A traditional Chinese medicine composition for blackening and growing hair, and its preparation method and application

Abstract

The present invention belongs to the field of cosmetics technology and provides a traditional Chinese medicine composition for blackening and growing hair, as well as its preparation method and application. The traditional Chinese medicine composition comprises the following raw materials by weight: 3-6 parts of Polygonum multiflorum, 3-6 parts of Angelica sinensis, 2-5 parts of Oroxylum indicum, and 3-5 parts of Lycium barbarum. The composition has the effect of alleviating the growth environment of hair follicles and promoting the production of melanin by melanocytes. Furthermore, the composition utilizes nanotechnology to facilitate scalp absorption. The preparation method of the traditional Chinese medicine composition of the present invention is simple and convenient, and is conducive to large-scale production and application.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics, and in particular to a traditional Chinese medicine composition for blackening and growing hair, and a preparation method and application thereof. Background Art

[0002] Gray hair, also known as gray hair or white hair, is a common physiological phenomenon, typically caused by a decrease in melanin. Melanin is a pigment produced in hair follicles that gives hair its natural color. With aging, melanin levels gradually decrease, causing hair to gray. In addition to age, genetics also play a role in the appearance of gray hair.

[0003] While gray hair is somewhat inevitable, there are ways to slow its onset or treat any gray hair that has already appeared.

[0004] Chinese patent CN105853276A discloses a natural herbal formula for blackening and delaying graying hair. The formula is made from 14 parts of processed Polygonum multiflorum, 2 parts of black sesame, 10 parts of Morus alba, 8 parts of Angelica sinensis, 5 parts of Platycladus orientalis leaves, 7 parts of Ecliptae chinensis, 6 parts of Platycladus arvense seeds, 4 parts of Menthol, 8 parts of Scutellaria baicalensis, 16 parts of Cuscuta australis, 2 parts of Angelica dahurica, 4 parts of Sargassum, 9 parts of Salvia miltiorrhiza, 9 parts of Ligustrum lucidum fruit, 16 parts of stir-fried Vitex rotundifolia fruit, 10 parts of Lycium barbarum fruit, 5 parts of Astragalus membranaceus, 3 parts of Smilax glabra, and 15 parts of Ginseng. This method uses numerous raw materials and has a complex preparation process. It has a blackening effect but no hair growth effect.

[0005] Chinese patent CN106977997A discloses a disease-preventing and health-promoting nanocoating using nanopowders from a secret traditional Chinese medicine formula for black hair. The ingredients, including Morinda officinalis, Cistanche deserticola, Asparagus cochinchinensis, Schisandra chinensis, Platycladus orientalis seeds, Achyranthes bidentata, Cuscuta australis, Polygala tenuifolia, Dendrobium candidum, Chinese yam, Saposhnikovia divaricata, Poria cocos, Panax ginseng, Rehmannia root, Rubus idaeus, Scutellaria baicalensis, Dioscorea paniculata, Acanthopanax acanthopanax bark, Tianxiong root, Chuanduan root, Photinia raphanus, Eucommia bark, Cnidium monnieri, and Aquilaria sinensis, are nano-pulverized. This method is complex, has high raw material costs, and offers limited hair-blackening and growth results. Summary of the Invention

[0006] The present invention aims to solve the technical problems of complex preparation process and general hair blackening and growth effect in the prior art, and provides a traditional Chinese medicine composition for hair blackening and growth, a preparation method and application thereof.

[0007] The technical solution adopted in the present invention is:

[0008] In a first aspect, the present invention provides a traditional Chinese medicine composition for blackening and growing hair.

[0009] Specifically, a traditional Chinese medicine composition for hair growth and development includes Polygonum multiflorum, Angelica sinensis, Oroxylum indica and Black Wolfberry;

[0010] Specifically, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 3-6 parts of Polygonum multiflorum, 3-6 parts of Angelica sinensis, 2-5 parts of Oroxylum indicum and 3-5 parts of Black Wolfberry;

[0011] Specifically, the traditional Chinese medicine composition comprises one or more of the following raw materials in parts by weight: 2-4 parts of Ligustrum lucidum fruit, 3-5 parts of ginseng, 3-5 parts of Platycladus orientalis leaves, and 3-4 parts of mint;

[0012] The traditional Chinese medicine composition for hair growth and development comprises Polygonum multiflorum, Chinese angelica, Oroxylum indica, black wolfberry, Ligustrum lucidum fruit, ginseng root, Platycladus orientalis leaf and mint;

[0013] The Chinese medicine composition is composed of the following raw materials in weight fractions: 3-6 parts of Polygonum multiflorum, 3-6 parts of Angelica sinensis, 2-5 parts of Oroxylum indicum, 3-5 parts of Black Wolfberry, 2-4 parts of Ligustrum lucidum, 3-5 parts of Ginseng, 3-5 parts of Platycladus orientalis leaves and 3-4 parts of Mint;

[0014] Preferably, the traditional Chinese medicine composition is composed of the following raw materials in weight fractions: 5 parts of Polygonum multiflorum, 5 parts of Angelica sinensis, 4 parts of Oroxylum indicum, 5 parts of Lycium barbarum, 3 parts of Ligustrum lucidum, 4 parts of Ginseng, 5 parts of Platycladus orientalis leaves and 3 parts of Mint;

[0015] Specifically, the angelica root can be selected from angelica root and angelica leaf;

[0016] Preferably, the angelica is angelica root;

[0017] Specifically, the Oroxylum indicum can be selected from Oroxylum indicum flowers, Oroxylum indicum roots and Oroxylum indicum bark;

[0018] Preferably, the Oroxylum butterfly is an Oroxylum butterfly root;

[0019] Specifically, the raw materials of the traditional Chinese medicine composition are in the form of Chinese herbal medicine plants and / or Chinese herbal medicine extracts.

[0020] The efficacy of each component of the traditional Chinese medicine composition for blackening and growing hair of the present invention is as follows:

[0021] Polygonum multiflorum is rich in nutrients such as amino acids, vitamins and minerals, which can promote the nutrition of hair follicles and enhance the growth ability of hair, thereby effectively promoting hair growth; it can nourish hair, replenish the nutrients needed by hair, improve hair dryness, frizziness and other problems, and make hair softer, smoother and more elastic; in addition, Polygonum multiflorum contains antioxidants, which can help protect hair from damage by free radicals, slow down the aging process of hair, and keep hair healthy, and it can also stimulate the production of melanin, help delay the process of hair graying, and maintain black hair color.

[0022] Dong quai has a certain effect on darkening hair. It contains active ingredients that can promote blood circulation to the hair follicles in the scalp, increasing nutrient supply to the follicles and helping to promote hair growth. Furthermore, it is rich in vitamins and minerals that nourish the hair, enhancing its health, elasticity, and quality. Furthermore, the antioxidant and anti-inflammatory properties of dong quai root can reduce hair loss and maintain healthy hair.

[0023] The main components of Oroxylum indicum are flavonoids, which have certain effects in anti-inflammatory, anti-allergic and other aspects.

[0024] Black goji berries have multiple benefits for dark hair. First, they are rich in nutrients that promote hair follicle growth and development, making hair thicker. Second, black goji berries have antioxidant, anti-inflammatory, and anti-aging properties, which can help reduce hair loss.

[0025] Ligustrum lucidum fruit is rich in vitamins C and E, which promote hair growth and enhance hair health. Its active ingredients also possess antioxidant and anti-inflammatory properties, helping to reduce hair loss and maintain healthy hair. Ligustrum lucidum fruit can also improve scalp condition, reducing dandruff and scalp inflammation, promoting hair growth and darkening.

[0026] Ginseng offers numerous benefits for hair health. It's rich in nutrients, such as ginsenosides, ginsenosides, and polysaccharides, which help nourish the hair and scalp, promote hair growth, and strengthen hair. It also possesses antioxidant and anti-inflammatory properties, helping to reduce hair damage and inflammation, protecting hair from environmental damage. Furthermore, ginseng can improve blood circulation to the scalp, improving scalp health and thus helping to prevent hair loss.

[0027] Platycladus orientalis leaves have the effects of cooling blood and stopping bleeding, removing phlegm and relieving coughs, and promoting hair growth and darkening. Studies have found that the resinous components in Platycladus orientalis leaves significantly activate TYR, making them the active ingredients in its hair-darkening effects. Furthermore, the petroleum ether fraction of Platycladus orientalis leaves has the highest activation rate for TYR, and this activation rate is concentration-dependent, with high concentrations upregulating TYR activity.

[0028] The main effects of mint include dispelling wind-heat, clearing the head and eyes, promoting qi circulation, soothing the liver, relieving sore throat, and clearing rashes. Its mechanism of action on black hair is mainly reflected in its ability to increase TYR activity, upregulate TYRmRNA levels, and promote melanocyte proliferation.

[0029] In a second aspect, the present invention provides a method for preparing a traditional Chinese medicine composition for blackening and growing hair, the preparation method comprising the following steps:

[0030] (1) The raw materials and water were mixed in a mass ratio of 1:3, heated at 90-110°C, and then paved and allowed to stand to obtain a sample solution;

[0031] (2) placing the sample solution obtained in step (1) in a vacuum microwave apparatus and heating it at 75-95° C. for 160-200 seconds to obtain a microwave-pretreated sample;

[0032] (3) The microwave pretreated sample obtained in step (2) was dissolved in 100 mL of ultrapure water by stirring to prepare a nanospray drying preparatory solution;

[0033] (4) The nano spray drying preliminary solution obtained in step (3) is atomized by a nano spray dryer and dried into nanoparticles at 60-80° C. in a spray drying cylinder.

[0034] Specifically, in step (1), when the raw material is in the form of a Chinese herbal medicine plant, it needs to be crushed;

[0035] The water in step (1) can be selected from distilled water and ultrapure water;

[0036] The vacuum microwave equipment in step (2) has a vacuum degree of 70 KPa.

[0037] In a third aspect, the present invention provides a use of a traditional Chinese medicine composition for blackening and growing hair in preparing cosmetics for blackening and growing hair.

[0038] In a fourth aspect, the present invention provides a cosmetic for blackening and growing hair;

[0039] Specifically, the cosmetics include a traditional Chinese medicine composition and excipients acceptable in cosmetics;

[0040] Specifically, the cosmetic is a product for use on the hair and / or scalp;

[0041] The mass percentage of the traditional Chinese medicine composition in the cosmetic is 0.01-10%;

[0042] Preferably, the mass percentage of the traditional Chinese medicine composition in the cosmetic is 0.01-2%;

[0043] The auxiliary materials include one or more of emulsifiers, thickeners, moisturizers, oils, chelating agents, and preservatives.

[0044] The emulsifier comprises one or more of organic olive oil emulsifying wax, self-emulsifying monoglyceride, methyl glucoside sesquistearate and glyceryl monostearate; and / or

[0045] The thickening agent comprises one or more of carbomer, polyacrylate cross-polymer and hydroxyethyl cellulose; and / or

[0046] The moisturizing agent comprises one or more of glycerin, propylene glycol and sodium hyaluronate; and / or

[0047] The oil comprises one or more of caprylic triglyceride, capric triglyceride, shea butter, isohexadecane, isopropyl palmitate, dimethicone and cyclopentasiloxane; and / or

[0048] The chelating agent includes disodium edetate; and / or

[0049] The preservatives include one or more of phenoxyethanol, methylparaben, propylparaben and sodium benzoate.

[0050] Compared with the prior art, the present invention has the following beneficial effects:

[0051] (1) The hair darkening and growth promoting composition provided by the present invention comprises a variety of plant ingredients, which are highly active, less irritating, and can effectively nourish hair follicles, activate tyrosinase, and promote the growth of melanocytes;

[0052] (2) The preparation method of the hair blackening and growth composition of the present invention is simple and convenient, which is conducive to large-scale production and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 The activity of the composition in Example 3 on B16 cells was measured, where ns indicates no statistical significance, * indicates p<0.05, with statistical significance; ** indicates p<0.01, with statistical significance; *** indicates p<0.001, with statistical significance; **** indicates p<0.0001, with statistical significance;

[0054] Figure 2 The effect of the composition of Example 3 on the melanin content of B16 melanoma cells, where ns indicates no statistical significance, * indicates p<0.05, there is a statistical difference; ** indicates p<0.01, there is a statistical difference; *** indicates p<0.001, there is a statistical difference; **** indicates p<0.0001, there is a statistical difference;

[0055] Figure 3 The effect of the composition of Example 3 on the tyrosinase activity of B16 melanoma cells, where ns indicates no statistical significance, * indicates p<0.05, with statistical significance; ** indicates p<0.01, with statistical significance; *** indicates p<0.001, with statistical significance; **** indicates p<0.0001, with statistical significance;

[0056] Figure 4 This is a photo of Volunteer 1's hair before using the composition;

[0057] Figure 5 Photo of Volunteer 1's hair after using the composition;

[0058] Figure 6Photo of volunteer 2's hair before using the composition;

[0059] Figure 7 This is a photo of volunteer 2's hair after using the composition. DETAILED DESCRIPTION

[0060] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only some embodiments of the present invention, rather than all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work are within the scope of protection of the present invention. The materials, reagents, etc. used, unless otherwise specified, are reagents and materials available from commercial sources.

[0061] Source of raw materials: Guangzhou Baizhitang Nanobiotechnology Co., Ltd.

[0062] 1. Example and comparative example

[0063] Preparation method of traditional Chinese medicine composition for blackening and growing hair

[0064] The raw materials were crushed and mixed with ultrapure water at a material-to-liquid ratio of 1:3. Stirring was continued at 100°C for 60 minutes. The samples were then allowed to stand for a period of time to allow the solution to fully penetrate the tissue. The fully infiltrated samples were placed in a vacuum microwave apparatus at 70 kPa and heated at 85°C for 180 seconds. The microwave-pretreated samples were dissolved in 100 mL of ultrapure water, stirred and dissolved at 60°C, and thoroughly mixed to prepare a preparatory solution for nanospray drying. The preparatory solution was then atomized using a nanospray dryer and dried in a spray dryer at 70°C to form nanoparticles.

[0065] Example 1 A Chinese medicine composition for blackening and growing hair

[0066] 3 parts of Polygonum multiflorum, 3 parts of Angelica sinensis roots, 2 parts of Oroxylum indicum roots and 3 parts of black wolfberry, prepared as above.

[0067] Example 2 A Chinese medicine composition for black hair growth

[0068] 6 parts of Polygonum multiflorum, 6 parts of Angelica sinensis roots, 5 parts of Oroxylum indicum roots and 5 parts of Black Wolfberry, prepared as above.

[0069] Example 3 A Chinese medicine composition for black hair growth

[0070] 5 parts of Polygonum multiflorum, 5 parts of Angelica sinensis roots, 4 parts of Oroxylum indicum roots, 5 parts of black wolfberries, 3 parts of Ligustrum lucidum fruits, 4 parts of Panax ginseng roots, 5 parts of Platycladus orientalis leaves, and 3 parts of mint, and the preparation method is as described above.

[0071] Example 4 A Chinese medicine composition for black hair growth

[0072] 6 parts of Polygonum multiflorum, 6 parts of Angelica sinensis roots, 4 parts of Ligustrum lucidum fruits, 5 parts of Oroxylum indicum roots, 5 parts of Panax ginseng roots, 3 parts of black wolfberries, 4 parts of Platycladus orientalis leaves, and 4 parts of mint. The preparation method is as described above.

[0073] Example 5 A Chinese medicine composition for black hair growth

[0074] 3 parts of Polygonum multiflorum, 3 parts of Angelica sinensis roots, 2 parts of Oroxylum indicum roots, 3 parts of black wolfberries, 2 parts of Ligustrum lucidum fruits, 3 parts of Panax ginseng roots, 3 parts of Platycladus orientalis leaves, and 3 parts of mint, prepared as described above.

[0075] Comparative Example 1

[0076] A single component, Polygonum multiflorum, is used, and the preparation method is as described above.

[0077] Comparative Example 2

[0078] A single component, angelica root, is used, and the preparation method is as described above.

[0079] Comparative Example 3

[0080] A single component, Oroxylum indicum root, is used, and the preparation method is as described above.

[0081] Comparative Example 4

[0082] A single component, black wolfberry, is used, and the preparation method is as described above.

[0083] Comparative Example 5

[0084] The difference from Example 1 is that it does not contain Polygonum multiflorum, and includes the following raw materials: including the following raw materials: 3 parts of Angelica sinensis roots, 2 parts of Oroxylum indicum roots and 3 parts of black wolfberries, and the preparation method is as described above.

[0085] Comparative Example 6

[0086] The difference from Example 1 is that it does not contain angelica root, and includes the following raw materials: including the following raw materials: 3 parts of Polygonum multiflorum, 2 parts of Oroxylum indicum roots and 3 parts of black wolfberry, and the preparation method is as described above.

[0087] Comparative Example 7

[0088] The difference from Example 1 is that the oroxylum indicum root is not contained, and the following raw materials are included: 3 parts of Polygonum multiflorum, 3 parts of Angelica sinensis root and 3 parts of black wolfberry, and the preparation method is as described above.

[0089] Comparative Example 8

[0090] The difference from Example 1 is that black wolfberry is not contained, and the following raw materials are included: 3 parts of Polygonum multiflorum, 3 parts of Angelica sinensis roots and 2 parts of Oroxylum indicum roots, and the preparation method is as described above.

[0091] Comparative Example 9

[0092] 2 parts of Polygonum multiflorum, 2 parts of Angelica sinensis root, 1 part of Oroxylum indicum root and 2 parts of black wolfberry, prepared as above.

[0093] Comparative Example 10

[0094] 7 parts of Polygonum multiflorum, 7 parts of Angelica sinensis roots, 6 parts of Oroxylum indicum roots and 6 parts of Black Wolfberry, prepared as above.

[0095] Comparative Example 11

[0096] 10 parts of Polygonum multiflorum, 7 parts of Angelica sinensis roots, 6 parts of Oroxylum indicum roots, 2 parts of black wolfberries, 1 part of Ligustrum lucidum fruits, 2 parts of Panax ginseng roots, 2 parts of Platycladus orientalis leaves, and 2 parts of mint, prepared as described above.

[0097] Comparative Example 12

[0098] 2 parts of Polygonum multiflorum, 2 parts of Angelica sinensis roots, 1 part of Oroxylum indicum roots, 6 parts of black wolfberries, 5 parts of Ligustrum lucidum fruits, 5 parts of Panax ginseng roots, 6 parts of Platycladus orientalis leaves, and 5 parts of mint, prepared as described above.

[0099] Comparative Example 13

[0100] Without using nanotechnology, a traditional preparation method is adopted, and the rest is the same as in Example 3.

[0101] Crush the herbs in a grinder, add 75% ethanol, and ultrasonicate in an ultrasonic cleaner for 1 hour. Vacuum filter the extract using a Büchner funnel. Repeat the process once, mixing the first and second vacuum-filtered extracts. Vacuum evaporate the extract on a rotary evaporator until it becomes a thick paste. Pour the concentrated extract into an evaporating dish and dry it in an oven until it becomes a solid.

[0102] Comparative Example 14

[0103] According to the technical solution disclosed in Chinese patent CN105853276A, 14 parts of processed Polygonum multiflorum, 2 parts of black sesame, 10 parts of mulberry, 8 parts of angelica, 5 parts of Platycladus orientalis leaves, 7 parts of Ecliptae, 6 parts of Platycladi seeds, 4 parts of mint, 8 parts of Scutellaria, 16 parts of Cuscuta seeds, 2 parts of Angelica dahurica, 4 parts of Sargassum, 9 parts of Salvia miltiorrhiza, 9 parts of Ligustrum lucidum, 16 parts of stir-fried Vitex rotundifolia, 10 parts of Lycium barbarum, 5 parts of Astragalus, 3 parts of Smilax glabra, and 15 parts of ginseng were ground into powder using a grinder. Crush; weigh 10g of the crushed medicinal material and put it into a conical flask; add 100ml of 75% ethanol; ultrasonicate it in an ultrasonic cleaner for 1 hour; vacuum filter the extract using a Buchner funnel; add 100ml of 75% ethanol; ultrasonicate it in an ultrasonic cleaner again for 1 hour; vacuum filter the extract using a Buchner funnel and mix it with the first extract; rotary evaporate the extract in a rotary evaporator until it becomes a thick paste; pour the concentrated extract into an evaporating dish and dry it in an oven to form a solid.

[0104] Composition effect test

[0105] Part I: Experimental Results

[0106] Performance test 1: DPPH free radical scavenging rate test

[0107] Free radicals can damage hair follicle cells, affect normal hair growth, and cause hair loss and gray hair. Therefore, the free radical scavenging abilities of Examples 1-5 and Comparative Examples 1-12 were tested as follows:

[0108] The traditional Chinese medicine composition for blackening and growing hair prepared in the comparative example of the embodiment was diluted with deionized water to a concentration of 0.25 mg / mL; 100 μL of the sample solution was added to a 96-well plate, and 100 μL of a 0.2 mmol / L DPPH anhydrous ethanol solution was added. After shaking evenly, the mixture was placed in a dark place at 25° C. to react for 30 minutes, and the absorbance A1 of the mixture was measured at a wavelength of 517 nm; 100 μL of anhydrous ethanol was used instead of the DPPH anhydrous ethanol solution and the same operation was performed; 100 μL of anhydrous ethanol was used instead of the sample and the same operation was performed; forskolin solution was used as a positive control, three parallel experiments were set up for each group, and the average value was calculated. The free radical scavenging ability of the traditional Chinese medicine composition for blackening and growing hair in different groups was calculated according to the DPPH scavenging rate formula.

[0109] DPPH clearance rate = [1-(A1-A2) / A3] × 100%

[0110] Where:

[0111] A1 is the absorbance value of 100 μL DPPH working solution + 100 μL sample. A2 is the absorbance value of 100 μL anhydrous ethanol + 100 μL sample.

[0112] A3 is the absorbance value of 100μL DPPH working solution + 100μL anhydrous ethanol. The test results can be found in Table 1.

[0113] Table 1 Test results of free radical scavenging ability of Examples 1-5 and Comparative Examples 1-14

[0114]

[0115]

[0116] As can be seen from Table 1, the compositions obtained in Examples 1-5 all have good DPPH free radical scavenging ability, with the highest scavenging rate reaching 96.99%, which is much better than that of Comparative Example 14 in the prior art.

[0117] Comparative Examples 1-4 are the effects of a single component, and Comparative Examples 5-8 are the effects of a composition removing one of Polygonum multiflorum, Angelica root, Oroxylum root, and Black Wolfberry. The free radical scavenging effect of the composition obtained in Examples 1-5 is better than that in Comparative Examples 1-8, indicating that the components have a synergistic effect and are indispensable.

[0118] The free radical scavenging effect of the compositions obtained in Examples 1-5 is better than that of Comparative Examples 9-12. The specific composition ratio of the Chinese medicine composition of the present invention enables Examples 1-5 to have better DPPH free radical scavenging ability.

[0119] Compared with Comparative Example 13 which does not use nanotechnology, Examples 1-5 have excellent DPPH free radical scavenging ability.

[0120] Performance Test 2: Tyrosinase Activation Rate

[0121] Prepare 0.5 mg / mL L-tyrosine solution (solvent is phosphate buffer), prepare 35 mg / mL mushroom tyrosinase solution (solvent is phosphate buffer), and use propylene glycol as solvent to prepare 1 mg / mL test sample solution.

[0122] Prepare 4 groups of solutions according to the following proportions:

[0123] (1) 0.7 mL of L-tyrosine solution + 0.7 mL of pure propylene glycol + 0.6 mL of phosphate buffer;

[0124] (2) 0.7 mL pure propylene glycol + 1.3 mL phosphate buffer;

[0125] (3) 0.7 mL of L-tyrosine solution + 0.7 mL of propylene glycol solution of the test sample + 0.6 mL of phosphate buffer;

[0126] (4) 0.7 mL of propylene glycol solution of the sample to be tested + 0.6 mL of phosphate buffer;

[0127] After the above four groups of solutions were kept constant temperature in a 37°C water bath for 10 minutes, 0.3 mL of mushroom tyrosinase solution was added to each, and 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 calculated according to the formula. Wherein, (OD1-OD2) represents the change in OD value of the reaction solution under the condition of no activator, and (OD3-OD4) represents the change in OD value of the reaction solution under the condition of activator (Examples 1-5, Comparative Examples 1-14 substances). The calculation formula for tyrosinase activation rate is:

[0128] Tyrosinase activation rate = 100% × [(OD3-OD4)-(OD1-OD2)] / (OD1-OD2)

[0129] The test results can be found in Table 2

[0130] Table 2 Test results of tyrosinase activation rates of Examples 1-5 and Comparative Examples 1-14

[0131] example Tyrosinase activation rate / % Example 1 18.85 Example 2 19.78 Example 3 25.33 Example 4 23.21 Example 5 20.88 Comparative Example 1 12.55 Comparative Example 2 11.33 Comparative Example 3 15.76 Comparative Example 4 10.22 Comparative Example 5 14.33 Comparative Example 6 16.23 Comparative Example 7 15.25 Comparative Example 8 16.21 Comparative Example 9 16.56 Comparative Example 10 17.88 Comparative Example 11 18.23 Comparative Example 12 15.33 Comparative Example 13 16.25 Comparative Example 14 15.23

[0132] As can be seen from Table 2, the compositions obtained in Examples 1-5 all exhibited good tyrosinase activation, with the highest activation rate reaching 25%. The tyrosinase activation efficacy of the compositions obtained in Examples 1-5 was significantly improved compared to the efficacy of the individual components used in Comparative Examples 1-4, and was also significantly improved compared to the efficacy of the compositions obtained in Comparative Examples 5-8 in which any component was removed. The components exhibited a strong synergistic effect, and the absence of any of the components significantly reduced the efficacy.

[0133] The compositions obtained in Examples 1-5 have better effects than Example 13 which does not use nanotechnology; the compositions obtained in Examples 1-5 have better effects than Example 14 of the prior art; the compositions obtained in Examples 1-5 have better effects than compositions obtained in other proportions, and the Chinese medicine compositions of the present invention have better tyrosinase activation ability.

[0134] Part II Cell Viability Test

[0135] The composition of Example 3 was subjected to a cell activity evaluation experiment.

[0136] Example 3 Determination of the activity of the composition on B16 mouse melanoma cells.

[0137] The operation was divided into a blank group, a negative control group, and a sample group. Each group had 6 parallel wells, and the process was repeated three times. The blank group was given normal culture medium without cells. The negative control group was given cells and normal culture medium. The sample group was given cells and the composition of Example 1 at different concentration gradients. The density of B16 cells in good growth condition was adjusted to 10 5 / mL, added to 96-well plates at 100μL / well, and cultured in a 37°C, 5% CO2 constant temperature incubator for 24 hours. After the cell monolayer adheres to the wall, prepare the sample solutions of various concentrations and add them to 96-well plates at 100μL / well, and culture in a 37°C, 5% CO2 constant temperature incubator for another 24 hours. Discard the supernatant, add 100μL MTT (0.5mg / mL) to each well, add 100μL DMSO after 4 hours, and shake on a 37°C shaker for 10 minutes to dissolve the crystals. Check the cell activity at 492nm and express it as cell survival rate. The results are as follows Figure 1 The calculation formula is as follows:

[0138] Cell survival rate = (A1-A2) / (A3-A2)×100%

[0139] Where:

[0140] A1: absorbance value of the sample group;

[0141] A2: absorbance value of blank group;

[0142] A3: absorbance value of negative control group.

[0143] By Figure 1 As shown, compared with the blank group, the sample with a concentration of 2.5 mg / mL had a certain toxicity to the cells. The cell survival rate at concentrations of 1.25 mg / mL and below reached over 90%. Among them, the cell survival rate at a concentration of 625.00 μg / mL was almost the same as that of the control, with a survival rate of 104.32%, showing no significant difference. As the concentration decreased, the cell survival rate gradually increased, reaching a maximum of 122.03% at a concentration of 156.25 μg / mL. The concentration then remained stable, indicating that the sample with a concentration of 625.00 μg / mL and below was non-toxic to the cells and had a certain cell proliferation effect. In addition, when observing cell morphology, there was no significant change in cell morphology in the sample group compared to the control group, and the cells showed normal appearance and morphological characteristics. This shows that the composition of the present invention is non-toxic to the scalp and hair follicles at a certain concentration and also has a certain hair growth effect.

[0144] Example 3: Determination of the effect of the composition on the melanin content of B16 mouse melanoma cells.

[0145] The tyrosinase activity in B16 mouse melanoma cells was measured by NaOH lysis method. The density of B16 melanoma cell suspension was adjusted to 1.8×10 6 On the basis of 1 mL of complete culture medium in each well, the cells were inoculated into 6-well plates with 1 mL of cell solution per well to make the cell density 3×10 5 / well, placed in an incubator (37 ℃, 5% carbon dioxide) and cultured for 24h. After the cells adhered, 100μL of test solution of different concentrations was added to each well as the sample group (according to the above experimental results, the concentration below 625.00μg / mL was used for operation), and 1mL of culture medium without the sample to be tested was added to the blank control group and the negative control group. After continuing to culture in the incubator for 48, the culture medium was discarded and washed twice with 1mL PBS. Add 100μL of 10% mass fraction DMSO and 1mol / L NaOH solution (accurately weigh 2g NaOH and dissolve it in 50mL deionized water, take 36mL and add 4mL DMSO), place in an 80℃ water bath for 45min to completely dissolve the cell clumps. After appropriate shaking, the cell extracts of each group in the 6-well plate were transferred to a 96-well plate, and the OD at a wavelength of 405nm was measured with an enzyme marker. The results are as follows Figure 2 shown.

[0146] The formula for calculating melanin content is as follows:

[0147] Melanin content = (OD1-OD3) / (OD2-OD3)×100%

[0148] Where:

[0149] OD1: absorbance value of the sample group;

[0150] OD2: absorbance value of negative control group;

[0151] OD3: absorbance value of the blank group.

[0152] By Figure 2 As can be seen, the 1.25 mg / mL concentration of the sample was lower than the control compared to the blank group, likely because at this concentration, the sample caused some damage to the cells, resulting in a lower melanin content. At a sample concentration of 156.25 μg / mL, the multiple plant extract composition provided in Example 3 of the present invention increased the melanin content in B16 melanoma cells to 113.20%, with a statistically significant difference (P < 0.0001). These results suggest the potential role of the composition in Example 3 in promoting melanin production. Melanin production is directly related to hair color, so this finding suggests that the composition at this ratio can help increase hair pigmentation, darken hair, or improve hair color uniformity. This effect is beneficial for improving gray hair, slowing the graying process, and enhancing the overall appearance of hair.

[0153] Example 3: Determination of the effect of the composition on the tyrosinase production rate of B16 mouse melanoma cells.

[0154] Tyrosinase activity in B16 mouse melanoma cells was measured by L-DOPA oxidation. The density of B16 cells in good growth condition was adjusted to 10 5 100 μL / well of the 5% 5% PBS solution was added to a 96-well plate and incubated at 37°C in a 5% CO2 incubator for 24 hours. B16 mouse melanoma cells were divided into a negative control group, a sample group, and a blank group, with three parallel wells set up in each group. The experiment was repeated three times. After the cells attached, 100 μL of normal culture medium was added to the negative control group, and 100 μL of culture medium containing different concentrations of the composition of Example 1 was added to the sample group. After incubation for 48 hours, the supernatant was discarded, the cells were rinsed once with PBS, and 50 μL of 1% Triton-X100 aqueous solution was added to each well. The cells were quickly frozen at -80°C for 30 minutes, removed, and thawed at 37°C (approximately 20 minutes) to completely rupture the cells. Then, 100 μL / well of 0.1% L-DOPA solution (0.1% L-DOPA solution: 0.01 g of L-DOPA powder dissolved in 10 mL of PBS) was added and incubated at 37°C for 2 hours. The OD value was measured at 475 nm using an enzyme-labeled instrument. Figure 3 The formula for calculating tyrosinase activity in B16 melanoma cells is as follows:

[0155] Tyrosinase activity (%) = (OD1-OD3) / (OD2-OD3) × 100%

[0156] Where:

[0157] OD1: absorbance value of the sample group;

[0158] OD2: absorbance value of negative control group;

[0159] OD3: absorbance value of the blank group.

[0160] Depend on Figure 3 The results show that compared with the blank group, the tyrosinase activation rates at different concentrations were 106.40%, 118.53%, 109.43%, 104.22%, and 96.67%, respectively. Except for the last two concentrations, the other concentrations showed statistical differences (P<0.0001). Compared with the above-mentioned melanin experiment, it was found that the trend of change in melanin content was consistent with the trend of change in tyrosinase activity. At a concentration of 156.25 μg / mL, the tyrosinase activity was the highest. At 1.25 mg / mL, due to certain cytotoxicity, the tyrosinase activity was the lowest. Therefore, the mechanism of the black hair effect of the composition of the present invention is to promote tyrosinase activity, increase the content of melanin production, promote melanin production in hair follicles, and pigment the hair color to appear black.

[0161] Part III: Safety and efficacy testing of application formula

[0162] Example 3: Human hair darkening test of the composition.

[0163] Twenty volunteers aged 35-50 with gray hair were selected and smeared with the composition of Example 3 above, 3 mL per night, for 3 minutes. After 4 months of continuous use, the subjects were observed and scored, and the average score was calculated. The evaluation criteria for the observation and scoring were as follows:

[0164] The highest score is 5 points and the lowest score is 1 point.

[0165] 5 points: All new hair turns black;

[0166] 4 points: The roots of the new hair turn black and the part above the roots is black;

[0167] 3 points: The roots of the new hair are black, and the part above the roots is light black;

[0168] 2 points: The roots of the new hair are light black, and the part above the roots is basically white;

[0169] 1 point: No black color is observed at the root of the new hair or above. This score can be rounded to a non-integer value depending on the actual situation. The results are shown in Table 3.

[0170] Table 3 Volunteer usage ratings

[0171] volunteer Fraction volunteer Fraction 1 3.7 11 3.4 2 3.6 12 3.8 3 4.0 13 3.5 4 3.8 14 4.0 5 4.1 15 3.9 6 3.5 16 4.1 7 3.6 17 4.3 8 3.8 18 3.1 9 3.5 19 3.8 10 3.1 20 3.6

[0172] From Table 3 and Figure 4-Figure 7 The results show that the composition of Example 3 has an average score of 3.71. After continuous use, the composition of the present invention shows a good hair darkening effect, which can make the roots of white hair darken and the hair above the roots darken in 4 months.

[0173] These results demonstrate that the hair-blackening and hair-growth-promoting compositions of the present invention have multiple benefits on hair follicles and the scalp. They not only promote hair growth and nourish hair, but also stimulate melanin production in hair follicles, darkening hair color and making it appear healthier and more lustrous. The various natural plant compositions of the present invention are mild and non-irritating, leave a refreshing feel on the skin, and are highly safe for long-term use. Furthermore, the present invention utilizes nanotechnology to produce products with higher levels of active ingredients, resulting in better delivery and better absorption by hair follicles, maximizing the effects of promoting dark hair. Furthermore, the method for preparing the hair-blackening and hair-growth-promoting cosmetics provided by the present invention is simple and convenient, facilitating large-scale production and application.

[0174] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, without departing from the principles of the present invention, several modifications or equivalent substitutions can be made to the technical solution, and these modifications or equivalent substitutions should also be regarded as the scope of protection of the present invention.

Claims

1. A Chinese medicine composition for blackening and growing hair, characterized in that: The invention is composed of the following raw materials in parts by weight: 3-6 parts of Polygonum multiflorum, 3-6 parts of angelica root, 2-5 parts of Oroxylum indicum root, 3-5 parts of black wolfberry, 2-4 parts of Ligustrum lucidum fruit, 3-5 parts of ginseng, 3-5 parts of Platycladus orientalis leaf and 3-4 parts of mint.

2. The Chinese medicine composition according to claim 1, characterized in that The invention is composed of the following raw materials in parts by weight: 5 parts of Polygonum multiflorum, 5 parts of angelica root, 4 parts of Oroxylum indicum root, 5 parts of black wolfberry, 3 parts of Ligustrum lucidum fruit, 4 parts of ginseng, 5 parts of Platycladus orientalis leaf and 3 parts of mint.

3. A method for preparing the Chinese medicine composition according to any one of claims 1 to 2, characterized in that: The preparation method comprises the following steps: (1) Mix the raw materials with water, heat them, and then lay them aside to obtain a sample solution; (2) heating the sample solution obtained in step (1) by microwave heating to obtain a microwave pretreated sample; (3) stirring and dissolving the microwave pretreated sample obtained in step (2) in ultrapure water to prepare a nanospray drying preparatory solution; (4) Atomizing and drying the nano spray drying preliminary solution obtained in step (3) to obtain nanoparticles, that is, obtaining the traditional Chinese medicine composition.

4. Use of the traditional Chinese medicine composition according to any one of claims 1 to 2 or the traditional Chinese medicine composition prepared by the preparation method according to claim 3 in the preparation of cosmetics.

5. A cosmetic for blackening and growing hair, characterized in that: The cosmetic is prepared from the Chinese medicine composition according to any one of claims 1 to 2 or the Chinese medicine composition prepared by the preparation method according to claim 3 and excipients acceptable in cosmetics.

6. The cosmetic according to claim 5, characterized in that The auxiliary material is selected from one or more of an emulsifier, a thickener, a moisturizer, a grease, a chelating agent and a preservative.

7. The cosmetic according to claim 6, characterized in that The emulsifier is selected from one or more of organic olive oil emulsifying wax, self-emulsifying monoglyceride, methyl glucoside sesquistearate and glyceryl monostearate; and / or The thickener is selected from one or more of carbomer, polyacrylate cross-linked polymer and hydroxyethyl cellulose; and / or The moisturizing agent is selected from one or more of glycerin, propylene glycol and sodium hyaluronate; and / or The oil is selected from one or more of caprylic triglyceride, capric triglyceride, shea butter, isohexadecane, isopropyl palmitate, dimethicone and cyclopentasiloxane; and / or The chelating agent is selected from disodium edetate; and / or The preservative is selected from one or more of phenoxyethanol, methylparaben, propylparaben and sodium benzoate.

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