure at 25 MPa, using 10 vol% ethanol as a cosolvent; setting the CO2 flow at 10 mL / min; extraction time: 30 minutes; collecting the extracted components, removing the residues using high-speed centrifugation (10000 rpm), and concentrating the extraction liquid using a rotary evaporator; drying the concentrated extract into powder through freeze-drying, thereby obtaining the tea leaf extract.

[0058] In the present application, the preparation method of the liquorice is as follows:

[0059] S1: loading liquorice powder into an extraction tank; using supercritical CO2 and 5 vol% ethanol as a cosolvent to carry out extraction at 30 DEG C and 25 MPa for 60 min; setting the CO2 flow at 10 mL / min; collecting and concentrating the extraction liquid; drying the concentrated extract into powder through freeze-drying;

[0060] S2 In the powder obtained in step S1, 5 times volume of distilled water is added, and the mixture is stirred well to disperse the powder in the water. Under stirring, 1 / 8 volume of aqueous potassium hydroxide solution (10 wt%) is slowly added.

[0061] S3 After stirring at 40°C for 2h, the excess water is removed by concentration under reduced pressure, and the glycyrrhiza extract is obtained by drying.

[0062] In the present application, the preparation method of the oldenlandia diffusa extract is as follows:

[0063] (1) Sample preparation: The oldenlandia diffusa sample is ground to fine particles

[0064] (2) The oldenlandia diffusa powder is added to 10 times volume of deionized water, and the mixture is heated and stirred at 80°C for 1-2h to extract the extract;

[0065] (3) After the extraction is completed, the extract is cooled to room temperature and filtered to remove the plant residues;

[0066] (4) The filtered extract is placed in a rotary evaporator, and the extract is concentrated to 1 / 5 volume under reduced pressure at 40°C. The concentrated extract is freeze-dried.

[0067] As an embodiment of the present application, the repair antioxidant composition further comprises honey.

[0068] The present application provides a skin care product comprising the repair antioxidant composition.

[0069] As an embodiment of the present application, the skin care product comprises the following raw materials:

[0070] Glycerol: 4 parts

[0071] p-Hydroxyacetophenone: 0.3 parts

[0072] Carbomer: 0.6 parts

[0073] Arginine: 0.6 parts

[0074] The repair antioxidant composition: 10-15 parts

[0075] 1,2-Hexanediol: 0.5 parts

[0076] Water is added to 100 parts.

[0077] The preparation method is as follows:

[0078] Water, glycerol, p-hydroxyacetophenone, carbomer are mixed and heated to 75-80 degrees, then homogenized for 2-3 minutes until completely dissolved, start to cool down to below 45 degrees, add arginine, slowly stir until transparent, add the remaining raw materials, stir evenly.

[0079] Anemarrhena has antioxidant and whitening effects on the skin, but its main effects are Anemarrhena polysaccharide and Anemarrhena saponin, both of which have certain challenges in skin absorption; at the same time, Anemarrhena extract has high requirements for acid-base value; therefore, the present application uses Auricularia auricular to ferment Anemarrhena; at the same time, Glycyrrhiza glabra and Serspentia aculeata extract are added, which, in combination with Anemarrhena and tea, can effectively solve the irritability of Glycyrrhiza glabra extract and Serspentia aculeata extract; at the same time, after the fermented Anemarrhena extract is mixed with other components, it is also not easy to produce precipitation. Saponin compounds (such as Anemarrhena saponin in Anemarrhena) are usually more easily dissolved in acidic conditions, but their solubility will decrease in alkaline conditions, and they may form precipitates. Glycyrrhizic acid (the main component in Glycyrrhiza glabra) is relatively stable in neutral or slightly alkaline environments, but it may also precipitate in extreme acidic or alkaline environments. At the same time, the combination or coordination between phenolic compounds, flavonoids, saponins and other components may lead to the formation of some insoluble complexes. For example, flavonoids in Serspentia aculeata may interact with certain components in Glycyrrhiza glabra, thereby affecting their solubility. The method used in the present application can effectively avoid the above-mentioned phenomena.

[0080] The present application will be further explained and described in conjunction with specific embodiments.

[0081] Example 1

[0082] The present embodiment provides a kind of repair antioxidant composition, the composition includes Anemarrhena extract, tea extract, Glycyrrhiza glabra extract, Serspentia aculeata extract, honey;And mass ratio is: 3:10:6:2:1.

[0083] The preparation method of the Anemarrhena extract is as follows:

[0084] S01 strain activation: Auricularia auricular is activated on PDA plate solid culture medium, and cultured at 28 DEG C for 7 days until obvious colonies grow;

[0085] S02 symbiotic bacteria activation: yeast is cultured in YPD medium for 24 hours for standby;

[0086] S03 preparation of liquid seed solution: the activated Auricularia auricular is inoculated into liquid culture medium, and cultured in a constant temperature shaker at 28 DEG C, 150 rpm for 48 hours to prepare 10 7 CFU / mL of Auricularia auricular strain liquid.

[0087] S04 preliminary liquid fermentation: add Anemarrhena asphodeloides powder to the liquid medium, adjust pH to 5.5, inoculate 5% of the Chrysosporium sp. seed liquid; the fermentation conditions are set as aerobic fermentation at 28℃, 150rpm for 3 days;

[0088] S05 intermediate addition of symbiotic bacteria: when the liquid fermentation is carried out for 3 days, add green tea powder and add the yeast seed liquid (inoculate 3%), continue to culture for 3-4 days;

[0089] S06 transfer to solid-state fermentation: adsorb the fermentation liquid on wheat bran, place in a constant humidity incubator for static fermentation at 25℃ for 7 days;

[0090] S07 obtain the final fermentation ingredient: after mixing and stirring the whole fermentation product of step S06 with the same volume of deionized water, then centrifugal separation to take the supernatant, then low-temperature reduced pressure concentration, freeze-drying, the Anemarrhena asphodeloides extract can be obtained.

[0091] The components of the liquid medium in S03 and S04 include:

[0092] glucose 25g / L, peptone 8g / L, yeast extract 2.5g / L, ammonium nitrate 0.5g / L, magnesium sulfate heptahydrate 0.5g / L, vitamin B1 0.5g / L, gum arabic 0.05g / L, water 1L.

[0093] The Chrysosporium sp. is Eurotium cristatum, China Industrial Microbial Culture Collection Center.

[0094] The yeast is Rhodosporidium toruloides, China Industrial Microbial Culture Collection Center.

[0095] The mass ratio of Anemarrhena asphodeloides powder and green tea powder is 3:1, and both are passed through 200 meshes.

[0096] In this embodiment, the tea leaves are a mixture of black tea and white tea, and the mass ratio is 2:3. The method for extracting tea leaves is as follows:

[0097] Crushing: crush the black tea leaves / white tea leaves, pass through 100 meshes;

[0098] Suspend the crushed tea leaf powder in 10 times the mass of a buffer solution at 50℃ (0.1M citric acid-phosphate buffer, pH 4.5-5.5), add cellulase and pectinase (total mass 8%, mass ratio 3:5), and perform enzymatic hydrolysis for 30 minutes;

[0099] Ultrasonic extraction: The enzyme-treated tea leaf suspension was placed in an ultrasonic extraction instrument, and treated at a frequency of 40 kHz and a temperature of 50°C for 30 minutes.

[0100] Microwave extraction: The solution was immediately transferred to a microwave extraction device, heated at a power of 300 W for 5 minutes; then, using CO2 as a supercritical fluid, the temperature was set to 35°C, the pressure was set to 25 MPa, and 10 vol% ethanol was used as a cosolvent; the CO2 flow rate was set to 10 mL / min; the extraction time was 30 minutes; the extracted components were collected, and the residue was removed by high-speed centrifugation (10000 rpm); and the extracted solution was concentrated by using a rotary evaporator; the concentrated extract was dried into a powder by freeze-drying, thereby obtaining the tea leaf extract.

[0101] In this embodiment, the preparation method of the liquorice is as follows:

[0102] S1: The liquorice powder was loaded into an extraction tank; supercritical CO2 and 5 vol% ethanol were used as cosolvents to extract the powder at 30°C and 25 MPa for 60 min; the CO2 flow rate was set to 10 mL / min; the extracted solution was collected and concentrated; and the concentrated extract was dried into a powder by freeze-drying;

[0103] S2: The powder obtained in step S1 was added with 5 times the volume of distilled water, and a stirrer was used to fully stir and disperse the powder in the water; under stirring, 1 / 8 times the volume of a potassium hydroxide aqueous solution (10 wt%) was slowly added.

[0104] S3: After continuous stirring at 40°C for 2 h, the excess water was removed by concentration under reduced pressure; and the liquorice extract was obtained by drying.

[0105] In this embodiment, the preparation method of the white flower snake tongue grass extract is as follows:

[0106] (1) Sample preparation: The white flower snake tongue grass sample was crushed into fine particles

[0107] (2) The white flower snake tongue grass powder was added with 10 times the amount of deionized water, and the mixture was stirred and heated at 80°C for 1-2 hours to obtain an extract;

[0108] (3) After the extraction was completed, the extract was cooled to room temperature, filtered, and the plant residue was removed;

[0109] (4) The filtered extract was placed in a rotary evaporator, concentrated under reduced pressure at 40°C, and the extract was concentrated to 1 / 5 times the volume; and the concentrated extract was freeze-dried.

[0110] The present embodiment also provides a skin care product, and the preparation raw materials of the skin care product comprise:

[0111] Glycerol: 4 parts

[0112] p-hydroxyacetophenone: 0.3 parts

[0113] carbomer: 0.6 parts

[0114] arginine: 0.6 parts

[0115] the repair antioxidant composition: 10-15 parts

[0116] 1,2-hexanediol: 0.5 parts

[0117] water is added to 100 parts.

[0118] The preparation method is as follows:

[0119] After mixing and stirring the water, glycerol, p-hydroxyacetophenone, and carbomer and heating to 75-80 degrees, homogenize for 2-3 minutes until completely dissolved, start cooling to below 45 degrees, add arginine, slowly stir until transparent, add the remaining raw materials, and stir uniformly.

[0120] Example 2

[0121] The example provides a repair antioxidant composition, which comprises Anemarrhena asphodeloides extract, tea extract, Glycyrrhiza glabra extract, Oldenlandia diffusa extract, and honey; and the mass ratio is 3:10:6:2:1.

[0122] The preparation of the Anemarrhena asphodeloides extract is as follows: mix Anemarrhena asphodeloides powder and green tea powder according to a mass ratio of 3:1, then perform water extraction according to a solid-liquid ratio of 1:10, at a temperature of 60°C for 2 hours, and then perform vacuum distillation and concentration to the same volume as in Example 1.

[0123] In this example, the tea is a mixture of black tea and white tea, and the mass ratio is 2:3. The extraction method of the tea is as follows:

[0124] Crushing: crush the black tea leaves / white tea leaves to pass through a 100-mesh sieve;

[0125] Suspend the crushed tea powder in 10 times the mass of a buffer solution at 50°C (0.1M citric acid-phosphate buffer, pH 4.5-5.5), add cellulase and pectinase (total mass 8%, mass ratio 3:5), and perform enzymatic treatment for 30 minutes;

[0126] Ultrasonic extraction: place the enzyme-treated tea suspension in an ultrasonic extractor, treat at a frequency of 40kHz and a temperature of 50°C for 30 minutes.

[0127] Microwave extraction: immediately transfer the solution into a microwave extraction device, heat for 5 minutes at 300 W power; then use CO2 as supercritical fluid, set the temperature at 35℃, the pressure at 25 MPa, use 10 vol% ethanol as cosolvent; set the CO2 flow at 10 mL / min; extraction time: 30 minutes; collect the extracted ingredients, remove the residue by high-speed centrifugation (10000 rpm), and concentrate the extract by using a rotary evaporator; dry the concentrated extract into powder by freeze-drying, and the tea leaf extract is obtained.

[0128] In this embodiment, the preparation method of the liquorice is as follows:

[0129] S1: fill the liquorice powder into an extraction tank; use supercritical CO2 and 5 vol% ethanol as cosolvent to extract at 30℃ and 25 MPa for 60 min; set the CO2 flow at 10 mL / min; collect and concentrate the extract; dry the concentrated extract into powder by freeze-drying;

[0130] S2: add 5 times the volume of distilled water to the powder obtained in step S1, and fully stir to disperse in water by using a stirrer; slowly add 1 / 8 volume of potassium hydroxide aqueous solution (10 wt%) under stirring.

[0131] S3: concentrate under reduced pressure after continuous stirring at 40℃ for 2 h to remove excess water; dry to obtain the liquorice extract.

[0132] In this embodiment, the preparation method of the extract of Oldenlandia diffusa is as follows:

[0133] (1) sample preparation: crush the Oldenlandia diffusa sample into fine particles

[0134] (2) add 10 times the amount of deionized water to the Oldenlandia diffusa powder, and heat the extract by stirring at 80℃ for 1-2 hours;

[0135] (3) after the extraction is completed, cool to room temperature, filter the extract, and remove the plant residue;

[0136] (4) place the filtered extract into a rotary evaporator, concentrate the extract by 1 / 5 volume under reduced pressure at 40℃, and freeze-dry the concentrated liquid.

[0137] This embodiment also provides a skin care product, and the preparation raw materials thereof comprise:

[0138] Glycerol: 4 parts

[0139] p-Hydroxyacetophenone: 0.3 parts

[0140] Carbomer: 0.6 parts

[0141] Arginine: 0.6 parts

[0142] The repair antioxidant composition: 10-15 parts

[0143] 1,2-hexanediol: 0.5 parts

[0144] Water is added to 100 parts.

[0145] The preparation method is as follows:

[0146] After the water, glycerol, p-hydroxyacetophenone, carbomer are mixed and heated to 75-80 degrees, homogenize for 2-3 minutes to completely dissolve, start to cool to below 45 degrees, add arginine, slowly stir to transparent, add the remaining raw materials, stir evenly.

[0147] Example 3

[0148] The example provides a repair antioxidant composition, the composition comprises Anemarrhena asphodeloides extract, tea extract, liquorice extract, oldenlandia diffusa willd extract, honey; and the mass ratio is: 3:10:6:2:1.

[0149] The preparation method of the Anemarrhena asphodeloides extract is as follows:

[0150] S01 Yeast is cultured in YPD medium for 24 hours for standby;

[0151] S02 Preliminary liquid fermentation: add Anemarrhena asphodeloides powder to the liquid medium, adjust pH to 5.5, inoculate 5% yeast; the fermentation condition is set as aerobic fermentation at 28℃, 150rpm for 3 days;

[0152] S03 Mid-term culture: when the liquid fermentation is carried out to the third day, add green tea powder, continue to culture for 3-4 days;

[0153] S04 Transfer to solid state fermentation: adsorb the fermentation liquid on wheat bran, place in a constant humidity incubator for static fermentation at 25℃ for 7 days;

[0154] S05 Obtain the final fermentation ingredient: mix and stir the whole fermentation product of step S06 and the same volume of deionized water after soaking, then centrifuge to take the supernatant, then low-temperature reduce-pressure concentration, freeze-drying, to obtain the Anemarrhena asphodeloides extract.

[0155] The composition of the liquid medium in S02 comprises:

[0156] Glucose 25g / L, peptone 8g / L, yeast extract 2.5g / L, ammonium nitrate 0.5g / L, magnesium sulfate heptahydrate 0.5g / L, vitamin B1 0.5g / L, gum arabic 0.05g / L, water 1L.

[0157] wherein the yeast is Rhodosporidium toruloides, China General Microbiological Culture Collection Center.

[0158] wherein the mass ratio of the Anemarrhena asphodeloides L. powder and the green tea powder is 3:1, and both are passed through a 200-mesh sieve.

[0159] In this embodiment, the tea leaves are a mixture of black tea and white tea, and the mass ratio is 2:3. The tea leaf extraction method is as follows:

[0160] Crushing: crush the black tea leaves / white tea leaves and pass them through a 100-mesh sieve;

[0161] Suspension: suspend the crushed tea leaf powder in 10 times the mass of a buffer solution at 50°C (0.1M citric acid-phosphate buffer, pH 4.5-5.5), and add cellulase and pectinase (total mass 8%, mass ratio 3:5) to perform enzymatic hydrolysis for 30 minutes.

[0162] Ultrasonic extraction: place the tea leaf suspension after enzymatic hydrolysis in an ultrasonic extraction instrument, and treat it at a frequency of 40 kHz and a temperature of 50°C for 30 minutes.

[0163] Microwave extraction: immediately transfer the solution to a microwave extraction device, heat it at a power of 300W for 5 minutes; then use CO2 as a supercritical fluid, set the temperature to 35°C and the pressure to 25MPa, use 10vol% ethanol as a cosolvent; set the CO2 flow rate to 10mL / min; extraction time: 30 minutes; collect the extracted components, remove the residue using high-speed centrifugation (10000rpm), and concentrate the extract using a rotary evaporator; dry the concentrated extract into a powder by freeze-drying to obtain the tea leaf extract.

[0164] In this embodiment, the preparation method of the liquorice is as follows:

[0165] S1: load the liquorice powder into an extraction tank; use supercritical CO2 and 5vol% ethanol as a cosolvent to extract at 30°C and 25MPa for 60min; set the CO2 flow rate to 10mL / min; collect and concentrate the extract; dry the concentrated extract into a powder by freeze-drying;

[0166] S2: add 5 times the volume of distilled water to the powder obtained in step S1, and use a stirrer to fully stir and disperse it in the water; under stirring, slowly add 1 / 8 the volume of a potassium hydroxide aqueous solution (10wt%).

[0167] S3: after continuous stirring at 40°C for 2h, concentrate under reduced pressure to remove excess water; dry to obtain the liquorice extract.

[0168] In this embodiment, the preparation method of the extract of Oldenlandia diffusa is as follows:

[0169] (1) Sample preparation: Oldenlandia diffusa sample is crushed into fine particles

[0170] (2) Add 10 times the amount of deionized water to the Oldenlandia diffusa powder, and stir and heat the extract at 80°C for 1-2 hours;

[0171] (3) After the extraction is completed, cool to room temperature and filter the extract to remove the plant residues;

[0172] (4) Place the filtered extract in a rotary evaporator, and concentrate the extract to 1 / 5 of the volume under reduced pressure at 40°C. Freeze-dry the concentrated solution.

[0173] This embodiment also provides a skin care product, the preparation raw materials of which comprise:

[0174] Glycerin: 4 parts

[0175] 4-Hydroxyacetophenone: 0.3 parts

[0176] Carbomer: 0.6 parts

[0177] Arginine: 0.6 parts

[0178] The repair antioxidant composition: 10-15 parts

[0179] 1,2-Hexanediol: 0.5 parts

[0180] Water is added to 100 parts.

[0181] The preparation method is as follows:

[0182] Mix and stir water, glycerin, 4-hydroxyacetophenone, and carbomer, heat to 75-80°C, homogenize for 2-3 minutes until completely dissolved, start to cool to below 45°C, add arginine, slowly stir until transparent, add the remaining raw materials, and stir uniformly.

[0183] Example 4

[0184] This embodiment provides a repair antioxidant composition, which comprises Anemarrhena asphodeloides extract, tea extract, Glycyrrhiza glabra extract, Oldenlandia diffusa extract, and honey, and the mass ratio is 3:10:6:2:1.

[0185] The preparation method of the Anemarrhena asphodeloides extract is as follows:

[0186] S01 strain activation: activate the S01 strain on PDA plate solid culture medium, and culture at 28°C for 7 days until obvious colonies are formed;

[0187] S02Symbiotic bacteria activation: Yeast was cultured in YPD medium for 24 hours, ready for use;

[0188] S03Preparation of liquid seed solution: The activated Aureobasidium pullulans was inoculated into liquid medium and cultured in a constant temperature shaker at 28℃, 150 rpm for 48 hours to prepare 10 7 CFU / mL Aureobasidium pullulans seed solution.

[0189] S04Preliminary liquid fermentation: The Anemarrhena rhizoma powder was added to the liquid medium, the pH was adjusted to 5.5, and 5% Aureobasidium pullulans seed solution was inoculated; the fermentation conditions were set as aerobic fermentation at 28℃, 150 rpm for 3 days;

[0190] S05Intermediate addition of symbiotic bacteria: When the liquid fermentation was carried out to the 3rd day, yeast was added (inoculated 3%), and continued to culture for 3-4 days;

[0191] S06Transition to solid-state fermentation: The fermentation liquid was adsorbed on wheat bran and placed in a constant humidity incubator for static fermentation at 25℃ for 7 days;

[0192] S07Obtaining the final fermentation component: After the whole fermentation product of step S06 and the same volume of deionized water were mixed, stirred and soaked, then centrifuged to take the supernatant, then low-temperature reduced pressure concentration, freeze-drying, the Anemarrhena rhizoma extract was obtained.

[0193] The components of the liquid medium in S03 and S04 include:

[0194] Glucose 25g / L, peptone 8g / L, yeast extract 2.5g / L, ammonium nitrate 0.5g / L, magnesium sulfate heptahydrate 0.5g / L, vitamin B1 0.5g / L, gum arabic 0.05g / L, water 1L.

[0195] Among them, the Aureobasidium pullulans is Eurotium cristatum, China Industrial Microbial Culture Collection Center.

[0196] The yeast is Rhodosporidium toruloides, China Industrial Microbial Culture Collection Center.

[0197] The mass ratio of Anemarrhena rhizoma powder and green tea powder is 3:1, and both are passed through 200 meshes.

[0198] In this embodiment, the tea leaves are a mixture of black tea and white tea, and the mass ratio is 2:3. The method for extracting tea leaves is as follows:

[0199] Crushing: Crush the black tea leaves / white tea leaves and pass through 100 meshes;

[0200] The pulverized tea powder is suspended in 10 times the mass of a buffer solution at 50°C (0.1 M citric acid-phosphate buffer, pH 4.5-5.5), and cellulase and pectinase are added (total mass 8%, mass ratio 3:5) to perform enzymatic treatment for 30 minutes;

[0201] Ultrasonic extraction: the tea leaf suspension after enzymatic treatment is placed in an ultrasonic extraction instrument, and treated at a frequency of 40 kHz and a temperature of 50°C for 30 minutes.

[0202] Microwave extraction: the solution is immediately transferred to a microwave extraction device, heated at a power of 300 W for 5 minutes; then using CO2 as a supercritical fluid, the temperature is set to 35°C, the pressure is set to 25 MPa, and 10 vol% ethanol is used as a cosolvent; the CO2 flow rate is set to 10 mL / min; the extraction time is 30 minutes; the extracted components are collected, and the residue is removed by high-speed centrifugation (10000 rpm); the extracted solution is concentrated using a rotary evaporator; and the concentrated extract is dried into a powder by freeze-drying to obtain the tea extract.

[0203] In this embodiment, the preparation method of the liquorice is as follows:

[0204] S1: The liquorice powder is loaded into an extraction tank; supercritical CO2 and 5 vol% ethanol are used as cosolvents to perform extraction at 30°C and 25 MPa for 60 min; the CO2 flow rate is set to 10 mL / min; the extracted solution is collected and concentrated; and the concentrated extract is dried into a powder by freeze-drying;

[0205] S2: The powder obtained in step S1 is added with 5 times the volume of distilled water, and a stirrer is used to fully stir and disperse it in the water; under stirring, 1 / 8 the volume of a potassium hydroxide aqueous solution (10 wt%) is slowly added.

[0206] S3: After continuous stirring at 40°C for 2 h, the excess water is removed by concentration under reduced pressure; and the liquorice extract is obtained by drying.

[0207] In this embodiment, the preparation method of the white flower snake tongue grass extract is as follows:

[0208] (1) Sample preparation: the white flower snake tongue grass sample is pulverized into fine particles

[0209] (2) The white flower snake tongue grass powder is added with 10 times the amount of deionized water, and the extract is stirred and heated at 80°C for 1-2 hours;

[0210] (3) After the extraction is completed, the extract is cooled to room temperature, filtered, and the plant residue is removed;

[0211] (4) The filtered extract is placed in a rotary evaporator, and concentrated under reduced pressure at 40°C. The extract is concentrated to 1 / 5 of the original volume, and the concentrated solution is freeze-dried.

[0212] The present embodiment also provides a skin care product, the raw materials for preparing which comprise:

[0213] Glycerin: 4 parts

[0214] p-Hydroxyacetophenone: 0.3 parts

[0215] Carbomer: 0.6 parts

[0216] Arginine: 0.6 parts

[0217] The repair antioxidant composition: 10-15 parts

[0218] 1,2-Hexanediol: 0.5 parts

[0219] Water is added to 100 parts.

[0220] The preparation method is as follows:

[0221] After the water, glycerin, p-hydroxyacetophenone, and carbomer are mixed, heated, and stirred to 75-80°C, they are homogenized for 2-3 minutes until completely dissolved, and then cooled to below 45°C. Arginine is added, slowly stirred until transparent, and then the remaining raw materials are added and stirred until uniform.

[0222] Example 5

[0223] The present embodiment provides a repair antioxidant composition, which comprises Anemarrhena asphodeloides extract, Glycyrrhiza glabra extract, Spermacoce latifolia extract, and honey, and has a mass ratio of 3:6:2:1.

[0224] The preparation method of the Anemarrhena asphodeloides extract is as follows:

[0225] S01 Strain activation: The Chrysosporium sp. is activated on PDA plate solid medium, and cultured at 28°C for 7 days until obvious colonies are formed;

[0226] S02 Symbiotic bacteria activation: The yeast is cultured in YPD medium for 24 hours and reserved;

[0227] S03 Preparation of liquid seed solution: The activated Chrysosporium sp. is inoculated into liquid medium, and cultured in a constant-temperature shaker at 28°C and 150 rpm for 48 hours to prepare 10 7 CFU / mL of Chrysosporium sp. seed solution.

[0228] S04 Preliminary liquid fermentation: The Anemarrhena asphodeloides powder is added to the liquid medium, the pH is adjusted to 5.5, and 5% Chrysosporium sp. seed solution is inoculated. The fermentation conditions are set to aerobic fermentation at 28°C and 150 rpm for 3 days.

[0229] S05 mid-stage addition of symbiotic bacteria: when the liquid fermentation is carried out to the third day, green tea powder and yeast strain liquid (inoculated 3%) are added, and the culture is continued for 3-4 days;

[0230] S06 transfer to solid-state fermentation: the fermentation liquid is adsorbed on wheat bran, and is placed in a constant humidity incubator for static fermentation at 25°C for 7 days;

[0231] S07 obtain final fermentation ingredients: after the whole fermentation product of step S06 is mixed with the same volume of deionized water and stirred and soaked, the upper clear liquid is then separated by centrifugation, followed by low-temperature and reduced-pressure concentration, and freeze-drying, to obtain the extract of Anemarrhena asphodeloides Bunge.

[0232] The components of the liquid culture medium in S03 and S04 include:

[0233] Glucose 25 g / L, peptone 8 g / L, yeast extract 2.5 g / L, ammonium nitrate 0.5 g / L, magnesium sulfate heptahydrate 0.5 g / L, vitamin B1 0.5 g / L, gum arabic 0.05 g / L, water 1 L.

[0234] Among them, the golden flower fungus is Eurotium cristatum, China Industrial Microbial Strain Preservation and Management Center.

[0235] The yeast is Rhodosporidium toruloides, China Industrial Microbial Strain Preservation and Management Center.

[0236] The mass ratio of Anemarrhena asphodeloides Bunge powder and green tea powder is 3:1, and both are passed through 200 meshes.

[0237] In this embodiment, the preparation method of Glycyrrhiza glabra L. is as follows:

[0238] S1: Glycyrrhiza glabra L. powder is loaded into an extraction tank; supercritical CO2 and 5 vol% ethanol are used as cosolvents for extraction at 30°C and 25 MPa for 60 min; the CO2 flow rate is set to 10 mL / min; the extraction liquid is collected and concentrated; the concentrated extract is dried into powder by freeze-drying;

[0239] S2: 5 times the volume of distilled water is added to the powder obtained in step S1, and a stirrer is used to fully stir and disperse it in water; under stirring, 1 / 8 volume of potassium hydroxide aqueous solution (10 wt%) is slowly added.

[0240] S3: after continuous stirring at 40°C for 2 h, the excess water is removed by concentration under reduced pressure; the Glycyrrhiza glabra L. extract is obtained by drying.

[0241] In this embodiment, the preparation method of the extract of Oldenlandia diffusa is as follows:

[0242] (1) Sample preparation: Oldenlandia diffusa sample is crushed into fine particles

[0243] (2) Add 10 times the amount of deionized water to the Oldenlandia diffusa powder, and stir and heat the extract at 80°C for 1-2 hours;

[0244] (3) After the extraction is completed, cool to room temperature and filter the extract to remove the plant residues;

[0245] (4) Put the filtered extract into a rotary evaporator, and concentrate the extract under reduced pressure at 40°C to 1 / 5 of the volume. Freeze-dry the concentrated solution.

[0246] This embodiment also provides a skin care product, the preparation raw materials of which comprise:

[0247] Glycerin: 4 parts

[0248] 4-Hydroxyacetophenone: 0.3 parts

[0249] Carbomer: 0.6 parts

[0250] Arginine: 0.6 parts

[0251] The repair antioxidant composition: 10-15 parts

[0252] 1,2-Hexanediol: 0.5 parts

[0253] Add water to 100 parts.

[0254] The preparation method is as follows:

[0255] Mix and stir water, glycerin, 4-hydroxyacetophenone, and carbomer, heat to 75-80°C, and homogenize for 2-3 minutes until completely dissolved. Start cooling to below 45°C, slowly stir to transparent, add the remaining raw materials, and stir uniformly.

[0256] Example 6

[0257] This embodiment provides a repair antioxidant composition, which comprises Anemarrhena asphodeloides extract, tea extract, Glycyrrhiza glabra extract, Oldenlandia diffusa extract, and honey, and the mass ratio is 3:10:6:2:1.

[0258] The preparation method of the Anemarrhena asphodeloides extract is as follows:

[0259] S01 strain activation: activate the golden flower fungus on PDA flat solid culture medium, cultivate at 28°C for 7 days, until obvious colonies grow;

[0260] S02Symbiotic bacteria activation: Yeast was cultured in YPD medium for 24 hours, ready for use;

[0261] S03Preparation of liquid seed solution: The activated Aureobasidium pullulans was inoculated into liquid medium and cultured in a constant temperature shaker at 28℃, 150 rpm for 48 hours to prepare 10 7 CFU / mL Aureobasidium pullulans seed solution.

[0262] S04Preliminary liquid fermentation: Add Anemarrhena asphodeloides powder to the liquid medium, adjust the pH to 5.5, inoculate 5% Aureobasidium pullulans seed solution; The fermentation conditions are set as aerobic fermentation at 28℃, 150 rpm for 3 days;

[0263] S05Intermediate addition of symbiotic bacteria: When the liquid fermentation is carried out to the 3rd day, add green tea powder and inoculate 3% yeast seed solution, continue to culture for 3-4 days;

[0264] S06Transition to solid state fermentation: The fermentation liquid is adsorbed on wheat bran and placed in a constant humidity incubator for static fermentation at 25℃ for 7 days;

[0265] S07Obtain the final fermentation ingredient: After mixing and stirring the whole fermentation product of step S06 with the same volume of deionized water, then centrifugal separation to take the supernatant, then low-temperature vacuum concentration, freeze-drying, the Anemarrhena asphodeloides extract can be obtained.

[0266] The composition of the liquid medium in S03 and S04 includes:

[0267] Glucose 25g / L, peptone 8g / L, yeast extract 2.5g / L, ammonium nitrate 0.5g / L, magnesium sulfate heptahydrate 0.5g / L, vitamin B1 0.5g / L, gum arabic 0.05g / L, water 1L.

[0268] Among them, the Aureobasidium pullulans is Eurotium cristatum, China Industrial Microbial Strain Preservation and Management Center.

[0269] The yeast is Rhodosporidium toruloides, China Industrial Microbial Strain Preservation and Management Center.

[0270] The mass ratio of Anemarrhena asphodeloides powder and green tea powder is 3:1, both of which are passed through 200 meshes.

[0271] In this embodiment, the tea leaves are a mixture of black tea and white tea with a mass ratio of 2:3, and the tea extraction method is as follows:

[0272] Crushing: Crush the black tea leaves / white tea leaves and pass them through 100 meshes;

[0273] The pulverized tea powder was suspended in 10 times the mass of a buffer solution at 50°C (0.1 M citric acid-phosphate buffer, pH 4.5-5.5), cellulase and pectinase were added (total mass 8%, mass ratio 3:5), and enzymatic treatment was performed for 30 minutes;

[0274] Ultrasonic extraction: The tea leaf suspension after enzymatic treatment was placed in an ultrasonic extraction instrument, and treated at a frequency of 40 kHz and a temperature of 50°C for 30 minutes.

[0275] Microwave extraction: The solution was immediately transferred to a microwave extraction device, heated at a power of 300 W for 5 minutes; then using CO2 as a supercritical fluid, the temperature was set to 35°C, the pressure was set to 25 MPa, and 10 vol% ethanol was used as a cosolvent; the CO2 flow rate was set to 10 mL / min; the extraction time was 30 minutes; the extracted components were collected, and the residue was removed using high-speed centrifugation (10000 rpm); the extracted solution was concentrated using a rotary evaporator; and the concentrated extract was dried into a powder by freeze-drying, thereby obtaining the tea leaf extract.

[0276] In this embodiment, the preparation method of the liquorice is as follows:

[0277] S1: The liquorice powder was loaded into an extraction tank; supercritical CO2 and 5 vol% ethanol were used as cosolvents to extract at 30°C and 25 MPa for 60 min; the CO2 flow rate was set to 10 mL / min; the extracted solution was collected and concentrated; the concentrated extract was dried into a powder by freeze-drying; and the liquorice extract was obtained.

[0278] In this embodiment, the preparation method of the white flower snake tongue grass extract is as follows:

[0279] (1) Sample preparation: The white flower snake tongue grass sample was pulverized into fine particles

[0280] (2) The white flower snake tongue grass powder was added to 10 times the amount of deionized water, and the extract was stirred and heated at 80°C for 1-2 hours;

[0281] (3) After the extraction was completed, the extract was cooled to room temperature and filtered to remove the plant residue;

[0282] (4) The filtered extract was placed in a rotary evaporator, and the extract was concentrated to 1 / 5 of the volume under reduced pressure at 40°C; and the concentrated solution was freeze-dried.

[0283] The embodiment also provides a skin care product, and the preparation raw materials thereof comprise:

[0284] Glycerol: 4 parts

[0285] p-Hydroxyacetophenone: 0.3 parts

[0286] Carbomer: 0.6 parts

[0287] Arginine: 0.6 parts

[0288] The repair antioxidant composition: 10-15 parts

[0289] 1,2-hexanediol: 0.5 parts

[0290] Water is added to 100 parts.

[0291] The preparation method is as follows:

[0292] After mixing and stirring the water, glycerol, p-hydroxyacetophenone, and carbomer and heating to 75-80 degrees, homogenize for 2-3 minutes until completely dissolved, start cooling to below 45 degrees, add arginine, slowly stir until transparent, add the remaining raw materials, and stir evenly.

[0293] Example 7

[0294] The example provides a repair antioxidant composition, which comprises Anemarrhena asphodeloides extract, tea extract, Glycyrrhiza glabra extract, Oldenlandia diffusa extract, and honey; and the mass ratio is 3:10:6:2:1.

[0295] The preparation method of the Anemarrhena asphodeloides extract, tea extract, Glycyrrhiza glabra extract, and Oldenlandia diffusa extract is as follows:

[0296] S01Mix Anemarrhena asphodeloides, tea, Glycyrrhiza glabra, and Oldenlandia diffusa and pulverize to fine particles;

[0297] S02Add 10 times the amount of deionized water, and stir and heat the extract at 80°C for 1-2 hours;

[0298] S03After the extraction is completed, cool to room temperature, filter the extract, and remove the plant residues;

[0299] S04Put the filtered extract into a rotary evaporator, concentrate the extract under reduced pressure at 40°C to 1 / 5 of the volume, and freeze-dry the concentrated liquid.

[0300] The example also provides a skin care product, which is prepared from the following raw materials:

[0301] Glycerol: 4 parts

[0302] p-hydroxyacetophenone: 0.3 parts

[0303] Carbomer: 0.6 parts

[0304] Arginine: 0.6 parts

[0305] The repair antioxidant composition: 10-15 parts

[0306] 1,2-hexanediol: 0.5 parts

[0307] Water is added to 100 parts.

[0308] The preparation method is as follows:

[0309] After water, glycerol, p-hydroxyacetophenone, carbomer are mixed and heated to 75-80 degrees, homogenize for 2-3 minutes until completely dissolved, start to cool down to below 45 degrees, add arginine, slowly stir until transparent, add the remaining raw materials, stir evenly.

[0310] Performance test

[0311] Test 1: The products prepared in Examples 1-7 were placed at 5°C, 25°C, and 40°C for 90 days, and the product properties were observed. The test results are shown in Table 1.

[0312] Table 1 Test results of performance test 1

[0313] Example 5℃ 25℃ 40℃ Example 1 No change No change No change Example 2 Partial haze No change Partial effervescence Example 3 No change No change Partial effervescence Example 4 Layering Partial haze Layering Example 5 Layering Layering Layering Example 6 Layering Layering Layering Example 7 Layering Layering Layering

[0314] Test 2: Elastase inhibition rate

[0315] Fermentation filtrate: diluted with pure water to a sample concentration of 5%;

[0316] Positive control (epigallocatechin gallate): diluted with water to a positive control concentration of 0.1%;

[0317] Negative control: pure water.

[0318] Test operation steps:

[0319] Design sample groups, sample background groups, solvent groups, and solvent background groups, and set up 3 parallel groups for each group. Add different reagent solutions to a 96-well plate, shake gently, incubate at 25°C for 15 min, then place it in an enzyme marker, and measure the absorbance at 410 nm.

[0320] Calculation formula: Elastase inhibition rate (%) = (1-(C-D) / (A-B)) x 100

[0321] Wherein, A is the absorbance of the reaction solution without sample; B is the absorbance of the reaction solution without sample and enzyme; C is the absorbance of the reaction solution containing sample and enzyme; D is the absorbance of the reaction solution containing sample and not containing enzyme.

[0322] Statistical analysis software is SPSS, and the comparison between the test sample, the positive control and the negative control in terms of elastase inhibition rate is made by independent sample t test. The above statistical analysis is a two-tailed test, and the significance level is a=0.05, P>0.05, indicating that there is no significant difference between the two groups; P<0.05, indicating that there is a significant difference between the two groups.

[0323] Table 2 Test results table

[0324] Name Test result (%) P value Example 1 68.243±0.153 ＜0.05 Example 2 60.642±0.275 ＜0.05 Example 3 65.269±0.361 ＜0.05 Example 4 62.163±0.637 ＜0.05 Example 5 53.283±0.163 ＜0.05 Example 6 58.053±0.227 ＜0.05 Example 7 51.141±0.226 ＜0.05 Negative control -8.127±1.326 / Positive control 70.321±7.356 ＜0.05

[0325] The elastase inhibition rate is retained to three decimal places, and P<0.05 indicates that the sample and the positive control have a significant difference compared with the negative control.

[0326] Test 3: difference in transepidermal water loss rate

[0327] 25-50 year-old healthy subjects (all subjects have the characteristics of sensitive skin, such as easy allergy, redness, etc.), 7 men and 35 women, with an average age of 37.32 years.

[0328] Use part: forearm

[0329] Method of use: when used on the forearm, the test sample is single-coated in an amount of (2.0±0.1) mg / cm 2 , and a latex glove is used to evenly spread the test sample in the test area.

[0330] Test steps

[0331] The subject does not touch the water on the inner part of the arm before visiting

[0332] Before the experiment, the subject needs to clean the arm with a dry tissue paper;

[0333] Mark 2 measurement areas on the inner side of the subject's left or right arm, each with an area of 3 cm*3 cm, and the distance between the areas is at least 1 cm;

[0334] The sample area and the negative control area are randomly distributed in the designated area, ensuring that the position of each area is statistically balanced: the subject exposes the arm, and rests in an environment with a temperature of 21±1℃ and a relative humidity of 50±10% RH for 30 minutes; apply the inducer (0.1% histamine solution) to the sample area and the negative area for 10 minutes, and test the transepidermal water loss rate of the designated area using Tewameter and the hemoglobin of the designated area using Mexamter before the end of the application and before using the sample (Timm);

[0335] Use the sample according to the sample use method on the designated sample area;

[0336] After the application, the subjects expose their arms in an environment with a temperature of 21 ± 1 °C and a relative humidity of 50 ± 10% RH for 1 h of rest; 1 h later (T1), the transepidermal water loss rate is measured on the calibration area using the Tewameter test.

[0337] Difference in the transepidermal water loss rate of the skin

[0338] Courage + Khazaka percutaneous water loss and water evaporation heat loss test probe Tewameter TM300 for detecting the transepidermal water loss rate of the skin.

[0339] Table 3 Comparison of the transepidermal water loss rate before and after

[0340]

[0341]

[0342] Test 4: Anti-inflammatory test

[0343] This test uses lipopolysaccharide (LPS) to stimulate macrophage Raw264.7 to establish an inflammation model, and the soothing effect of the sample is evaluated by detecting the change in the secretion amount of the relevant inflammatory factors after the sample is applied

[0344] The cells used in this test are macrophage Raw264.7 (mouse macrophage cell line), which are purchased from the Chinese Academy of Sciences Cell Bank.

[0345] Cell inoculation: inoculate cells into a 6-well plate at a seeding density of 6.4*10 5 The cells are incubated in an incubator (37 °C, 5% CO2) overnight.

[0346] 2. Solution preparation: prepare the test substance working solution according to the experimental design (Table 4).

[0347] Table 4

[0348]

[0349] Dosing: when the plating rate of the cells in the 6-well plate reaches 40% to 60%, group dosing is performed, with a dosing amount of 1.8 mL per well, and 3 replicate wells are set for each group. The incubator (37 °C, 5% CO2) is continued to be cultured for 2 h.

[0350] LPS stimulation: after 2 h of culture, 200 uL of LPS working solution prepared from the corresponding test substance working solution is added to the dosed well plate according to the experimental design, the well plate is shaken left and right to mix the drug in the well plate, the final concentration of LPS is 1 ug / mL, and the incubator (37 °C, 5% CO2) is continued to be cultured for 22 h.

[0351] Sample collection: After the incubation, collect the cell culture supernatant in an EP tube (Note: the amount of sample collected is determined according to the detection index), and after collection, store the sample in a -80°C freezer for frozen preservation. The detection of IL-1α content is carried out according to the operating instructions of the Mouse IL-1α ELISA kit. The detection of PGE2 content is carried out according to the operating instructions of the PGE2 ELISA kit.

[0352] Table 5 test results

[0353]

[0354]

[0355] Significance is indicated by #, p-value < 0.05 is indicated by #, p-value < 0.01 is indicated by ##. The significance of the sample group, PC group and NC group is indicated by *, p-value < 0.05 is indicated by *, p-value < 0.01 is indicated by **.

[0356] Finally, it should be noted that: the above embodiments are only used to illustrate the technical solutions of the present application, and not to limit it; although the present application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that: it can still modify the technical solutions recorded in the foregoing embodiments, or make equivalent replacement for part or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. An anti-inflammatory repair composition characterized in that, The composition comprises an extract of Anemarrhena asphodeloides obtained by fermenting Anemarrhena asphodeloides with Mycorrhiza glabra. The preparation method of the Anemarrhena asphodeloides extract is as follows: S01 Strain Activation: Activate the golden flower fungus on PDA plate solid medium and culture at 28℃ for 7 days until obvious colonies grow. S02 Symbiotic activation: Yeast cells are cultured in YPD medium for 24 hours and then ready for use; S03 Preparation of liquid seed culture: The activated strain is inoculated into a liquid culture medium containing glucose and yeast extract, and cultured in a constant temperature shaker at 28°C and 150 rpm for 48 hours to prepare the seed culture; S04 Preliminary liquid fermentation: Add Anemarrhena asphodeloides powder to the liquid culture medium, adjust the pH to 5.5, and inoculate with 5% Aureobasidium aureum inoculum solution; set the fermentation conditions to aerobic fermentation at 28℃ and 150 rpm for 3 days; S05 Mid-term addition of symbiotic bacteria: When the liquid fermentation is in progress for 3 days, add the activated yeast and continue to culture for 3-4 days; S06 Transfer to solid-state fermentation: Adsorb the fermentation broth onto wheat bran and place it in a constant humidity incubator for static fermentation at 25°C for 7 days; S07 Obtain the final fermentation components: After mixing and stirring the whole fermentation product from step S06 with the same volume of deionized water, centrifuge to separate the supernatant, concentrate under low temperature and reduced pressure, and freeze-dry to obtain the Anemarrhena asphodeloides extract.

2. A repair antioxidant composition characterized in that, The composition comprises an extract of Anemarrhena asphodeloides obtained by fermenting Anemarrhena asphodeloides with Mycorrhiza glabra. The preparation method of the Anemarrhena asphodeloides extract is as follows: S01 Strain Activation: Activate the golden flower fungus on PDA plate solid medium and culture at 28℃ for 7 days until obvious colonies grow. S02 Symbiotic activation: Yeast cells are cultured in YPD medium for 24 hours and then ready for use; S03 Preparation of liquid seed culture: The activated strain is inoculated into a liquid culture medium containing glucose and yeast extract, and cultured in a constant temperature shaker at 28°C and 150 rpm for 48 hours to prepare the seed culture; S04 Preliminary liquid fermentation: Add Anemarrhena asphodeloides powder to the liquid culture medium, adjust the pH to 5.5, and inoculate with 5% Aureobasidium aureum inoculum solution; set the fermentation conditions to aerobic fermentation at 28℃ and 150 rpm for 3 days; S05 Mid-term addition of symbiotic bacteria: When the liquid fermentation is in progress for 3 days, add the activated yeast and continue to culture for 3-4 days; S06 Transfer to solid-state fermentation: Adsorb the fermentation broth onto wheat bran and place it in a constant humidity incubator for static fermentation at 25°C for 7 days; S07 Obtain the final fermentation components: After mixing and stirring the whole fermentation product from step S06 with the same volume of deionized water, centrifuge to separate the supernatant, concentrate under low temperature and reduced pressure, and freeze-dry to obtain the Anemarrhena asphodeloides extract.

3. The composition of claim 2, wherein, The composition also includes tea extract, licorice root extract, and oldenlandia diffusa extract.

4. The composition of claim 3, wherein, The mass ratio of Anemarrhena asphodeloides extract, tea extract, Glycyrrhiza glabra extract, and Hedyotis diffusa extract is 1~3:5~15:2~8:1~3.

5. The composition of claim 3, wherein The mass ratio of Anemarrhena asphodeloides extract, tea extract, Glycyrrhiza glabra extract, and Hedyotis diffusa extract is 3:10:6:

2.

6. The composition of claim 5, wherein, The liquid culture medium in S04 comprises: Glucose 20-30 g / L, peptone 5-10 g / L, yeast extract 2-5 g / L, ammonium nitrate 1-2 g / L, magnesium sulfate heptahydrate 0.5 g / L, vitamin B1 0.5 g / L, gum arabic 0.05 g / L, water 1 L.

7. A skin care product, characterized by, The composition of any one of claims 2-6.

8. The skin care product according to claim 7, characterized in that The raw materials for preparing the same comprise: Glycerin: 4 parts p-Hydroxyacetophenone: 0.3 parts Carbomer: 0.6 parts Arginine: 0.6 parts The repair antioxidant composition: 10-15 parts 1,2-Hexanediol: 0.5 parts Water is added to 100 parts.

9. The composition according to any one of claims 2 to 6, characterized in that, Applied to the field of daily chemicals.

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