A composition with black hair efficacy and its use

Abstract

This invention belongs to the technical field of daily-use shampoo and hair care raw materials, specifically relating to a composition with hair-darkening effects and its application. The composition with hair-darkening effects provided by this invention is formulated from a compound of Platycladus orientalis leaf extract, Polygonum multiflorum extract, Eclipta prostrata leaf extract, Gynostemma pentaphyllum extract, palmitoyl tetrapeptide-10, L-arginine, and decapeptide-4. By adjusting the types and amounts of each component, the interaction between the components is improved, thereby increasing the relative activity of tyrosinase and the relative content of melanin, indicating that the components have a synergistic effect and excellent hair-darkening efficacy. The hair-darkening composition obtained by this invention is applied to the preparation of shampoo, which exhibits excellent hair-darkening effects and high safety, and has broad market application value in the field of hair-darkening or hair-protecting products.

Description

Technical Field

[0001] This invention belongs to the technical field of daily shampoo and hair care raw materials, specifically relating to a composition with hair-darkening effects and its application. Background Technology

[0002] Hair is part of the human body, an organism composed of cells. With societal development and a faster pace of life, people face increasing work and life pressures, leading to a decline in immunity and an increase in gray hair. Aside from genetic factors, graying hair is mainly related to declining bodily functions, heredity, mental stress, excessive mental exertion, and malnutrition.

[0003] Currently, drugs used to promote the reversal of gray hair color include minoxidil and finasteride. However, these drugs often have certain side effects, such as scalp allergies, itching, and dandruff, making them unsuitable for all patients, and long-term use may lead to dependence. Compared with Western medicine, natural medicines, herbal medicines, and traditional Chinese medicine formulas are gradually gaining attention in the field of hair darkening. Many Chinese medicinal herbs, such as angelica, black sesame, he shou wu, and mulberry, have long been used in dietary therapy or topical formulas for hair growth and darkening, with a long history of traditional application. However, their effects are often unsatisfactory, and they are expensive and require extremely long treatment courses.

[0004] Therefore, there is an urgent need to develop a well-matched composition that has excellent hair-darkening effects and is safe and non-irritating, in order to meet market demand. Summary of the Invention

[0005] In view of the above-mentioned defects or improvement needs of the prior art, the present invention provides a composition with hair-darkening effect and its application, solving the problems of strong irritation and poor white-to-black effect of existing hair-darkening products.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides a composition having hair-darkening effects, comprising, by weight, the following components: Platycladus orientalis leaf extract: 10-15 parts; Polygonum multiflorum extract: 13-17 parts; Eclipta prostrata leaf extract: 21-25 parts; Gynostemma pentaphyllum extract: 22-27 parts; Palmitoyl tetrapeptide-10: 7-9 parts; L-arginine: 6-9 parts; Decapeptide-4: 9-13 parts.

[0007] The extract of Platycladus orientalis leaves mainly contains flavonoids and resins such as quercetin and kaempferol. The resins have an activating effect on tyrosinase, which is the effective component for its hair-darkening effect. In addition, the extract of Platycladus orientalis leaves also has the effects of cooling blood and stopping bleeding, and improving the scalp microenvironment.

[0008] Polygonum multiflorum extract contains abundant lecithin, emodin, rhein, and other components, which can promote the proliferation of melanocytes, improve cell activity, and have significant anti-aging, immunity-enhancing, lipid-lowering, antibacterial, liver and kidney tonifying, essence and blood nourishing, and hair darkening effects.

[0009] The extract of *Eclipta prostrata* leaves is rich in terpenoids such as pentacyclic triterpenes, pentacyclic triterpenoid saponins, diterpenes, and sesquiterpenes. In addition, it also contains active ingredients such as aromatic heterocyclic compounds and steroidal alkaloids, which can improve cell activity and have excellent hair-darkening effects.

[0010] The main active ingredient in Gynostemma pentaphyllum extract is gypenosides, which have a variety of medicinal effects, including sedation, hypnosis, relieving tension, anti-allergy, hair darkening, blood pressure reduction, and treatment of migraines.

[0011] Palmitoyl tetrapeptide-10 is a small molecule polypeptide that can stimulate the production of natural hair pigment, gradually restoring natural hair color, and also resist hair aging and prevent gray hair.

[0012] L-arginine, also known as protein amino acid, is a small molecule amino acid with antioxidant properties. Its small molecular weight can reduce sebum secretion, thus having a certain oil-controlling effect, and it also has good permeability.

[0013] Decapeptide-4 is a synthetic peptide that can stimulate the proliferation of collagen, elastin, and hyaluronic acid, improve the scalp's moisture content and hydration, and penetrate deep into the scalp to prevent hair loss and promote hair growth.

[0014] Preferably, the composition comprises, by weight, the following components: Platycladus orientalis leaf extract: 13 parts; Polygonum multiflorum extract: 15 parts; Eclipta prostrata leaf extract: 23 parts; Gynostemma pentaphyllum extract: 25 parts; Palmitoyl tetrapeptide-10: 8 parts; L-arginine: 7 parts; Decapeptide-4: 11 parts.

[0015] Secondly, the present invention provides the use of the composition described in the first aspect in the preparation of a hair care product with hair-darkening effects.

[0016] Preferably, the hair care product is one of shampoo, scalp care solution, or conditioner.

[0017] Preferably, the amount of the composition added to the shampoo and conditioner is 0.5wt%-5wt%.

[0018] Thirdly, the present invention provides a shampoo with hair-darkening effects, wherein the shampoo contains the composition described in the first aspect.

[0019] Preferably, the amount of the composition added to the shampoo is 1wt%-4wt%.

[0020] Preferably, the shampoo further contains at least one of the following: surfactant, hair conditioner, pH adjuster, moisturizer, antioxidant, thickener, chelating agent, emulsifying stabilizer, fragrance, and solvent.

[0021] The beneficial effects of this invention are: This invention provides a composition with hair-darkening effects. By adjusting the types and amounts of each component, the interaction between the components is improved. The composition prepared by combining Platycladus orientalis leaf extract, Polygonum multiflorum extract, Eclipta prostrata leaf extract, Gynostemma pentaphyllum extract, palmitoyl tetrapeptide-10, L-arginine and decapeptide-4 can increase the relative activity of tyrosinase and the relative content of melanin, indicating that the components have a synergistic effect and have excellent hair-darkening effects.

[0022] The hair-darkening composition obtained by this invention is applied to the preparation of shampoo. The shampoo has excellent hair-darkening effect and high safety, and has broad market application value in the field of hair-darkening and hair care products. Detailed Implementation

[0023] To further illustrate the technical means and effects of the present invention in achieving the intended purpose, the following detailed description of the specific embodiments, structure, features and effects of the present invention, in conjunction with the composition, is provided below.

[0024] For the following compositions, unless otherwise specified, experimental methods are generally performed under standard conditions or as recommended by the manufacturer. Unless otherwise specified, all materials and reagents used are commercially available.

[0025] The sources of some of the raw materials for this invention are as follows: The Polygonum multiflorum extract was prepared according to the preparation method of Chinese Patent CN121003578B, "A process for separating and purifying active ingredients of Polygonum multiflorum". The leaves of Platycladus orientalis were purchased from Yulin Hongjian Chinese Medicinal Materials Co., Ltd. The leaves of *Eclipta prostrata* were purchased from Hongjian Chinese Medicinal Materials Co., Ltd. in Yulin City, and the commercial name is *Eclipta prostrata*. Gynostemma pentaphyllum was purchased from Yulin Hongjian Chinese Medicinal Materials Co., Ltd. Palmitoyl tetrapeptide-10 was purchased from Guangzhou Huilingdun Trading Co., Ltd. L-arginine was purchased from Guangzhou Huilingdun Trading Co., Ltd. Decapeptide-4 was purchased from Guangzhou Huilingdun Trading Co., Ltd.

[0026] The method for preparing the Platycladus orientalis leaf extract described in this invention includes the following steps: (1) Take 10 kg of dried arborvitae leaves, add 9 times the total weight of the arborvitae leaves in purified water, and soak in a sealed container at room temperature for 30 min; (2) After heating to 75℃ at a rotation speed of 25 r / min, extract for 3h at an ultrasonic frequency of 50Hz and an ultrasonic power of 1500W. (3) After extraction, while still hot, use a 100-mesh filter for coarse filtration, collect the filtrate and let it stand in a sealed container at room temperature for 24 hours. (4) Take the supernatant after standing, centrifuge and filter it using a 1000-mesh filter bag at 1650 r / min for 30 min, and collect the filtrate; (5) After sterilizing the filtrate at 95℃ for 30 min, cool it down to 45℃, add 5% glycerol and 0.6% phenoxyethanol of the total mass of the filtrate, stir until completely mixed, filter with a 0.22μm microporous membrane, discharge, seal and store to obtain the Platycladus orientalis leaf extract.

[0027] The preparation method of the *Eclipta prostrata* leaf extract described in this invention includes the following steps: (1) Take 10 kg of dried Eclipta prostrata leaves, add 9 times the weight of Eclipta prostrata leaves in purified water, and soak in a sealed container at room temperature for 30 min; (2) After heating to 75℃ at a rotation speed of 25 r / min, extract for 3h at an ultrasonic frequency of 50Hz and an ultrasonic power of 1500W. (3) After extraction, while still hot, use a 100-mesh filter for coarse filtration, collect the filtrate and let it stand in a sealed container at room temperature for 24 hours. (4) Take the supernatant after standing, centrifuge and filter it using a 1000-mesh filter bag at 1650 r / min for 30 min, and collect the filtrate; (5) After sterilizing the filtrate at 95℃ for 30 min, cool it down to 45℃, add 5% glycerol and 0.6% phenoxyethanol of the total mass of the filtrate, stir until completely mixed, filter with a 0.22μm microporous membrane, discharge, seal and store to obtain the guarana leaf extract.

[0028] The preparation method of Gynostemma pentaphyllum extract described in this invention includes the following steps: (1) Take 10 kg of dried Gynostemma pentaphyllum, add 9 times the weight of Gynostemma pentaphyllum in purified water, and soak in a sealed container at room temperature for 30 min; (2) After heating to 85℃ at a rotation speed of 25 r / min, extract for 3h at an ultrasonic frequency of 50Hz and an ultrasonic power of 1500W. (3) After extraction, while still hot, use a 100-mesh filter for coarse filtration, collect the filtrate and let it stand in a sealed container at room temperature for 24 hours. (4) Take the supernatant after standing, centrifuge and filter it using a 1000-mesh filter bag at 1650 r / min for 30 min, and collect the filtrate; (5) After sterilizing the filtrate at 95℃ for 30 min, cool it down to 45℃, add 5% glycerol and 0.6% phenoxyethanol of the total mass of the filtrate, stir until completely mixed, filter with a 0.22μm microporous membrane, discharge, seal and store to obtain the Gynostemma pentaphyllum extract.

[0029] The component composition (parts by weight) of the compositions in Examples 1-3 of this invention is shown in Table 1 below. Each component was weighed according to the formula amount, mixed and homogenized to obtain each composition.

[0030] Table 1. Components and their weight parts of the compositions in Examples 1-3 Example Platycladus orientalis leaf extract (parts by weight) Polygonum multiflorum extract (parts by weight) Eclipta prostrata leaf extract (parts by weight) Gynostemma pentaphyllum extract (parts by weight) Palmitoyl Tetrapeptide-10 (parts by weight) L-Arginine (parts by weight) Decapeptide-4 (parts by weight) 1 13 15 23 25 8 7 11 2 10 13 21 22 7 6 9 3 15 17 25 27 9 9 13 Comparative Example 1 Compared with Example 1, the difference is that no eclipta prostrata leaf extract was added in Comparative Example 1, and the missing amount was made up by Gynostemma pentaphyllum extract and palmitoyl tetrapeptide-10 in a weight ratio of 25:8. All other conditions and preparation methods are the same as in Example 1.

[0031] Comparative Example 2 Compared with Example 1, the difference is that Gynostemma pentaphyllum extract was not added in Comparative Example 2, and the missing amount was made up with Eclipta prostrata leaf extract and palmitoyl tetrapeptide-10 in a weight ratio of 23:8. All other conditions and preparation methods were the same as in Example 1.

[0032] Comparative Example 3 Compared with Example 1, the difference is that palmitoyl tetrapeptide-10 was not added in Comparative Example 3, and the missing amount was made up by extracts of Eclipta prostrata and Gynostemma pentaphyllum in a weight ratio of 23:25. All other conditions and preparation methods are the same as in Example 1.

[0033] Comparative Example 4 Compared with Example 1, the difference is that L-arginine was not added in Comparative Example 4, and the missing amount was made up with decapeptide-4. All other conditions and preparation methods are the same as in Example 1.

[0034] Comparative Example 5 Compared with Example 1, the difference is that decapeptide-4 was not added in Comparative Example 5, and the missing amount was made up with L-arginine. All other conditions and preparation methods are the same as in Example 1.

[0035] Comparative Example 6 Compared with Example 1, the difference is that L-arginine in Comparative Example 6 is replaced with an equal amount of lysine, while other conditions and preparation methods are the same as in Example 1.

[0036] Comparative Example 7 Compared with Example 1, the difference is that the weight ratio of Eclipta prostrata leaf extract, Gynostemma pentaphyllum extract and palmitoyl tetrapeptide-10 in Comparative Example 7 is 8:25:23, while other conditions and preparation methods are the same as in Example 1.

[0037] Comparative Example 8 Compared with Example 1, the difference is that the weight ratio of L-arginine to decapeptide-4 in Comparative Example 8 is 13:7, while other conditions and preparation methods are the same as in Example 1.

[0038] Example 1 of efficacy test: Composition test The formation of gray hair is closely related to the decrease in melanin content and insufficient tyrosinase activity. Therefore, this efficacy test case measures the composition prepared in Examples 1-3 and Comparative Examples 1-8 of this application to measure B16 cell viability, relative intracellular tyrosinase activity and relative melanin content, thereby evaluating the hair-darkening effect of the composition.

[0039] Test objective: To investigate the effects of the composition on cell viability, tyrosinase activity, and melanin content using mouse melanoma B16 cells (which are structurally similar to human melanocytes and are commonly used cell models in pigment-related studies). Experimental materials: Cells: Mouse melanoma B16 cells; Reagents: DMEM medium, CCK8 kit, PBS buffer containing 1% Triton X100, 2 mmol / L L-DOPA solution (prepared with 0.1 mol / L PBS buffer at pH 6.8), 1 mol / L NaOH solution (containing 10% dimethyl sulfoxide), 0.25% trypsin-EDTA; Sample preparation for the experimental group: The compositions of Examples 1-3 and Comparative Examples 1-8 were diluted with DMEM medium to a sample solution with a mass fraction of 0.4%; Cell culture: B16 cells were placed in a special culture medium and cultured at 37°C and 5% CO2. Cells in the logarithmic growth phase were used for experiments. 1) B16 cell viability assay: Resuspend cells in DMEM medium and adjust cell concentration to 1×10⁶. 5Cells / mL were seeded at 100 μL per well in a 96-well plate and cultured at 37℃ and 5% CO2 for 24 h. The culture medium was removed and the plates were washed twice with PBS buffer. 100 μL of each experimental group sample was added to each well, and 100 μL of DMEM culture medium was added to each well of the blank control group. Each group was set up in 3 replicates and cultured for another 48 h. 10 μL of CCK8 solution was added to each well and incubated for 0.5 h. After shaking for 1 min with a microplate reader, the absorbance was measured at 450 nm. ; In the formula: the absorbance of the experimental group is the average absorbance of each group in the experimental group; the absorbance of the blank group is the average absorbance of the blank group. 2) Tyrosinase activity assay: Resuspend cells in DMEM medium and adjust cell concentration to 1×10⁻⁶. 5 Cells were seeded at 100 μL per well in a 96-well plate and cultured at 37°C and 5% CO2 for 24 h. The culture medium was removed, and the cells were washed twice with PBS buffer. 100 μL of each experimental group sample was added to each well, and 100 μL of DMEM culture medium was added to each well of the blank control group. Each group was divided into 3 replicates, and the cells were cultured for another 48 h. After culture, the culture medium was removed, and the cells were washed twice with PBS buffer. 150 μL of PBS buffer containing 1% Triton X100 was added to each well, and the cells were frozen at -80°C for 40 min, thawed at room temperature, and the freeze-thaw cycle was repeated 3 times to lyse the cells. 100 μL of the supernatant was centrifuged and transferred to a new 96-well plate. 60 μL of 2 mmol / L L-DOPA solution was added, and the cells were incubated at 37°C in the dark for 20 min. The absorbance was measured at 475 nm. ; In the formula: the absorbance of the experimental group is the average absorbance of each group in the experimental group; the absorbance of the blank group is the average absorbance of the blank group. 3) Melanin content determination: Resuspend cells in DMEM medium and adjust cell concentration to 1×10⁻⁶. 5 Cells / mL were seeded at 1.5 mL per well in 6-well plates and cultured at 37°C and 5% CO2 for 24 h. The culture medium was removed, and the cells were washed twice with PBS buffer. 1.5 mL of each experimental group sample was added, and 1.5 mL of DMEM medium was added to the blank control group. Three replicates were set up for each group. After 48 h of further culture, the culture medium was removed, and the cells were washed twice with PBS buffer. 1.5 mL of 0.25% trypsin-EDTA was added to each well for digestion, and the cell slurry was collected, centrifuged, and the supernatant was discarded. 200 μL of 1 mol / L NaOH solution (containing 10% dimethyl sulfoxide) was added to each well, and the cells were incubated at 80°C for 1 h. The absorbance was measured at 405 nm. ; In the formula: the absorbance of the experimental group is the average absorbance of each group in the experimental group; the absorbance of the blank group is the average absorbance of the blank group. Table 2 Test data of the composition Group Relative cell viability (%) Relative tyrosinase activity (%) Relative melanin content (%) Example 1 99.37 142.78 147.62 Example 2 95.81 140.65 145.79 Example 3 97.63 139.20 144.16 Comparative Example 1 96.42 120.67 127.24 Comparative Example 2 93.28 122.74 130.37 Comparative Example 3 98.34 121.36 128.45 Comparative Example 4 94.96 127.25 134.19 Comparative Example 5 99.15 126.53 133.40 Comparative Example 6 97.60 130.21 136.86 Comparative Example 7 95.13 134.49 139.25 Comparative Example 8 96.38 137.64 141.57 As shown in Table 2, the relative viability of B16 cells in Examples 1-3 and Comparative Examples 1-8 is ≥93%, indicating that the composition prepared in this invention is non-toxic to B16 cells and has good safety.

[0040] Compared with Comparative Examples 1-3, the relative activity of tyrosinase and the relative content of melanin in Example 1 were both increased, indicating that the extracts of Eclipta prostrata, Gynostemma pentaphyllum, and palmitoyl tetrapeptide-10 had a synergistic effect and had a hair-darkening effect. Compared with Comparative Examples 4-6, the relative activity of tyrosinase and the relative content of melanin in Example 1 were both increased, indicating that L-arginine and decapeptide-4 synergistically enhanced tyrosinase activity and melanin content.

[0041] The difference between Examples 1-3 and Comparative Examples 7 and 8 lies in the different weight ratios of *Eclipta prostrata* extract, *Gynostemma pentaphyllum* extract, palmitoyl tetrapeptide-10, L-arginine, and decapeptide-4. Based on the data from Examples 1-3 and Comparative Examples 7-8 in Table 2, the composition exhibits better effects when the weight ratio of *Eclipta prostrata* extract, *Gynostemma pentaphyllum* extract, and palmitoyl tetrapeptide-10 is (21-25):(22-27):(7-9), and the weight ratio of L-arginine and decapeptide-4 is (6-9):(9-13). Furthermore, the composition shows the best effect when the weight ratio of *Eclipta prostrata* extract, *Gynostemma pentaphyllum* extract, and palmitoyl tetrapeptide-10 is 23:25:8, and the weight ratio of L-arginine and decapeptide-4 is 7:11.

[0042] Application example: A shampoo with hair-darkening properties The compositions prepared in Examples 1-3 and Comparative Examples 1-8 were added to shampoos to obtain shampoos of Application Examples 1-3 and Comparative Application Examples 1-8, respectively.

[0043] The hair-darkening shampoo comprises the following components in weight percentages: 4% composition, 1.2% lauryl hydroxysulfonate betaine, 2% cocamidopropyl betaine, 0.6% disodium cocoamphodiacetate, 1.5% decyl glucoside, 0.03% sodium salicylate, 0.1% sodium citrate, 0.2% citric acid, 1.1% 1,2-hexanediol, 0.7% butylene glycol, 1% sodium chloride, 1% cetearyl alcohol, 0.1% disodium EDTA, 1.5% xanthan gum, 0.2% vitamin C, with the balance being deionized water.

[0044] The preparation method of the hair-darkening shampoo specifically includes the following steps: A1. Heat water to 90°C, then add xanthan gum, lauryl hydroxysulfonate betaine, cocamidopropyl betaine, disodium cocoamphodiacetate, decyl glucoside, sodium chloride and cetearyl alcohol in sequence. Mix and stir homogenize at 8000 r / min to obtain mixture A for later use. A2. Dissolve the composition in water until homogeneous to obtain mixture B; A3. Mixture A and mixture B are homogenized at 80℃ and 5500 r / min to obtain mixture C, and the mixture is kept at this temperature for 35 min. A4. Cool mixture C to 60°C, add disodium EDTA, 1,2-hexanediol and butylene glycol and stir until homogeneous. Then cool to 35°C, add vitamin C in sequence and stir until homogeneous at 150 r / min. Finally, add sodium salicylate, sodium citrate and citric acid to adjust the pH value to obtain the shampoo.

[0045] Blank application example: Compared with application examples 1-3, the difference is that the shampoo in the blank application example does not contain a composition with hair-darkening effect, and uses an equal amount of deionized water instead of the composition with hair-darkening effect. The preparation method is the same as that in application examples 1-3.

[0046] Example 2: Shampoo Efficacy Test The shampoos prepared in Application Examples 1-3, Comparative Application Examples 1-8, and Blank Application Examples were used as samples to test their hair-darkening effect.

[0047] The experiment used SPF-grade male C57 mice, weighing 18-20g; Model establishment: On day 1 of the experiment, mice were anesthetized by intraperitoneal injection of 4% chloral hydrate, weighed, and injected at a rate of 0.1 mL / 10g body weight. After anesthesia, the long hair on the back of the mice was shaved gently with an electric razor to avoid skin damage. Then, 1 mL of 6% sodium sulfide was drawn up with a 1 mL syringe and evenly applied to the back of the mice. After 2.5 minutes, it was rinsed off with warm water. At this point, the hair on the back of the mice was completely removed, and the skin was intact. Starting on day 7 of the experiment, 4% hydroquinone (1 g of sodium carboxymethyl cellulose and 4 g of hydroquinone dissolved in 100 mL of deionized water) was drawn up with a 1 mL syringe and applied to the center of the hair removal area of ​​each mouse in a 2×2 cm zone. 2 Apply the solution until the entire area is wetted, keep it for 5 minutes, and then put the mouse back into the cage; apply it once in the morning and once in the evening; after 30 days of modeling, when the white hair on the back of the mouse accounts for more than 80% of the experimental area, the modeling is completed. Grouping and administration: Mice that successfully developed the model were divided into 12 groups of 10 mice each; a blank control group was applied; the other 11 groups were experimental groups, and the mice were applied with the shampoos prepared in Examples 1-3 and Examples 1-8, respectively. Each group was applied with 0.05g of the corresponding shampoo, and the mice were gently massaged with their fingers for 1 minute until rich foam was produced, ensuring that the shampoo was in full contact with the skin. The shampoo was left on for 5 minutes, rinsed with warm water, and dried with sterile filter paper. The mice were treated twice a day, morning and evening, and the gray hair turning black was observed. Detection indicators: After the experiment, the improvement effect of each experimental group was observed, and the criteria are shown in Table 3 (5 points in total): "5" represents the observation area (2×2cm at the center of the hair removal area of ​​each mouse). 2 "4" indicates that the new hair in the observation area is >75% black; "3" indicates that the new hair in the observation area is >25% black; "2" indicates that the new hair in the observation area is <25% black; "1" indicates that the hair in the observation area is completely white or has scattered black hair. Experimental data are expressed as mean ± standard deviation (Mean ± SD), and the experimental results are shown in Table 4. The scoring criteria are as follows: Table 3 Scoring Criteria score Skin reaction 1 The observation area was entirely white or had only a few new black hairs. 2 Black hair in the observation area <25% 3 New black hair growth in the observation area >25% 4 New black hair growth in the observation area > 50% 5 In the observation area, >75% of the newborn hair is black. Table 4 Scoring Data Group score Blank control group 1.91±0.16 Application Example 1 <![CDATA[4.63±0.12 * <!-- 7 -->]]> Application Example 2 <![CDATA[4.45±0.30 * ]]> Application Example 3 <![CDATA[4.57±0.28 * ]]> Comparative Application Example 1 <![CDATA[2.41±0.17 *▲ ]]> Comparative Application Example 2 <![CDATA[2.68±0.31 *▲ ]]> Comparative Application Example 3 <![CDATA[2.53±0.15 *▲ ]]> Comparative Application Example 4 <![CDATA[3.09±0.32 *▲ ]]> Comparative Application Example 5 <![CDATA[2.97±0.29 *▲ ]]> Comparative Application Example 6 <![CDATA[3.46±0.37 *▲ ]]> Comparative Application Example 7 <![CDATA[3.85±0.33 *▲ ]]> Comparative Application Example 8 <![CDATA[4.01±0.26 *▲ ]]> Note:" * "This indicates that the scores of Application Examples 1-3 and Comparative Application Examples 1-8 were compared with those of the blank control group, with P < 0.05." ▲ "" indicates that the scores of application examples 1-8 are compared with those of application example 1, and P < 0.05.

[0048] The specific results are shown in Table 4: Compared with the blank control group, the scores of Application Examples 1-3 and Comparative Application Examples 1-8 showed significant improvement, indicating that the composition prepared in this application has excellent hair-darkening effect when applied to the prepared shampoo.

[0049] Compared with comparative application examples 1-3, the scores of application example 1 group were all higher, indicating that there is a synergistic effect between the extract of *Eclipta prostrata* leaf, extract of *Gynostemma pentaphyllum*, and palmitoyl tetrapeptide-10, which has the effect of blackening hair; compared with comparative application examples 4-6, the scores of application example 1 group were all higher, indicating that the extract of *Eclipta prostrata* leaf, extract of *Gynostemma pentaphyllum*, and palmitoyl tetrapeptide-10 are all indispensable.

[0050] The difference between Application Examples 1-3 and Comparative Application Examples 7-8 lies in the different weight ratios of *Eclipta prostrata* extract, *Gynostemma pentaphyllum* extract, palmitoyl tetrapeptide-10, L-arginine, and decapeptide-4. Based on the data from Application Examples 1-3 and Comparative Application Examples 7-8 in Table 4, the shampoo showed better effects when the weight ratio of *Eclipta prostrata* extract, *Gynostemma pentaphyllum* extract, and palmitoyl tetrapeptide-10 was (21-25):(22-27):(7-9) and the weight ratio of L-arginine and decapeptide-4 was (6-9):(9-13). Furthermore, the shampoo showed the best effects when the weight ratio of *Eclipta prostrata* extract, *Gynostemma pentaphyllum* extract, and palmitoyl tetrapeptide-10 was 23:25:8 and the weight ratio of L-arginine and decapeptide-4 was 7:11.

[0051] Example 3: Human Skin Patch Test Shampoos prepared using Application Examples 1-3 and the blank application example were used as samples to verify their safety on human skin. In accordance with the "Cosmetic Safety Technical Specifications" for human skin patch testing, 40 volunteers aged 18-50 years were recruited. The test environment was a temperature of (21±1)℃ and a humidity of (50±10)%, with 10 volunteers per group. Volunteers washed their forearms with water. After 5-10 minutes, when the moisture had evaporated, the testers applied 0.025 mg of each sample to each chamber of a 10-well patch applicator. The inner sides of both forearms of the volunteers were selected, avoiding the skin at the joints of the front and back of the hands. Each arm was covered with a patch applicator and marked accordingly. The patch application lasted for 24 hours. After removing the patch applicator and wiping away any remaining sample, the skin reaction was observed after the indentation disappeared. If the result was negative, observations were repeated at 24 and 48 hours after the patch test. The test results were recorded according to the adverse skin reaction grading standards in Table 5. Table 5. Adverse skin reaction levels Rating levels Skin reaction 0 negative reaction 1 Suspicious reaction, only slight erythema 2 Weak positive reaction (erythema reaction): erythema, infiltration, edema, and papules may be present. 3 Strong positive reaction (herpes reaction): erythema, infiltration, edema, papules; the reaction may extend beyond the test area. 4 Extremely strong positive reaction (confluent herpes simplex reaction): obvious erythema, severe infiltration, edema, confluent herpes simplex; reaction extends beyond the test area. After testing, the shampoos prepared in Application Examples 1-3 and the blank application examples of this invention all showed negative reactions after human patch testing, proving their safety and effectiveness.

[0052] The above description is merely a preferred composition of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with preferred compositions, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-disclosed technical content to create equivalent compositions without departing from the scope of the present invention. Any simple modifications, equivalent changes and alterations made to the above compositions based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A composition having hair-darkening effects, characterized in that, By weight, it includes the following components: Platycladus orientalis leaf extract: 10-15 parts; Polygonum multiflorum extract: 13-17 parts; Eclipta prostrata leaf extract: 21-25 parts; Gynostemma pentaphyllum extract: 22-27 parts; Palmitoyl tetrapeptide-10: 7-9 parts; L-arginine: 6-9 parts; Decapeptide-4: 9-13 parts.

2. The composition according to claim 1, characterized in that, By weight, it includes the following components: Platycladus orientalis leaf extract: 13 parts; Polygonum multiflorum extract: 15 parts; Eclipta prostrata leaf extract: 23 parts; Gynostemma pentaphyllum extract: 25 parts; Palmitoyl tetrapeptide-10: 8 parts; L-arginine: 7 parts; Decapeptide-4: 11 parts.

3. The use of the composition of claim 1 or 2 in the preparation of a hair care product with hair-darkening effect.

4. The application as described in claim 3, characterized in that, The hair care product is one of the following: shampoo, scalp care solution, or conditioner.

5. The application as described in claim 4, characterized in that, The composition is added to the shampoo and body care product at a rate of 0.5wt%-5wt%.

6. A shampoo with hair-darkening effects, characterized in that, The shampoo contains the composition according to claim 1 or 2.

7. The shampoo as described in claim 6, characterized in that, The composition is added to the shampoo in an amount of 1wt%-4wt%.

8. The shampoo as described in claim 6 or 7, characterized in that, The shampoo also contains at least one of the following: surfactant, hair conditioner, pH adjuster, moisturizer, antioxidant, thickener, chelating agent, emulsifying stabilizer, fragrance, and solvent.