A composition for preventing hair loss and promoting hair growth, and its preparation method and application

Abstract

The present invention discloses a composition for preventing hair loss and promoting hair growth, as well as its preparation method and application, specifically belonging to the technical field of compounding cosmetic raw materials. The composition comprises the following components in parts by weight: 12-16 parts of Polygonum multiflorum root extract, 9-13 parts of Zingiber officinale root extract, 0.7-1.3 parts of Cynanchum fasciatum root extract, and 1-2 parts of Astragalus sinensis extract. The Polygonum multiflorum root extract, the Zingiber officinale root extract, the Cynanchum fasciatus root extract, and the Astragalus sinensis extract in the composition provided by the present invention act synergistically through multiple mechanisms to inhibit 5α-reductase activity, reduce DHT damage to hair follicles, promote hair follicle angiogenesis, and delay hair follicle degeneration. The shampoo containing the composition has excellent anti-hair loss and hair growth efficacy.

Description

Technical Field

[0001] The present invention belongs to the technical field of compounding cosmetic raw materials, and in particular relates to a composition for preventing hair loss and promoting hair growth, and a preparation method and application thereof. Background Art

[0002] Currently, hair loss prevention and growth products are primarily categorized as chemical medications (such as minoxidil and finasteride) and natural plant extracts. While chemical medications have some efficacy, long-term use can be associated with side effects such as scalp irritation and sexual dysfunction, and carries the risk of recurrence of hair loss after discontinuation. Natural plant extracts, on the other hand, are becoming a research hotspot due to their safety and widespread availability.

[0003] In the existing technology, a variety of plant extracts have been used in the field of preventing hair loss and promoting hair growth. For example, Polygonum multiflorum root extract: Studies have shown that it can reduce the production of dihydrotestosterone (DHT) by inhibiting the activity of 5α-reductase, while promoting the proliferation of hair follicle cells. However, the effect of a single ingredient is limited. Ginger root extract: Contains ingredients such as gingerol, which can improve scalp microcirculation, but its inhibitory effect on androgenic alopecia is weak when used alone. Other plant ingredients: such as Platycladus orientalis leaves and ginseng, although they have certain hair growth effects, are mostly due to rough extraction methods or unreasonable compounding ratios, resulting in insufficient synergistic effects and unstable actual application effects.

[0004] In response to the above problems, there is an urgent need to develop an efficient and safe anti-hair loss and growth composition that enhances efficacy through the synergistic effect of multiple ingredients and optimizes the extraction process to maximize the retention of active ingredients. Summary of the Invention

[0005] In response to the shortcomings of the existing technology, the present invention significantly improves the comprehensive effect of antioxidant and inhibition of DHT production by compounding Polygonum multiflorum root extract, ginger root extract, Cynanchum wilfordii root extract and Astragalus membranaceus extract, and combining innovative processes such as ultrasound-assisted extraction and enzymatic purification, filling the technical gap in the existing technology of insufficient synergistic effect of natural ingredients and low process efficiency.

[0006] To achieve the above objectives, the present invention discloses the following technical solutions:

[0007] In a first aspect, the present invention provides a composition for preventing hair loss and promoting hair growth, comprising the following components in parts by mass:

[0008] 12-16 parts of Polygonum multiflorum root extract;

[0009] 9-13 parts of ginger root extract;

[0010] 0.7-1.3 parts of Cynanchum chinense root extract;

[0011] 1-2 parts of Chinese milk vetch extract.

[0012] Preferably, the preparation method of the Chinese astragalus extract comprises the following steps:

[0013] 1-1. After drying, crush and sieve the Chinese milk vetch to obtain Chinese milk vetch powder;

[0014] 1-2. Soak the Chinese milk vetch powder in a composite solvent of ethanol and ethyl acetate and ultrasonically treat it at 40-50°C and 25-40kHz for 50-60min;

[0015] 1-3. The ultrasonic mixture was centrifuged at 6000-10000 r / min for 10-20 min to remove the solid residue and collect the supernatant. The supernatant was distilled under reduced pressure using a vacuum rotary evaporator and concentrated to a non-irritating odor to obtain an extract;

[0016] 1-4. Place the extract in an oven at 80-90°C and dry for 30-40 minutes. After drying, crush and grind the extract and pass it through a sieve of ≥120 mesh to obtain the Chinese milk vetch extract.

[0017] Further preferably, the composite solvent in step 1-2 is prepared by mixing ethanol and ethyl acetate in a volume ratio of 6:4 to 8:2.

[0018] More preferably, the conditions for the reduced pressure distillation in step 1-3 are a temperature of 40 to 50° C. and a pressure of 0.03 to 0.05 MPa.

[0019] Preferably, the preparation method of the Cynanchum truncatum root extract comprises the following steps:

[0020] 2-1. Dry the roots of Cynanchum chinense at 40-50°C to constant weight, and grind through a 30-mesh sieve to obtain Cynanchum chinense root powder;

[0021] 2-2. Take Cynanchum root powder, add deionized water at a material-liquid ratio of 1:(10-20) g / mL, stir and disperse, adjust the pH to 4.5-5.5, add 3-5% of the total weight of the material and water, shake in a constant temperature water bath at 45-55°C for 2-4 hours, and terminate the enzymatic hydrolysis in a 90°C water bath for 10-15 minutes. Cool to room temperature to obtain an enzymatic solution, in which the enzyme complex contains mannanase, pectinase, and acid protease;

[0022] 2-3. Ultrasonicate the hydrolyzate at 50-60°C and a frequency of 25-40 kHz for 20-30 minutes. Add a certain amount of ethanol to the sonicated hydrolyzate to a final concentration of 60-70 v / v%. Extract under reflux in a water bath at 70-80°C three times for 1 hour each time. Combine the extracts and centrifuge them at 4000-5000 r / min for 15-20 minutes. Retain the supernatant and concentrate under reduced pressure until it is alcohol-free to obtain a crude extract.

[0023] 2-4. The crude extract was passed through a D101 macroporous resin chromatography column at a flow rate of 0.1-0.5 BV / h and an eluent of 60-70 v / v% ethanol aqueous solution at a flow rate of 0.1-0.5 BV / h to obtain a purified solution. The purified solution was concentrated under reduced pressure until there was no alcohol taste to obtain a concentrated solution. The concentrated solution was dried at low temperature into a powder to obtain a Cynanchum root extract.

[0024] Further preferably, the complex enzyme has a mannanase activity of 500-600 U / g, a pectinase activity of 1000-1500 U / g, and an acid protease activity of 200-300 U / g.

[0025] In a second aspect, the present invention provides the use of the composition for preventing hair loss and promoting hair growth described in the first aspect in the preparation of a hair care product having the efficacy of promoting hair growth and / or strengthening hair and preventing hair loss.

[0026] In a third aspect, the present invention provides an anti-hair loss and hair growth shampoo, wherein the shampoo comprises the anti-hair loss and hair growth composition according to the first aspect;

[0027] The amount of the anti-hair loss and hair growth composition added to shampoo is 0.1-1 wt %.

[0028] Preferably, the shampoo further comprises a surfactant, a preservative, a moisturizer, a fragrance, an anti-dandruff agent, a thickener, a pH regulator and a solvent.

[0029] In a fourth aspect, the present invention provides a method for preparing the anti-hair loss and hair growth shampoo according to the third aspect, comprising the following steps:

[0030] The anti-hair loss and hair growth composition, a surfactant, a preservative, a moisturizer, a fragrance, an anti-dandruff agent, a thickener and a solvent are stirred and mixed evenly to obtain an anti-hair loss and hair growth shampoo.

[0031] In the present invention:

[0032] Polygonum multiflorum root extract effectively promotes hair growth, shifting hair follicle cells from the resting phase to the growing phase. It also upregulates anti-apoptotic genes, downregulates apoptosis-related proteins and androgen receptors, and enhances mitochondrial enzyme activity, exerting an anti-apoptotic effect and delaying hair follicle degeneration. It also inhibits 5α-reductase activity, reduces dihydrotestosterone production, and mitigates the negative effects of androgens on hair follicles, providing a preventative and therapeutic effect on androgenic alopecia.

[0033] Ginger root extract contains active ingredients such as gingerol, which can promote blood circulation in the scalp, enable hair follicles to obtain more nutrients, and create good conditions for hair growth. It can also reduce inflammatory reactions such as redness and swelling of the scalp caused by irritation by inhibiting the expression of inflammatory factors. At the same time, ingredients such as gingerol can scavenge free radicals, exert antioxidant and anti-skin aging effects, and optimize the scalp environment.

[0034] The root extract of Cynanchum chinense is rich in flavonoids. The present invention further purifies the crude extract to increase the content of total flavonoids in the root extract of Cynanchum chinense, thereby improving the efficacy of the root extract of Cynanchum chinense in anti-inflammatory, antibacterial, antioxidant, and hair follicle cell growth promotion.

[0035] Milk vetch extract contains various amino acids, flavonoids, and other ingredients that have a nourishing and conditioning effect on the hair and scalp. Research has found that combining Polygonum multiflorum root extract, Zingiber officinale root extract, Cynanchum tigrinum root extract, and Milk vetch extract has a more effective effect on inhibiting 5α-reductase activity and improving hair follicle cell growth markers.

[0036] Beneficial effects of the present invention:

[0037] 1. The composition provided by the present invention comprises Polygonum multiflorum root extract, Zingiber officinale root extract, Cynanchum tigrinum root extract, and Astragalus sinensis root extract, which work synergistically through multiple mechanisms to inhibit 5α-reductase, reduce DHT damage to hair follicles, promote angiogenesis in hair follicles, and delay hair follicle degeneration.

[0038] 2. The anti-hair loss and hair growth shampoo provided by the present invention has excellent anti-hair loss and hair growth efficacy, and has significant effects compared to the control product. DETAILED DESCRIPTION

[0039] To make the objectives, technical solutions, and advantages of the present invention more clearly understood, the present invention is further described in detail below with reference to specific embodiments. However, the following embodiments are merely preferred embodiments of the present invention and are not exhaustive. For the sake of clarity, not all features of the actual embodiments are described.

[0040] Based on the embodiments in the implementation manner, other embodiments obtained by those skilled in the art without making any creative work shall all fall within the protection scope of the present invention.

[0041] In the present invention

[0042] Polygonum multiflorum root extract: purchased from Xi'an Guohao Biotechnology Co., Ltd.;

[0043] Ginger root extract: purchased from Xi'an Guohao Biotechnology Co., Ltd.;

[0044] Tiger orchid root: the root of the tiger orchid plant (CYMBIDIUM GRANDIFLORUM) of the Orchidaceae family;

[0045] Milk vetch: also known as red flower grass seed, it is a plant of the genus Milkvetch (ASTRAGALUS SINICUS) of the genus Astragalus in the leguminosae family.

[0046] Preparation of Chinese Milkvetch Extract

[0047] Step 1-1. The fresh whole herb of Chinese milk vetch was dried in a constant temperature drying oven to constant weight and then ground into powder and passed through a 30-mesh sieve to obtain Chinese milk vetch powder to improve the subsequent extraction efficiency;

[0048] Step 1-2. Soaking the astragalus powder in a 6:4 volume ratio of ethanol and ethyl acetate composite solvent, wherein the astragalus powder to solvent ratio is 1:10 g / mL, and ultrasonically treating at 50°C and 28 kHz frequency for 50 minutes to promote the release of the active substance;

[0049] Step 1-3. The sonicated mixture was centrifuged at 10,000 rpm for 10 min to remove the solid residue and collect the supernatant. The supernatant was subjected to reduced pressure distillation using a vacuum rotary evaporator at a temperature of 50°C and a pressure of 0.03 MPa, and concentrated until there was no irritating odor to obtain an extract;

[0050] Step 1-4. Place the extract in an oven at 80° C. and dry for 40 minutes. After drying, crush and grind the extract and pass it through a sieve with a mesh size of ≥120 to obtain the Chinese milk vetch extract.

[0051] Preparation of Cynanchum truncatum root extract

[0052] Step 2-1. Take fresh Cynanchum root and rinse with water to remove impurities, drain the water, place in a 50 ° C oven to dry to constant weight, grind through a 30 mesh sieve to obtain Cynanchum root powder;

[0053] Step 2-2. Weigh the Cynanchum root powder, add deionized water at a material-liquid ratio of 1:10 g / mL, stir and disperse, adjust the pH to 4.5-5.5, add 5% of the total mass of the material and water, shake in a constant temperature water bath at 45°C for 4 hours, terminate the enzymatic hydrolysis in a 90°C water bath for 15 minutes, and cool to room temperature to obtain an enzymatic solution, wherein the mannanase activity of the complex enzyme is 500 U / g, the pectinase activity is 1000 U / g, and the acid protease activity is 200 U / g;

[0054] Step 2-3. The enzymatic hydrolysate was transferred to an ultrasonic kettle and sonicated at 60°C and 28 kHz for 20 min. A certain amount of ethanol was added to the sonicated enzymatic hydrolysate to a final concentration of 60 v / v%. The extract was refluxed in a water bath at 80°C for three times, each for 1 h. The extracts were combined and centrifuged at 5000 rpm for 20 min. The supernatant was retained and concentrated under reduced pressure until free of alcohol to obtain a crude extract.

[0055] Step 2-4. The crude extract was passed through a D101 macroporous resin chromatography column with a sample flow rate of 0.15 BV / h and an eluent of 70 v / v% ethanol aqueous solution at a flow rate of 0.15 BV / h to obtain a purified solution after elution. The purified solution was concentrated under reduced pressure until there was no alcohol taste to obtain a concentrated solution. The concentrated solution was dried at low temperature into a powder to obtain a Cynanchum root extract.

[0056] Anti-hair loss and hair growth composition

[0057] The raw materials are accurately weighed according to the mass ratio in Table 1, and the anti-hair loss and hair growth combination is obtained after compounding.

[0058] Table 1

[0059] Raw material name Composition 1 Composition 2 Composition 3 Polygonum multiflorum root extract 12 14 16 Ginger root extract 9 11 13 Tiger orchid root extract 0.7 1 1.3 Chinese Milkvetch Extract 1 1.5 2

[0060] In order to verify the functional relationship between the components in the composition, the components were omitted and the ratios were adjusted based on the formula of composition 2, as shown in Table 2:

[0061] Table 2

[0062] Raw material name Composition 4 Composition 5 Composition 6 Composition 7 Composition 8 Polygonum multiflorum root extract - 14 14 14 1 Ginger root extract 11 - 11 11 1 Tiger orchid root extract 1 1 - 1 1 Chinese Milkvetch Extract 1.5 1.5 1.5 - 1

[0063] Note: “-” in the table means no additive.

[0064] Composition performance test

[0065] Before conducting the following test experiments, the above-mentioned compositions 1-8 prepared according to the corresponding mass ratio were respectively taken, and a certain amount of solvent was added for dilution, specifically 10wt% of the composition, 1.3wt% of 1,2-hexanediol, 2% of glycerol, and the balance was deionized water. After mixing evenly at 5000r / min, composition solutions 1-8 were obtained.

[0066] 1 DPPH free radical scavenging experiment

[0067] Test samples 1-8: The composition solutions 1-8 prepared above were respectively taken and mixed with deionized water at a mass ratio of 1:4 to obtain test samples 1-8 containing 20 wt % of the composition solution.

[0068] Experimental plan: DPPH was prepared with anhydrous ethanol to prepare 2×10 -4 mol / L DPPH solution, take several 2 mL portions of the test sample, DPPH solution, and anhydrous ethanol, mix 2 mL of the test sample and 2 mL of DPPH solution, and place them at room temperature for 30 minutes. Measure the absorbance at a wavelength of 517 nm to obtain Ai;

[0069] Mix 2 mL of the test sample and 2 mL of anhydrous ethanol and measure the absorbance as Aj according to the above method;

[0070] Mix 2 mL of DPPH solution and 2 mL of anhydrous ethanol and measure the absorbance as Ac according to the above method.

[0071] Each sample was measured three times in parallel and the average value was taken.

[0072] The scavenging rate of DPPH free radicals of each test sample was calculated according to the following formula and recorded in Table 3 below.

[0073] Clearance rate (%) = (1-(Ai-Aj) / Ac) × 100%.

[0074] Where: Ai is the absorbance of a mixture of 2 mL of test sample and 2 mL of DPPH solution; Aj is the absorbance of a mixture of 2 mL of test sample and 2 mL of anhydrous ethanol; Ac is the absorbance of a mixture of 2 mL of DPPH solution and 2 mL of anhydrous ethanol.

[0075] Table 3 DPPH clearance rate

[0076] Group DPPH clearance rate / % Test sample 1 (corresponding to composition 1) 76.31 Test sample 2 (corresponding to composition 2) 78.24 Test sample 3 (corresponding to composition 3) 78.09 Test sample 4 (corresponding to composition 4) 62.40 Test sample 5 (corresponding to composition 5) 64.72 Test sample 6 (corresponding to composition 6) 59.86 Test sample 7 (corresponding to composition 7) 69.26 Test sample 8 (corresponding to composition 8) 73.88

[0077] Result Analysis

[0078] As shown in Table 3, compositions 1-3 exhibited superior antioxidant efficacy, while compositions 4-7 lacked the following: Polygonum multiflorum root extract, Zingiber officinale root extract, Cynanchum cyrtonema root extract, and Astragalus chinensis extract, respectively, resulting in a decrease in the antioxidant performance of the overall formulation. Although composition 8 contained Polygonum multiflorum root extract, Zingiber officinale root extract, Cynanchum cyrtonema root extract, and Astragalus chinensis extract, the mass ratio was not within the optimal range claimed by the present invention, resulting in an inferior antioxidant effect to compositions 1-3.

[0079] 2 5α-reductase inhibition assay

[0080] 2.1 Preparation of 5α-reductase extract

[0081] 2.1.1 Animal Preparation: Six healthy male Sprague-Dawley rats were fasted for 12 h and then anesthetized and sacrificed. The rats were quickly transferred to a 4°C dissection table, and the prostate tissue was completely dissected, minced, and accurately weighed.

[0082] 2.1.2 5α-Reductase Extraction: Prostate tissue was mixed with pre-chilled buffer containing 0.32 mol / L sucrose, 0.1 mmol / L dithiothreitol, 1 mmol / L EDTA, and 0.2 mol / L PBS at a mass / volume ratio of 1:5 (pH 6.5). The tissue was quickly homogenized in a homogenizer. Centrifuge at 13,000 rpm for 10 min at 4°C. The upper lipid layer and precipitate were discarded, and the clear supernatant was collected.

[0083] 2.1.3 Storage of enzyme extract: dilute the supernatant to 20 mL with buffer, dispense into pre-cooled EP tubes, and store at -80°C until use.

[0084] 2.2 5α-reductase activity assay

[0085] 2.2.1 Test samples: The composition solutions 1-8 prepared above were taken respectively, and mixed with deionized water at a mass ratio of 1:4 to obtain test samples 1-8 containing 20 wt % of the composition solution.

[0086] 2.2.2 Reaction System: Add 0.5 mL of phosphate buffer, 0.2 mL of the test sample, 200 μL of a 300 mg / L testosterone solution, and 200 μL of a 0.8 g / L NADPH (reduced coenzyme II) solution to a stoppered test tube. Finally, add 0.5 mL of the prepared 5α-reductase extract. Incubate at 37°C for 30 min.

[0087] 2.2.3 Control settings: Positive control: 0.05 mg / L finasteride solution instead of the test sample; Enzyme reaction tube: Contains only enzyme extract and substrate (no test sample); Blank tube: No enzyme extract (replaced with an equal volume of buffer).

[0088] 2.2.4 Detection: After the reaction is complete, add 3 mL of dichloromethane to stop the reaction and extract the testosterone. Add 0.25 mL of 100 mg / L propyl hydroxybenzoate as an internal standard and centrifuge at 5000 rpm for 10 min. Discard the upper aqueous phase, remove approximately 1 mL of the organic phase, evaporate to dryness, and dissolve the residue in 1.5 mL of methanol. Aspirate 10 μL of the residue and determine the residual testosterone content by HPLC. HPLC conditions: C18 reversed-phase column (4.6 × 250 mm, 5 μm); mobile phase: methanol / water (70:30, v / v); flow rate: 1.0 mL / min; detection wavelength: 254 nm; injection volume: 10 μL.

[0089] The 5α-reductase inhibition rate was calculated according to the following formula:

[0090] ;

[0091] Where: C is the ratio of the testosterone peak area to the internal standard peak area.

[0092] 2.3 Test Results

[0093] Table 4 Inhibitory effect on 5α-reductase

[0094] Group 5α-reductase inhibition rate / % Positive control group 93.66 Test sample 1 (corresponding to composition 1) 68.25 Test sample 2 (corresponding to composition 2) 69.71 Test sample 3 (corresponding to composition 3) 70.28 Test sample 4 (corresponding to composition 4) 40.67 Test sample 5 (corresponding to composition 5) 38.31 Test sample 6 (corresponding to composition 6) 52.94 Test sample 7 (corresponding to composition 7) 51.45 Test sample 8 (corresponding to composition 8) 56.34

[0095] 2.4 Results Analysis

[0096] According to the results in Table 4, the inhibition rates of compositions 1-3 (test samples 1-3) were relatively high, reaching 68.25%, 69.71%, and 70.28%, respectively, which was 70% of the performance of the positive control finasteride. Finasteride is a recognized strong inhibitor of 5α-reductase. Therefore, the composition provided in this application has excellent anti-hair loss and hair growth effects.

[0097] 3 Human hair follicle dermal papilla cells

[0098] 3.1 Experimental Principle

[0099] VEGF is highly expressed in hair follicle neoplastic cells, promoting angiogenesis around the follicles and inducing the anagen phase. High expression of TGF-β1 inhibits hair follicle development, leading to epithelial cell apoptosis and a shift in the hair follicle cycle from anagen to catagen. ELISA was used to measure the levels of VEGF and TGF-β1 in the supernatant of HDPCs after co-culture of the test samples with them to evaluate their efficacy in promoting hair growth.

[0100] 3.2 Cell culture system

[0101] Human hair follicle dermal papilla cells (HDPCs) were cultured in DMEM complete medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin. Cells were routinely subcultured in a 37°C, 5% v / v CO2 incubator.

[0102] 3.3 Experimental group design

[0103] Dermal papilla cells were collected at 5×10 4 The cells were seeded at a density of 100 μg / well in a 96-well plate and incubated for 24 h. The culture medium solutions corresponding to the following experimental groups, positive control group and blank control group were added for co-culture for 24 h. Subsequent ELISA detection was performed 24 h after treatment. Three parallel treatments were performed in each group, and the results were averaged.

[0104] Experimental group: The composition solutions 1-8 prepared above were prepared with DMEM to prepare DMEM culture medium solutions containing 10 wt% of the composition solution;

[0105] Positive control group: DMEM medium solution containing 0.1 μmol / L minoxidil;

[0106] Blank control group: DMEM medium solution without any reagent added;

[0107] Three replicate wells were set up in each group as parallel experiments.

[0108] 3.4 Detection methods

[0109] ELISA was used to quantitatively analyze the expression levels of vascular endothelial growth factor (VEGF) and transforming growth factor-β1 (TGF-β1) in the cell culture supernatant. The specific procedures strictly followed the kit instructions. The final test data were the average of three independent experiments. The results are detailed in Table 5.

[0110] Table 5 VEGF and TGF-β1 content

[0111] Group VEGF content (pg / mL) TGF-β1 content (pg / mL) Blank control group 623.75 743.28 Positive control group 807.36 627.53 Test sample 1 (corresponding to composition 1) 772.15 708.64 Test sample 2 (corresponding to composition 2) 797.08 686.37 Test sample 3 (corresponding to composition 3) 786.93 698.15 Test sample 4 (corresponding to composition 4) 648.31 735.91 Test sample 5 (corresponding to composition 5) 689.57 740.82 Test sample 6 (corresponding to composition 6) 714.89 723.49 Test sample 7 (corresponding to composition 7) 732.06 733.66 Test sample 8 (corresponding to composition 8) 739.42 715.20

[0112] 3.5 Results Analysis

[0113] The results in Table 5 show that Composition 2 had the highest VEGF content, reaching 797.08 pg / mL, and the lowest TGF-β1 content, 686.37 pg / mL, indicating its optimal potential for promoting hair follicle growth. Composition 4, lacking Polygonum multiflorum root extract, significantly reduced its VEGF content to 648.31 pg / mL, demonstrating the key role of Polygonum multiflorum root extract in upregulating VEGF expression and promoting hair follicle angiogenesis.

[0114] From the overall results, the Polygonum multiflorum root extract, Zingiber officinale root extract, Cynanchum wilfordii root extract, and Astragalus membranaceus extract provided by the present invention jointly promoted VEGF expression, inhibited the TGF-β1 signaling pathway, and optimized the hair follicle growth environment.

[0115] Preparation of anti-hair loss and hair growth shampoo

[0116] Accurately weigh the raw materials according to the mass percentage in Table 6;

[0117] Step 3-1. Heat component A to 85°C and stir until completely dissolved and transparent to obtain solution A;

[0118] Step 3-2. Then, the temperature of solution A was lowered to 55°C, and the raw materials in component B were added to solution A in sequence, and stirred at 300 rpm for 30 min to obtain solution AB;

[0119] Step 3-3. Cool solution AB to 45°C and add pre-mixed components C, D, E and F, stir evenly at a speed of 200 r / min, adjust the pH to 6.5-7.0 with citric acid, stir and cool to room temperature to obtain an anti-hair loss shampoo.

[0120] Table 6 Mass percentage of shampoo raw materials

[0121]

[0122] Human efficacy experiments

[0123] 1 Materials and Methods

[0124] 1.1 Test product

[0125] 1.1.1 Test product: the anti-hair loss and hair growth shampoo prepared in Application Example 2 above.

[0126] 1.1.2 Control product: A product based on the corresponding test product matrix that does not contain the anti-hair loss ingredients and is tested in parallel with the test product.

[0127] 1.1.3 Washout products: Same as the control product. Both the experimental and control groups use the control product during the washout period.

[0128] 1.2 Instruments

[0129] 1.2.1 Professional digital camera: Pixels not less than 15 million. Parameters such as aperture, ISO, and focal length must remain consistent throughout the test.

[0130] 1.2.2 Dermatoscope: LED light source, wavelength 450-750nm, color temperature 6500-8000K, illumination not less than 540 lx, magnification ≥20 times, detection diameter not less than 1.0cm or area not less than 0.8cm 2 .

[0131] 1.2.3 Comb: The teeth should have a moderate density (tooth spacing 0.9-1.1 mm), a tooth length of 2.0-3.0 cm, and a comb length of no less than 10 cm (excluding the handle). Combs of the same specifications and material must be used throughout the test. After each use, they should be disinfected in accordance with the relevant requirements of the Technical Specifications for Disinfection in Medical Institutions (WS-T367-2012).

[0132] 1.3 Subjects

[0133] The experimental group consisted of 30 subjects, including 4 males and 26 females, ranging in age from 24 to 58 years old, with an average age of 42.5±8.1 years old, who met the voluntary inclusion criteria.

[0134] The control group consisted of 30 subjects, including 3 males and 27 females, ranging in age from 27 to 60 years old, with an average age of 46.1±9.4 years old, who met the voluntary inclusion criteria.

[0135] 1.4 Environmental Conditions: The test results should be observed in a constant environment with a temperature of 21±1°C and a relative humidity of 50+10%RH. Visual evaluation should also be conducted under constant lighting conditions (fluorescent lamps or LED lighting with a color temperature of 5500-6500K). All subjects should adapt to this environmental condition for at least 30 minutes before evaluation and testing.

[0136] 1.5 Detection Methods

[0137] 1.5.1 Tests shall be conducted in accordance with the specific requirements for the "Safety Technical Specifications for Cosmetics" (2015 edition) for the prevention of hair loss. Subjects shall be screened using the 60-comb method, and those with at least 10 hair loss counts shall be considered qualified.

[0138] 1.5.2 Qualified subjects will undergo a 2-week washout period. The products used during this period are as described in 1.1.3. After the washout period, the subjects will be combed 60 times again. Only those with a hair loss count of more than 10 will be able to enter the formal trial.

[0139] 1.5.3 Subjects enrolled in the formal trial will be divided into a test product group and a control product group using a stratified randomization method to ensure balance of important factors that may affect the trial results (gender, age, hair length, severity of hair loss, etc.).

[0140] 1.5.4 Before using the product, the enrolled subjects will be assessed for baseline hair values, including hair loss counting, hair density assessment, and imaging, and the results will be recorded. Four, eight, and 12 weeks after product use, visual assessment and instrument testing will be conducted again, and the results will be recorded according to the hair density grading standards in the current valid technical specifications.

[0141] 1.5.4.1 Hair loss count: During each visit, trained staff will comb the subject's hair using the 60-comb method, count the number of hair loss, and record the number.

[0142] 1.5.4.2 Local Hair Density Assessment: A clipping area measuring at least 1.5 cm × 1.5 cm (temporal to the vertex) should be fixed on the subject's head. Position the area carefully and ensure consistent hair removal at each visit, with hair remaining no longer than 1 mm. During image acquisition, the operator should position the subject in a comfortable position and place the dermatoscope directly in the center of the clipped area to capture a localized image of the hair. The dermatoscope lens should be in full contact with the scalp and maintained perpendicular to the scalp during the image acquisition. The clarity of the captured image should be checked. The number and density of localized hairs should be counted using image analysis software or manual counting, and recorded.

[0143] 1.6 Statistical methods:

[0144] Statistical analysis software was used to perform statistical analysis of the data. Measurement data were expressed as mean + standard deviation, and a normal distribution test was performed. If the data met the normal distribution requirements, a paired t-test was used for comparison before and after the data were used; otherwise, a two-related sample rank sum test was used. For comparisons of rank data before and after use, a two-related sample rank sum test was used; and for comparisons between the test product and the control group, an independent sample t-test or rank sum test was used. All statistical analyses were two-tailed tests, with a significance level of a = 0.05.

[0145] 1.7 Test evaluation method:

[0146] According to the data analysis results, at any visit time point during the trial, if there is no significant increase in the hair loss count before and after the use of the test product, or the difference in hair loss count before and after (hair loss count at a certain visit time point after product use - hair loss count before product use) is significantly lower than that of the control group (P < 0.05); or if there is no significant decrease in hair density (local hair density) or the difference in hair density before and after (hair density at a certain visit time point after product use - hair density before product use) is significantly higher than that of the control group (P < 0.05), then the test product is deemed to have anti-hair loss efficacy; otherwise, the test product is deemed to have no anti-hair loss efficacy.

[0147] 2 Experimental Results

[0148] Table 7 Difference in hair loss counts before and after product use (mean + standard deviation)

[0149]

[0150] Note: Compared with the control product group, “*”: P < 0.05, “**”: 0.001 ≤ P < 0.01, “***”: P < 0.001.

[0151] Table 8 Difference in local hair density before and after product use (mean + standard deviation)

[0152]

[0153] Note: Compared with the control product group, “*”: P < 0.05, “**”: 0.001 ≤ P < 0.01, “***”: P < 0.001.

[0154] 3 Experimental Conclusions

[0155] According to the data analysis results, the difference in hair loss counts 12 weeks after product use was significantly lower than that of the control group (p < 0.05), and the difference in local hair density 12 weeks after product use was significantly higher than that of the control group (p < 0.05), which indicates that the product has the effect of preventing hair loss and promoting hair growth.

[0156] Although specific embodiments of the present invention have been described above, those skilled in the art will appreciate that these are merely illustrative and that the scope of the present invention is defined by the appended claims. Those skilled in the art may make various changes or modifications to these embodiments without departing from the principles and essence of the present invention, and such changes and modifications are intended to fall within the scope of the present invention.

Claims

1. A composition for preventing hair loss and promoting hair growth, characterized in that: The composition consists of the following components in parts by mass: 12-16 parts of Polygonum multiflorum root extract; 9-13 parts of ginger root extract; 0.7-1.3 parts of Cynanchum chinense root extract; 1-2 parts of Chinese milk vetch extract; The preparation method of the Chinese astragalus extract comprises the following steps: 1-1. After drying, crush and sieve the Chinese milk vetch to obtain Chinese milk vetch powder; 1-2. Soak the Chinese milk vetch powder in a composite solvent of ethanol and ethyl acetate and ultrasonically treat it at 40-50°C and 25-40kHz for 50-60min; 1-3. The ultrasonic mixture was centrifuged at 6000-10000 r / min for 10-20 min to remove the solid residue and collect the supernatant. The supernatant was distilled under reduced pressure using a vacuum rotary evaporator and concentrated to a non-irritating odor to obtain an extract; 1-4. Place the extract in an oven at 80-90°C and dry for 30-40 minutes. After drying, crush and grind it through a 120-mesh sieve to obtain the Chinese milk vetch extract. The composite solvent in step 1-2 is prepared by mixing ethanol and ethyl acetate in a volume ratio of 6:4 to 8:2; The preparation method of the Cynanchum root extract comprises the following steps: 2-1. Dry the roots of Cynanchum chinense at 40-50°C to constant weight, and grind through a 30-mesh sieve to obtain Cynanchum chinense root powder; 2-2. Take Cynanchum root powder, add deionized water at a material-liquid ratio of 1:(10-20) g / mL, stir and disperse, adjust the pH to 4.5-5.5, add 3-5% of the total weight of the material and water, shake in a constant temperature water bath at 45-55°C for 2-4 hours, and terminate the enzymatic hydrolysis in a 90°C water bath for 10-15 minutes. Cool to room temperature to obtain an enzymatic solution, in which the enzyme complex contains mannanase, pectinase, and acid protease; 2-3. Ultrasonicate the hydrolyzate at 50-60°C and a frequency of 25-40 kHz for 20-30 minutes. Add a certain amount of ethanol to the sonicated hydrolyzate to a final concentration of 60-70 v / v%. Extract under reflux in a water bath at 70-80°C three times for 1 hour each time. Combine the extracts and centrifuge them at 4000-5000 r / min for 15-20 minutes. Retain the supernatant and concentrate under reduced pressure until it is alcohol-free to obtain a crude extract. 2-4. The crude extract was passed through a D101 macroporous resin chromatography column at a flow rate of 0.1-0.5 BV / h. The eluent was an ethanol aqueous solution with a concentration of 60-70 v / v% at a flow rate of 0.1-0.5 BV / h to obtain a purified solution after elution. The purified solution was concentrated under reduced pressure until there was no alcohol taste to obtain a concentrate, which was then dried at low temperature to a powder to obtain a Cynanchum root extract. The complex enzyme has an enzymatic activity of mannanase of 500-600 U / g, an enzymatic activity of pectinase of 1000-1500 U / g, and an enzymatic activity of acid protease of 200-300 U / g.

2. The composition according to claim 1, characterized in that The conditions for the reduced pressure distillation in steps 1-3 are a temperature of 40-50° C. and a pressure of 0.03-0.05 MPa.

3. Use of the composition for preventing hair loss and promoting hair growth according to claim 1 or 2 in the preparation of a hair care product having the efficacy of promoting hair growth and / or strengthening hair and preventing hair loss.

4. An anti-hair loss and hair growth shampoo, characterized in that: The shampoo contains the anti-hair loss and hair growth composition according to claim 1 or 2; The amount of the anti-hair loss and hair growth composition added to shampoo is 0.1-1 wt %.

5. The anti-hair loss and hair growth shampoo according to claim 4, characterized in that: The shampoo also contains a surfactant, a preservative, a moisturizer, a fragrance, an anti-dandruff agent, a thickener, a pH regulator and a solvent.

6. The method for preparing the anti-hair loss and hair growth shampoo according to claim 5, characterized in that: The following steps are involved: The anti-hair loss and hair growth composition, a surfactant, a preservative, a moisturizer, a fragrance, an anti-dandruff agent, a thickener and a solvent are stirred and mixed evenly to obtain an anti-hair loss and hair growth shampoo.

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