Red pomegranate tea fermentation composition capable of resisting oxidation, relieving and resisting inflammation and application of red pomegranate tea fermentation composition

Abstract

The invention relates to a red pomegranate tea fermentation composition capable of resisting oxidation, relieving and resisting inflammation and application, and belongs to the technical field of cosmetics. The pomegranate tea fermentation composition is prepared from the following components in parts by weight: 3 to 15 parts of black tea fermentation filtrate, 1 to 10 parts of pomegranate extract and 0.01 to 0.5 part of recombinant collagen. The pomegranate tea fermentation composition has the following effects: firstly, the safety of a product is improved, the product tends to a skin weak acid environment, and the pH value of the recombinant collagen is controlled between 6.5 and 7.5 due to the fact that the recombinant collagen contains various amino acid residues which have different functional groups; even if the pomegranate is slightly acidic, the recombinant collagen region is neutral, the skin is in a weakly acidic environment after the three are combined, and the allergy risk is reduced; secondly, the anti-oxidation effect is enhanced, and the damage of free radicals to the skin can be more effectively resisted; and thirdly, the composition has a soothing effect and can relieve discomfort of the skin, so that the skin is more comfortable.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and particularly to a pomegranate tea ferment composition with antioxidant, soothing and anti-inflammatory effects and its application. Background Art

[0002] Black tea ferment contains a large amount of tea polyphenols, which are powerful antioxidants that can effectively scavenge free radicals in the body and protect the skin from oxidative damage. Moreover, during the fermentation process of black tea, substances such as theaflavins and thearubigins are produced, which have various biological activities such as antioxidant and antibacterial effects, and also have a certain protective effect on the skin. The tea leaves of black tea have excellent quality and contain rich components such as tea polyphenols, theaflavins and thearubigins, which are ideal raw materials for making black tea ferment. Coupled with the unique fermentation process, Keemun black tea has stronger biological activities in aspects such as antioxidant, antibacterial and antiviral, and is very suitable for making black tea ferment. Currently, during the application of black tea in cosmetics, it will undergo high-temperature treatments such as heating and sterilization. If the control is improper or too high, the antioxidant substances in the black tea fermentation filtrate will undergo thermal decomposition or denaturation, resulting in a decline in its antioxidant performance and affecting its skin care efficacy in cosmetics.

[0003] Pomegranate is rich in various nutrients such as vitamin C, vitamin E, folic acid and potassium. Among them, vitamin C and vitamin E can effectively fight free radicals and protect cells from oxidative damage. In addition, pomegranate also contains rich polyphenolic compounds such as tannic acid and anthocyanins, which also have powerful antioxidant effects. However, pomegranate contains various organic acids such as punicic acid and citric acid. The physiological environment of normal skin cells is weakly acidic (pH between 4.5 and 6.5). When these organic acids are applied to cosmetics and act on the skin, they will change the acidity of the skin surface, reduce the pH value of the skin surface and tend to be acidic, which is likely to cause discomfort to the skin.

[0004] Recombinant collagen refers to collagen that is recombinantly transformed from the human collagen gene using synthetic biology technology and, after high-density fermentation, achieves 100% consistency with the amino acid sequence of human collagen. In overcoming the drawbacks of animal-derived collagen, it is not only less likely to be rejected, but is also more easily absorbed by the body and is safe. Recombinant collagen achieves anti-aging by directly supplementing collagen, saving the loss of collagen and effectively reducing wrinkles and fine lines. Although recombinant collagen is similar to human collagen in structure and function, some impurities will be introduced or its structure will be changed during the production process, thus leading to the generation of immunogenicity. If recombinant collagen is recognized as a foreign substance by the human immune system, it is likely to trigger an immune response, resulting in adverse reactions such as allergies. Summary of the Invention

[0005] The object of the present invention is to overcome the deficiencies of the prior art and provide a pomegranate tea fermentation composition with the advantages of antioxidant, soothing and anti-inflammatory effects and its application.

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

[0007] In the first aspect, the present invention provides a pomegranate tea fermentation composition, comprising the following components in parts by weight: 3-15 parts of black tea fermentation filtrate, 1-10 parts of pomegranate extract, and 0.01-0.5 parts of recombinant collagen.

[0008] By selecting specific component ratios, the obtained pomegranate tea fermentation composition of the present invention has the advantages of synergistic antioxidant, soothing and anti-inflammatory effects. In addition, recombinant collagen contains a variety of amino acid residues, and these amino acid residues carry different functional groups. For example, amino groups and carboxyl groups can regulate the PH through the processes of protonation and deprotonation, and the PH of recombinant collagen is controlled between 6.5 and 7.5; even though pomegranates are acidic, but the recombinant collagen is neutral, and the skin returns to a weakly acidic environment after the three are combined, reducing the risk of allergies.

[0009] As a preferred embodiment of the pomegranate tea fermentation composition of the present invention, the pomegranate tea fermentation composition comprises the following components in parts by weight: 10 parts of black tea fermentation filtrate, 5 parts of pomegranate extract, and 0.1 part of recombinant collagen. Under the preferred ratio, the pomegranate tea fermentation composition of the present invention has better technical effects of antioxidant, soothing and anti-inflammatory effects.

[0010] Furthermore, the black tea fermentation filtrate of the present invention is a fermentation product filtrate of fermenting black tea with yeast and / or acetic acid bacteria, and the INCI name is yeast / acetic acid bacteria / black tea fermentation product.

[0011] The black tea used in the present invention is Keemun black tea. The tea polyphenols in Keemun black tea and rooibos tea extract are detected by the method of GB / T 8313-2018 "Method for the determination of tea polyphenols and catechins in tea". The detection results show that the tea polyphenol content of Keemun black tea is 1109 ppm, while the tea polyphenol content of rooibos tea extract is 76 ppm, proving that Keemun black tea has stronger antioxidant ability.

[0012] Furthermore, the pomegranate extract of the present invention is an extract of the fermentation product of Lactobacillus / Pomegranate (PUNICA GRANATUM) fruit.

[0013] The effect of using lactobacillus to ferment pomegranate fruit is better, because its pomegranate fruit is rich in simple sugars such as glucose and fructose, which can provide a fast and available energy source for the growth and fermentation of lactobacillus, and can make lactobacillus reproduce and produce acid quickly, which is conducive to shortening the fermentation cycle; while pomegranate roots contain more polysaccharides such as lignin and cellulose, which have complex structures and are difficult for lactobacillus to directly use, and cannot provide fast and effective energy and nutritional support for the fermentation of lactobacillus. Not only that, the use of lactobacillus for fermentation has a strong acid-producing ability, which can quickly reduce the pH value during the fermentation of pomegranate fruit, create an acidic environment, inhibit the growth of harmful microorganisms, and ensure the safety and stability of the fermentation process.

[0014] The dansyl chloride method was used to test that the cell renewal rate of 5% (w / v) lactobacillus pomegranate fruit fermentation product was 26%, which was equivalent to the cell renewal rate of 4% (w / v) glycolic acid.

[0015] In a second aspect, the present invention provides the use of the pomegranate tea ferment composition in the preparation of antioxidant, soothing and / or anti-inflammatory products.

[0016] In a third aspect, the present invention provides a cosmetic, characterized in that the cosmetic contains the pomegranate tea ferment composition.

[0017] Furthermore, the cosmetic contains 4.01 to 25.5 parts of the pomegranate tea ferment composition and 74.5 to 95.99 parts of cosmetic accessories.

[0018] As a preferred embodiment of the pomegranate tea fermentation composition of the present invention, the cosmetic contains 15.1 parts of the copper peptide tea extract composition and 84.9 parts of cosmetic accessories.

[0019] Furthermore, the cosmetic auxiliary material includes at least one of an emollient, a dispersant, a humectant, a preservative, an emulsion stabilizer, a soothing agent, a thickener, a pH adjuster and a solvent.

[0020] Further, the moisturizer includes glycerin; the chelating agent includes disodium EDTA; the soothing agent includes allantoin; the preservative includes parahydroxyacetophenone and / or phenoxyethanol; the emollient includes at least one of polydimethylsiloxane, propylene glycol dicaprylate / dicaprate, ethylhexyl palmitate, C10-18 fatty acid triglycerides, C13-14 isoparaffin and avocado butter; the pH adjuster includes aminomethyl propanol; the thickener includes carbomer and / or polyacrylamide; the solvent includes water; the emulsifier includes at least one of glyceryl stearate PEG-100 stearate, cetearyl alcohol, C14-22 alcohol C12-20 alkyl glucoside and laureth-7.

[0021] As a more preferred embodiment of the pomegranate tea fermentation composition of the present invention, the cosmetic adjuvants include the following components in parts by weight: 72.44 parts of water, 5 parts of glycerol, 0.05 part of disodium EDTA, 0.15 part of carbomer, 1 part of propylene glycol dicaprylate / dicaprate, 3 parts of ethylhexyl palmitate, 0.1 part of allantoin, 1 part of C10-18 triglycerides, 1.8 parts of cetearyl alcohol, 1 part of glyceryl stearate PEG-100 stearate, 1.2 parts of C14-22 alcohol C12-20 alkyl glucoside, 1 part of shea butter, 1 part of polydimethylsiloxane, 0.5 part of p-hydroxyacetophenone, 0.05 part of polyacrylamide, 0.5 part of phenoxyethanol, 0.03 part of C13-14 isoparaffin, 0.02 part of laureth-7, and 0.06 part of aminomethyl propanol.

[0022] Furthermore, the cosmetic includes at least one of cleansing products, lotions, creams, and facial masks.

[0023] Fourthly, the present invention provides a method for preparing the cosmetic, which is characterized by comprising the following steps:

[0024] S1: Homogenize the raw materials in phase B at 2500 rpm for 4 to 10 minutes and stir at 1200 rpm for 2 to 5 minutes to completely dissolve and uniformly disperse the components of the raw materials;

[0025] S2: Mix the raw materials in phase A with the raw materials in phase B treated in step S1, keep stirring at 800 to 1000 rpm, and homogenize at 1200 rpm for 1 to 3 minutes; adjust the temperature to 80 to 85 °C and stir until dissolved uniformly;

[0026] S3: Mix the raw materials in phase C with the product treated in step S2, homogenize at 2500 to 2800 rpm for 2 to 5 minutes and stir at 1200 rpm for 2 to 5 minutes to completely dissolve and uniformly stir; pay attention to observing the appearance after the raw materials are completely dissolved and stirred uniformly. After there is no oil flower, adjust the temperature to 50 to 55 °C;

[0027] S4: Mix the raw materials in phase D with the product treated in step S3, stir at 800 to 1400 rpm for 4 to 10 minutes to completely dissolve and uniformly stir, add the raw materials in phase E, and stir until completely dissolved and uniformly stirred; adjust the temperature to 45 °C, add the pomegranate tea fermentation composition, and stir evenly; adjust the temperature to 38 °C and filter to obtain the cosmetic;

[0028] The raw materials in phase A include at least one of glycerol, disodium EDTA, carbomer, water, and allantoin;

[0029] The raw material of Phase B includes at least one of dicaprylyl carbonate, ethylhexyl palmitate, triglycerides of C10-18 fatty acids, cetearyl alcohol, glyceryl stearate PEG-100 stearate, C14-22 alcohol C12-20 alkyl glucoside, shea butter, and dimethicone;

[0030] The raw material of Phase C includes at least one of water, polyacrylamide, C13-14 isoparaffin, and laureth-7;

[0031] The raw material of Phase D includes glycerol and / or hydroxyacetophenone;

[0032] The raw material of Phase E includes aminomethylpropanol and / or water.

[0033] Furthermore, first mix the raw materials in Phase A, Phase C, Phase D, and / or Phase E, disperse them evenly, and then carry out the preparation.

[0034] Furthermore, in step S4, filter through 190-210 mesh, preferably 200 mesh.

[0035] Furthermore, step S4 also includes inspecting, filling, and packaging the filtered cosmetics to obtain the final commercially available cosmetic product.

[0036] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0037] Black tea, as a natural ingredient, is non-toxic, non-irritating, and non-sensitizing to the skin and has antioxidant effects; the vitamins in pomegranate also have strong antioxidant effects; recombinant collagen has a strong anti-wrinkle effect; when the present invention combines black tea fermentation filtrate, pomegranate extract, and recombinant collagen, the following effects are achieved: First, the safety of the combined product is improved, and the product tends to be in a weakly acidic environment for the skin. Since recombinant collagen contains various amino acid residues, these amino acid residues carry different functional groups, such as amino and carboxyl groups, which can regulate the PH through the processes of protonation and deprotonation. The PH of recombinant collagen is controlled between 6.5 and 7.5. Even though pomegranate is slightly acidic, recombinant collagen is between neutral. After the three are combined, the skin returns to a weakly acidic environment, reducing the risk of allergy; Second, the antioxidant effect is enhanced, and it can more effectively combat the damage of free radicals to the skin; Third, the compounded soothing effect can relieve the discomfort of the skin and make the skin more comfortable. This combination brings new possibilities for the application of cosmetics and is expected to bring a better skin care experience to consumers. Description of the Drawings

[0038] Figure 1 It is a cream prepared from the pomegranate recombinant collagen tea fermentation composition of the embodiment of the present invention. Detailed Embodiments

[0039] To better illustrate the objectives, technical solutions, 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 sources unless otherwise specified.

[0040] The black tea fermentation filtrate, with the INCI name of Saccharomyces / Xylacetic Acid Bacteria / Black Tea Fermentation Product, was purchased from Guangzhou Xiyuan Biotechnology Co., Ltd.

[0041] The pomegranate extract, namely Lactobacillus / Pomegranate (PUNICA GRANATUM) Fruit Fermentation Product Extract, was purchased from Huzhou Zhenlu Bioproducts Co., Ltd.

[0042] The recombinant collagen was purchased from Xi'an Giant Biogene Technology Co., Ltd.

[0043] The preparation method of the orange fruit extract is as follows:

[0044] (1) Pretreatment: Crush the orange pulp to obtain pulp.

[0045] (2) Enzymatic hydrolysis: Add pectinase and cellulase to the pulp. The mass of pectinase and cellulase is the same, and the total mass of pectinase and cellulase is 0.1% - 0.3% (w / w) of the pulp. Hydrolyze at 40°C for 1 - 2 h to break the cell wall and improve the release of active ingredients, obtaining the enzymolysis product.

[0046] (3) Extraction: Add 50% - 70% (v / v) ethanol aqueous solution to the enzymolysis product in step (2). The ratio of the enzymolysis product to the ethanol aqueous solution is 1 g:(3 - 5) mL. Stir and extract at 50 - 60°C for 1 - 2 h to obtain the crude orange fruit extract.

[0047] (4) Filtration: Filter the crude orange fruit extract with a gauze or filter to remove the residue, obtaining the orange fruit filtrate.

[0048] (5) Concentration: Transfer the orange fruit filtrate to a rotary evaporator and concentrate it under reduced pressure at 40°C - 50°C to remove ethanol, obtaining the orange fruit extract.

[0049] The preparation method of the rooibos tea extract is as follows:

[0050] (1) Pretreatment: Remove the impurities from the rooibos tea leaves and crush them to obtain tea powder.

[0051] (2) Extraction: Put the tea powder into an extraction container and add 60% - 80% (v / v) ethanol aqueous solution. The ratio of the tea powder to the ethanol aqueous solution is 1 g:(10 - 15) mL. Stir and extract at 50°C - 70°C for 2 - 3 times, 1 - 2 h each time, obtaining the tea extract.

[0052] (3) Filtration: Filter the tea extract with filter paper or a filter screen to remove tea residues and obtain a tea filtrate.

[0053] (4) Concentration: Transfer the tea filtrate to a rotary evaporator and concentrate it at 40°C - 50°C under reduced pressure to remove ethanol and obtain a concentrated tea solution.

[0054] (5) Drying: Dry the concentrated tea solution into powder by spray drying or freeze drying to obtain rooibos tea extract, and collect and store it.

[0055] Examples 1 - 11

[0056] The composition components of the red pomegranate recombinant collagen tea fermentation composition in Examples 1 - 11 are shown in Table 1 by weight parts.

[0057] Table 1

[0058]

[0059] Comparative Examples 1 - 12

[0060] The composition components of the red pomegranate recombinant collagen tea fermentation composition in Comparative Examples 1 - 12 are shown in Table 2 by weight parts.

[0061] Table 2

[0062]

[0063]

[0064] Application Examples 1 - 11

[0065] Prepare a cosmetic (cream) according to the red pomegranate recombinant collagen tea fermentation composition in Examples 1 - 11 and the application, as Figure 1 shown.

[0066] I. Cosmetic formula:

[0067] The cosmetic consists of 5% glycerol, 0.05% disodium EDTA, 0.15% carbomer, 1% propylene glycol dicaprylate / dicaprate, 3% ethylhexyl palmitate, 0.1% allantoin, 1% triglycerides of C10 - 18 fatty acids, 1.8% cetearyl alcohol, 1% glyceryl stearate PEG - 100 stearate, 1.2% C14 - 22 alcohol C12 - 20 alkyl glucoside, 1% shea (BUTYROSPERMUM PARKII) fruit butter, 1% polydimethylsiloxane, 0.5% p - hydroxyacetophenone, 0.05% polyacrylamide, 0.5% phenoxyethanol, 0.03% C13 - 14 isoparaffin, 0.02% laureth - 7, 0.06% aminomethylpropanol, water and the ectoine tea fermentation composition, with water making up the balance, and the above are weight percentages.

[0068] Among the cosmetics of Application Examples 1 to 11, the weight percentages of the black tea fermentation product filtrate, pomegranate extract, and recombinant collagen in the red pomegranate recombinant collagen tea fermentation composition are shown in Table 3.

[0069] Table 3

[0070]

[0071] II. Cosmetic preparation method:

[0072] 1. Pretreatment

[0073] (1) Pretreat the raw materials of Phase A: Disperse glycerol, disodium EDTA, carbomer, water, and allantoin evenly and set aside.

[0074] (2) Pretreat the raw materials of Phase C: Disperse water, polyacrylamide, C13-14 isoparaffin, and laureth-7 evenly and set aside.

[0075] (3) Pretreat the raw materials of Phase D: Disperse glycerol and p-hydroxyacetophenone evenly and set aside.

[0076] (4) Pretreat the raw materials of Phase E: Disperse aminomethylpropanol and water in advance and set aside.

[0077] 2. Preparation

[0078] (1) Add dipropylene glycol dicaprylate / dicaprate, ethylhexyl palmitate, C10-18 triglyceride fatty acids, cetearyl alcohol, glyceryl stearate PEG-100 stearate, C14-22 alcohol C12-20 alkyl glucoside, shea butter (BUTYROSPERMUM PARKII) fruit butter, and polydimethylsiloxane in the raw materials of Phase B to the emulsifying pan in sequence, homogenize at 2500 rpm for 4 - 10 min, stir at 1200 rpm for 2 - 5 min to completely dissolve and disperse all components of the raw materials evenly.

[0079] (2) Mix the pretreated raw materials of Phase A with the raw materials of Phase B processed in step (1), keep stirring at 800 - 1000 rpm, homogenize at 1200 rpm for 1 - 3 min; heat up to 80 - 85 °C and stir until dissolved evenly.

[0080] (3) Mix the pretreated raw materials of Phase C with the raw materials processed in step (2), homogenize at 2500 - 2800 rpm for 2 - 5 min, stir at 1200 rpm for 2 - 5 min, stir until completely dissolved evenly, pay attention to observing the appearance after the raw materials are completely dissolved evenly, and after there is no oil flower, cool down to 50 - 55 °C.

[0081] (4) Mix the pretreated raw material of Phase D with the raw material processed in step (3), stir at 800 - 1400 rpm for 4 - 10 min until completely and evenly dissolved, add the pretreated raw material of Phase E, and stir until completely and evenly dissolved; cool down to 45°C, then add the pomegranate recombinant collagen tea fermentation composition to make all components of the raw materials completely and evenly stirred.

[0082] (5) Send the product processed in step (4) for in-process inspection; after passing the inspection, cool the temperature to 38°C and filter through a 200-mesh sieve for discharging.

[0083] (6) Let the discharged product of step (5) stand still for semi-finished product inspection.

[0084] (7) After the semi-finished product inspection in step (6) is qualified, carry out filling and packaging to obtain the finished cosmetic product.

[0085] (8) Inspect the finished cosmetic product.

[0086] Apply Comparative Examples 1 - 12

[0087] In the cosmetics applying Comparative Examples 1 - 12, the weight percentages of the filtrate of black tea fermentation product, pomegranate extract and recombinant collagen in the pomegranate recombinant collagen tea fermentation composition, as well as the weight percentages of orange fruit extract and rooibos tea extract are shown in Table 4. Other formulations and preparation methods are the same as those in Examples 1 - 12.

[0088] Among them, in Applied Comparative Example 11, the orange fruit extract contains various proanthocyanidin substances, which can effectively scavenge various free radicals, protect skin cells from oxidative stress damage, and thus slow down skin aging. However, the maximum addition amount of the orange fruit extract in the specified leave-on products is 2.95%, so its addition amount of 2% is selected to replace the pomegranate extract as a comparative example.

[0089] Since the filtrate of black tea fermentation product of the present invention is prepared from Keemun black tea as the raw material, in Applied Comparative Example 12, the rooibos tea extract is used to replace the filtrate of black tea fermentation product as a comparative example.

[0090] Table 4

[0091]

[0092] Test Example 1: Determination of Physicochemical Indexes

[0093] 1. Test samples: Creams prepared from the pomegranate recombinant collagen tea fermentation compositions of Examples 1 - 11 and Comparative Examples 1 - 12.

[0094] 2. Test the pH value with a pH tester; measure the conductivity color with a conductivity tester; pour the sample into a transparent sample bottle to observe the color of the composition, and then measure the refractive index with a refractometer.

[0095] 3. The test results are shown in Table 5. The results indicate that the physical and chemical indexes and appearance indexes of the cream prepared from the pomegranate recombinant collagen tea fermentation composition prepared by the present invention all meet the standards in actual production.

[0096] Table 5

[0097]

[0098]

[0099] Test Example 2: Safety patch test

[0100] 1. Test sample: The cream prepared from the pomegranate recombinant collagen tea fermentation composition in Application Example 9.

[0101] 2. Subjects: A total of 30 people, 7 males and 23 females, aged 18 to 60 years old, with an average age of 48.27 ± 10.01 years old, meeting the volunteer selection criteria for subjects.

[0102] 3. The patch test method is as follows:

[0103] Select qualified patch test equipment. Using the closed patch test method, place the patch-type product containing the test sample (cut into the size of the patch tester, about 50 mm 2 ) in the patch tester, and apply it to the back of the subject with a low-allergy tape. After 24 hours, remove the test sample, and observe the skin reaction at 0.5, 24, and 48 hours after removing the test sample. At the same time, set a negative control (blank, without treatment). Record the results according to the skin reaction grading standard of the skin closed patch test in Chapter 7 of the "Technical Specifications for Cosmetics Safety" (2015 Edition).

[0104] 4. The results of the safety patch test are shown in Table 6. The results of the human skin patch test show that among the 30 subjects, 0 cases had skin adverse reactions, indicating that the cream prepared from the pomegranate recombinant collagen tea fermentation composition of the present invention has high safety.

[0105] Table 6

[0106]

[0107] Test Example 3: Antioxidant test (DPPH method)

[0108] 1. Test samples: The creams prepared in Application Examples 1 to 11 and Application Comparative Examples 1 to 11.

[0109] 2. Test principle: Based on the property that DPPH free radicals have a single electron, have a strong absorption at 517 nm, and their alcoholic solution is purple. When there is a free radical scavenger, due to the pairing of its single electron, its absorption gradually disappears, and the degree of fading has a quantitative relationship with the number of electrons it accepts. Therefore, rapid quantitative analysis can be carried out using a spectrophotometer.

[0110] 3. Reagents and materials:

[0111] (1) Reagents: 0.2 mM DPPH solution (1,1-diphenyl-2-trinitrophenylhydrazine, C 18 H 12 N5O6): Weigh 0.007875 g of DPPH powder and dissolve it in 100 mL of absolute ethanol to prepare 0.2 mM DPPH; VC solution (positive control group): Dilute VC successively with deionized water to VC solutions of 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, and place them in brown bottles for later use.

[0112] (2) Instruments and equipment: Analytical balance (accurate to 0.001 g), ultrasonic cleaner, ultraviolet-visible spectrophotometer.

[0113] 4. Test procedures:

[0114] Divide into four sample tubes and add samples successively according to Table 7.

[0115] Positive control group: Set up a vitamin C standard control tube, and add 4.5 mL of DPPH and 0.5 mL of vitamin C solution to each tube.

[0116] After mixing the solutions in each tube, react them in the dark at room temperature for 30 min. Zero the spectrophotometer with distilled water at a wavelength of 517 nm, measure the absorbance, and record the data (note that in data processing, the absolute difference between two independent measurement results obtained under repeatability conditions shall not exceed 10% of the arithmetic mean).

[0117] Calculate the scavenging rate (P) according to Equation (1):

[0118]

[0119] In Equation (1), T: absorbance value of the sample tube, that is, the absorbance value of the solution after the test sample reacts with DPPH; T0: absorbance value of the test sample background; C: absorbance value of the DPPH tube, that is, the absorbance value of the DPPH solution without adding the test sample; C0: absorbance value of the solvent (absolute ethanol) background.

[0120] Table 7

[0121] Reagent T - Sample tube <![CDATA[T0 - Sample blank tube]]> C - Control tube <![CDATA[C0—Control blank]]> Test sample (mL) 2 2 / / Absolute ethanol (mL) / 2 2 4 DPPH solution (mL) 2 / 2 /

[0122] 5. The test results are shown in Table 8. Verified by the antioxidant experiment, the pomegranate recombinant collagen tea fermentation composition prepared in the examples of the present invention all has good effects of scavenging free radicals and antioxidant. Among them, the pomegranate recombinant collagen tea fermentation composition prepared in Example 9 has the best antioxidant effect. The increase in the content of the black tea fermentation filtrate helps its synergistic effect with other components and improves the antioxidant efficacy of the pomegranate recombinant collagen tea fermentation composition. After replacing the pomegranate extract with the orange fruit extract in Application Comparative Example 11, the DPPH scavenging rate is only 17.54±1.25%, and after replacing the black tea fermentation product filtrate with the rooibos tea extract in Application Comparative Example 12, the DPPH scavenging rate is only 17.29±2.07%, which is lower than that of Application Example 9 and lower than that of Application Comparative Example 1 or 2 without adding the black tea fermentation product filtrate or the pomegranate extract.

[0123] Table 8

[0124]

[0125]

[0126] Test Example 4: Hyaluronidase Test

[0127] 1. Experimental principle: Hyaluronidase is an endogenous hexosaminidase that causes the decomposition of large-molecular hyaluronic acid. Under the action of hyaluronidase, large-molecular hyaluronic acid decomposes to produce N-acetylglucosamine. Hyaluronidase can hydrolyze the 1,4-glycosidic bond between β-N-acetylglucosamine and D-glucuronic acid in hyaluronic acid to obtain β-N-acetylglucosamine. β-N-acetylglucosamine can condense with acetylacetone to form a chromogen, 2-methyl-3-acetylpyrrole derivative, under alkaline conditions. The chromogen reacts with dimethylaminobenzaldehyde in concentrated hydrochloric acid-ethanol to produce color. Therefore, measuring the content of N-acetylglucosamine in the reaction system can indirectly reflect the activity of hyaluronidase. Hyaluronidase has a strong correlation with inflammation, so inhibiting the activity of hyaluronidase is used as an index for studying anti-inflammatory effects.

[0128] 2. Solution preparation

[0129] (1) Acetic acid-sodium acetate buffer solution (pH = 5.6): Measure 1.155 mL of glacial acetic acid and dilute it to 100 mL and mix well to prepare acetic acid solution; Weigh 2.72 g of sodium acetate, dissolve it in water and make up the volume to 100 mL to prepare sodium acetate solution; Add 4.8 mL of acetic acid solution to 45.2 mL of sodium acetate solution, mix well and make up the volume to 100 mL to prepare acetic acid-sodium acetate buffer solution (pH = 5.6).

[0130] (2) Sodium hyaluronate solution: Weigh 0.0050 g of sodium hyaluronate and dissolve it in 10 mL of acetate-sodium acetate buffer solution (pH = 5.6) to prepare a sodium hyaluronate solution with a mass concentration of 0.5 mg / mL.

[0131] (3) Ehrlich's reagent: Weigh 0.8 g of p-dimethylaminobenzaldehyde and dissolve it in 15 mL of concentrated hydrochloric acid and 15 mL of absolute ethanol, and store it in the dark.

[0132] (4) CaCl₂ solution: Weigh 2.8 g of anhydrous CaCl₂ and dissolve it in 10 mL of deionized water, and ultrasonicate until completely dissolved.

[0133] (5) NaOH solution: Weigh 0.16 g of NaOH and dissolve it in 10 mL of deionized water, and ultrasonicate until completely dissolved.

[0134] (6) Hyaluronidase solution: Weigh 0.0100 g of hyaluronidase and dissolve it in 4 mL of acetate-sodium acetate buffer solution to prepare a hyaluronidase solution with a working concentration of 1250 U / mL, which is prepared before use.

[0135] (7) Acetylacetone solution: Measure 3.5 mL of acetylacetone and dissolve it in 50 mL of sodium carbonate solution (1.0 mol / L) to prepare an acetylacetone solution, which is prepared before use.

[0136] 3. Sample addition

[0137] Add samples sequentially according to Table 9 below:

[0138] Table 9

[0139]

[0140]

[0141] 4. Data analysis

[0142] (1) The inhibition rate calculation formula is shown in Equation (2):

[0143]

[0144] In Equation (2):

[0145] A: Absorbance value of the control solution (using acetate-sodium acetate buffer solution instead of the sample solution);

[0146] B: Absorbance value of the blank control solution (using acetate-sodium acetate buffer solution instead of the sample solution and enzyme solution);

[0147] C: Absorbance value of the test sample solution;

[0148] D: Absorbance value of the sample blank solution (acetic acid-sodium acetate buffer instead of enzyme solution).

[0149] 5. Data processing, and the test results are shown in Table 10. The red pomegranate recombinant collagen tea fermentation composition has significant hyaluronidase inhibitory activity, that is, it has good soothing efficacy. The optimal weight ratio is 10 parts of tea fermentation filtrate, 5 parts of pomegranate extract, and 0.1 part of recombinant collagen. After replacing the pomegranate extract with orange fruit extract in Application Comparative Example 11, the hyaluronidase inhibition rate is only 10.33%, which is lower than that of Application Example 9 and lower than that of Application Comparative Example 2 without adding pomegranate extract; after replacing the tea fermentation product filtrate with rooibos tea extract in Application Comparative Example 12, the hyaluronidase inhibition rate is only 10.74%, which is lower than that of Application Example 9 and is basically the same as that of Application Comparative Example 1 without adding tea fermentation product filtrate.

[0150] Table 10

[0151]

[0152]

[0153] 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. A pomegranate tea fermentation composition, characterized in that: The invention comprises the following components in parts by weight: 3 to 15 parts of black tea fermentation filtrate, 1 to 10 parts of pomegranate extract and 0.01 to 0.5 parts of recombinant collagen.

2. The pomegranate tea fermentation composition according to claim 1, characterized in that The invention comprises the following components in parts by weight: 10 parts of black tea fermentation filtrate, 5 parts of pomegranate extract and 0.1 parts of recombinant collagen.

3. Use of the pomegranate tea ferment composition according to claim 1 or 2 in the preparation of antioxidant, soothing and / or anti-inflammatory products.

4. A cosmetic, characterized in that: The cosmetic contains the pomegranate tea ferment composition according to claim 1 or 2.

5. The cosmetic according to claim 4, characterized in that: The cosmetic contains 4.01 to 25.5 parts of the pomegranate tea ferment composition and 74.5 to 95.99 parts of cosmetic auxiliary materials.

6. The cosmetic according to claim 5, characterized in that The cosmetic contains 15.1 parts of the pomegranate tea ferment composition and 84.9 parts of cosmetic accessories.

7. The cosmetic according to claim 5, characterized in that The cosmetic auxiliary material includes at least one of an emollient, a dispersant, a humectant, a preservative, an emulsion stabilizer, a soothing agent, a thickener, a pH regulator and a solvent.

8. The cosmetic according to any one of claims 7, characterized in that: The moisturizer includes glycerin; the chelating agent includes disodium EDTA; the soothing agent includes allantoin; the preservative includes parahydroxyacetophenone and / or phenoxyethanol; the emollient includes at least one of polydimethylsiloxane, propylene glycol dicaprylate / dicaprate, ethylhexyl palmitate, C10-18 fatty acid triglycerides, C13-14 isoparaffin and shea butter; the pH adjuster includes aminomethyl propanol; the thickener includes carbomer and / or polyacrylamide; the solvent includes water; the emulsion stabilizer includes at least one of glyceryl stearate PEG-100 stearate, cetearyl alcohol, C14-22 alcohol C12-20 alkyl glucoside and laureth-7.

9. The cosmetic according to claim 4, characterized in that: The cosmetics include at least one of cleansing type, water type, cream type and mask type.

10. The method for preparing a cosmetic according to any one of claims 4 to 9, characterized in that: The following steps are involved: S1: Homogenize the raw material of phase B at 2500rpm for 4-10min and stir at 1200rpm for 2-5min to completely dissolve the components of the raw material and disperse them evenly; S2: Mix the raw material of phase A with the raw material of phase B processed in step S1, keep stirring at 800-1000 rpm, and homogenize at 1200 rpm for 1-3 min; adjust the temperature to 80-85°C, and stir to dissolve evenly; S3: Mix the raw material of phase C with the product treated in step S2, homogenize at 2500-2800 rpm for 2-5 min, stir at 1200 rpm for 2-5 min, stir until the mixture is evenly dissolved, observe the appearance of the raw material after it is evenly dissolved, and adjust the temperature to 50-55° C. after there is no oil droplets; S4: Mix the raw material of phase D with the product treated in step S3, stir at 800-1400 rpm for 4-10 min, stir and dissolve evenly, add the raw material of phase E, stir and dissolve evenly; adjust the temperature to 45° C., add the pomegranate tea ferment composition of claim 1 or 2, stir evenly; adjust the temperature to 38° C., filter, and obtain the cosmetic; The phase A raw material includes at least one of glycerin, disodium EDTA, carbomer, water and allantoin; The phase B raw materials include at least one of propylene glycol dicaprylate / dicaprate, ethylhexyl palmitate, C10-18 fatty acid triglycerides, cetearyl alcohol, glyceryl stearate PEG-100 stearate, C14-22 alcohol C12-20 alkyl glucoside, shea butter and dimethicone; The C phase raw material includes at least one of water, polyacrylamide, C13-14 isoparaffin and laureth-7; The phase D raw material includes glycerol and / or p-hydroxyacetophenone; The E phase raw material includes aminomethyl propanol and / or water.

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