A tea fermentation composition for promoting double protein penetration and its application

Abstract

The present invention relates to the field of cosmetics, and in particular to a tea fermentation composition and application that promotes the penetration of dual proteins. The tea fermentation composition of the present invention comprises black tea fermentation filtrate, whey protein, and recombinant collagen. The black tea fermentation filtrate used in the present invention contains more inositol and small molecule biological penetration-promoting polypeptides that promote skin penetration, which can effectively improve the antioxidant, anti-aging, and repair capabilities. After the black tea fermentation filtrate is compounded with whey protein and recombinant collagen, it can help solve the problem of the difficulty of dual protein penetration, thereby improving the utilization rate of dual protein in cosmetics, making it play its maximum effect, and achieving synergistic effect. The tea fermentation composition obtained by the present invention is added to cosmetics to greatly enhance the antioxidant, repair, anti-wrinkle and other effects.

Description

Technical Field

[0001] The present invention relates to the field of cosmetics, and in particular to a tea fermentation composition for promoting dual protein penetration and an application thereof. Background Art

[0002] Whey protein is a globular protein mixture extracted from milk. It has the characteristics of high nutritional value, easy digestion and absorption, and contains a variety of active ingredients. It is recognized as one of the high-quality protein supplements for the human body and is called the king of protein. It is commonly used as a nutritional supplement. Its molecular weight is mostly 14,000-18,000 Daltons, which makes it difficult to absorb when used in cosmetics, resulting in waste.

[0003] Recombinant collagen (human-like collagen) refers to collagen that is 100% identical to the amino acid sequence of human collagen obtained by recombinantly transforming human collagen genes using synthetic biology technology and then fermenting them at high density. It overcomes the drawbacks of animal-derived collagen, is not easily rejected, is more easily absorbed by the body, and is safe. Recombinant collagen achieves anti-aging effects by directly supplementing collagen, saving collagen loss and effectively fighting wrinkles and lightening lines. However, recombinant collagen also has the same problems as whey protein. Because its molecular weight is mostly 30,000-80,000 Daltons, it is not absorbed after application and can only stay on the surface of the skin, causing waste.

[0004] The above two proteins are used in skin care and have powerful multiple benefits such as repair, anti-wrinkle, and firming. However, due to their large molecular weight, they are difficult to absorb when added to skin care products, and their transdermal absorption is poor, which prevents them from exerting their maximum effectiveness. Currently, there are two ways to solve this problem in the industry: the first is to develop collagen with a smaller molecular weight, but this technology has a long R&D cycle and high uncertainty; the second is to achieve skin penetration and absorption through penetration ingredients or technologies. There are three common types of penetration enhancers: the first is chemical penetration enhancers such as ethanol, fatty acids, fatty alcohols, and azones; the second is biological penetration enhancers such as short peptides with less than 30 amino acid residues, which can promote peptides, proteins, nucleic acids, nanoparticles, and other bioactive molecules to penetrate cell membranes or skin and exert biological effects; the third is natural active penetration enhancers such as inositol, terpenes, mints, and piperine. Currently, all three types of penetration enhancers can effectively promote skin absorption. Summary of the Invention

[0005] The purpose of the present invention is to overcome the shortcomings of the existing technology and provide a tea fermentation composition, which can make the double protein more easily permeable and absorbable, thereby improving its anti-aging effect.

[0006] To achieve the above object, the technical solution adopted by the present invention is:

[0007] In a first aspect, the present invention provides a tea fermentation composition comprising the following components in parts by weight: 3-12 parts of black tea fermentation filtrate and 1.51-4.95 parts of double protein; the double protein comprises whey protein and recombinant collagen.

[0008] Tea, as a natural ingredient, is non-toxic, non-irritating, and non-sensitizing to the skin. It takes effect quickly, has regenerative activity, and lasts for a predictable period of time. It promotes penetration while preventing the loss of body components. When applied to the skin, the skin's barrier function can be quickly and completely restored. The black tea fermentation filtrate used in this invention is a fermentation filtrate product of dual yeast (Saccharomyces cerevisiae and Acetobacter xylinum) fermentation. It is rich in substances such as inositol and small molecule biological penetration-promoting polypeptides that promote skin penetration. Molecules with a molecular weight of less than 1000 Daltons (especially small molecules less than 500 Daltons) account for 66.65% of the black tea fermentation filtrate, which helps promote the penetration of synergistic ingredients. Whey protein is extracted from milk and can supplement the nutrients needed by humans. Recombinant collagen is obtained through biosynthesis and can directly replenish collagen, achieving anti-aging effects. The present invention has found through research that the combination of whey protein and recombinant collagen has a synergistically enhanced anti-wrinkle effect. After the black tea fermentation filtrate is combined with whey protein and recombinant collagen, the black tea fermentation filtrate can be used as a penetration enhancer to help solve the problem of the difficulty of penetration of the dual proteins, thereby improving the utilization rate of the dual proteins in cosmetics, enabling them to exert their maximum efficacy and achieve synergistic enhancement.

[0009] Preferably, in the double protein, the weight ratio of whey protein to recombinant collagen is whey protein: recombinant collagen = (1.5-4.5): (0.01-0.45).

[0010] Preferably, the tea fermentation composition comprises the following components in parts by weight: 7-12 parts of black tea fermentation filtrate, 2-4.5 parts of whey protein, and 0.05-0.45 parts of recombinant collagen.

[0011] The weight ratio of black tea fermentation filtrate, whey protein and recombinant collagen is within the above range, and has excellent antioxidant, repair, anti-wrinkle and other effects.

[0012] More preferably, the tea fermentation composition comprises the following components in parts by weight: 10 parts of black tea fermentation filtrate, 3 parts of whey protein, and 0.1 parts of recombinant collagen.

[0013] When the weight ratio of black tea fermentation filtrate, whey protein and recombinant collagen is within the above-mentioned specific range, it has the best antioxidant, repair and anti-wrinkle effects.

[0014] In a second aspect, the present invention provides use of the tea fermentation composition in cosmetics.

[0015] In a third aspect, the present invention provides a cosmetic comprising the above-mentioned tea fermentation composition.

[0016] The tea fermentation composition of the present invention can be added to cosmetics to effectively improve the anti-oxidation, anti-aging, and repairing effects on the skin, and is more conducive to the comprehensiveness and effectiveness of the skin effects.

[0017] Preferably, the tea fermentation composition is added in an amount of 4.51%-16.95% in cosmetics.

[0018] More preferably, the tea fermentation composition is added in an amount of 13.1% in cosmetics.

[0019] Preferably, the dosage form of the cosmetic includes water, lotion, and cream.

[0020] Preferably, the cosmetics further include excipients acceptable in daily chemical products.

[0021] Preferably, the auxiliary material includes at least one of an emollient, a dispersant, a humectant, a preservative, a thickener, and a pH regulator.

[0022] The beneficial effects of the present invention are:

[0023] Tea, as a natural ingredient, is non-toxic, non-irritating, and non-sensitizing to the skin. It takes effect quickly, has renewable activity, and a predictable duration. It prevents the loss of components in the body while promoting penetration. When applied to the skin, the skin's barrier function can be quickly and completely restored. The black tea fermentation filtrate used in the present invention is a product of double yeast fermentation, and is rich in substances such as inositol and small molecule biological penetration-promoting polypeptides that promote skin penetration. After being compounded with whey protein and recombinant collagen, the black tea fermentation filtrate can be used as a penetration enhancer to help solve the problem of the difficulty of double protein penetration, thereby improving the utilization rate of double protein in cosmetics, allowing it to exert its maximum efficacy and achieve synergistic enhancement.

[0024] The tea fermentation composition of the present invention can be added to cosmetics to effectively improve the anti-oxidation, anti-aging, and repairing effects on the skin, and is more conducive to the comprehensiveness and effectiveness of the skin effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is the tea fermentation composition prepared in Example 1.

[0026] Figure 2 This is the tea fermentation composition prepared in Example 10.

[0027] Figure 3 Schematic diagram of the Franz diffusion cell. DETAILED DESCRIPTION

[0028] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0029] The black tea fermentation filtrate used in the present invention was purchased from Guangzhou Xiyuan Biological Co., Ltd.

[0030] The whey protein used in the present invention was purchased from Huzhou Zhenlu Biological Products Co., Ltd.

[0031] The recombinant collagen used in the present invention was purchased from Xi'an Juzi Bio-Gene Technology Co., Ltd.

[0032] The appearance of the tea fermentation composition prepared by the present invention is shown in FIG. Figure 1-2 As shown, Example 1 and Example 10 are taken as examples.

[0033] Examples 1-12:

[0034] An embodiment of the tea fermentation composition of the present invention; the components by weight of the tea fermentation composition are shown in Table 1.

[0035] Table 1 Components by weight of the tea fermentation composition described in each example

[0036]

[0037] Comparative Examples 1-6:

[0038] Comparative Example of the tea fermentation composition of the present invention; the components by weight of the tea fermentation composition are shown in Table 2.

[0039] Table 2 Components by weight of the tea fermentation compositions of each comparative example

[0040]

[0041] Among them, the only difference between Comparative Examples 1-3 and Example 10 is that: Comparative Example 1 does not add black tea fermentation product filtrate; Comparative Example 2 does not add whey protein; Comparative Example 3 does not add recombinant collagen; and the missing components are replaced with equal weight parts of water.

[0042] The only difference between Comparative Examples 5 and 6 and Example 10 is that in Comparative Example 5, the black tea fermentation product filtrate is replaced by an equal weight portion of green tea fermentation filtrate (purchased from Shanghai Ruidian Biotechnology Co., Ltd.); and in Comparative Example 6, the whey protein is replaced by an equal weight portion of fibrin.

[0043] Application Examples 1-12:

[0044] Application examples of the cosmetics of the present invention; the cosmetics are specifically essence water; the cosmetics include a tea fermentation composition. In each application example, the mass percentage of the tea fermentation composition in the cosmetics is shown in Table 3.

[0045] Table 3 Content of tea fermentation composition added in each application example

[0046] Application Examples Tea ferment composition containing Addition amount of tea fermentation composition Application Example 1 Example 1 4.51％ Application Example 2 Example 2 7.05％ Application Example 3 Example 3 8.1％ Application Example 4 Example 4 9.2％ Application Example 5 Example 5 9.05％ Application Example 6 Example 6 10.1％ Application Example 7 Example 7 11.2％ Application Example 8 Example 8 12.05％ Application Example 9 Example 9 14.2％ Application Example 10 Example 10 13.1％ Application Example 11 Example 11 16.95％ Application Example 12 Example 12 13.95％

[0047] The formula of the cosmetic is shown in Table 4.

[0048] Table 4 Cosmetic formula

[0049]

[0050]

[0051] The preparation method of the cosmetic comprises the following steps:

[0052] (1) Add part of the water to the emulsifier and homogenize at 800 rpm. Slowly add carbomer and then add ammonium acryloyldimethyltaurate / VP copolymer. Homogenize at 2200 rpm for 3-5 minutes. Then, stir at 1200 rpm until completely dissolved. Then add polyethylene glycol-8, allantoin, sodium hyaluronate, and cetearyl olivate. Heat to 80-85°C, stir at 1200 rpm, and homogenize at 2200 rpm for 3-5 minutes to completely dissolve and disperse the material evenly.

[0053] (2) Add pre-dispersed polydimethylsiloxane and polydimethylsiloxane alcohol, homogenize at 2200 rpm for 3-5 minutes; stir at 1200-2200 rpm for 3 minutes, keep warm and stir for 10-15 minutes (observe the oil dispersion and increase the homogenization and stirring time appropriately), and cool down;

[0054] (3) When the temperature drops to 60°C, add pre-dispersed butanediol and p-hydroxyacetophenone;

[0055] (4) Cooling to 45°C, adding pre-dispersed glycerin, phenoxyethanol, 1,3-propylene glycol, tea fermentation composition and remaining water, stirring at 1000-2000 rpm to completely stir the material uniformly;

[0056] (5) Sending the product for inspection at the central control; after passing the inspection, the temperature is lowered to 38°C and the product is discharged through a 200-mesh filter to obtain the cosmetics;

[0057] (6) After discharging, the material is allowed to stand and the semi-finished product is inspected;

[0058] (7) After passing the inspection, filling and packaging;

[0059] (8) Inspection of finished products.

[0060] Application Comparative Example 1:

[0061] An application comparison example of the cosmetics of the present invention; the cosmetics are specifically an essence water; the difference between this application comparison example and application example 10 is only that: the tea ferment composition of Example 10 contained in application example 10 is replaced with an equal amount of the tea ferment composition of comparison example 1, and the remaining components are the same as application example 10.

[0062] The preparation method of the cosmetic is the same as that in the application example.

[0063] Application Comparative Example 2:

[0064] An application comparison example of the cosmetics of the present invention; the cosmetics are specifically an essence water; the difference between this application comparison example and application example 10 is only that: the tea ferment composition of Example 10 contained in application example 10 is replaced with an equal amount of the tea ferment composition of comparison example 2, and the remaining components are the same as application example 10.

[0065] The preparation method of the cosmetic is the same as that of the application example.

[0066] Application Comparative Example 3:

[0067] An application comparison example of the cosmetics of the present invention; the cosmetics are specifically an essence water; the difference between this application comparison example and application example 10 is only that: the tea ferment composition of Example 10 contained in application example 10 is replaced with an equal amount of the tea ferment composition of comparison example 3, and the remaining components are the same as application example 10.

[0068] The preparation method of the cosmetic is the same as that in the application example.

[0069] Application Comparative Example 4:

[0070] An application comparison example of the cosmetics of the present invention; the cosmetics are specifically an essence water; the difference between this application comparison example and application example 10 is that: 13.1% of the tea ferment composition of Example 10 contained in application example 10 is replaced with 26.6% of the tea ferment composition of comparison example 4, and the remaining components are the same as application example 10.

[0071] The preparation method of the cosmetic is the same as that of the application example.

[0072] Application Comparative Example 5:

[0073] An application comparison example of the cosmetics described in the present invention; the cosmetics are specifically an essence water; the difference between this application comparison example and application example 10 is that the tea ferment composition of Example 10 contained in application example 10 is replaced with an equal amount of the tea ferment composition of comparison example 5, and the remaining components are the same as application example 10.

[0074] The preparation method of the cosmetic is the same as that in the application example.

[0075] Application Comparative Example 6:

[0076] An application comparison example of the cosmetics described in the present invention; the cosmetics are specifically an essence water; the difference between this application comparison example and application example 10 is that the tea ferment composition of Example 10 contained in application example 10 is replaced with an equal amount of the tea ferment composition of comparison example 6, and the remaining components are the same as application example 10.

[0077] The preparation method of the cosmetic is the same as that of the application example.

[0078] Application Comparative Example 7:

[0079] An application comparison example of the cosmetics described in the present invention; the cosmetics are specifically an essence water; the difference between this application comparison example and application example 10 is that: this application comparison example does not add the tea fermentation composition, but replaces it with an equal amount of water, and the remaining components are the same as application example 10.

[0080] The preparation method of the cosmetic is the same as that of Application Example 10.

[0081] Test Example 1: Penetration Test

[0082] Test samples: cosmetics (essence water) prepared in Application Examples 1-12 and Comparative Examples 1-7.

[0083] Test principle: Using a porcine skin model, a fluorescently labeled sample is administered and incubated for 8 hours. Frozen sections are then taken and photographed using an intelligent live cell imaging analyzer to evaluate the penetration effect. Principle: Fluorescein isothiocyanate (FITC) has the unique ability to permanently label biological molecules and can be used to detect or track the interaction between conjugates and other biological molecules. The isothiocyanate of FITC can react with the primary amine groups of proteins or antibodies, and small molecule drugs with NH groups can be fluorescently labeled with FITC. When irradiated with ultraviolet light or blue-violet light, FITC is stimulated to an excited state, and when it returns to its basic state from the excited state, it emits fluorescence. Protein fluorescent labeling technology utilizes the covalent binding of fluorescent materials to the groups of target molecules, and uses its fluorescent properties to provide information about the research object.

[0084] 1. Testing instruments and equipment: 1 / 10,000 balance, magnetic stirrer, pipette, water bath, refrigerator, ultrasonic analyzer, Franz diffusion cell transdermal absorption instrument, cryostat, intelligent living cell imaging analyzer.

[0085] 2. Raw materials: pig skin (Jingde Agricultural Products Sales Co., Ltd.).

[0086] 3. Detection method:

[0087] (3.1) Fix the skin: According to the Franz diffusion cell transdermal absorption test method ( Figure 3) Fix the skin between the supply chamber and the receiving chamber of the Franz diffusion cell, with the skin stratum corneum facing the supply chamber and the dermis layer facing the receiving chamber. Add an appropriate amount of physiological saline to the receiving chamber. After fixing the skin, remove bubbles to ensure that the skin dermis layer is in close contact with the receiving solution without bubbles.

[0088] (3.2) Drug administration: Use a dust-free cloth to absorb the physiological saline in the supply reservoir, add 0.5 mL of the experimental sample to the supply reservoir, seal the upper end of the supply reservoir with a parafilm, cover with an opaque bottle cover, and set the temperature and speed required for the experimental conditions.

[0089] (3.3) Sampling: At specific time points, the test skin was removed from the supply reservoir and washed with PBS to remove the test sample remaining in the stratum corneum. The skin was fixed in 4% paraformaldehyde, cleaned with PBS, and then vertically embedded in OCT. Full-thickness sections were sliced ​​using a freezing microtome, and tissue imaging was performed to observe the penetration of the sample's active ingredients in the skin.

[0090] 4. The fluorescence intensity test results are shown in Table 5.

[0091] Table 5

[0092] Test samples Fluorescence intensity value Application Example 1 2277.3 Application Example 2 3726.4 Application Example 3 3815.1 Application Example 4 3857.3 Application Example 5 4867.2 Application Example 6 4843.2 Application Example 7 4899.1 Application Example 8 5211.5 Application Example 9 5302.4 Application Example 10 5530.7 Application Example 11 5412.3 Application Example 12 5403.5 Comparative Application Example 1 1002.4 Application Comparative Example 2 1872.3 Application Comparative Example 3 1854.7 Comparative Application Example 4 1967.2 Comparative Application Example 5 1905.5 Application Comparative Example 6 1989.6 Application Comparative Example 7 354.3

[0093] The penetration enhancement experiment verified that the essence water added with the tea fermentation composition prepared in the application examples of the present invention all had a good penetration enhancement effect. Among them, the penetration enhancement effect of application example 10 was the best, indicating that the increase in the content of black tea fermentation filtrate helps it to synergize with other components and improve the penetration enhancement effect of the tea fermentation composition. Test Example 2: Antioxidant Test (DPPH Method)

[0094] Test samples: cosmetics (essence water) prepared in Application Examples 1-12 and Comparative Examples 1-7.

[0095] Test Principle: This test is based on the fact that the DPPH free radical has a single electron, exhibits strong absorption at 517nm, and exhibits a purple color in alcohol solutions. In the presence of a free radical scavenger, it pairs with the single electron in the DPPH free radical, causing its absorption to gradually disappear. The degree of color fading is quantitatively related to the number of electrons accepted, allowing for rapid quantitative analysis using a spectrophotometer.

[0096] 1. Reagents and materials:

[0097] (1.1) Reagents:

[0098] 0.2 mM DPPH solution (1,1-diphenyl-2-trinitrophenylhydrazine, C 18 H 12N5O6): Weigh 0.007875 g of DPPH powder and dissolve it in 100 mL of anhydrous ethanol to prepare a 0.2 mM DPPH solution.

[0099] Vitamin C standard control solution (positive control group): Dilute VC with deionized water to 0.001 mg / mL, 0.002 mg / mL, 0.004 mg / mL, 0.006 mg / mL, 0.008 mg / mL, 0.01 mg / mL, 0.02 mg / mL and 0.5 mg / mL in sequence and place in a brown bottle for later use.

[0100] (1.2) Instruments and equipment: analytical balance (accurate to 0.001 g), ultrasonic cleaning machine, UV-visible spectrophotometer.

[0101] 2. Test steps:

[0102] (2.1) Test four sample tubes and add samples in sequence according to Table 6.

[0103] Table 6

[0104] Reagents T-sample tube <![CDATA[T0 - Sample blank tube]]> C—Control tube <![CDATA[C0—Control blank]]> Test samples 2mL 2mL / / Anhydrous ethanol / 2mL 2mL 4mL DPPH solution 2mL / 2mL /

[0105] Positive control group: set up vitamin C standard control tubes, and add 4.5 mL of DPPH solution and 0.5 mL of vitamin C standard control solution to each tube.

[0106] (2.2) After mixing the solutions in each tube, allow them to react in the dark at room temperature for 30 minutes. Measure the absorbance at a wavelength of 517 nm using distilled water, adjusting the wavelength to zero, and record the data. (In data processing, ensure that the absolute difference between two independent measurements under repeatable conditions does not exceed 10% of the arithmetic mean.) Calculate the clearance (P) according to formula (1); the test results are shown in Table 7.

[0107]

[0108] In the above formula, T is the absorbance of the sample tube, i.e., the absorbance of the solution after the test sample reacts with DPPH; T0 is the background absorbance of the test sample; C is the absorbance of the DPPH tube, i.e., the absorbance of the DPPH solution when no test sample is added; C0 is the background absorbance of the solvent (anhydrous ethanol).

[0109] Table 7

[0110] Test samples DPPH clearance rate (%) Application Example 1 24.37±2.64 Application Example 2 33.88±2.36 Application Example 3 33.45±1.24 Application Example 4 33.32±0.73 Application Example 5 34.26±1.28 Application Example 6 33.56±2.34 Application Example 7 31.12±1.24 Application Example 8 40.29±3.35 Application Example 9 40.11±3.68 Application Example 10 43.21±1.27 Application Example 11 42.54±1.28 Application Example 12 34.92±2.35 Comparative Application Example 1 13.45±5.25 Application Comparative Example 2 21.36±4.57 Application Comparative Example 3 23.24±0.64 Comparative Application Example 4 21.76±3.56 Comparative Application Example 5 22.76±2.11 Application Comparative Example 6 22.34±1.02 Application Comparative Example 7 3.74±2.43

[0111] Antioxidant experiments confirmed that the tea fermentation composition-added essence waters prepared in the application examples of the present invention all exhibited excellent free radical scavenging and antioxidant effects. Application Example 10 achieved the highest antioxidant effect, demonstrating that the increased content of black tea fermentation filtrate contributes to synergistic effects with other components, enhancing the antioxidant efficacy of the tea fermentation composition.

[0112] Test Example 3: Elastase Test

[0113] Principle: Collagen forms collagen fibers and reticular fibers, while elastin forms elastic fibers. Together, these three fibers comprise the connective tissue of the skin, contributing to its plump and elastic structure. Elastase has the ability to break down multiple proteins, including collagen and elastin. Changes in elastin structure are closely associated with skin aging. A microplate reader is used to measure the inhibitory effect of the test sample on the catalytic activity of elastase on its substrate to determine whether the test sample has the ability to inhibit elastase.

[0114] 1. Reagents and materials: 30 U / mg elastase (porcine pancreas), N-succinyl-alanine-alanine-alanine-p-nitroaniline, tris(hydroxymethyl)aminomethane, concentrated hydrochloric acid, and EGCG.

[0115] 2. Instruments and equipment: analytical balance (accurate to 0.001g), ultrasonic cleaning machine, pipette, pipette, colorimetric tube, volumetric flask (10mL, 100mL).

[0116] 3. Determination steps:

[0117] (3.1) Solution preparation:

[0118] (3.1.1) Preparation of 0.2 mol / L Tris-HCl buffer at pH 8.0:

[0119] Solution A: Accurately weigh 4.8456 g of Tris particles, add 100 mL of deionized water, and stir to dissolve to obtain a 0.4 mol / L Tris solution.

[0120] Solution B: Take 3.34 mL of concentrated hydrochloric acid and dilute it with 96.66 mL of deionized water to obtain a 0.4 mol / L HCl solution.

[0121] Take 100 mL of solution A and 44.76 mL of solution B, mix them, adjust the pH to 8.0, and add deionized water to make up to 200 mL.

[0122] (3.1.2) Elastase solution: Prepare elastase solution using pH 8.0 Tris-HCl buffer. Take 0.033 g of elastase and add 10 mL of pH 8.0 Tris-HCl buffer to make a 100 U / mL elastase solution. Prepare immediately before use.

[0123] (3.1.3) Substrate solution: Weigh 1.8057 mg of N-succinyl-alanine-alanine-alanine-p-nitroaniline and dissolve it in 10 mL of pH 8.0 Tris-HCl buffer.

[0124] (3.1.4) Positive control EGCG solution: Weigh 0.001 g of EGCG powder and dissolve it in 1 mL of pH 8.0 Tris-HCl buffer to prepare a 1.0 mg / mL solution. Then dilute it with buffer to prepare solutions of 0.5, 0.1, 0.01, 0.005, 0.003, 0.002, and 0.001 mg / mL.

[0125] Sample solution: cosmetics (essence water) prepared in Examples 1-12 and Comparative Examples 1-8.

[0126] 5. Prepare the test solution:

[0127] Table 8

[0128]

[0129] Each solution was added to a 96-well plate in sequence according to Table 8, incubated at room temperature for 60 min, and the absorbance was measured at 410 nm using a microplate reader. The data were recorded and saved, and the elastase inhibition rate (%) was calculated according to formula (2).

[0130] Elastase inhibition rate (%) = [1-(OD A -OD B ) / (OD C -OD D )]×100%——Formula (2)

[0131] In the above formula, OD A is the absorbance value of the sample group (containing enzyme and substrate);

[0132] OD B is the absorbance value of the sample control group (containing enzyme but not substrate);

[0133] OD C is the absorbance value of the blank group (containing enzyme, substrate and no sample);

[0134] OD D is the absorbance value of the blank control group (containing enzyme but no substrate or sample).

[0135] 6. Precision: The absolute difference between two independent measurement results obtained under repeatability conditions shall not exceed 10% of the arithmetic mean.

[0136] Table 9

[0137]

[0138]

[0139] The test results are shown in Table 9. Compared to the comparative examples, the tea fermentation compositions prepared in the present examples showed significant inhibition of elastase activity, indicating excellent anti-wrinkle efficacy. Among them, the tea fermentation composition prepared in Example 10 showed the best anti-wrinkle effect. The increased content of black tea fermentation filtrate contributes to its synergistic effect with other components, enhancing the anti-wrinkle efficacy of the tea fermentation composition.

[0140] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A tea fermentation composition, characterized in that The invention comprises the following components in parts by weight: 3-12 parts of black tea fermentation filtrate and 1.51-4.95 parts of double protein; the double protein comprises whey protein and recombinant collagen; the weight ratio of the whey protein and the recombinant collagen is whey protein: recombinant collagen = (1.5-4.5): (0.01-0.45); the black tea fermentation filtrate is a fermentation filtrate product of yeast and Acetobacter xylinum fermentation.

2. The tea fermentation composition according to claim 1, wherein The invention comprises the following components in parts by weight: 7-12 parts of black tea fermentation filtrate, 2-4.5 parts of whey protein and 0.05-0.45 parts of recombinant collagen.

3. The tea fermentation composition according to claim 2, wherein The invention comprises the following components in parts by weight: 10 parts of black tea fermentation filtrate, 3 parts of whey protein and 0.1 parts of recombinant collagen.

4. Use of the tea fermentation composition according to any one of claims 1 to 3 in cosmetics.

5. A cosmetic, characterized in that: The invention comprises the tea fermentation composition as described in any one of claims 1 to 3.

6. The cosmetic according to claim 5, wherein The added amount of the tea fermentation composition in cosmetics is 4.51%-16.95%.

7. The cosmetic according to claim 6, wherein The addition amount of the tea fermentation composition in cosmetics is 13.1%.

8. The cosmetic according to claim 5, wherein The dosage forms of the cosmetics include water, lotion and cream.

9. The cosmetic according to claim 5, wherein The cosmetics also include auxiliary materials acceptable in daily chemical products.

Images