Hair care composition for improving tyrosinase activity and preparation method thereof

Abstract

The invention relates to a hair care composition capable of improving tyrosinase activity and a preparation method thereof.The hair care composition is prepared from ginkgo leaves, polygonum multiflorum, rhododendron simsii, glossy privet fruits, figs, angelica sinensis, astragalus membranaceus and radix sophorae flavescentis through enzymolysis and other processes. According to the hair care composition, the influence of oxidative stress on hair follicles is reduced by improving the activity of tyrosinase, and the hair is promoted to turn black from white.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and in particular to a hair care composition for improving tyrosinase activity and a preparation method thereof. Background Art

[0002] The formation of gray hair is the result of the combined effect of multiple factors. Genetic factors are one of the important reasons. If there is premature graying of hair in the family, the probability of "premature graying" in offspring will increase. Long-term mental stress, emotional tension, staying up late, etc. can lead to excessive activation of the sympathetic nerves, consume melanin stem cells in the hair follicles, and then lead to gray hair. Nutritional deficiency is also an important cause of gray hair. The lack of nutrients such as protein, trace elements (such as copper, zinc, iron), B vitamins and vitamin D can affect the production of melanin. In addition, some diseases such as abnormal thyroid function, anemia, etc., as well as unhealthy lifestyles such as smoking, drinking, excessive exposure to ultraviolet rays, picky eating, lack of exercise, etc., can also cause hair to turn gray. In the natural aging process, the function of melanocytes declines, and the hair will gradually turn gray.

[0003] Prior art CN202410972249.2 discloses a herbal hair loss prevention, hair growth, and hair blackening composition and preparation method. The composition combines Polygonum multiflorum, elm bark, ginger, alfalfa, gallnut, wolfberry, and witch hazel to create a hair composition that enhances tyrosinase activity to achieve a hair blackening effect. However, studies have shown that reduced catalase activity leads to the accumulation of hydrogen peroxide and the death of melanocytes in the hair bulb, resulting in hair graying. The above solution does not mention addressing the issue of hair graying by increasing catalase activity and reducing the effects of oxidative stress on hair follicles. Summary of the Invention

[0004] In view of this, a hair care composition for improving tyrosinase activity and a preparation method thereof are provided to solve the above problems.

[0005] To achieve the above object, the present invention adopts the following technical solutions:

[0006] In a first aspect, the present invention provides a hair care composition for improving the activity of tyrosinase, the hair care composition comprising a plant fermentation broth and a plant extract in a mass ratio of 1:0.8-1.5, the plant fermentation broth comprising the following raw materials in parts by weight: 10-15 parts of ginkgo leaves, 20-30 parts of polygonum multiflorum, 6-14 parts of alpine rose azalea flowers, 15-22 parts of privet fruits and 16-23 parts of figs, and the plant extract comprising the following raw materials in parts by weight: 3-7 parts of angelica sinensis, 8-15 parts of astragalus root and 10-20 parts of sophora flavescens.

[0007] In a second aspect, the present invention provides a method for preparing the hair care composition for improving tyrosinase activity as described in the first aspect, the preparation method comprising the following steps: uniformly mixing the plant fermentation liquid and the plant extract according to the formula amount to prepare the hair care composition.

[0008] Preferably, the method for preparing the plant fermentation liquid comprises the following steps:

[0009] (1) Grinding and sieving dried ginkgo leaves, Polygonum multiflorum, alpine rose azalea, Ligustrum lucidum fruit and fig respectively, and mixing them according to the formula amount to obtain a mixed powder;

[0010] (2) Mixing the mixed powder and deionized water, sterilizing, adding Lactobacillus plantarum and Lactobacillus rhamnosus, fermenting under anaerobic conditions at 28-35° C. for 120-144 hours, filtering, and sterilizing the filtrate to obtain the plant fermentation liquid.

[0011] Further preferably, in step (2), the total viable count of Lactobacillus plantarum and Lactobacillus rhamnosus is 1×10 9 -8×10 9 CFU / mL.

[0012] More preferably, the ratio of the number of live bacteria of Lactobacillus plantarum to Lactobacillus rhamnosus is 1:1-3.

[0013] Further preferably, in step (2), the material-liquid ratio of the mixed powder and deionized water is 1 g:5-10 mL.

[0014] Preferably, the method for preparing the plant extract comprises the following steps:

[0015] S1: Grinding and sieving the dried angelica, astragalus and sophora flavescens respectively, and mixing them according to the formula to obtain a mixed material;

[0016] S2: mixing the mixed material and the extraction solvent, performing ultrasonic extraction under the conditions of 40-50° C. and 180-220 W, filtering to obtain a filtrate, and obtaining the plant extract.

[0017] Further preferably, the extraction solvent is a low eutectic solvent and deionized water in a mass ratio of 1:0.3-0.4, and the low eutectic solvent comprises betaine and 1,3-propylene glycol in a molar ratio of 1:4.5-5.5.

[0018] Further preferably, in step S2, the material-liquid ratio of the mixture to the extraction solvent is 1 g:15-20 mL.

[0019] In a third aspect, the present invention provides use of the hair care composition for improving tyrosinase activity according to the first aspect in the preparation of hair cosmetics.

[0020] In a fourth aspect, according to the use described in the third aspect of the present invention, the hair cosmetic is any one of a hair care essence, a hair care essential oil, a hair care spray, a hair care lotion, a shampoo, a conditioner or a hair mask.

[0021] In the present invention:

[0022] The active ingredients in ginkgo biloba mainly include flavonoids, terpene lactones, and polysaccharides. These ingredients have antioxidant, blood circulation, and cell proliferation effects. In terms of promoting hair health, ginkgo biloba extract can increase blood circulation and have a good proliferation effect on hair papilla cells and epidermal cells, thereby supporting hair growth and melanin production. In addition, ginkgo biloba extract has the effect of nourishing hair and relieving scalp itching.

[0023] Polygonum multiflorum is rich in lecithin, polysaccharides, and various trace elements. These ingredients have multiple benefits: lecithin nourishes the scalp and hair follicles, promoting blood circulation and providing nutrients to the scalp; polysaccharides enhance scalp immunity and regulate the scalp's microenvironment; and trace elements such as zinc and iron contribute to melanin synthesis. Polygonum multiflorum also possesses antioxidant properties, reducing free radical damage to hair follicles, thereby improving hair health, reducing the occurrence of gray hair, and promoting dark, thick hair.

[0024] Alpine rose azalea is rich in active ingredients such as flavonoids, polyphenols, and amino acids. These ingredients have strong antioxidant properties that can effectively scavenge free radicals and reduce oxidative stress damage to hair follicle cells, thereby delaying the onset of gray hair. At the same time, its nutrients can deeply nourish the scalp and hair follicles, improve the scalp microenvironment, enhance hair follicle vitality, promote healthy hair growth, and make hair more supple and shiny.

[0025] Ligustrum lucidum fruit is rich in active ingredients such as oleanolic acid, ligustrin, polysaccharides, and various vitamins. In hair care products, oleanolic acid has anti-inflammatory and sebum-regulating properties, improving scalp health. Ligustrum lucidum and polysaccharides nourish the scalp, promote blood circulation, and provide nutrients to hair follicles, enhancing their vitality. Furthermore, its antioxidants scavenge free radicals, reduce oxidative damage, and protect hair follicle cells, thereby promoting dark, thick hair and alleviating dry, brittle, and flaky hair, leaving them healthy and lustrous.

[0026] Figs are rich in vitamin C, vitamin E, flavonoids, organic acids, and various minerals. In hair care products, the vitamin C and flavonoids possess potent antioxidant properties, effectively scavenging free radicals and reducing oxidative stress damage to hair follicle cells, thereby delaying the onset of gray hair. Organic acids regulate the scalp's acid-base balance, improving the scalp's microenvironment and inhibiting the growth of harmful microorganisms. Furthermore, the nutrients in figs nourish the scalp and hair follicles, replenishing essential nutrients for hair, making it smoother and more lustrous, reducing dryness and split ends, and promoting healthy hair growth.

[0027] Angelica sinensis is rich in active ingredients such as volatile oils, organic acids, polysaccharides, vitamins, and trace elements. In hair care products, its volatile oils promote blood circulation in the scalp, improving nutrient supply and boosting hair follicle vitality. Organic acids regulate the scalp's acid-base balance, maintaining a healthy microenvironment and reducing excessive or insufficient oil secretion. Polysaccharides and vitamins nourish scalp cells, providing essential nutrients and enhancing scalp immunity. Furthermore, angelica sinensis' antioxidants help scavenge free radicals and protect hair follicle cells from oxidative damage, thereby promoting dark, thick hair, reducing hair loss, and leaving hair healthier and more lustrous.

[0028] Astragalus root is rich in active ingredients such as flavonoids, polysaccharides, amino acids, and various trace elements. In hair care products, its flavonoids have antioxidant properties, scavenging free radicals and reducing oxidative damage to hair follicle cells. Polysaccharides can enhance scalp immunity, regulate the scalp microenvironment, and promote scalp health. Furthermore, astragalus root has the benefits of replenishing Qi and raising Yang, improving blood circulation in the scalp, providing adequate nutrition to hair follicles, and enhancing their vitality, thereby promoting thicker hair growth, reducing hair loss, and making hair more lustrous and smooth.

[0029] Sophora flavescens is rich in active ingredients such as matrine, oxymatrine, and flavonoids. In hair care products, matrine has significant antibacterial and anti-inflammatory properties, effectively inhibiting the growth of harmful microorganisms on the scalp and reducing inflammatory issues such as itching, redness, and swelling. Its flavonoids possess antioxidant properties, scavenging free radicals and protecting hair follicle cells from oxidative damage. Furthermore, Sophora flavescens can regulate scalp oil secretion, balance the scalp environment, and alleviate scalp conditions such as seborrheic dermatitis, thereby providing a healthy environment for hair growth, reducing hair loss, and leaving hair fresher and healthier.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] The present invention uses ginkgo leaves, polygonum multiflorum, alpine rose azalea leaves, privet fruit, fig, angelica, astragalus, and sophora flavescens as raw materials to prepare a hair care composition. The composition promotes melanin production in cells by increasing tyrosinase activity and reducing hydrogen peroxide accumulation in hair follicles, thereby achieving the effect of blackening white hair. In the present invention, ginkgo leaves, polygonum multiflorum, alpine rose azalea leaves, privet fruit, and fig are fermented and extracted using Lactobacillus plantarum and Lactobacillus rhamnosus; while angelica, astragalus, and sophora flavescens are extracted using a deep eutectic solvent. The resulting plant fermentation broth and plant extract contain more active ingredients that promote the blackening of white hair. By selecting a rational ratio of fermentation strains during the fermentation and extraction process, the strains complement each other, fully utilizing the various components in the substrate. The plant fermentation broth contains more active ingredients that promote the blackening of white hair, further enhancing the hair-blackening effect of the hair care composition. Experiments show that the present invention uses a specific method to extract the raw materials, so that the composition contains more effective ingredients that can activate tyrosinase activity, promote melanin production and increase catalase activity, thereby enabling the composition to achieve better hair blackening effect. DETAILED DESCRIPTION

[0032] 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.

[0033] Unless otherwise specified, other materials and reagents used in the examples can be obtained from commercial sources.

[0034] The Lactobacillus plantarum (specification model: ATCC8014) and Lactobacillus rhamnosus (specification model: ATCC53103) in the present invention were purchased from Beijing Baozang Biotechnology Co., Ltd.

[0035] Example 1

[0036] A hair care composition for improving tyrosinase activity, the hair care composition comprising a plant fermentation liquid and a plant extract in a mass ratio of 1:0.8, the plant fermentation liquid comprising the following raw materials in parts by weight: 10 parts of ginkgo biloba leaves, 20 parts of polygonum multiflorum, 6 parts of alpine rose azalea flowers, 15 parts of privet fruits, and 16 parts of figs; the plant extract comprising the following raw materials in parts by weight: 3 parts of angelica sinensis, 8 parts of astragalus root, and 10 parts of sophora flavescens;

[0037] The preparation method of the plant fermentation liquid comprises the following steps:

[0038] (1) Grinding dried ginkgo leaves, Polygonum multiflorum, alpine rose azalea, Ligustrum lucidum fruit and fig respectively, passing through a 100-mesh sieve, and mixing according to the formula amount to obtain a mixed powder;

[0039] (2) The mixed powder and deionized water were mixed at a material-liquid ratio of 1 g:5 mL, 5% glucose was added according to the mass of the mixed material, and the mixture was sterilized. Then, Lactobacillus plantarum and Lactobacillus rhamnosus were added, and the mixture was fermented under anaerobic conditions at 28°C for 120 h, filtered, and the filtrate was sterilized to obtain the plant fermentation liquid; wherein the total viable count of Lactobacillus plantarum and Lactobacillus rhamnosus was 1×10 9 CFU / mL, the ratio of the number of viable Lactobacillus plantarum and Lactobacillus rhamnosus was 1:1;

[0040] The preparation method of the plant extract comprises the following steps:

[0041] S1: Grind the dried angelica, astragalus and sophora flavescens separately, pass through a 100-mesh sieve, and mix according to the formula to obtain a mixed material;

[0042] S2: mixing the mixed material and the extraction solvent at a material-liquid ratio of 1 g:15 mL, performing ultrasonic extraction at 40° C. and 180 W for 50 min, and filtering to obtain a filtrate; wherein the extraction solvent is a deep eutectic solvent and deionized water in a mass ratio of 1:0.3, and the deep eutectic solvent includes betaine and 1,3-propylene glycol in a molar ratio of 1:4.5, to obtain the plant extract;

[0043] The preparation method of the hair care composition comprises the following steps: mixing plant fermentation liquid and plant extract according to the formula amount to prepare the hair care composition.

[0044] Example 2

[0045] A hair care composition for improving tyrosinase activity, the hair care composition comprising a plant fermentation liquid and a plant extract in a mass ratio of 1:1.5, the plant fermentation liquid comprising the following raw materials in parts by weight: 15 parts of ginkgo biloba leaves, 30 parts of polygonum multiflorum, 14 parts of alpine rose azalea flowers, 22 parts of privet fruits, and 23 parts of figs; the plant extract comprising the following raw materials in parts by weight: 7 parts of angelica sinensis, 15 parts of astragalus root, and 20 parts of sophora flavescens;

[0046] The preparation method of the plant fermentation liquid comprises the following steps:

[0047] (1) Grinding dried ginkgo leaves, Polygonum multiflorum, alpine rose azalea, Ligustrum lucidum fruit and fig respectively, passing through a 100-mesh sieve, and mixing according to the formula amount to obtain a mixed powder;

[0048] (2) The mixed powder and deionized water were mixed at a material-liquid ratio of 1 g:10 mL, 5% glucose was added according to the mass of the mixed material, and the mixture was sterilized. Then, Lactobacillus plantarum and Lactobacillus rhamnosus were added, and the mixture was fermented under anaerobic conditions at 35°C for 144 h, filtered, and the filtrate was sterilized to obtain the plant fermentation liquid; wherein the total viable count of Lactobacillus plantarum and Lactobacillus rhamnosus was 8×109 CFU / mL, the ratio of the number of viable Lactobacillus plantarum and Lactobacillus rhamnosus was 1:3;

[0049] The preparation method of the plant extract comprises the following steps:

[0050] S1: Grind the dried angelica, astragalus and sophora flavescens separately, pass through a 100-mesh sieve, and mix according to the formula to obtain a mixed material;

[0051] S2: mixing the mixed material and the extraction solvent at a material-liquid ratio of 1 g:20 mL, performing ultrasonic extraction at 50° C. and 220 W for 60 min, and filtering to obtain a filtrate; wherein the extraction solvent is a deep eutectic solvent and deionized water in a mass ratio of 1:0.4, and the deep eutectic solvent includes betaine and 1,3-propylene glycol in a molar ratio of 1:5.5, to obtain the plant extract;

[0052] The preparation method of the hair care composition comprises the following steps: mixing plant fermentation liquid and plant extract according to the formula amount to prepare the hair care composition.

[0053] Example 3

[0054] A hair care composition for improving tyrosinase activity, the hair care composition comprising a plant fermentation liquid and a plant extract in a mass ratio of 1:1.1, the plant fermentation liquid comprising the following raw materials in parts by weight: 13 parts of ginkgo biloba leaves, 25 parts of polygonum multiflorum, 11 parts of alpine rose azalea flowers, 19 parts of privet fruits, and 20 parts of figs; the plant extract comprising the following raw materials in parts by weight: 5 parts of angelica sinensis, 11 parts of astragalus root, and 15 parts of sophora flavescens;

[0055] The preparation method of the plant fermentation liquid comprises the following steps:

[0056] (1) Grinding dried ginkgo leaves, Polygonum multiflorum, alpine rose azalea, Ligustrum lucidum fruit and fig respectively, passing through a 100-mesh sieve, and mixing according to the formula amount to obtain a mixed powder;

[0057] (2) The mixed powder and deionized water were mixed at a material-liquid ratio of 1 g:8 mL, 5% glucose was added according to the mass of the mixed material, and the mixture was sterilized. Then, Lactobacillus plantarum and Lactobacillus rhamnosus were added, and the mixture was fermented under anaerobic conditions at 30°C for 136 h, filtered, and the filtrate was sterilized to obtain the plant fermentation liquid; wherein the total viable count of Lactobacillus plantarum and Lactobacillus rhamnosus was 6×10 9 CFU / mL, the ratio of the number of viable Lactobacillus plantarum and Lactobacillus rhamnosus was 1:2;

[0058] The preparation method of the plant extract comprises the following steps:

[0059] S1: Grind the dried angelica, astragalus and sophora flavescens separately, pass through a 100-mesh sieve, and mix according to the formula to obtain a mixed material;

[0060] S2: mixing the mixed material and the extraction solvent at a material-liquid ratio of 1 g:20 mL, performing ultrasonic extraction at 45° C. and 200 W for 55 min, and filtering to obtain a filtrate; wherein the extraction solvent is a deep eutectic solvent and deionized water at a mass ratio of 1:0.35, and the deep eutectic solvent comprises betaine and 1,3-propylene glycol at a molar ratio of 1:5, to obtain the plant extract;

[0061] The preparation method of the hair care composition comprises the following steps: mixing plant fermentation liquid and plant extract according to the formula amount to prepare the hair care composition.

[0062] Comparative Example 1

[0063] Compared with Example 3, the hair care composition in Comparative Example 1 consists of plant fermentation liquid and plant extract in a mass ratio of 1:2, and the remaining steps and process parameters are the same as those in Example 3.

[0064] Comparative Example 2

[0065] Compared with Example 3, the hair care composition in Comparative Example 2 consists of plant fermentation liquid and plant extract in a mass ratio of 1:0.5, and the remaining steps and process parameters are the same as those in Example 3.

[0066] Comparative Example 3

[0067] Compared with Example 3, the preparation method of the plant fermentation liquid in Comparative Example 3 was adjusted, and the specific preparation steps are as follows:

[0068] (1) Grinding dried ginkgo leaves, Polygonum multiflorum, alpine rose azalea, Ligustrum lucidum fruit and fig respectively, passing through a 100-mesh sieve, and mixing according to the formula to obtain a mixed material;

[0069] (2) The mixed material and the extraction solvent were mixed at a material-liquid ratio of 1 g:20 mL, ultrasonically extracted at 45°C and 200 W for 55 min, and filtered to obtain a filtrate; wherein the extraction solvent was a low eutectic solvent and deionized water in a mass ratio of 1:0.35, and the low eutectic solvent included betaine and 1,3-propylene glycol in a molar ratio of 1:5, to obtain the plant extract 1.

[0070] The preparation method of the hair care composition comprises the following steps: mixing the plant extract 1 and the plant extract in a mass ratio of 1:1.1 to prepare the hair care composition.

[0071] The remaining steps and process parameters are the same as in Example 3.

[0072] Comparative Example 4

[0073] Compared with Example 3, the preparation method of the plant extract in Comparative Example 4 was adjusted, and the specific preparation steps were as follows:

[0074] (1) Grinding the dried angelica, astragalus and sophora flavescens separately, passing through a 100-mesh sieve, and mixing according to the formula to obtain a mixed material;

[0075] (2) The mixed material and deionized water were mixed at a material-liquid ratio of 1 g:8 mL, 5% glucose was added according to the mass of the mixed material, and the mixture was sterilized. Then, Lactobacillus plantarum and Lactobacillus rhamnosus were added, and the mixture was fermented under anaerobic conditions at 30° C. for 136 h, filtered, and the filtrate was sterilized to obtain the plant fermentation liquid 1; wherein the total viable count of Lactobacillus plantarum and Lactobacillus rhamnosus was 6×10 9 CFU / mL, the ratio of the number of live bacteria of Lactobacillus plantarum to Lactobacillus rhamnosus was 1:2.

[0076] The preparation method of the hair care composition comprises the following steps: mixing plant fermentation liquid and plant fermentation liquid 1 at a mass ratio of 1:1.1 to prepare the hair care composition.

[0077] The remaining steps and process parameters are the same as in Example 3.

[0078] Comparative Example 5

[0079] Compared with Example 3, the preparation method of the plant fermentation liquid and the plant extract in Comparative Example 5 was adjusted as follows:

[0080] A hair care composition for improving tyrosinase activity, the hair care composition comprising a plant fermentation liquid and a plant extract in a mass ratio of 1:1.1, the plant fermentation liquid comprising the following raw materials in parts by weight: 5 parts of angelica sinensis, 11 parts of astragalus root, and 15 parts of sophora flavescens; the plant extract comprising the following raw materials in parts by weight: 13 parts of ginkgo biloba leaves, 25 parts of polygonum multiflorum, 11 parts of alpine rose azalea flowers, 19 parts of privet fruits, and 20 parts of figs;

[0081] The preparation method of the plant fermentation liquid comprises the following steps:

[0082] (1) Grind the dried angelica, astragalus and sophora flavescens separately, pass through a 100-mesh sieve, and mix according to the formula to obtain a mixed powder;

[0083] (2) The mixed powder and deionized water were mixed at a material-liquid ratio of 1 g:8 mL, 5% glucose was added according to the mass of the mixed material, and the mixture was sterilized. Then, Lactobacillus plantarum and Lactobacillus rhamnosus were added, and the mixture was fermented under anaerobic conditions at 30°C for 136 h, filtered, and the filtrate was sterilized to obtain the plant fermentation liquid; wherein the total viable count of Lactobacillus plantarum and Lactobacillus rhamnosus was 2×10 8CFU / mL, the ratio of the number of viable Lactobacillus plantarum and Lactobacillus rhamnosus was 1:2;

[0084] The preparation method of the plant extract comprises the following steps:

[0085] S1: Grind the dried ginkgo leaves, polygonum multiflorum, alpine rose azalea, privet fruit and fig respectively, pass through a 100-mesh sieve, and mix according to the formula to obtain a mixed material;

[0086] S2: mixing the mixed material and the extraction solvent at a material-liquid ratio of 1 g:20 mL, performing ultrasonic extraction at 45° C. and 200 W for 55 min, and filtering to obtain a filtrate; wherein the extraction solvent is a deep eutectic solvent and deionized water at a mass ratio of 1:0.35, and the deep eutectic solvent comprises betaine and 1,3-propylene glycol at a molar ratio of 1:5, to obtain the plant extract;

[0087] The preparation method of the hair care composition comprises the following steps: mixing plant fermentation liquid and plant extract according to the formula amount to prepare the hair care composition.

[0088] Comparative Example 6

[0089] Compared with Example 3, in the preparation of the plant fermentation liquid in Comparative Example 6, only Lactobacillus plantarum was added for fermentation, and the number of viable Lactobacillus plantarum was 6×10 9 CFU / mL, and the remaining steps and process parameters were the same as those in Example 3.

[0090] Comparative Example 7

[0091] Compared with Example 3, in the preparation of the plant fermentation liquid in Comparative Example 7, only Lactobacillus rhamnosus was added for fermentation, and the viable cell count of Lactobacillus rhamnosus was 6×10 9 CFU / mL, and the remaining steps and process parameters were the same as those in Example 3.

[0092] Comparative Example 8

[0093] Compared with Example 3, the ratio of the number of live bacteria of Lactobacillus plantarum to Lactobacillus rhamnosus in the preparation of the plant fermentation liquid in Comparative Example 8 was 1:0.5, and the remaining steps and process parameters were the same as those in Example 3.

[0094] Comparative Example 9

[0095] Compared with Example 3, the ratio of the number of live bacteria of Lactobacillus plantarum to Lactobacillus rhamnosus in the preparation of the plant fermentation broth in Comparative Example 9 was 1:3.5, and the remaining steps and process parameters were the same as those in Example 3.

[0096] Test Example 1

[0097] Tyrosinase is a key rate-limiting enzyme in melanin synthesis, and its activation rate is closely related to melanin production. By measuring the tyrosinase activation rate of a hair care composition and regulating tyrosinase activity, melanin production can be effectively regulated. Therefore, by testing the tyrosinase activation effect of the hair care compositions prepared in Examples 1-3 and Comparative Examples 1-9, the hair darkening effect of the hair care compositions was determined based on the tyrosinase activation rate and the increase in melanin content.

[0098] The tyrosinase activation rate detection method is as follows:

[0099] (1) Preparation of solution

[0100] A 1 g / L L-dopa solution, a 100 U / mL tyrosinase solution, and a 0.2 mg / mL hair care composition (the hair care compositions in Examples 1-3 and Comparative Examples 1-9) test solution were prepared using a 0.2 mol / L, pH 6.8 phosphate buffer solution.

[0101] (2) Prepare sample solutions numbered A, B, C, and D

[0102] A: Add 40 μL of L-dopa solution, 40 μL of phosphate buffer solution (0.2 mol / L, pH 6.8), and 40 μL of tyrosinase solution to a 96-well plate and incubate in a 30°C water bath for 40 min.

[0103] B: Add 40 μL of L-dopa solution and 80 μL of phosphate buffer solution (concentration: 0.2 mol / L, pH: 6.8) to a 96-well plate and incubate in a 30°C water bath for 40 min.

[0104] C: Add 40 μL of L-dopa solution, 40 μL of test solution, and 40 μL of tyrosinase solution to a 96-well plate and incubate in a 30°C water bath for 40 min.

[0105] D: Add 40 μL of L-dopa solution, 40 μL of test solution, and 40 μL of phosphate buffer solution (0.2 mol / L, pH 6.8) to a 96-well plate and incubate in a 30°C water bath for 40 min.

[0106] After the reaction of sample solutions A, B, C, and D was completed, the absorbance was detected at 490 nm.

[0107] Tyrosinase activation rate = [(CD)-(AB)] / (AB)×100%

[0108] The method for detecting the growth rate of melanin content is as follows:

[0109] (1) Preparation of solution

[0110] A 1 g / L L-dopa solution, a 100 U / mL tyrosinase solution, and a test solution containing the hair care compositions prepared in Examples 1-3 and Comparative Examples 1-9 at a concentration of 2 mg / mL were prepared using a 0.2 mol / L phosphate buffer solution at a pH of 6.8.

[0111] (2) Prepare sample solutions numbered A and B

[0112] A: Add 40 μL of L-dopa solution, 40 μL of test solution, and 40 μL of tyrosinase solution to a 96-well plate and incubate in a 37°C water bath for 60 min.

[0113] B: Add 40 μL of L-dopa solution, 40 μL of phosphate buffer solution (0.2 mol / L, pH 6.8), and 40 μL of tyrosinase solution to a 96-well plate and incubate in a 37°C water bath for 60 min.

[0114] After the reaction of sample solutions A and B is completed, 0.1 mol / L hydrochloric acid solution is added to terminate the reaction, the supernatant is discarded by centrifugation, and the solution is washed with deionized water. 100 μL of 1 mol / L sodium hydroxide solution (containing 10% DMSO) is added to each well, and the absorbance is detected at 400 nm.

[0115] Melanin content growth rate = (A / B-1) × 100%

[0116] Table 1 Test results of tyrosinase activation rate and melanin content growth rate

[0117]

[0118]

[0119] Experimental results show that the hair care composition prepared from ginkgo leaves, polygonum multiflorum, alpine rose azalea leaves, privet fruits, figs, angelica, astragalus and sophora flavescens as raw materials can significantly increase tyrosinase activity and promote melanin production.

[0120] Compared with Example 3, the changes in the dosage ratio of the plant fermentation liquid and the plant extract in Comparative Examples 1 and 2 resulted in a decrease in the tyrosinase activation rate and the growth rate of melanin content. This indicates that the rational ratio of the active ingredients in the composition of the present invention can achieve component complementarity and enhance the hair darkening effect of the hair care composition.

[0121] Compared with Example 3, the preparation methods of the raw materials in the compositions of Comparative Examples 3-5 were changed respectively, resulting in a decrease in the tyrosinase activation rate and the growth rate of melanin content. This shows that the present invention adopts a specific method to extract the raw materials, so that the composition contains more active ingredients that can activate tyrosinase activity and promote melanin production, thereby achieving a better hair darkening effect.

[0122] Compared with Example 3, in Comparative Examples 6-9, varying the fermentation bacteria species and viable bacterial count ratio in the plant fermentation broth, respectively, resulted in decreased tyrosinase activation rates and melanin content growth rates. This indicates that the present invention, by screening the fermentation bacteria species and viable bacterial count ratio, can improve the utilization efficiency of the fermentation substrate and the content of active ingredients in the plant fermentation broth, thereby enabling the composition to achieve a better hair darkening effect.

[0123] Test Example 2

[0124] Oxidative stress can impair the antioxidant defense mechanisms of melanocytes in hair follicles, thereby affecting melanin production. Therefore, the effects of the hair care compositions prepared in Examples 1-3 and Comparative Examples 1-9 on catalase activity were tested, and the hair darkening effect of the hair care compositions was determined by using catalase activity.

[0125] The method for determining catalase activity is as follows:

[0126] (1) Preparation of solution

[0127] Catalase solution: Prepare catalase solution with deionized water to a concentration of 0.1 mg / mL.

[0128] (2) Preparation of experimental and control group solutions

[0129] Experimental group solution: 0.2 mL of the enzyme solution and 0.1 mL of the hair care composition (the hair care composition of Examples 1-3 and Comparative Examples 1-9) were mixed, respectively, and allowed to stand at 25°C for 15 minutes. 20 mL of deionized water and 3 mL of a 2% hydrogen peroxide solution were added, and the mixture was allowed to stand at 25°C for 15 minutes. 10 mL of a 20% sulfuric acid solution was added to terminate the reaction, and titration was performed with a 0.2 mol / L potassium permanganate standard solution until the solution turned pink and did not fade for 30 seconds, which was the titration endpoint. The volume of the potassium permanganate standard solution consumed was recorded. The determination was repeated three times, and the average value was calculated and recorded as V1.

[0130] Control solution: Mix 0.2 mL of enzyme solution and 0.1 mL of deionized water, and place at 25°C for 15 min. Add 20 mL of deionized water and 3 mL of 2% hydrogen peroxide solution, and place at 25°C for 15 min. Add 10 mL of 20% sulfuric acid solution to terminate the reaction, and titrate with 0.2 mol / L potassium permanganate standard solution. Titrate until the solution turns pink and does not fade for 30 seconds, which is the titration end point. Record the volume of potassium permanganate standard solution consumed. Perform three parallel determinations and calculate the average value, which is recorded as V0.

[0131] Enzyme activity (U / ml) = [(V0-V1) × 0.2 × 2.5 × df] / (T) × 0.2

[0132] Wherein: 0.2 is the concentration of potassium permanganate standard solution; 2.5 means that 2.5 mol of hydrogen peroxide is required for every 1 mol of potassium permanganate consumed; df is the dilution factor of catalase; T is the reaction time (min); 0.2 is the amount of catalase solution used in the reaction system.

[0133] Table 2 Catalase activity test results

[0134]

[0135]

[0136] Test results demonstrate that the hair care composition prepared by the present invention enhances catalase activity. Catalase decomposes hydrogen peroxide, reducing its inhibitory effect on tyrosinase, thereby maintaining normal melanin synthesis. More importantly, catalase protects the survival and function of melanocytes by scavenging hydrogen peroxide, thereby reducing oxidative stress damage to melanocytes in hair follicles and enhancing the effectiveness of the hair care composition.

[0137] Compared to Example 3, changes in the dosage ratio of the plant fermentation broth and plant extract in Comparative Examples 1 and 2 resulted in decreased catalase activity. This demonstrates that the rational ratio of the active ingredients in the present composition can achieve complementary effects, increasing catalase activity in hair follicles and reducing oxidative stress, thereby protecting melanocytes and melanocyte stem cells within the hair follicles and further enhancing the hair-blackening effect of the hair care composition.

[0138] Compared with Example 3, Comparative Examples 3-5 change the preparation method of the raw materials in the composition, resulting in a decrease in catalase activity. This shows that the present invention uses a specific method to extract the raw materials, so that the composition contains more active ingredients that increase catalase activity, thereby achieving a better hair darkening effect.

[0139] Compared with Example 3, in Comparative Examples 6-9, the fermentation bacteria species and viable bacterial count ratio in the plant fermentation broth were changed, resulting in a decrease in catalase activity. This indicates that by screening the fermentation bacteria species and viable bacterial count ratio in the plant fermentation broth of the present invention, the utilization efficiency of the fermentation substrate and the content of the catalase active ingredient in the plant fermentation broth can be improved, thereby enabling the composition to achieve a better hair darkening effect.

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

Claims

1. A hair care composition for improving tyrosinase activity, characterized in that: The invention comprises a plant fermentation liquid and a plant extract in a mass ratio of 1:0.8-1.

5. The plant fermentation liquid comprises the following raw materials in parts by weight: 10-15 parts of ginkgo leaves, 20-30 parts of polygonum multiflorum, 6-14 parts of alpine rose azalea flowers, 15-22 parts of privet fruits and 16-23 parts of figs; and the plant extract comprises the following raw materials in parts by weight: 3-7 parts of angelica sinensis, 8-15 parts of astragalus root and 10-20 parts of sophora flavescens.

2. The method for preparing a hair care composition for improving tyrosinase activity according to claim 1, wherein: The preparation method comprises the following steps: uniformly mixing the plant fermentation liquid and the plant extract according to the formula amount to prepare the hair care composition.

3. The method for preparing a hair care composition for improving tyrosinase activity according to claim 2, wherein: The preparation method of the plant fermentation liquid comprises the following steps: (1) Grinding and sieving dried ginkgo leaves, Polygonum multiflorum, alpine rose azalea, Ligustrum lucidum fruit and fig respectively, and mixing them according to the formula amount to obtain a mixed powder; (2) Mixing the mixed powder and deionized water, sterilizing, adding Lactobacillus plantarum and Lactobacillus rhamnosus, fermenting under anaerobic conditions at 28-35° C. for 120-144 hours, filtering, and sterilizing the filtrate to obtain the plant fermentation liquid.

4. The method for preparing a hair care composition for improving tyrosinase activity according to claim 3, wherein: In step (2), the total viable count of Lactobacillus plantarum and Lactobacillus rhamnosus is 1×10 9 -8×10 9 CFU / mL; the material-liquid ratio of the mixed powder and deionized water is 1g:5-10mL.

5. The method for preparing a hair care composition for improving tyrosinase activity according to claim 4, wherein: The ratio of the number of live bacteria of Lactobacillus plantarum to Lactobacillus rhamnosus is 1:1-3.

6. The method for preparing a hair care composition for improving tyrosinase activity according to claim 2, wherein: The preparation method of the plant extract comprises the following steps: S1: Grinding and sieving the dried angelica, astragalus and sophora flavescens respectively, and mixing them according to the formula to obtain a mixed material; S2: mixing the mixed material and the extraction solvent, performing ultrasonic extraction for 50-60 min at 40-50° C. and 180-220 W, filtering to obtain a filtrate, and obtaining the plant extract.

7. The method for preparing a hair care composition for improving tyrosinase activity according to claim 6, wherein: The extraction solvent is a low eutectic solvent and deionized water in a mass ratio of 1:0.3-0.4, and the low eutectic solvent includes betaine and 1,3-propylene glycol in a molar ratio of 1:4.5-5.

5.

8. The method for preparing a hair care composition for improving tyrosinase activity according to claim 6, wherein: In step S2, the material-liquid ratio of the mixture to the extraction solvent is 1 g: 15-20 mL.

9. Use of the hair care composition for improving tyrosinase activity according to claim 1 in preparing hair cosmetics.

10. The use according to claim 9, characterized in that The hair cosmetic is any one of a hair care essence, a hair care essential oil, a hair care spray, a hair care lotion, a shampoo, a hair conditioner or a hair mask.

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