Tea-protease composition with mild exfoliating and whitening effects as well as preparation method and application of tea-protease composition

Abstract

The invention relates to a tea-protease composition with mild exfoliating and whitening effects as well as a preparation method and application thereof, and belongs to the technical field of daily chemical products. The invention provides a tea-protease composition which comprises the following components in parts by mass: 0.5-2.0 parts of black tea fermentation liquor and 0.1-6.0 parts of protease, and the protease comprises papain and lipase. The black tea fermentation liquor, the papain and the lipase are matched to form the tea-protease composition, the black tea fermentation liquor and the lipase can relieve irritation brought by papaya protein, and meanwhile, the papain, the lipase and the black tea fermentation liquor can synergistically improve the exfoliating and whitening effects of the tea-protease composition; the dual effects of mildly exfoliating and whitening are realized.

Description

Technical Field

[0001] The present invention relates to the technical field of daily chemical products, and in particular to a tea-protease composition with mild exfoliation and whitening effects, and a preparation method and application thereof. Background Art

[0002] The normal metabolism of the skin's stratum corneum is crucial to maintaining skin health. With aging and the influence of environmental factors, keratin metabolism is prone to abnormalities, leading to problems such as keratin accumulation and dull skin tone. Although traditional exfoliating products (such as chemical exfoliants such as fructose and salicylic acid or physical scrubs) are effective, they require tolerance to use and have a high operating threshold. If not used carefully, they can easily burn the skin. Long-term use may damage the skin's barrier function and cause problems such as sensitivity and stinging. At the same time, the market demand for skin care products that combine exfoliation and whitening effects with high mildness is growing.

[0003] Therefore, it is urgent to develop a composition that can gently exfoliate and make the skin white and transparent. Summary of the Invention

[0004] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a tea-protease composition with mild exfoliation and whitening effects, as well as a preparation method and application thereof.

[0005] To achieve the above object, the technical solution adopted by the present invention is:

[0006] In a first aspect, the present invention provides a tea-protease composition with mild exfoliation and whitening effects, comprising the following components in parts by weight: 0.5-2.0 parts of black tea fermentation broth, 0.1-6.0 parts of protease, wherein the protease includes papain and lipase.

[0007] Black tea fermentation liquid contains tea polyphenols, theaflavins and other antioxidant ingredients, which can effectively scavenge free radicals, reduce oxidative damage, delay skin aging, and keep the skin elastic; papain can decompose aging keratinocytes on the skin surface, making the skin smoother and more delicate, and promote skin metabolism, but if the concentration is too high or the action time is too long, it may cause irritation to the skin, causing redness, stinging, etc.; lipase can decompose excess oil on the skin surface and regulate skin oil secretion. For oily skin, it can effectively reduce oiliness and keep the skin fresh.

[0008] The present invention combines black tea fermentation liquor with papain and lipase to form a tea-protease composition. The black tea fermentation liquor and lipase can alleviate the irritation caused by papain. At the same time, papain, lipase and black tea fermentation liquor can synergistically improve the exfoliating and whitening effects of the tea-protease composition, achieving the dual effects of mild exfoliation and whitening.

[0009] As a preferred embodiment of the tea-protease composition of the present invention, the tea-protease composition comprises the following components in parts by weight: 1.0-2.0 parts of black tea fermentation broth, 1.1-5.1 parts of protease, wherein the protease comprises papain and lipase.

[0010] In a preferred embodiment of the tea-protease composition of the present invention, the mass ratio of papain to lipase in the protease is papain:lipase = (10-650):1. Experiments have demonstrated that when the mass ratio of papain to lipase is within a specific range, the tea-protease composition exhibits enhanced whitening and exfoliation effects.

[0011] In a preferred embodiment of the tea-protease composition of the present invention, the mass ratio of papain to lipase in the protease is papain:lipase = (100-500):1. Experiments have demonstrated that within a specific mass ratio of papain to lipase, the tea-protease composition exhibits enhanced whitening and exfoliation properties.

[0012] In a preferred embodiment of the tea-protease composition of the present invention, the mass ratio of papain to lipase in the protease is papain:lipase = (200-400):1. Experiments have demonstrated that within a specific mass ratio of papain to lipase, the tea-protease composition exhibits enhanced whitening and exfoliation properties.

[0013] In a preferred embodiment of the tea-protease composition of the present invention, the mass ratio of papain to lipase in the protease is 300:1. Experiments have demonstrated that within a specific mass ratio of papain to lipase, the tea-protease composition exhibits enhanced whitening and exfoliation properties.

[0014] As a preferred embodiment of the tea-protease composition of the present invention, the black tea fermentation broth is primarily prepared by fermenting a black tea aqueous extract with Lactobacillus plantarum and Lactobacillus fermentum. By incorporating the black tea fermentation broth, which exhibits excellent free radical scavenging and anti-wrinkle properties, as one of the active ingredients of the tea-protease composition, the present invention effectively enhances the whitening efficacy of the tea-protease composition.

[0015] As a preferred embodiment of the tea-protease composition of the present invention, the black tea aqueous extract is obtained by extracting black tea powder with water at 70° C. for 10 minutes, shaking once every 5 minutes, centrifuging, extracting twice, and combining the two extracts.

[0016] As a preferred embodiment of the tea-protease composition of the present invention, the inoculation amounts of Lactobacillus plantarum and Lactobacillus fermentum are 4% (v / v) respectively.

[0017] As a preferred embodiment of the tea-protease composition of the present invention, 0.5% polyphenol oxidase and tannase, 5% spirulina powder, 4% glucose and 1.5% peptone are added to the black tea fermentation broth during the fermentation process.

[0018] In a second aspect, the present invention provides use of the tea-protease composition in the preparation of daily chemical products.

[0019] The present invention verifies through experiments that the tea-protease composition has excellent whitening and mild exfoliating effects, and can be used in various daily chemical products with exfoliating and / or whitening effects.

[0020] As a preferred embodiment of the application of the present invention, the daily chemical product includes at least one of detergents, cosmetics, oral hygiene products, and cosmetics and toiletries for animals.

[0021] As a preferred embodiment of the application of the present invention, the dosage form of the cosmetic includes at least one of cream, lotion, water, gel, oil, powder, block powder or solid, patch, film and freeze-dried.

[0022] In a third aspect, the present invention provides a mild, whitening exfoliant, comprising the following components by weight: 2-4 parts of the aforementioned tea-protease composition, 0.1-0.5 parts of a thickener, 0.3-0.6 parts of a preservative, 0.05-0.2 parts of a chelating agent, and 90-99.9 parts of a matrix component. The present invention combines the tea-protease composition with other excipients to form an exfoliant. Experimental testing has shown that combining the tea-protease composition with other excipients to form an exfoliant further enhances the exfoliating and whitening efficacy of the tea-protease composition.

[0023] As a preferred embodiment of the exfoliant of the present invention, the exfoliant comprises the following components in parts by mass: 3.1 parts of the above-mentioned tea-protease composition, 0.2 parts of a thickener, 0.5 parts of a preservative, 0.1 parts of a chelating agent, and 90-99.9 parts of a matrix component.

[0024] As a preferred embodiment of the exfoliant of the present invention, the thickener includes but is not limited to at least one of carbomer, xanthan gum, agar and gelatin.

[0025] As a preferred embodiment of the exfoliant of the present invention, the preservative includes but is not limited to at least one of phenoxyethanol, chlorphenesin, p-hydroxyphenylethanol, etc.

[0026] As a preferred embodiment of the exfoliant of the present invention, the chelating agent includes but is not limited to disodium EDTA.

[0027] As a preferred embodiment of the exfoliant of the present invention, the base component of the exfoliant includes at least one of glycerin, butylene glycol and water.

[0028] As a preferred embodiment of the exfoliant of the present invention, the mass ratio of glycerol to butylene glycol in the matrix component is glycerol:butylene glycol = (3.0-7.0): (2.0-5.0).

[0029] As a preferred embodiment of the exfoliant of the present invention, the mass ratio of glycerin to butylene glycol in the matrix component is glycerin:butylene glycol = 5.0:3.0.

[0030] As a preferred embodiment of the exfoliant of the present invention, when the base components of the exfoliant are glycerin, butylene glycol and water, the mass parts of glycerin are 3.0-7.0, the mass parts of butylene glycol are 2.0-5.0, and the water is supplemented to 90-99.9 parts by mass.

[0031] In a fourth aspect, the present invention provides a method for preparing the above-mentioned exfoliant, comprising the following steps:

[0032] (1) Mixing the substances contained in the matrix component at 60-70° C. to obtain a compound A;

[0033] (2) mixing the thickener with the compound A obtained in step (1) at 60-70° C. and 300-500 rpm to obtain an aqueous phase;

[0034] (3) mixing the aqueous phase obtained in step (2) with the tea-protease composition at 40-45° C. and 150-250 rpm, and then homogenizing at 40-45° C. and 1500-2500 rpm for 3-4 minutes to obtain a complex B;

[0035] (4) The complex B obtained in step (3) is mixed with a preservative and a chelating agent, the pH value is adjusted to 5.5-6.5, and the mixture is filtered when the temperature is 23-27° C. to obtain an exfoliant.

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] The present invention combines black tea fermentation liquor with papain and lipase to form a tea-protease composition. The black tea fermentation liquor and lipase can alleviate the irritation caused by papain. At the same time, papain, lipase and black tea fermentation liquor can synergistically improve the exfoliating and whitening effects of the tea-protease composition, achieving the dual effects of mild exfoliation and whitening. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 The results of the exfoliation efficacy test of different tea-protease compositions and exfoliants in Example 1 of the present invention are as follows;

[0039] Figure 2 These are the results of tyrosinase inhibition rate determination of different tea-protease compositions and enzyme removal in Example 2 of the present invention. DETAILED DESCRIPTION

[0040] 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.

[0041] Unless otherwise specified, other materials, reagents, etc. used in the Examples, Comparative Examples, and Effect Examples can be obtained from commercial sources.

[0042] The black tea fermentation liquid is prepared by fermenting a black tea water extract with Lactobacillus plantarum and Lactobacillus fermentum. The specific operation steps are shown in the optimal fermentation process of Example 2 and Example 3 disclosed in the invention patent CN 118844517 A, which specifically includes the following steps:

[0043] (1) Preparation of black tea extract

[0044] Weigh 5 g of black tea and grind it into a powder with a fineness of 80 mesh. Add pure water to the black tea powder at a solid-liquid ratio of 1:40 g / mL, extract at 70°C for 10 min, shake once every 5 min, centrifuge at 3000 r / min for 10 min, extract twice, combine the two extracts, and filter sterilize the black tea extract using a 0.22 μm filter membrane.

[0045] (2) Lactic acid bacteria activation and seed culture

[0046] Refrigerated Lactobacillus plantarum dy-1 (purchased from CGMCC No. 6016) and Lactobacillus fermentum (purchased from Shanghai Xuanke Biotechnology Co., Ltd. CECT5716) were streaked on MRS solid culture medium and cultured at 35°C for 48 h. Single colonies were picked and transferred to fresh MRS liquid culture medium conical flasks and cultured at 35°C for 24 h. This process was repeated twice to obtain secondary seed solutions of Lactobacillus plantarum and Lactobacillus fermentum.

[0047] The formula of MRS liquid culture medium is: 20g glucose, 10g beef extract, 10g peptone, 5g yeast extract, 5g anhydrous sodium acetate, 2g dipotassium hydrogen phosphate, 2g diammonium hydrogen citrate, 1mL Tween-80, 0.2g magnesium sulfate, 0.05g manganese sulfate, adjust the pH to 6.2-6.4, and sterilize at 115°C for 15min. The formula of MRS solid culture medium is to add 2% agar powder to MRS liquid culture medium.

[0048] (3) Compound lactic acid bacteria black tea fermentation

[0049] The seed liquid of activated Lactobacillus plantarum and Lactobacillus fermentum were inoculated into 10 mL of the black tea extract obtained in step (1) at an inoculum amount of 4% of the volume of the black tea extract, and then 0.5% of polyphenol oxidase and tannase, 5% spirulina powder, 4% glucose, and 1.5% peptone were added respectively, and fermented at 35° C. for 48 hours.

[0050] (4) After the fermentation culture is completed, the fermentation culture medium is filtered using a 12 μm filter plate, and the obtained filtrate is sterilized at 85° C. for 30 min to obtain the black tea fermentation liquid.

[0051] 0.05mol / L PBS (pH=6.8) was prepared by mixing 50mL of solution A and 50mL of solution B. Solution A was a 0.05mol / L Na2HPO4 solution, and solution B was a 0.05mol / L KH2PO4 solution.

[0052] The tyrosinase solution is a 100 U / mL tyrosinase PBS solution, and the PBS is 0.05 mol / L PBS (pH=6.8), which is prepared before use.

[0053] The levodopa solution was a 1 mg / mL levodopa PBS solution, and the PBS was 0.05 mol / L PBS (pH=6.8), which was stored in the dark.

[0054] The BCA protein concentration determination kit was purchased from Nanjing Novozymes Biotechnology Co., Ltd. with the catalog number E112.

[0055] Examples 1-6 and Comparative Examples 1-5

[0056] Examples 1-6 and Comparative Examples 1-5 respectively provide a tea-protease composition with mild exfoliation and whitening effects. The components and amounts of the tea-protease composition are shown in Table 1.

[0057] Table 1 Components and dosages of different tea-protease compositions (parts by mass)

[0058]

[0059] Comparative Example 6

[0060] Comparative Example 6 provides a tea-protease composition with mild exfoliation and whitening effects. The components and amounts of the tea-protease composition are similar to those of Example 1, except that the black tea fermentation broth is replaced by a black tea extract, which is the black tea extract obtained in step (1) of the black tea fermentation broth preparation process. The remaining components and amounts remain unchanged.

[0061] Comparative Example 7

[0062] Comparative Example 7 provides a tea-protease composition with mild exfoliation and whitening effects. The components and amounts of the tea-protease composition are similar to those of Example 1, except that papain in the protease is replaced by superoxide dismutase, and the other components and amounts remain unchanged.

[0063] Examples 7-11 and Comparative Examples 8-10

[0064] Examples 7-11 and Comparative Examples 8-10 respectively provide a mild and whitening exfoliant and a preparation method thereof. The components and amounts of the exfoliant are shown in Table 2. The preparation method comprises the following steps:

[0065] (1) mixing glycerol, butylene glycol, and water in the matrix components at 60-70° C. to obtain a compound A;

[0066] (2) mixing the thickener with the compound A obtained in step (1) at 60-70° C. and 300-500 rpm to obtain an aqueous phase;

[0067] (3) mixing the aqueous phase obtained in step (2) with the tea-protease composition at 40-45° C. and 150-250 rpm, and then homogenizing at 40-45° C. and 1500-2500 rpm for 3-4 minutes to obtain a complex B;

[0068] (4) The complex B obtained in step (3) is mixed with a preservative and a chelating agent, the pH value is adjusted to 5.5-6.5, and the mixture is filtered when the temperature is 23-27° C. to obtain an exfoliant.

[0069] Table 2 Components and dosage of different exfoliants (parts by mass)

[0070]

[0071] Comparative Example 12

[0072] Comparative Example 12 provides a mild, whitening exfoliant and a preparation method thereof. The components and amounts of the exfoliant are similar to those of Example 7, except that the tea-protease composition obtained in Example 1 used in Example 7 is replaced by the tea-protease composition obtained in Comparative Example 5. The remaining components and amounts remain unchanged, and the preparation method is the same as that of Example 7.

[0073] Effect Example 1

[0074] The tea-protease compositions or exfoliants obtained in Examples 1-13 and Comparative Examples 1-12 were subjected to irritation tests, and the chicken embryo chorioallantoic membrane test was performed in accordance with the group standard "T / SHRH 011-2018". The chicken embryos used were Bailaihang chickens purchased from Xinxin Dahua Agricultural Poultry and Egg Company, with the experimental animal production license number SCXK (Guangdong) 2023-0019. The irritation scores are shown in Table 3, and the results are shown in Table 4.

[0075] Table 3 Irritation score

[0076]

[0077] Table 4 Test results of chicken embryo chorioallantoic membrane with different tea-protease compositions or exfoliants

[0078]

[0079] As shown in Table 4, the tea-protease compositions and exfoliants obtained in the Examples were non-irritating, indicating that the tea-protease compositions and exfoliants of the present invention are mild and non-irritating products. Compared with the Examples, the tea-protease compositions obtained in Comparative Examples 1-4 and Comparative Example 7 exhibited some irritation. This suggests that only the combination of black tea fermentation broth, papain, and lipase can impart a mild effect to the tea-protease compositions. If any one component is missing or replaced with a component with similar efficacy, the mild and non-irritating effect cannot be achieved.

[0080] Effect Example 2

[0081] The tea-protease compositions or exfoliants obtained in Examples 1-13 and Comparative Examples 1-12 were tested for exfoliation and whitening efficacy. The specific protocol is as follows:

[0082] 1. Exfoliation test.

[0083] The stratum corneum is the outermost layer of the epidermis, primarily composed of 10-20 layers of flat, anucleated, dead keratinocytes. Normally, as the keratinocytes in the basal layer of the epidermis differentiate and migrate upward, the outermost layer of keratin gradually sheds. Protein is the primary component of the stratum corneum. Since the amount of shed keratinocytes is proportional to the amount of shed protein, quantitative analysis of shed protein can be used to assess the exfoliating efficacy of a sample.

[0084] Fresh pig abdominal skin was selected as the test site. The interval between each test area was not less than 1 cm. 1 g of the test substance was taken to exfoliate the test area. After exfoliation, the detached keratinocytes were collected using an EP tube, rinsed three times with phosphate buffer, centrifuged, and the supernatant discarded. Lysis buffer was added to the EP tube, and the tube was allowed to stand for 40 minutes and centrifuged. The supernatant was retained and protein quantification was performed using the BCA method according to the instructions of the BCA protein concentration assay kit. The assay results are shown in Table 5.

[0085] The standard curve equation constructed by the BCA method is y=0.718x+0.1646.

[0086] Table 5 Exfoliation efficacy test results of different tea-protease compositions or keratin

[0087]

[0088]

[0089] As shown in Table 5 and Figure 1 As shown, compared with the blank control group, the protein concentration of the embodiment is higher, indicating that the tea-protease composition and keratin of the present invention have good exfoliating effect.

[0090] Compared with Example 1, the protein concentration of Comparative Examples 1-7 is lower, indicating that when there is only one component in the tea-protease composition, when the black tea fermentation broth and protease in the tea-protease composition are replaced with other similar components, and when the dosage of each component of the tea-protease composition is inappropriate, a good exfoliating effect cannot be achieved. This shows that the tea-protease composition of the present invention can only achieve a good exfoliating effect under specific types and specific ratios.

[0091] Compared with Example 8, the protein concentration of Comparative Examples 8-12 is lower, indicating that the tea-protease composition needs to be added in a specific amount to achieve good exfoliation effect. If other exfoliating products are added, the exfoliation effect of the tea-protease composition may not be achieved.

[0092] 2. Whitening effect.

[0093] The test solution was prepared according to Table 6. The test sample, 0.05 M PBS buffer (pH 6.8), and tyrosinase solution were added to the test tube in sequence. The test tube was placed in a 37°C water bath for 10 minutes. Then, 2 mL of levodopa solution was added. After reacting for 5 minutes, the absorbance was measured at 475 nm. The absolute difference between two independent determination results obtained under repeatability conditions should not exceed 10% of the arithmetic mean. The tyrosinase inhibition rate (%) was calculated according to formula (1). The results are shown in Tables 7 and Figure 2 .

[0094] Table 6 Reaction solution composition (mL)

[0095]

[0096] Tyrosinase inhibition rate (%) = [1-(OD C -OD D ) / (OD A -OD B )]×100%——Formula (1)

[0097] In the above formula, OD A is the absorbance value of the enzyme solution control group; OD B is the absorbance value of the enzyme solution blank group; OD C is the absorbance value of the sample group; OD D is the absorbance value of the blank control of the sample group.

[0098] Table 7 Tyrosinase inhibition test results of different tea-protease compositions or exfoliants

[0099] Group Tyrosinase inhibition rate (%) Group Tyrosinase inhibition rate (%) Example 1 71.563 Comparative Example 1 19.572 Example 2 77.567 Comparative Example 2 17.336 Example 3 91.765 Comparative Example 3 19.993 Example 4 76.132 Comparative Example 4 55.687 Example 5 65.332 Comparative Example 5 57.543 Example 6 61.029 Comparative Example 6 53.657 Example 7 92.543 Comparative Example 7 53.212 Example 8 87.561 Comparative Example 8 9.653 Example 9 72.727 Comparative Example 9 48.776 Example 10 86.112 Comparative Example 10 43.895 Example 11 85.774 Comparative Example 11 34.872 Example 12 85.365 Comparative Example 12 28.980 Example 13 84.652

[0100] As shown in Table 7 and Figure 2 As shown, compared with the blank control group, the tyrosinase inhibition rate of the embodiment is higher, indicating that the tea-protease composition and keratin of the present invention both have good whitening effects.

[0101] Compared with Example 1, the tyrosinase in Comparative Examples 1-7 is lower, indicating that when there is only one component in the tea-protease composition, when the black tea fermentation broth and protease in the tea-protease composition are replaced with other similar components, and when the dosage of each component of the tea-protease composition is inappropriate, a good whitening effect cannot be achieved. This shows that the tea-protease composition of the present invention can only achieve a good whitening effect under specific types and specific ratios.

[0102] Compared with Example 8, the tyrosinase inhibition rate of Comparative Examples 8-12 is lower, indicating that the tea-protease composition needs to be added in a specific amount to achieve good exfoliation effect. If other exfoliating products are added, the whitening effect of the tea-protease composition may not be achieved.

[0103] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A tea-protease composition with mild exfoliation and whitening effects, characterized in that: The invention comprises the following components in parts by mass: 0.5-2.0 parts of black tea fermentation liquor and 0.1-6.0 parts of protease, wherein the protease comprises papain and lipase.

2. The tea-protease composition according to claim 1, wherein The mass ratio of papain to lipase in the protease is papain:lipase=(10-650):

1.

3. The tea-protease composition according to claim 2, wherein The mass ratio of papain to lipase in the protease is papain:lipase=(100-500):

1.

4. The tea-protease composition according to claim 1, wherein The black tea fermentation liquid is mainly prepared by fermenting black tea water extract with Lactobacillus plantarum and Lactobacillus fermentum.

5. Use of the tea-protease composition according to any one of claims 1 to 4 in the preparation of daily chemical products.

6. The use according to claim 5, characterized in that The daily chemical products include at least one of cleaning products, cosmetics, oral cleaning products and cosmetic toiletries for animals.

7. A mild, whitening exfoliant, characterized by: The exfoliant comprises the following components in parts by mass: 2-4 parts of the tea-protease composition according to any one of claims 1 to 4, 0.1-0.5 parts of a thickener, 0.3-0.6 parts of a preservative, 0.05-0.2 parts of a chelating agent, and 90-99.9 parts of a matrix component.

8. The exfoliant according to claim 7, wherein The base component of the exfoliant includes at least one of glycerin, butylene glycol and water.

9. The exfoliant according to claim 8, wherein The matrix component comprises glycerol and butylene glycol, and the mass ratio of glycerol to butylene glycol is glycerol:butylene glycol=(3.0-7.0):(2.0-5.0).

10. The method for preparing the exfoliant according to any one of claims 7 to 9, characterized in that: The following steps are involved: (1) Mixing the substances contained in the matrix component at 60-70° C. to obtain a compound A; (2) mixing the thickener with the compound A obtained in step (1) at 60-70° C. and 300-500 rpm to obtain an aqueous phase; (3) mixing the aqueous phase obtained in step (2) with the tea-protease composition according to any one of claims 1 to 4 at 40-45° C. and 150-250 rpm, and then homogenizing at 40-45° C. and 1500-2500 rpm for 3-4 minutes to obtain a complex B; (4) The complex B obtained in step (3) is mixed with a preservative and a chelating agent, the pH value is adjusted to 5.5-6.5, and the mixture is filtered when the temperature is 23-27° C. to obtain an exfoliant.

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