Hair-blacking composition for promoting melanin secretion as well as preparation method and application of hair-blacking composition

Abstract

The invention discloses a hair-blacking composition for promoting melanin secretion as well as a preparation method and application thereof. The hair-blacking composition is prepared from the following raw materials in parts by weight: 10 to 20 parts of coleus forskohlii extract, 1 to 5 parts of tetrahydropiperine, 1 to 10 parts of phenylalanine, 1 to 10 parts of copper compound, 1 to 10 parts of zinc compound, 5 to 25 parts of solubilizer and the balance of water, wherein the total amount is 100 parts. The components in the hair-blacking composition for promoting melanin secretion are safe, effective, mild and non-irritant, after the hair-blacking composition is used for a period of time, black hair increase and new black hair can be observed, and the hair-blacking composition has an excellent hair-blacking effect.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and in particular to a black hair composition for promoting melanin secretion, a preparation method and application thereof. Background Art

[0002] Gray hair in old age is a normal aging phenomenon. With age, the tyrosinase activity in melanocytes is progressively lost, resulting in the depletion of melanin in the hair shaft. In recent years, premature graying of hair in young children has received increasing attention from the public. Factors such as genetics, stress, atmospheric pollutants, and ultraviolet rays can all induce premature graying of hair in young children. Among them, atmospheric pollutants and ultraviolet light factors can easily cause oxidative stress and genotoxicity, leading to gray hair. In addition, stress can also cause nerve activation of the sympathetic nervous system and deplete the melanocyte stem cells in the hair follicles, causing the hair to turn white.

[0003] Melanin formation is highly correlated with the activities of three enzymes in melanocytes: tyrosinase, dopa isomerase (TRP-2), and 5,6-dihydroxyindolecarboxylic acid, or DHICA oxidase (TRP-1). These three enzymes regulate melanin secretion in melanocytes by acting alone or synergistically. Tyrosinase is the most critical rate-limiting enzyme in melanin synthesis, mainly involved in the conversion of tyrosine to dopa and dopaquinone. Its expression and activity determine melanin production. In addition, TRP-2, as a dopachrome tautomerase, maintains the phenolic component in melanin, while TRP-1 can oxidize the 5,6-dihydroxyindolecarboxylic acid produced by TRP-2, ultimately forming true melanin.

[0004] Currently, most products and patents are mainly aimed at improving the activity of tyrosinase. For example, the patent document with patent number CN110840883B discloses a hair blackening composition and its application, and proposes a technical solution containing psoralen and forskolin. The two can synergistically promote the cell proliferation of melanocytes, the activity of tyrosinase and the production of melanin, and can be used to treat gray hair. It is also commonly used in clinical practice to treat vitiligo. However, psoralen is banned in cosmetics. Therefore, it is of great significance to find a more effective and safe hair blackening solution. Summary of the Invention

[0005] The main purpose of the present invention is to provide a hair-blackening composition that promotes melanin secretion, and its preparation method and application, aiming to solve the technical problem that existing hair-blackening solutions are not safe and effective enough.

[0006] In order to achieve the above-mentioned purpose, the first aspect of the present invention proposes a black hair composition for promoting melanin secretion, comprising the following raw materials in parts by weight: 10-20 parts of Coleus forskohlii extract, 1-5 parts of tetrahydropiperine, 1-10 parts of phenylalanine, 1-10 parts of copper compounds, 1-10 parts of zinc compounds, 5-25 parts of solubilizers, and water to make up the balance to 100 parts.

[0007] Furthermore, the invention comprises the following raw materials in parts by weight: 14-18 parts of Coleus forskohlii extract, 3-5 parts of tetrahydropiperine, 4-6 parts of phenylalanine, 3-8 parts of copper compound, 3-8 parts of zinc compound, 15-25 parts of solubilizer and the balance of water.

[0008] Furthermore, the Coleus forskohlii extract is obtained by extracting Coleus forskohlii through supercritical CO2 and then including it with hydroxypropyl-β-cyclodextrin.

[0009] Furthermore, the preparation process of the Coleus forskohlii flower extract is as follows: crushing and sieving the Coleus forskohlii flower, using 95% ethanol as an entrainer in an amount of 0.1 to 0.5 times the amount of the Coleus forskohlii flower, the extraction temperature is 35 to 55° C., the extraction pressure is 20 to 40 MPa, the CO2 flow rate is 20 to 35 L / h, the extraction time is 3 hours, the temperature of separation kettle 1 is 30 to 40° C., the pressure is 8 to 10 MPa, the temperature of separation kettle 2 is 28 to 35° C., the pressure is 5 to 8 MPa, and after reducing pressure to recover the entrainer, a Coleus forskohlii flower extract is obtained, and the Coleus forskohlii flower extract and hydroxypropyl-β-cyclodextrin are added to a saturated hydroxypropyl-β-cyclodextrin solution at a ratio of 1 ml:3 to 4 g, and the mixture is included at 40° C. for 2 hours. After precipitation, filtration, washing with anhydrous ethanol, and drying, the extract is obtained.

[0010] Furthermore, the tetrahydropiperine is prepared by using pepper seed powder as a raw material, extracting it by reflux using methanol, and concentrating it in vacuum to obtain pepper oleoresin, which is dissolved in an isopropanol:n-hexane = 1:1 solution at 50-60°C, cooled to room temperature and filtered to obtain a precipitated wet solid, which is dissolved in methanol, crystallized at 50°C, filtered, concentrated to 1 / 4 of the volume of methanol, cooled to 10°C and filtered to obtain piperine crystals, which are then subjected to a hydrogenation reaction to obtain the product.

[0011] Furthermore, the mass fraction ratio of the tetrahydropiperine to the solubilizer is 1:5.

[0012] Furthermore, the copper compound is one or more of copper salts and copper ion complexes.

[0013] Another aspect of the present invention provides a method for preparing a black hair composition that promotes melanin secretion, comprising the following steps:

[0014] S1: pre-dispersing the solubilizing agent and the tetrahydropiperine and dissolving them at 40-50° C.;

[0015] S2: slowly adding the Coleus forskohlii extract, phenylalanine, copper compound, and zinc compound into 40° C. water and stirring to dissolve;

[0016] S3: Combine the above components and stir until dissolved to obtain the hair-blackening composition that promotes melanin secretion.

[0017] The present invention also provides the use of the hair-blackening composition that promotes melanin secretion in the preparation of hair-blackening products.

[0018] The present invention also provides a black hair product, comprising the black hair composition that promotes melanin secretion, water, ethanol, a moisturizer and a preservative;

[0019] Wherein, the mass percentage of the black hair composition that promotes melanin secretion in the black hair product is 0.5-5%.

[0020] Beneficial effects:

[0021] The black hair composition for promoting melanin secretion of the present invention works synergistically against different targets. The key catalytic site of tyrosinase contains histidine residues, which can bind to copper ions in copper compounds, which is beneficial to improving the activity of tyrosinase; piperine can effectively improve the expression of TRP-1 and thus improve the production of melanin. At the same time, tetrahydropiperine also has a penetration-promoting property, which can enhance the penetration of the active ingredient; and the zinc ions in the zinc compound are also the key metal ions for the enzymatic activity of TRP-2, which can effectively improve its activity; forskolin in the Coleus forskohlii extract is a natural diterpenoid compound that can quickly increase the concentration of intracellular cyclic adenosine monophosphate (cAMP) and enhance the activity of protein kinase A (PKA), activating the AC-cAMP-PKA signaling pathway. When added to the cell culture medium, it can increase the intracellular cAMP level, activate the cAMP-PKA signaling pathway, induce melanocyte division, and thus promote the production of melanin; phenylalanine can be converted into tyrosine through the action of phenylalanine hydroxylase in the body, and then finally form melanin through the action of various melanin-related enzymes, achieving multi-target coverage and playing the role of preventing gray hair and black hair.

[0022] The Coleus forskohlii flower extract of the present invention retains the main active ingredients, and the use of hydroxypropyl-β-cyclodextrin for inclusion can effectively enhance the water solubility of the active ingredients, which is conducive to application in various formulas and has the effect of highly effective hair blackening.

[0023] The ingredients in the hair-blackening composition for promoting melanin secretion of the present invention are safe and effective, mild and non-irritating. After using it for a period of time, an increase in black hair and new black hair can be observed, and it has an excellent hair-blackening effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 Microscopic examination of the hair of a test subject before and after using the black hair essence prepared in Example 1, before use (left) and after one month of use (right);

[0025] Figure 2The following are comparison pictures of a tester's hair before and after using the black hair essence prepared in Example 1, before use (left) and after one month of use (right).

[0026] The purpose, features and advantages of the present invention will be further described with reference to the accompanying drawings and in conjunction with the embodiments. DETAILED DESCRIPTION

[0027] The experimental methods in the following examples of the present invention, where specific conditions are not specified, are generally carried out under conventional conditions or conditions recommended by the manufacturers. The various commonly used chemical reagents used in the examples are all commercially available products.

[0028] Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those commonly understood by those skilled in the art. The terms used in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0029] The terms "comprise," "comprising," and "having," and any variations thereof, are intended to cover non-exclusive inclusions. For example, a process, method, apparatus, product, or device comprising a series of steps is not limited to the listed steps or modules but may optionally include steps not listed, or other steps inherent to the process, method, product, or device.

[0030] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention is further described in detail below in conjunction with specific embodiments. It should be understood that these descriptions are merely exemplary and are not intended to limit the scope of the present invention. In addition, in the following description, descriptions of well-known structures and technologies are omitted to avoid unnecessary confusion of the concept of the present invention.

[0031] The present invention is further described in the following examples, but these examples are not intended to limit the scope of protection of the present invention.

[0032] One embodiment of the present invention provides a black hair composition for promoting melanin secretion, comprising the following raw materials in parts by weight: 10-20 parts of Coleus forskohlii extract, 1-5 parts of tetrahydropiperine, 1-10 parts of phenylalanine, 1-10 parts of copper compounds, 1-10 parts of zinc compounds, 5-25 parts of solubilizers, and water to make up the balance to 100 parts.

[0033] The present invention synergistically acts on different targets. The key catalytic site of tyrosinase contains histidine residues, which can bind to copper ions in copper compounds, thereby increasing the activity of tyrosinase. Piperine can effectively increase the expression of TRP-1, thereby increasing the production of melanin. At the same time, tetrahydropiperine also has a penetration-promoting property, which can enhance the penetration of the active ingredient. The zinc ion in the zinc compound is also a key metal ion for the enzymatic activity of TRP-2, which can effectively increase its activity. Forskolin in the Coleus forskohlii extract is a natural diterpenoid compound that can rapidly increase the concentration of intracellular cyclic adenosine monophosphate (cAMP) and enhance the activity of protein kinase A (PKA), activating the AC-cAMP-PKA signaling pathway. When added to cell culture medium, it can increase the intracellular cAMP level, activate the cAMP-PKA signaling pathway, induce melanocyte division, and thus promote the production of melanin. Phenylalanine can be converted into tyrosine in the body through the action of phenylalanine hydroxylase, and then finally form melanin through the action of various melanin-related enzymes, achieving multi-target coverage and having the effects of preventing gray hair and blackening hair.

[0034] The Coleus forskohlii flower extract of the present invention retains the main active ingredients, and the use of hydroxypropyl-β-cyclodextrin for inclusion can effectively enhance the water solubility of the active ingredients, which is conducive to application in various formulas and has the effect of highly effective hair blackening.

[0035] The ingredients in the hair-blackening composition for promoting melanin secretion of the present invention are safe and effective, mild and non-irritating. After using it for a period of time, volunteers can observe an increase in black hair and new black hair, and it has an excellent hair-blackening effect.

[0036] In the above embodiment, the raw material of the Coleus forskohlii extract is a plant of the genus Coleus Lour. of the family Labiatae, also known as Coleus forskohlii, Coleus forskohlii, and Coleus forskohlii; the whole herb of Coleus forskohlii is often used in Indian Ayurveda for decoction to treat colds, coughs, and respiratory diseases. Coleus forskohlii is a traditional Chinese medicine and is included in the 2005 edition (Volume 1) of the Yunnan Province Traditional Chinese Medicine Standards. Its traditional Chinese medicine name is "Coleus forskohlii". The main active ingredients, forskolin and isoforskolin, have excellent antioxidant properties and have the ability to scavenge DPPH free radicals, ·OH free radicals, and react with Fe 2+ Chelating ability.

[0037] The copper compound can be any compound that can provide copper ions. Preferably, the copper compound is one or more of tripeptide-1 copper, EDTA-copper disodium, PCA copper, alanine / histidine / lysine polypeptide copper hydrochloride, yeast / copper fermentation product, copper sulfate, copper gluconate, copper aspartate, copper acetate, acetylmethionine copper, and the like.

[0038] Preferably, the zinc compound is one or more of DNA zinc, PCA zinc, glycinate zinc, yeast / zinc fermentation product, zinc sulfate, zinc gluconate, zinc lactate, zinc aspartate, and the like.

[0039] Preferably, the solubilizer is one or more of propylene glycol, ethanol, 1,3-butylene glycol, polyethylene glycol-400, PVP K30, hydroxypropyl-β-cyclodextrin, β-cyclodextrin, PEG-40 hydrogenated castor oil, and lecithin.

[0040] Preferably, the mass fraction ratio of the tetrahydropiperine to the solubilizer is 1:5, which can maximize the solubility of tetrahydropiperine and ensure uniform dispersion and good solubility in subsequent processes or applications.

[0041] Preferably, the extraction site of the Coleus forskohlii extract is the root or stem of Coleus forskohlii.

[0042] Another embodiment of the present invention provides a method for preparing a black hair composition that promotes melanin secretion, comprising the following steps:

[0043] S1: pre-dispersing the solubilizing agent and the tetrahydropiperine and dissolving them at 40-50° C.;

[0044] S2: slowly adding the Coleus forskohlii extract, phenylalanine, copper compound, and zinc compound into 40° C. water and stirring to dissolve;

[0045] S3: Combine the above components and stir until dissolved to obtain the hair-blackening composition that promotes melanin secretion.

[0046] The present invention also provides the use of the hair-blackening composition that promotes melanin secretion in the preparation of hair-blackening products.

[0047] The present invention also provides a black hair product, comprising the black hair composition that promotes melanin secretion, water, ethanol, a moisturizer and a preservative;

[0048] The hair-blackening composition that promotes melanin secretion accounts for 0.5-5% of the hair-blackening product by mass. The hair-blackening product can be formulated in the form of solutions, suspensions, emulsions, pastes, gels, creams, powders, soaps, sprays, oils, mousses, cleansers, and the like. Preferably, the hair-blackening composition that promotes melanin secretion is used in an amount of 1-5% in rinse-off products and 0.5-2% in leave-on products.

[0049] The technical solutions of the present invention are further illustrated by specific examples below. Those skilled in the art should understand that the examples are only for helping to understand the present invention and should not be regarded as specific limitations of the present invention.

[0050] The following raw material sources are for illustrative purposes only:

[0051] Coleus forskohlii: Yongji Dayun Commercial and Trade Management Department;

[0052] Tetrahydropiperine: Sami-Sabinsa Group Limited;

[0053] Hydroxypropyl-β-cyclodextrin: Mengzhou Huaxing Biochemical Co., Ltd.;

[0054] Other raw materials can be purchased through regular commercial channels.

[0055] Example 1

[0056] (1) Formula: 160 g of Coleus forskohlii extract, 40 g of tetrahydropiperine, 50 g of phenylalanine, 60 g of tripeptide-1 copper, 60 g of PCA zinc, 200 g of propylene glycol, and water to make up the balance to 1000 g.

[0057] (2) Preparation process: crush the Coleus forskohlii flower through a 20-mesh sieve, use 95% ethanol as an entrainer in an amount of 0.35 times the amount of the Coleus forskohlii flower, the extraction temperature is 40°C, the extraction pressure is 31 MPa, the CO2 flow rate is 25 L / h, the extraction time is 3 hours, the temperature of separation kettle 1 is 35°C, the pressure is 9 MPa, the temperature of separation kettle 2 is 30°C, the pressure is 6 MPa, and the entrainer ethanol is recovered under reduced pressure to obtain the Coleus forskohlii flower extract, the Coleus forskohlii flower extract and hydroxypropyl-β-cyclodextrin are added to a saturated hydroxypropyl-β-cyclodextrin solution at a ratio of 1 ml:3.5 g, and the mixture is included at 40°C for 2 hours. After precipitation, filtration, washing with anhydrous ethanol, and drying, the Coleus forskohlii flower extract is obtained;

[0058] (3) Pepper seed powder is used as raw material, extracted with methanol under reflux, and concentrated under vacuum to obtain pepper oleoresin, which is dissolved in a solution of isopropyl alcohol:n-hexane = 1:1 at 50-60°C, cooled to room temperature and filtered to obtain a precipitated wet solid, which is dissolved in methanol, crystallized at 50°C, filtered, concentrated to 1 / 4 of the volume of methanol, cooled to 10°C and filtered to obtain piperine crystals, which are then hydrogenated to obtain tetrahydropiperine.

[0059] (4) S1: pre-dispersing the solubilizer and the tetrahydropiperine and dissolving them at 40-50° C.;

[0060] S2: slowly adding the Coleus forskohlii extract, phenylalanine, copper compound, and zinc compound into 40° C. water and stirring to dissolve;

[0061] S3: Combine the above components and stir until dissolved to obtain a hair-blackening composition that promotes melanin secretion.

[0062] Examples 2 to 5

[0063] The preparation process of this embodiment is the same as that of Example 1, and the composition of each component in the composition is shown in Table 1:

[0064] raw material Example 2 Example 3 Example 4 Example 5 Coleus Forskohlii Extract 100g 200g 140g 180g Tetrahydropiperine 50g 10g 50g 30g Phenylalanine 10g 100g 40g 60g Tripeptide-1 Copper 100g 10g 80g 30g PCA Zinc 10g 100g 30g 80g Propylene glycol 250g 50g 250g 150g water To 1000g To 1000g To 1000g To 1000g

[0065] Example 6

[0066] The components of the black hair composition for promoting melanin secretion in this embodiment are the same as those in Example 1, the difference being the preparation process of the Coleus forskohlii extract:

[0067] The preparation process of the Coleus forskohlii extract comprises the following steps: grinding the Coleus forskohlii root through a 20-mesh sieve, using 95% ethanol as an entrainer in an amount of 0.1 times the amount of the Coleus forskohlii, setting the extraction temperature at 55°C, the extraction pressure at 20 MPa, the CO2 flow rate at 35 L / h, the extraction time at 3 hours, setting the temperature of a separation kettle 1 at 30°C and the pressure at 10 MPa, setting the temperature of a separation kettle 2 at 28°C and the pressure at 8 MPa, recovering the entrainer ethanol under reduced pressure to obtain a Coleus forskohlii extract, adding the Coleus forskohlii extract to a saturated hydroxypropyl-β-cyclodextrin solution at a ratio of 1 ml:3 g of hydroxypropyl-β-cyclodextrin, carrying out inclusion reaction at 40°C for 2 hours, precipitating, filtering, washing with anhydrous ethanol, and drying to obtain the Coleus forskohlii extract.

[0068] Example 7

[0069] The components of the black hair composition for promoting melanin secretion in this embodiment are the same as those in Example 1, the difference being the preparation process of the Coleus forskohlii extract:

[0070] The preparation process of the Coleus forskohlii extract comprises the following steps: grinding the Coleus forskohlii root through a 20-mesh sieve, using 95% ethanol as an entrainer in an amount 0.5 times that of the Coleus forskohlii, setting the extraction temperature at 35°C, the extraction pressure at 40 MPa, the CO2 flow rate at 205 L / h, the extraction time at 3 hours, setting the temperature of separation kettle 1 at 40°C and the pressure at 8 MPa, setting the temperature of separation kettle 2 at 35°C and the pressure at 8 MPa, recovering the entrainer ethanol under reduced pressure to obtain a Coleus forskohlii extract, adding the Coleus forskohlii extract to a saturated hydroxypropyl-β-cyclodextrin solution at a ratio of 1 ml:4 g of hydroxypropyl-β-cyclodextrin, carrying out inclusion reaction at 40°C for 2 hours, precipitating, filtering, washing with anhydrous ethanol, and then drying to obtain the Coleus forskohlii extract.

[0071] Example 8

[0072] The difference between this comparative example and Example 1 is that tripeptide-1 copper is replaced by PCA copper.

[0073] Example 9

[0074] The difference between this comparative example and Example 1 is that PCA zinc is replaced by DNA zinc.

[0075] Comparative Example 1

[0076] The difference between this comparative example and Example 1 is that the hair-blackening composition does not contain the Coleus forskohlii root extract.

[0077] Comparative Example 2

[0078] The difference between this comparative example and Example 1 is that the hair blackening composition does not contain tetrahydropiperine.

[0079] Comparative Example 3

[0080] The difference between this comparative example and Example 1 is that the hair-blackening composition does not contain phenylalanine.

[0081] Comparative Example 4

[0082] The difference between this comparative example and Example 1 is that the hair-blackening composition does not contain tripeptide-1 copper.

[0083] Comparative Example 5

[0084] The difference between this comparative example and Example 1 is that the hair-blackening composition does not contain PCA zinc.

[0085] Comparative Example 6

[0086] The difference between this comparative example and Example 1 is that the formula of the black hair composition of this comparative example is: 50g of Coleus forskohlii extract, 100g of tetrahydropiperine, 5g of phenylalanine, 150g of copper compound, 5g of zinc compound, 300g of solubilizer, and water to make up the balance to 1000g.

[0087] Comparative Example 7

[0088] The difference between this comparative example and Example 1 is that the formula of the black hair composition of this comparative example is: 250g of Coleus forskohlii extract, 5g of tetrahydropiperine, 150g of phenylalanine, 5g of copper compound, 150g of zinc compound, 25g of solubilizer, and water to make up the balance to 1000g.

[0089] Comparative Example 8

[0090] The difference between this comparative example and Example 1 is that the extraction process of the Coleus forskohlii extract is conventional 30 times solid-liquid ratio boiling water reflux extraction for 1.5h, repeated twice and the combined filtrate is concentrated to the same weight of extract.

[0091] Comparative Example 9

[0092] The difference between this comparative example and Example 1 is that the Coleus forskohlii flower extract is not subjected to inclusion complexation with hydroxypropyl-β-cyclodextrin, and the Coleus forskohlii flower extract is directly filtered to remove water-insoluble matter to obtain the extract.

[0093] Experiment 1: Safety Test

[0094] (1) Test samples: The sample compositions obtained in Examples 1 to 9 and Comparative Example 6 were prepared into corresponding aqueous solutions at a mass fraction of 5%.

[0095] (2) Test population: 300 healthy subjects aged 20 to 50 years with no history of skin allergies were selected and randomly divided into 10 groups, with 30 people in each group.

[0096] (3) Test method: First, clean the skin of the test area with physiological saline and wait for it to dry for 5 minutes; select a qualified patch tester and use a closed patch test method. Take about 0.02g of the solution sample prepared in Examples 1 to 3 into the patch tester respectively, and apply it to the inner side of the subject's arm with special adhesive tape. The test area is prohibited from contacting water, exercising, scratching, etc. during the test. After 48 hours, remove the tester and mark it. After 30 minutes, when the indentation disappears, make a judgment under sufficient light. Record the results with reference to the skin reaction grade standard in the "Cosmetic Safety Technical Specifications 2015 Edition". The skin reaction grade standard is shown in Table 2 below.

[0097] (4) Test results: as shown in Table 3.

[0098] Table 2 Skin reaction grading standards

[0099]

[0100] Table 3 Safety test results

[0101]

[0102]

[0103] As can be seen from the results in Table 3, the skin patch test results of the hair-blackening compositions prepared in Examples 1 to 9 of the present invention at a usage amount of 5% by mass were all negative, indicating that the hair-blackening compositions prepared in Examples 1 to 9 of the present invention have no potential adverse reactions on human skin. In the test of Comparative Example 6, there were 3 suspicious reactions, 2 weak positive reactions and 2 strong positive reactions, which was mainly due to the irritation caused by tetrahydropiperine. Tetrahydropiperine has a penetration-promoting effect, and excessive addition will cause skin irritation. Therefore, from the above results, it can be seen that the composition of the present invention has the advantages of high safety and mild effect.

[0104] Experiment 2: In vitro testing

[0105] Tyrosinase activity enhancement rate: prepare 0.5 mg / mL L-tyrosine solution (solvent is phosphate buffer) and 35 mg / mL mushroom tyrosinase solution (solvent is phosphate buffer). Use propylene glycol as solvent to prepare 1 mg / mL test sample solution (test the black hair composition that promotes melanin secretion prepared in Examples 1 to 9 and the samples prepared in Comparative Examples 1 to 9 respectively) as a comparative test group.

[0106] Prepare 4 groups of solutions according to the following proportions:

[0107] A: 0.7 mL of L-tyrosine solution + 0.7 mL of pure propylene glycol + 0.6 mL of phosphate buffer;

[0108] B: 0.7 mL pure propylene glycol + 1.3 mL phosphate buffer;

[0109] C: 0.7 mL of L-tyrosine solution + 0.7 mL of propylene glycol solution of the test sample + 0.6 mL of phosphate buffer;

[0110] D: 0.7 mL of propylene glycol solution of the sample to be tested + 1.3 mL of phosphate buffer;

[0111] After the above four groups of solutions were kept in a 37°C water bath for 10 minutes, 0.3 mL of tyrosinase solution was added to each of them. After reacting for 5 minutes, the solution was quickly transferred to a cuvette and the absorbance (OD value) was measured at 475 nm. The OD value was recorded and the tyrosinase activation rate was calculated according to the formula = 100% × ((OD C -OD D )-(OD A -OD B )) / (OD A -OD B ); among them, (OD A -OD B ) represents the change in OD value of the reaction solution under the condition of no test sample, (OD C -OD D ) represents the change in OD value of the reaction solution under the conditions of the test samples (the black hair compositions provided in Examples 1 to 7 and Comparative Examples 1 to 9).

[0112] Melanin content change test: The density of B16 melanoma cell suspension was adjusted to 1×105 cells / mL, and 1 mL was inoculated into each well of a 12-well plate to make the cell density 5×10 4Each well of the experimental group and the zero-adjustment group was added with 1 mL of 1 μM α-MSH solution prepared with RPMI 1640 medium. The blank control group was added with 1 mL of medium without α-MSH. After 36 hours of incubation in an incubator, the cells were washed twice with 1 mL of PBS. 100 μL of 1N NaOH solution containing 10% DMSO (prepared with PBS) was added to each well. After 1 hour of water bath at 80°C, the cell extracts of each group in the 12-well plate were transferred to a 96-well plate. The absorbance OD value of each well was measured at 475 nm on a microplate reader. The formula for calculating the intracellular melanin content is: melanin content = 100% × (OD1-OD3) / (OD2-OD3), where OD1 is the absorbance value of the experimental group, OD2 is the absorbance value of the blank control group, and OD3 is the absorbance value of the zero-adjustment group.

[0113] According to the above experimental method, the effects of the black hair compositions provided in Examples 1 to 9 and Comparative Examples 1 to 9 on the tyrosinase activity enhancement rate and melanin content change were tested respectively. The specific test data are shown in Table 4.

[0114] Table 4 Tyrosinase activity enhancement rate and melanin content change results

[0115]

[0116]

[0117] According to the data in Table 4, the hair blackening composition provided by Examples 1 to 9 of the present invention has an improvement rate of tyrosinase activity of 13.2 to 29.6%, and increases the rate of change of melanin content in B16 melanoma cells to 9.24 to 13.20%. From the comparison of the tyrosinase improvement rate and the melanin content change rate in Examples 1 to 9, it can be seen that it not only promotes the production of melanin by improving tyrosinase, but also the various components of the composition of the present invention work synergistically with each other to promote melanin production, cover the target more comprehensively, and has good application prospects for hair blackening.

[0118] It can be seen from Examples 1 to 5, 8 and Comparative Examples 4, 6, and 7 that when the content of tripeptide-1 copper in the composition changes greatly, the tyrosinase activity is most affected, which may be because the coordinated copper ions in tripeptide-1 copper can interact with the histidine structure in tyrosinase to enhance the tyrosinase activity; According to the comparison between Example 1 and Comparative Examples 1 to 5, it can be seen that when any component in the composition is missing, the tyrosinase activity enhancement rate and the change in melanin content will decrease to a certain extent, which is mainly due to the different targets of the components in the composition. In the copper compound represented by tripeptide-1 copper, the tyrosinase activity enhancement rate and the melanin content change will decrease to a certain extent. It contains copper ions. The key catalytic site of tyrosinase is histidine residues, which will bind to copper ions. Therefore, it can help to enhance the activity of tyrosinase. Piperine can effectively enhance the expression of TRP-1 and thus enhance the production of melanin. In addition, zinc ions are the key metal ions for the enzyme activity of TRP-2 and can effectively enhance its activity. Forskolin, the active substance in the Coleus forskohlii extract, can induce the division of melanocytes, thereby promoting the production of melanin. Phenylalanine can be converted into tyrosine in the body through the action of phenylalanine hydroxylase, and then finally form melanin through the action of various melanin-related enzymes.

[0119] Experiment 3: Hair Blackening Effect Test

[0120] In this experiment, the black hair compositions prepared in Examples 1 to 3 and the black hair essences prepared from the samples in Comparative Examples 2 and 8 were tested for their black hair effects. The formula of the black hair essence is shown in Table 5. The raw materials in the formula can be directly mixed and stirred evenly at room temperature. The group without the composition is set as the conventional group.

[0121] Test method: 130 volunteers aged 18-65 years (with similar gray hair problems) were randomly divided into 13 groups, with 10 people in each group, for a 6-month hair care program. The volunteers received hair care every 3 days, and their gray hair rates were tested before, 3 months after, and 6 months after the hair care program. The gray hair rate was measured using a sensory evaluation method performed by professional salon assessors combined with a hair microscope. The experimental results are shown in Table 6 and Figure 2 , after 4 weeks of testing, some white hair turned black (refer to Figure 1 ).

[0122] Table 5 Black hair essence formula

[0123]

[0124]

[0125] Table 6 Evaluation effect of dark hair using Examples 1 to 3, Comparative Examples 1 to 9 and conventional group

[0126]

[0127] As can be seen from the results in Table 6, Examples 1 to 3 have good anti-gray hair and black hair effects. After using the essence containing the black hair composition, the regeneration of gray hair can be effectively inhibited, the hair can be gradually blackened, and the gray hair rate can be reduced. According to Example 3 and Comparative Example 2, it can be seen that when the tetrahydropiperine content is too low or lacking, the onset speed is significantly lower than that of Example 1. From Comparative Example 8, it can be seen that when the preparation process of the Coleus forskohlii extract is replaced or hydroxypropyl-β-cyclodextrin inclusion is not used, the use effect will be significantly reduced, which is sufficient to prove that the Coleus forskohlii extraction process of the present invention can effectively retain its effective ingredients.

[0128] The above description is only a preferred embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structure or equivalent process transformation made by using the contents of the present invention description and drawings, or directly or indirectly applied in other related technical fields, are also included in the patent protection scope of the present invention.

Claims

1. A black hair composition for promoting melanin secretion, characterized in that: The invention comprises the following raw materials in parts by weight: 10-20 parts of Coleus forskohlii extract, 1-5 parts of tetrahydropiperine, 1-10 parts of phenylalanine, 1-10 parts of copper compound, 1-10 parts of zinc compound, 5-25 parts of solubilizer, and the balance of water to 100 parts.

2. The black hair composition for promoting melanin secretion according to claim 1, characterized in that The invention comprises the following raw materials in parts by weight: 14-18 parts of Coleus forskohlii extract, 3-5 parts of tetrahydropiperine, 4-6 parts of phenylalanine, 3-8 parts of copper compound, 3-8 parts of zinc compound, 15-25 parts of solubilizer and the balance of water.

3. The composition for promoting melanin secretion in black hair according to claim 1 or 2, characterized in that: The Coleus forskohlii extract is obtained by extracting Coleus forskohlii through supercritical CO2 and then inclusion complexing with hydroxypropyl-β-cyclodextrin.

4. The composition for promoting melanin secretion in black hair according to claim 3, characterized in that The preparation process of the Coleus forskohlii flower extract comprises the following steps: crushing and sieving the Coleus forskohlii flower, using 95% ethanol as an entrainer in an amount of 0.1 to 0.5 times the amount of the Coleus forskohlii flower, setting the extraction temperature to 35 to 55° C., the extraction pressure to 20 to 40 MPa, the CO2 flow rate to 20 to 35 L / h, and the extraction time to 3 hours; setting the temperature of a separation kettle 1 to 30 to 40° C. and the pressure to 8 to 10 MPa, setting the temperature of a separation kettle 2 to 28 to 35° C. and the pressure to 5 to 8 MPa; recovering the entrainer under reduced pressure to obtain a Coleus forskohlii flower extract; adding the Coleus forskohlii flower extract and hydroxypropyl-β-cyclodextrin to a saturated hydroxypropyl-β-cyclodextrin solution at a ratio of 1 ml:3 to 4 g; carrying out inclusion reaction at 40° C. for 2 hours; precipitating, filtering, washing with anhydrous ethanol, and drying to obtain the extract.

5. The composition for promoting melanin secretion in black hair according to claim 1 or 2, characterized in that: The tetrahydropiperine is prepared by using pepper seed powder as a raw material, extracting it by reflux using methanol, and concentrating it in vacuum to obtain pepper oleoresin, which is then dissolved in a solution of isopropyl alcohol:n-hexane = 1:1 at 50-60°C, cooled to room temperature, and filtered to obtain a precipitated wet solid, which is dissolved in methanol, crystallized at 50°C, filtered, concentrated to 1 / 4 of the volume of methanol, cooled to 10°C, and filtered to obtain piperine crystals, which are then subjected to a hydrogenation reaction to obtain the obtained product.

6. The composition for promoting melanin secretion in black hair according to claim 5, characterized in that: The mass fraction ratio of the tetrahydropiperine to the solubilizer is 1:

5.

7. The composition for black hair that promotes melanin secretion according to claims 1 to 2, characterized in that: The copper compound is one or more of copper salts and copper ion complexes.

8. A method for preparing a black hair composition that promotes melanin secretion according to any one of claims 1 to 7, characterized in that: The following steps are involved: S1: pre-dispersing the solubilizing agent and the tetrahydropiperine and dissolving them at 40-50° C.; S2: slowly adding the Coleus forskohlii extract, phenylalanine, copper compound, and zinc compound into 40° C. water and stirring to dissolve; S3: Combine the above components and stir until dissolved to obtain the hair-blackening composition that promotes melanin secretion.

9. Use of the black hair composition that promotes melanin secretion according to any one of claims 1 to 7 in the preparation of black hair products.

10. A black hair product, characterized in that: A composition for promoting melanin secretion in black hair according to any one of claims 1 to 7, further comprising water, ethanol, a moisturizer and a preservative; Wherein, the mass percentage of the black hair composition that promotes melanin secretion in the black hair product is 0.5-5%.

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