Ectoin tea fermentation composition with anti-aging and repairing functions and application of Ectoin tea fermentation composition

Abstract

The invention relates to an Ectoin tea fermentation composition with anti-aging and repairing functions and application, and belongs to the technical field of cosmetics. The Ectoin tea fermentation composition disclosed by the invention is prepared from the following components in parts by weight: 1 to 10 parts of saccharomycetes / acetobacter xylinum / black tea fermentation products, 0.5 to 2 parts of Ectoin and 0.05 to 0.15 part of carnosine. The Ectoin tea fermentation composition is high in ultraviolet protection capacity, good in skin deep cell repairing effect, comprehensive in function and safe, has the efficient anti-aging and efficient repairing effects after being scientifically proportioned, has the synergistic effect, meets the standard in physical and chemical indexes and appearance indexes, is good in temperature change stability and has the good application prospect. And long-term preservation and use are facilitated.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and in particular to an ectoin tea fermentation composition with anti-photoaging repair and its application. Background Art

[0002] The fermentation product of yeast / acetic acid bacteria / black tea is rich in antioxidant components such as tea polyphenols. Under ultraviolet irradiation, the skin will produce a large number of free radicals, which are one of the important factors leading to skin aging. They will attack various biomolecules in skin cells, such as DNA, proteins, and lipids. The antioxidant component tea polyphenols in the fermentation product of yeast / acetic acid bacteria / black tea can scavenge free radicals before they cause damage to the skin. Ultraviolet rays are the main cause of photoaging, especially UVA and UVB, which can penetrate the skin surface, damage collagen and elastic fibers, and cause skin wrinkles, relaxation, and pigmentation problems. Although the fermentation product of yeast / acetic acid bacteria / black tea has beneficial effects on the skin such as antioxidant effects, its ability to protect against ultraviolet rays is limited.

[0003] Ectoin is a component with strong protective ability. In terms of anti-photoaging repair, it can protect skin cells from environmental stresses such as ultraviolet rays, high temperature, and dryness. However, ectoin mainly forms a protective effect on the skin surface, and there is still room for improvement in the cell repair and anti-photoaging effects in the deep layer of the skin.

[0004] Carnosine is mainly used for anti-glycation. During photoaging, the glycation reaction in the skin will accelerate. Advanced glycation end products (AGEs) will make collagen and elastic fibers hard and brittle, lose elasticity, and cause skin relaxation and wrinkles. Carnosine can prevent sugars from binding to proteins, reduce the formation of AGEs, and thus effectively avoid the occurrence of skin relaxation and wrinkles. Although carnosine performs well in anti-glycation, it is difficult for carnosine alone to comprehensively address all aspects of photoaging. For example, for the oxidative damage of skin cells caused by already generated free radicals, or skin inflammation caused by ultraviolet rays, carnosine has no direct repair or defense effect, and it needs to be used in combination with other components with antioxidant, anti-inflammatory, and other effects to provide more comprehensive anti-photoaging protection. Summary of the Invention

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and provide an ectoin tea fermentation composition with anti-photoaging repair and its application.

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

[0007] In a first aspect, the present invention provides an ectoine tea fermentation composition. By weight, the ectoine tea fermentation composition contains 1-10 parts of yeast / acetic acid bacteria / black tea fermentation product, 0.5-2 parts of ectoine, and 0.05-0.15 parts of carnosine.

[0008] Yeast / acetic acid bacteria / black tea fermentation product, ectoine, and carnosine each have their own advantages. Yeast / acetic acid bacteria / black tea fermentation product has antioxidant active ingredients such as phenolic compounds. Ectoine can protect skin cells from various environmental stress damages, reduce water loss in cells, maintain the stability of the intracellular environment, and thus indirectly play an antioxidant role by reducing the rate of free radical generation; carnosine can prevent glycosylation reactions. After the three are proportioned, they have the efficacy of high-efficiency anti-photoaging and high-efficiency repair, with a more comprehensive anti-photoaging effect and a synergistic effect, bringing new possibilities for the anti-photoaging application of cosmetics.

[0009] Preferably, by weight, the ectoine tea fermentation composition contains 10 parts of yeast / acetic acid bacteria / black tea fermentation product, 1.5 parts of ectoine, and 0.15 parts of carnosine. The ectoine tea fermentation composition with the above-mentioned ratio has the best antioxidant, anti-photoaging, and skin repair effects.

[0010] Further, the yeast / acetic acid bacteria / black tea fermentation product is a fermentation product filtrate obtained by fermenting black tea with yeast and / or acetic acid bacteria.

[0011] In a second aspect, the present invention provides the application of the above-mentioned ectoine tea fermentation composition in the preparation of anti-aging products and / or skin damage repair products.

[0012] Further, the anti-aging includes at least one of antioxidant, anti-photoaging, and anti-glycation.

[0013] Further, the products include cosmetics.

[0014] Further, the cosmetics include at least one of cleansing cosmetics, lotion cosmetics, cream cosmetics, and facial mask cosmetics.

[0015] In a third aspect, the present invention provides a cosmetic, which contains the above-mentioned ectoine tea fermentation composition and cosmetic auxiliaries.

[0016] Further, by weight, the cosmetic contains 1.55-12.15 parts of ectoine tea fermentation composition and 87.85-98.45 parts of cosmetic auxiliaries.

[0017] Preferably, by weight, the cosmetic contains 11.65 parts of ectoine tea fermentation composition and 88.35 parts of cosmetic auxiliaries.

[0018] Further, the cosmetic adjuvant includes at least one of an emollient, a dispersant, a humectant, a preservative, an emulsifier, a thickener, a soother, a pH regulator, and a solvent.

[0019] Further, the emollient includes polydimethylsiloxane and / or polydimethylsiloxanol; the thickener includes acrylate / C10-30 alkyl acrylate cross-linked polymer and / or sodium acrylate / sodium acryloyldimethyltaurate copolymer; the humectant includes at least one of glycerin, sodium hyaluronate, butanediol, and 1,3-propanediol; the soother includes allantoin; the emulsifier includes cetearyl olivate and / or sorbitan olivate; the preservative includes p-hydroxyacetophenone and / or phenoxyethanol; the pH regulator includes aminomethylpropanol; the solvent includes water.

[0020] In a specific embodiment of the invention, by weight, the cosmetic adjuvant contains 0.60 part of polydimethylsiloxane, 0.40 part of polydimethylsiloxanol, 0.15 part of acrylate / C10-30 alkyl acrylate cross-linked polymer, 4.10 parts of glycerin, 0.10 part of allantoin, 0.20 part of sodium hyaluronate, 0.20 part of cetearyl olivate, 0.10 part of sorbitan olivate, 0.15 part of sodium acrylate / sodium acryloyldimethyltaurate copolymer, 3.00 parts of butanediol, 0.40 part of p-hydroxyacetophenone, 0.10 part of phenoxyethanol, 0.40 part of 1,3-propanediol, 0.10 part of aminomethylpropanol, and 78.35 parts of water.

[0021] Fourthly, the present invention provides a preparation method of the cosmetic, which is characterized by comprising the following steps:

[0022] S1: Stir the raw materials of phase B evenly at 800-1200 rpm, homogenize at 2000-2500 rpm for 1-3 min, adjust the temperature to 80-85 °C, and keep warm for 10-15 min until the raw materials are completely dissolved and there is no gel-like substance;

[0023] S2: Stir the raw materials of phase A evenly at 800-1200 rpm, adjust the temperature to 80-85 °C, homogenize at 800-1200 rpm for 5-8 min, and stir until completely dissolved;

[0024] S3: Adjust the temperature of the phase B raw materials treated in step S1 to 50-55 °C, slowly add the phase C raw materials, stir evenly at 800-1200 rpm, add the phase A raw materials treated in step S2, homogenize at 800-1200 rpm for 5-8 min, and stir at 800-1200 rpm until all the raw materials are completely dissolved;

[0025] S4: Adjust the temperature of the raw materials processed in step S3 to 40 - 45°C, add the ectoine tea fermentation composition as described in claim 1 or 2, stir at 800 - 1200 rpm for 3 - 8 min until evenly mixed, add an appropriate amount of water, and continue to stir at a speed of 800 - 1200 rpm until all raw materials are completely emulsified and there are no oil flowers on the surface;

[0026] S5: Adjust the stirring speed in step S4 to 600 - 1000 rpm, stir evenly, adjust the temperature to 35 - 40°C, and filter to obtain the cosmetic;

[0027] The raw materials of phase A include polydimethylsiloxane and / or polydimethylsiloxanol;

[0028] The raw materials of phase B include at least one of water, acrylate / C10 - 30 alkyl acrylate cross - linked polymer, glycerin, allantoin, sodium hyaluronate, cetearyl olivate, sorbitan olivate, and sodium acrylate / sodium acryloyldimethyl taurate copolymer;

[0029] The raw materials of phase C include at least one of butanediol, p - hydroxyacetophenone, 1,3 - propanediol, phenoxyethanol, aminomethylpropanol, and water.

[0030] Furthermore, the components in the raw materials of phase A, phase B, or phase C can be first mixed and stirred evenly before preparing the cosmetic.

[0031] Furthermore, in step S4, adjust the temperature of the raw materials processed in step S3 to 45°C.

[0032] Furthermore, in step S5, adjust the temperature to 38°C.

[0033] Furthermore, in step S5, filter through a 80 - 100 mesh sieve, preferably 100 mesh.

[0034] Furthermore, step S5 also includes inspecting, filling, and packaging the filtered essence water to obtain the final commercially available essence water product.

[0035] Compared with the prior art, the beneficial effects of the present invention are:

[0036] The present invention conducts research on anti-photoaging repair, and provides an ectoin tea fermentation composition that comprehensively utilizes yeast / Acetobacter xylinum / black tea fermentation products, ectoin, and carnosine, solves the deficiencies of each component in the prior art, and develops a skin care product with strong ultraviolet protection ability, good repair effect on deep skin cells, comprehensive antiglycation function, and safety. It is verified that the scientific ratio of the three has the efficacy of high-efficiency anti-photoaging and high-efficiency repair, and has a synergistic effect, bringing new possibilities for the application of cosmetics. Moreover, the physical and chemical indexes and appearance indexes of the product meet the standards, and it has good thermostability and is suitable for long-term storage and use.

[0037] Carnosine can prevent the damage of glycation to the skin internally, and the yeast / Acetobacter xylinum / black tea fermentation products play an antioxidant and anti-inflammatory role externally, jointly resisting photoaging. After ultraviolet irradiation, the anti-inflammatory effect of the yeast / Acetobacter xylinum / black tea fermentation products can reduce the skin's inflammatory response, while ectoin can help repair damaged cell structures. The synergistic effect of the three can greatly improve the effect of anti-photoaging repair. Description of the Drawings

[0038] Figure 1 It is the essence water sample containing the ectoin tea fermentation composition prepared in Application Example 10. Detailed Embodiments

[0039] To better illustrate the purpose, technical solution, and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments. Other materials, reagents, etc. used in the embodiments can be obtained from commercial channels without special instructions.

[0040] The yeast / Aceobacter xylinum / black tea fermentation product of the present invention is purchased from Bozhou Mingqi Biotechnology Co., Ltd.

[0041] The ectoin of the present invention is purchased from Kelaide (Xi'an) Medical Technology Co., Ltd.

[0042] The carnosine of the present invention is purchased from Shenzhen Vicky Technology Co., Ltd.

[0043] The green tea fermentation filtrate of the present invention is purchased from Bozhou Mingqi Biotechnology Co., Ltd.

[0044] Examples 1-12, Comparative Examples 1-7

[0045] Table 1 Formulation table of the ectoin tea fermentation composition of Examples 1-12 (parts by weight)

[0046]

[0047] Table 2 Formulation table of the ectoin tea fermentation composition of Comparative Examples 1-7 (parts by weight)

[0048]

[0049] Application Examples 1 - 12, Application Comparative Examples 1 - 7

[0050] Prepare essence water from the ectoine tea fermentation composition according to Examples 1 - 12 and Comparative Examples 1 - 7.

[0051] I. Essence water formula:

[0052] The essence water consists of 0.60% polydimethylsiloxane, 0.40% polydimethylsiloxanol, 0.15% acrylate / C10 - 30 alkanol acrylate cross - linked polymer, 4.10% glycerin, 0.10% allantoin, 0.20% sodium hyaluronate, 0.20% cetearyl olivate, 0.10% sorbitan olivate, 0.15% sodium acrylate / sodium acryloyldimethyl taurate copolymer, 3.00% butanediol, 0.40% p - hydroxyacetophenone, 0.10% phenoxyethanol, 0.40% 1,3 - propanediol, 0.10% aminomethylpropanol, water and the ectoine tea fermentation composition. The water makes up the balance, and the above are weight percentages.

[0053] Among them, in the essence water of Application Examples 1 - 12, the weight percentages of yeast / Acetobacter xylinum / black tea fermentation product, ectoine and carnosine in the ectoine tea fermentation composition are shown in Table 3; in the essence water of Application Comparative Examples 1 - 7, the weight percentages of yeast / Acetobacter xylinum / black tea fermentation product, ectoine and carnosine in the ectoine tea fermentation composition, as well as the weight percentage of green tea fermentation filtrate, are shown in Table 4. In Application Comparative Example 7, the green tea fermentation filtrate is used instead of the yeast / Acetobacter xylinum / black tea fermentation product.

[0054] Table 3 Formulation table of the ectoine tea fermentation composition in Application Examples 1 - 12 (weight percentage)

[0055]

[0056] Table 4 Formulation table of the ectoine tea fermentation composition in Application Comparative Examples 1 - 7 (weight percentage)

[0057]

[0058] II. Essence water preparation method:

[0059] 1. Pretreatment

[0060] (1) C - phase pretreatment: Add butanediol, p - hydroxyacetophenone, 1,3 - propanediol, phenoxyethanol, aminomethylpropanol and water into a container in sequence, stir evenly, and set aside. The pretreatment is the raw material of the C - phase.

[0061] (2)Pre-treatment of Phase A: Mix polydimethylsiloxane and polydimethylsiloxanol evenly and set aside for use as the raw material for Phase A after pre-treatment.

[0062] (3)Pre-treatment of Phase B: Add water, acrylate / C10-30 alkyl acrylate cross-linked polymer, glycerol, allantoin, sodium hyaluronate, cetearyl olivate, sorbitan olivate, and sodium acrylate / sodium acryloyldimethyltaurate copolymer into a container in sequence, stir evenly, set aside for use as the raw material for Phase B after pre-treatment.

[0063] 2. Preparation

[0064] (1)Treatment of Phase B: Add the pre-treated raw material of Phase B into an emulsifying pot, set the stirring speed at 800 - 1200 rpm, stir evenly, homogenize at 2000 - 2500 rpm for 1 - 3 min, heat up to 80 - 85 °C, and keep warm for 10 - 15 min until the raw materials are completely dissolved without any gel-like substances.

[0065] (2)Treatment of Phase A: Add the pre-treated raw material of Phase A into an oil-phase pot, set the stirring speed at 800 - 1200 rpm, heat up to 80 - 85 °C, homogenize at 800 - 1200 rpm for 5 - 8 min, and stir until completely dissolved.

[0066] (3)Mixing and stirring: Cool the raw material of Phase B treated in step (1) to 50 - 55 °C, slowly add the pre-treated raw material of Phase C, keep the stirring speed at 800 - 1200 rpm, after stirring evenly, add the raw material of Phase A treated in step (2), homogenize at 800 - 1200 rpm for 5 - 8 min, and stir at 800 - 1200 rpm until completely dissolved.

[0067] (4)Cooling and stirring: Continue to cool the raw material treated in step (3) to 45 °C, add ectoin tea fermentation composition, stir at 800 - 1200 rpm for 3 - 8 min, after stirring evenly, add an appropriate amount of water, and continue to stir at a speed of 800 - 1200 rpm until completely emulsified and there is no oil film on the surface.

[0068] (5)Further stirring: Adjust the stirring speed in step (4) to 600 - 1000 rpm, after stirring evenly, continue to cool down to 38 °C.

[0069] (6)Filtration and discharging: Conduct in-process inspection on the raw material treated in step (5), after passing the inspection, filter and discharge it with a 100-mesh filter bag.

[0070] (7)Standing and inspection: Let the product treated in step (6) stand and conduct semi-finished product inspection.

[0071] (8)Filling, Packaging and Final Product Inspection: After the semi-finished product in step (7) passes the inspection, filling and packaging are carried out, and finally the finished essence water is inspected.

[0072] Test Example 1: Determination of Physical and Chemical Indexes

[0073] 1. Test Samples: Three batches of essence water samples prepared in Application Example 10 ( Figure 1 ).

[0074] 2. Test Methods: The pH value is tested by a pH tester; the conductivity color is determined by a conductivity tester; the essence water sample to be tested is poured into a transparent sample bottle to observe the color of the composition, and then the refractive index is determined by a refractometer.

[0075] 3. Experimental Results: As shown in Table 5, the physical and chemical indexes and appearance indexes of the essence water prepared from the ectoine tea ferment composition of the present invention meet the standards in actual production.

[0076] Table 5

[0077]

[0078] Test Example 2: Stability Test

[0079] 1. Test Purpose: This stability test aims to comprehensively evaluate the stability performance of the essence water samples prepared in Application Example 10 under different environmental conditions and time spans, including but not limited to changes in physical properties, etc., to determine that the product can maintain its effectiveness, safety and reliability within the expected service life, and provide key data support and quality assurance basis for the research and development, production, storage, transportation and application of the product.

[0080] 2. Test Samples: Representative essence water samples prepared in Application Example 10 are selected, and the essence water samples should be prepared according to the established production process and quality standards to ensure their consistency and comparability.

[0081] 3. Test Items and Methods

[0082] (1) High-temperature Stability Test:

[0083] The essence water samples are respectively placed in an incubator at 30 °C, 40 °C and 50 °C and stored continuously for 3 months. During the storage period, the samples are taken out every 14 days to observe whether there are phenomena such as discoloration, deformation, melting, sublimation and volatilization in their appearance.

[0084] (2) Variable-temperature Stability Test

[0085] Fluctuate frequently between 0°C and 50°C (temperature-changing group 1): Within 24 hours, let the essence water sample fluctuate frequently between 0°C and 50°C. Repeat this operation for 15 to 30 days. After restoring to room temperature, observe its stability to simulate the actual usage situation of the product alternating under different temperature environments.

[0086] Store in cycles at 40°C and -5°C (temperature-changing group 2): Place the essence water sample in a constant temperature incubator pre-adjusted to 40°C ± 2°C. After storing for 24 hours, take it out and transfer it to a refrigerator pre-adjusted to -5°C ± 2°C. After storing for 24 hours, take it out. Alternate with an interval of 24 hours and repeat this cycle 5 to 7 times. After restoring to room temperature, observe and conduct tests according to the standard, and compare and evaluate the results with the initial state.

[0087] 4. Test results

[0088] As shown in Table 6, after the essence water sample prepared in Example 10 underwent high-temperature (40°C and -50°C, 3 months) and temperature-changing (0°C to 50°C cycle, 3 months) tests, it presented stable results in many aspects, showing the advantages of the formulation design of the essence water sample in terms of temperature-changing stability.

[0089] Table 6

[0090]

[0091]

[0092] Test Example 3: Antioxidant test (DPPH method)

[0093] 1. Test samples: Essence water samples prepared in Examples 1 to 12 and Comparative Examples 1 to 7.

[0094] 2. Test principle:

[0095] DPPH (1,1-diphenyl-2-picrylhydrazyl) is a stable free radical, which is dark purple in solution and has a characteristic absorption peak at 517 nm. Antioxidants can provide hydrogen atoms or electrons to pair with the unpaired electrons of DPPH free radicals, reducing them to stable DPPH-H molecules, resulting in a decrease in the concentration of DPPH free radicals in the solution and a weakening of the absorbance at 517 nm. By measuring the change in the absorbance of the solution at 517 nm before and after the reaction with a spectrophotometer, the scavenging rate is calculated using the scavenging rate formula to evaluate the antioxidant ability of the antioxidant. The degree of decrease in absorbance is proportional to the concentration and antioxidant ability of the antioxidant.

[0096] 3. Reagents and materials:

[0097] (1) Main reagents: 1,1-diphenyl-2-trinitrophenylhydrazine (DPPH, C 18 H12 NSO 6 ) and absolute ethanol.

[0098] (2) Instruments and equipment: Electronic balance (brand: Sartiorius, model: BSA-224S), UV-visible spectrophotometer (brand: HITATACHI, model: U2910).

[0099] 4. Test steps:

[0100] (1) Solution preparation: Mix DPPH and absolute ethanol to prepare a 0.2 mM DPPH ethanol solution.

[0101] (2) Add samples successively according to Table 7 below:

[0102] Table 7

[0103]

[0104]

[0105] (3) After mixing the reaction solutions in each test tube, react them in the dark at room temperature for 30 min, transfer them into 3-cm cuvettes, and measure the absorbance of each group at 517 nm, and record the data.

[0106] (4) Data analysis

[0107] The inhibition rate calculation formula is:

[0108]

[0109] In the formula:

[0110] T: Absorbance value of the sample tube, that is, the absorbance value of the solution after the sample reacts with DPPH;

[0111] T 0 : Absorbance value of the sample background;

[0112] C: Absorbance value of the DPPH tube, that is, the absorbance value of the DPPH solution without adding the sample;

[0113] C 0 : Absorbance value of the solvent background.

[0114] 5. Test results

[0115] As shown in Table 8, verified by antioxidant experiments, the essence water samples containing ectoine tea fermentation composition prepared in Application Examples 1 to 12 of the present invention all have good effects of scavenging free radicals and antioxidant. Among them, the essence water sample prepared in Application Example 10 has the best antioxidant effect. With the increase of the contents of yeast / Acetobacter xylinum / black tea fermentation products, ectoine and carnosine, the three synergistically enhance the antioxidant effect of the cosmetic sample of the ectoine tea fermentation composition with anti-photoaging repair effect. After replacing the yeast / Acetobacter xylinum / black tea fermentation products with green tea fermentation filtrate in Comparative Example 7, the DPPH scavenging rate is only 10.29±0.38%, which is lower than that of Examples 1 to 12.

[0116] Table 8

[0117]

[0118]

[0119] Test Example 4: Test on the effect of UVB radiation on the SOD content in cells

[0120] 1. Test samples: Essence water samples prepared in Application Examples 1 to 12 and Comparative Examples 1 to 7.

[0121] 2. Experimental principle: Superoxide dismutase (SOD) is a key enzyme in the cell antioxidant defense system. It mainly reduces the accumulation of reactive oxygen free radicals in cells by catalyzing the dismutation reaction of superoxide anion free radicals, protecting cells from oxidative damage. The principle is that superoxide anion free radicals are converted into hydrogen peroxide and oxygen under the action of SOD. This reaction is an important biochemical reaction in cells. The content change of SOD in cells is measured by specific detection methods (such as colorimetry, chemiluminescence method and ELISA method, etc.) to judge the effect of UVB radiation on the SOD content in cells.

[0122] 3. Reagents and materials

[0123] DMEM medium (a medium containing various amino acids and glucose), fetal bovine serum, DMSO (dimethyl sulfoxide), superoxide dismutase (SOD) activity detection kit (brand: Solarbio, product number: BC0175).

[0124] 4. Experimental steps

[0125] (1) Cell culture: Prepare a cell suspension 24 h before the experiment. Inoculate the suspension of logarithmically growing HACAT cells (human immortalized keratinocytes) into a 96-well cell culture plate, with 5000 cells per well, and culture for 24 h. Culture conditions: 37°C; 5% CO 2 ; culture under saturated humidity conditions.

[0126] (2) Exposure: Discard the culture medium in the wells of step (1), add 100 μL of the test sample to each well, and culture for 24 h. Set up a normal cell group Control(-L), an ultraviolet stress group Control(+L), a positive control group, and a test sample group respectively.

[0127] (3) Sample collection: Collect the HACAT cells processed in step (2), perform ultrasonic disruption, centrifuge to obtain the supernatant, and use it fresh or store it at -20 °C for later use.

[0128] (4) Determination: Add the corresponding reagents to the 96-well plate in the order shown in Table 9, and operate according to the instructions of the superoxide dismutase (SOD) activity detection kit. Measure the absorbance value of each well at 560 nm using an enzyme-labeling instrument.

[0129] Table 9

[0130] Reagent A - Assay Tube A - Control Tube A - Blank Tube 1 A - Blank Tube 2 Sample (μL) 20 20 / / Reagent 1 (μL) 45 45 45 45 Reagent 2 Working Solution (μL) 20 / 20 / Reagent 3 (μL) 35 35 35 35 Deionized Water 70 90 90 110 Reagent 4 Working Solution (μL) 10 10 10 10

[0131] 5. Instruments and equipment: Multifunctional enzyme-labeling instrument (brand: BioTeTek, model Cytation 1), electronic balance (brand: Sartiorius, model BSA-224S).

[0132] 6. Data processing

[0133] The calculation formula for SOD activity is:

[0134]

[0135] In the formula:

[0136] △A blank: The difference in absorbance between A-blank tube 1 and A-blank tube 2;

[0137] △A determination: The difference in absorbance between A-determination tube and A-control tube;

[0138] W: Sample mass;

[0139] F: Sample dilution factor.

[0140] 7. Test results

[0141] As shown in Table 10, after calculation, the SOD activity of cells cultured normally decreased from 403.1 U / g to 92.4 U / g after UVA and VA illumination. After co-incubation with the test samples for 24 h, as shown in Table 10, the cosmetic samples containing the ectoine tea fermentation composition prepared in the examples of the present invention all had the effect of promoting SOD activity. Among them, in the optimal combination ratio, the weight ratio of yeast / Acetobacter xylinum / black tea fermentation product, ectoine and carnosine was yeast / Acetobacter xylinum / black tea fermentation product: ectoine: carnosine = 10:1.5:0.15, and the SOD activity increased to 233.7 U / g. After replacing the yeast / Acetobacter xylinum / black tea fermentation product with green tea fermentation filtrate in Comparative Example 7, the SOD activity was only 90.28 ± 1.27 U / g, which was lower than that of Examples 1 to 12.

[0142] Table 10

[0143] Sample SOD Activity (U / g) Sample SOD Activity (U / g) Example 1 183.2±4.42 Example 11 212.4±4.63 Example 2 191.8±5.12 Example 12 196.8±5.21 Example 3 196.6±4.67 Comparative Example 1 42.6±4.15 Example 4 198.3±6.33 Comparative Example 2 30.5±4.88 Example 5 200.6±5.87 Comparative Example 3 130.7±6.11 Example 6 203.7±4.37 Comparative Example 4 101.6±4.52 Example 7 205.8±4.54 Comparative Example 5 140.92±5.42 Example 8 219.4±5.47 Comparative Example 6 35.9±5.23 Example 9 225.8±5.83 Comparative Example 7 90.28±1.27 Example 10 233.7±6.43

[0144] Test Example 5: UV Damage Repair Test

[0145] 1. Test samples: Essence water samples prepared in Examples 1 to 12, Comparative Examples 1 to 7, Control group (negative control) and Positive control group (positive control).

[0146] 2. Experimental principle: Reactive oxygen species, ultraviolet irradiation, etc. can all cause DNA double-strand breaks, which are considered the most serious DNA damage. H2A histone family member X (H2AX) is one of the variants of chromosomal histone H2A. When DNA double-strands in cells are broken, H2AX will rapidly undergo phosphorylation modification to form phosphorylated H2AX (γ-H2AX). The formation of γ-H2AX is a rapid response to DNA damage. Therefore, γ-H2AX is an important DNA damage marker and is widely used in the research of DNA damage and apoptosis. The content level of γ-H2AX can effectively reflect the DNA damage situation. The stronger the fluorescence (green) intensity, the more serious the DNA damage.

[0147] 3. Instrument equipment: Laser confocal microscope (brand: LAICA; model: SP8), electronic balance (brand: Sartiorius; model: BSA-224S).

[0148] 4. Main reagents: DNA damage detection kit (γ-H2AX immunofluorescence method, manufacturer: Shanghai Beyotime Biotechnology Co., Ltd.; product number: C2035S).

[0149] (1) Control group (negative control)

[0150] Normal cell culture reagents are usually used without adding any factors that can cause DNA damage. For example, normal cell culture media (such as DMEM medium, RPMI - 1640 medium, etc., selected according to cell types) are used, which contain basic nutrients required for cell growth, such as glucose, amino acids, vitamins, and inorganic salts, and an appropriate amount of fetal bovine serum (FBS), generally 10% (v / v), is added to provide components such as growth factors for cell growth. In this way, it serves as a basic control, representing the γ - H2AX level in the state of undamaged cells.

[0151] (2) Positive control group (positive control)

[0152] Reagents that can clearly cause DNA damage are used.

[0153] Hydrogen peroxide (H 2 O 2 ) : It is a reactive oxygen species (ROS) reagent, and reactive oxygen species can cause DNA double - strand breaks. Cells can be treated with hydrogen peroxide solution at a final concentration of 100 - 500 μmol / L for a certain period of time (such as 1 - 2 h). Because hydrogen peroxide can generate hydroxyl radicals (·OH), these radicals will attack DNA molecules, resulting in DNA damage and thus inducing the production of γ - H2AX.

[0154] Ultraviolet irradiation (UV): Ultraviolet light is a common factor inducing DNA damage. Cells are exposed to a certain dose of ultraviolet light (such as 20 - 100 J / m 2 of UV - C irradiation) to simulate the damage of ultraviolet light to cell DNA. Ultraviolet light can cause pyrimidine bases (such as thymine) in DNA molecules to form pyrimidine dimers, resulting in the distortion of the DNA double - helix structure, and further causing damage such as DNA double - strand breaks, increasing the level of γ - H2AX.

[0155] 5. Test results

[0156] As shown in Table 11, DNA damage was caused to normally cultured cells by ultraviolet irradiation, and the fluorescence intensity increased from 6.01 to 85.10; after co - incubating with the test samples for 24 h, the fluorescence intensities of the samples in Examples 1 - 12 and Comparative Examples 1 - 6 all decreased to a certain extent; among them, the fluorescence intensity in Example 10 decreased from 85.10 to 15.95, which was the lowest. Therefore, at the detected concentration, the sample in Example 10 had the most significant effect of protecting DNA from damage. The yeast / Acetobacter xylinum / black tea fermentation product, ectoine, and carnosine had a synergistic effect. When replacing the yeast / Acetobacter xylinum / black tea fermentation product with green tea fermentation filtrate in Comparative Example 7, the fluorescence intensity was as high as 60.32 ± 0.62, which was higher than that of Examples 1 - 12.

[0157] Table 11

[0158]

[0159]

[0160] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the protection scope of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. An edodein tea fermentation composition, characterized in that: By weight, the ectoine tea fermentation composition contains 1 to 10 parts of yeast / acetobacillus xylinum / black tea fermentation products, 0.5 to 2 parts of ectoine and 0.05 to 0.15 parts of carnosine.

2. The edodein tea fermentation composition according to claim 1, characterized in that: The yeast / Acetobacter xylinum / black tea fermentation product is a fermentation product filtrate obtained by fermenting black tea with yeast and / or Acetobacter xylinum.

3. The edodein tea fermentation composition according to claim 1, characterized in that: The ectoine tea fermentation composition contains 10 parts of yeast / acetobacter xylinum / black tea fermentation products, 1.5 parts of ectoine and 0.15 parts of carnosine.

4. Use of the ectoine tea ferment composition according to any one of claims 1 to 3 in the preparation of anti-aging products and / or skin damage repair products.

5. The use according to claim 4, wherein the anti-aging effect comprises at least one of anti-oxidation, anti-photoaging and anti-glycation.

6. A cosmetic, characterized in that: The cosmetics contain the ectoine tea ferment composition according to any one of claims 1 to 3 and cosmetic accessories.

7. The cosmetic according to claim 5, characterized in that: By weight, the cosmetic contains 1.55 to 12.15 parts of the ectoine tea ferment composition and 87.85 to 98.45 parts of cosmetic accessories.

8. The cosmetic according to claim 6 or 7, characterized in that: The cosmetic auxiliary material includes at least one of an emollient, a dispersant, a humectant, a preservative, an emulsion stabilizer, a thickener, a soothing agent, a pH regulator and a solvent.

9. The cosmetic according to claim 8, characterized in that: The emollient includes polydimethylsiloxane and / or polydimethylsiloxane alcohol; the thickener includes acrylates / C10-30 alkyl acrylate crosspolymer and / or sodium acrylate / sodium acryloyldimethyl taurate copolymer; the moisturizer includes at least one of glycerin, sodium hyaluronate, butylene glycol and 1,3-propylene glycol; the soothing agent includes allantoin; the emulsion stabilizer includes cetearyl olivate and / or sorbitan olivate; the preservative includes p-hydroxyacetophenone and / or phenoxyethanol; the pH adjuster includes aminomethyl propanol; and the solvent includes water.

10. The method for preparing the cosmetic according to any one of claims 6 to 9, characterized in that: The following steps are involved: S1: Stir the raw material of phase B at 800-1200 rpm, homogenize at 2000-2500 rpm for 1-3 min, adjust the temperature to 80-85°C, and keep warm for 10-15 min until the raw material is completely dissolved and there is no gel-like substance; S2: Stir the raw materials of phase A at 800-1200 rpm, adjust the temperature to 80-85°C, homogenize at 800-1200 rpm for 5-8 minutes, and stir until completely dissolved; S3: Adjust the temperature of the phase B raw material treated in step S1 to 50-55°C, slowly add the phase C raw material, stir evenly at 800-1200rpm, add the phase A raw material treated in step S2, homogenize at 800-1200rpm for 5-8min, and stir at 800-1200rpm until all raw materials are completely dissolved; S4: Adjust the temperature of the raw materials treated in step S3 to 45°C, add the edodein tea fermentation composition according to any one of claims 1 to 3, stir at 800 to 1200 rpm for 3 to 8 minutes, stir evenly, add an appropriate amount of water, and continue stirring at a speed of 800 to 1200 rpm until all the raw materials are completely emulsified and no oil spots appear on the surface; S5: adjusting the stirring speed of step S4 to 600-1000 rpm, stirring evenly, adjusting the temperature to 38° C., filtering, and obtaining the cosmetic; The phase A raw material includes polydimethylsiloxane and / or polydimethylsiloxane alcohol; The phase B raw material includes at least one of water, acrylic acid (ester) / C10-30 alkyl acrylate cross-linked polymer, glycerin, allantoin, sodium hyaluronate, cetearyl olive oil ester, sorbitan olive oil ester and sodium acrylate / sodium acryloyldimethyl taurate copolymer; The C phase raw material includes at least one of butylene glycol, p-hydroxyacetophenone, 1,3-propylene glycol, phenoxyethanol, aminomethyl propanol and water.

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