Tea fermentation composition for promoting dual protein permeation and application

Abstract

The invention relates to the field of cosmetics, in particular to a tea fermentation composition for promoting dual protein permeation and application. The tea fermentation composition comprises black tea fermentation filtrate, whey protein and recombinant collagen. The black tea fermentation filtrate used in the invention contains more inositol and micromolecular biological permeation-promoting polypeptide for promoting skin permeation, and can effectively improve the antioxidant, anti-aging and repairing capabilities. After the black tea fermentation filtrate is compounded with the whey protein and the recombinant collagen, the problem that double proteins are difficult to permeate can be solved in an assisting manner, so that the utilization rate of the double proteins in cosmetics is increased, the double proteins can play the maximum effect, and synergistic interaction is realized. When the obtained tea fermentation composition is added into cosmetics, the effects of resisting oxidation, repairing, resisting wrinkles and the like can be greatly improved.

Description

Technical Field

[0001] The invention relates to the field of cosmetics, and in particular to a tea fermentation composition for promoting double protein penetration and 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 the collagen that is 100% consistent with the amino acid sequence of human collagen after the recombinant transformation of human collagen gene by synthetic biology technology and high-density fermentation. It overcomes the disadvantages of animal-derived collagen, is not easily rejected, is more easily absorbed by the body, and is safe. Recombinant collagen achieves anti-aging by directly supplementing collagen, saves collagen loss, and effectively resists wrinkles and lines. However, recombinant collagen also has the same problem as whey protein. Because the 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 for skin care, and have powerful repair, anti-wrinkle, firming and other multiple functions, but because of their large molecular weight, they are difficult to absorb when added to skin care products, and their transdermal absorption is poor, and they cannot exert their maximum effect. There are currently two ways in the industry to solve this problem: the first is to develop collagen with a smaller molecular weight, but this technology has a long development cycle and high uncertainty; the second is to achieve skin penetration and absorption through penetration-promoting 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 a class of short peptides with less than 30 amino acid residues, such as biological penetration-promoting polypeptides, which can promote polypeptides, 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. At present, all three 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 prior art and provide a tea fermentation composition, which can make the double protein easier to penetrate and absorb, 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, non-sensitizing to the skin, has a rapid onset, is regenerative and has a predictable duration. It prevents the loss of body components while promoting penetration. When applied to the skin, the barrier function of the skin can be quickly and completely restored. The black tea fermentation filtrate used in the present invention is a fermentation filtrate product of double yeast (yeast, acetobacter xylinum), which contains rich substances such as inositol and small molecule biological penetration-promoting polypeptides that promote skin penetration; the molecular range of the black tea fermentation filtrate with a molecular weight of less than 1000 Daltons (especially small molecules less than 500 Daltons) accounts for 66.65%, which helps to promote the penetration of synergistic ingredients. Whey protein is extracted from milk and can supplement the nutrients needed by people. Recombinant collagen is obtained by biosynthesis and can directly supplement collagen to achieve anti-aging effects. The present invention has found through research that the compounding of whey protein and recombinant collagen has a synergistically enhanced anti-wrinkle effect, and after the black tea fermentation filtrate is compounded with the whey protein and the recombinant collagen, the black tea fermentation filtrate can be used as a penetration enhancer to help solve the problem of the difficulty in penetration of the double proteins, thereby improving the utilization rate of the double proteins in cosmetics, allowing 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 specific range, it has the best antioxidant, repair and anti-wrinkle effects.

[0014] In a second aspect, the present invention provides application 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 ferment composition of the present invention is added to cosmetics to effectively improve the anti-oxidation, anti-aging, repair and other effects on the skin, and is more conducive to the comprehensiveness and effectiveness of the skin effect.

[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 also include auxiliary materials 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 adjuster.

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

[0023] As a natural ingredient, tea is non-toxic, non-irritating and non-sensitizing to the skin, has a rapid onset of action, is regenerative in activity and has a predictable duration, prevents the loss of components in the body while promoting penetration, and can be applied to the skin to quickly and completely restore the skin's barrier function. 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 penetration of the double protein, thereby improving the utilization rate of the double protein in cosmetics, allowing it to exert its maximum efficacy and achieve synergistic enhancement.

[0024] The tea ferment composition of the present invention is added to cosmetics to effectively improve the anti-oxidation, anti-aging, repair and other effects on the skin, and is more conducive to the comprehensiveness and effectiveness of the skin effect. 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 is 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 as follows Figure 1-2 As shown, Example 1 and Example 10 are taken as examples.

[0033] Embodiment 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 embodiment

[0036]

[0037] Comparative Examples 1-6:

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

[0039] Table 2 Components by weight of the tea fermentation composition 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; the missing components are replaced by equal weight portions 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 Embodiment 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, 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 olive oil ester, 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 the pre-dispersed glycerin, phenoxyethanol, 1,3-propylene glycol, tea fermentation composition and the 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 the product passes the inspection, the temperature is lowered to 38° C. and the product is discharged through a 200-mesh screen to obtain the cosmetic product;

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

[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 described in 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 fermentation composition of example 10 contained in application example 10 is replaced with an equal amount of the tea fermentation 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 of the application example.

[0063] Application Comparative Example 2:

[0064] 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 only that: the tea fermentation composition of example 10 contained in application example 10 is replaced with an equal amount of the tea fermentation 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 described in 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 fermentation composition of example 10 contained in application example 10 is replaced with an equal amount of the tea fermentation 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 of the application example.

[0069] Application Comparative Example 4:

[0070] 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: 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 of 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 other 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 pig skin model, the fluorescently labeled sample is administered, incubated for 8 hours, and frozen sections and intelligent live cell imaging analyzer are taken for characterization to evaluate its penetration effect. Principle: Fluorescein isothiocyanate (FITC) has a unique detection performance of permanently labeling 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 group 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 the basic state from the excited state, it emits fluorescence. Protein fluorescent labeling technology uses 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 instrument, Franz diffusion cell transdermal absorption instrument, cryostat, intelligent living cell imaging analyzer.

[0085] 2. Raw materials and auxiliary 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 stratum corneum facing the supply chamber and the dermis facing the receiving chamber, add an appropriate amount of physiological saline into the receiving chamber, and after fixing the skin, remove the bubbles to ensure that the dermis layer of the skin 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 pool, add 0.5 mL of the experimental sample to the supply pool, seal the upper end of the supply pool with a sealing film, cover it with an opaque bottle cover, and set the temperature and speed required by the experimental conditions.

[0089] (3.3) Sampling: At a specific time point, the tested 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, washed with PBS, and then vertically embedded in OCT. Full-thickness sections were cut using a freezing microtome and tissue imaging was performed to observe the penetration of the active ingredients in the sample 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 Application Comparative Example 1 1002.4 Application Comparative Example 2 1872.3 Application Comparative Example 3 1854.7 Application Comparative Example 4 1967.2 Application Comparative Example 5 1905.5 Application Comparative Example 6 1989.6 Application Comparative Example 7 354.3

[0093] The penetration-promoting 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-promoting effect. Among them, the penetration-promoting 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-promoting 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: The test is based on the characteristics of DPPH free radicals, which have a single electron, strong absorption at 517nm, and purple alcohol solution. When there is a free radical scavenger, it pairs with the single electron in the DPPH free radical, causing its absorption to gradually disappear. The degree of fading is quantitatively related to the number of electrons it accepts, so a spectrophotometer can be used for rapid quantitative analysis.

[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 4.5mL DPPH solution and 0.5mL vitamin C standard control solution into each tube.

[0106] (2.2) After mixing the solutions in each tube, react them at room temperature in the dark for 30 minutes, adjust the wavelength to zero with distilled water at 517nm, measure the absorbance with a spectrophotometer, and record the data (in data processing, it should be noted that the absolute difference between two independent measurement results obtained under repeatability conditions should not exceed 10% of the arithmetic mean). Calculate the clearance rate (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 Application Comparative Example 1 13.45±5.25 Application Comparative Example 2 21.36±4.57 Application Comparative Example 3 23.24±0.64 Application Comparative Example 4 21.76±3.56 Application Comparative Example 5 22.76±2.11 Application Comparative Example 6 22.34±1.02 Application Comparative Example 7 3.74±2.43

[0111] The antioxidant experiment verified that the essence water added with the tea fermentation composition prepared in the application examples of the present invention has good free radical scavenging and antioxidant effects. Among them, the antioxidant effect of application example 10 is the best, indicating that the increase in the content of black tea fermentation filtrate helps it to synergize with other components and improve the antioxidant effect of the tea fermentation composition.

[0112] Test Example 3: Elastase Test

[0113] Experimental principle: Collagen forms collagen fibers and reticular fibers, and elastin forms elastic fibers. These three fibers together form the connective tissue of the skin, which makes the skin plump and elastic. Elastase has the ability to decompose collagen, elastin and other proteins. The change in elastin structure is closely related to skin aging. The inhibitory effect of the test sample on the catalysis of elastase and its substrate is detected by an ELISA instrument 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, 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, stir and dissolve to obtain a 0.4 mol / L Tris solution;

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

[0121] Take 100 mL of liquid A and 44.76 mL of liquid 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 with pH 8.0 Tris-HCL buffer. Take 0.033 g of elastase and add 10 mL of pH 8.0 Tris-HCL buffer to prepare a 100 U / mL elastase solution. Prepare it 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, which was then diluted with the 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] According to Table 8, each solution was added to a 96-well plate in sequence, incubated at room temperature for 60 min, and the absorbance was detected at 410 nm using an ELISA reader. The data was 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 It 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 with the comparative example, the tea fermentation composition prepared in the embodiment of the present invention has significant activity of inhibiting elastase, that is, has good anti-wrinkle effect. Among them, the tea fermentation composition prepared in Example 10 has the best anti-wrinkle effect. The increase in the content of black tea fermentation filtrate helps it to synergize with other components and improve the anti-wrinkle effect of the tea fermentation composition.

[0140] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution 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.

2. The tea fermentation composition according to claim 1, characterized in that 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).

3. The tea fermentation composition according to claim 1, characterized in that 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.

4. The tea fermentation composition according to claim 3, characterized in that 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.

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

6. A cosmetic, characterized in that: The invention comprises the tea fermentation composition as described in any one of claims 1 to 4.

7. The cosmetic according to claim 6, characterized in that The tea fermentation composition is added in an amount of 4.51% to 16.95% in cosmetics.

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

9. The cosmetic according to claim 6, characterized in that The dosage forms of the cosmetics include water, milk and cream.

10. The cosmetic according to claim 6, characterized in that: The cosmetics also include auxiliary materials acceptable in daily chemical products.

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