Anti-alopecia hair-blacking liquid for nourishing white hair and black hair and preparation method of anti-alopecia hair-blacking liquid

Abstract

The invention relates to the technical field of hair treatment medicines, and particularly discloses an anti-alopecia hair-blacking liquid for nourishing white hair and blacking hair and a preparation method of the anti-alopecia hair-blacking liquid. The hair loss preventing and blackening liquid for white hair and black hair comprises the following raw materials in parts by mass: 5-10 parts of kaempferol, 1-5 parts of aromatic folic acid, 1-2 parts of lecithin, 1-2 parts of a surfactant, 40-60 parts of vegetable oil, 1-3 parts of blue copper peptide, 1-2 parts of dopamine hydrochloride, 0.01-0.1 part of 1-ethyl-(3-dimethylaminopropyl) carbodiimide and 1-2 parts of a traditional Chinese medicine extract. Hydrophilic and lipophilic active components are loaded at the same time, repair and development of melanocytes are effectively promoted, connection of hair papilla and blood vessels can be enhanced, the hair growing effect is remarkable, irritation is small, hair follicles can be deeply nourished, dependence cannot be generated, and the composition is suitable for large-scale application and popularization.

Description

Technical Field

[0001] The invention relates to the technical field of hair treatment drugs, and in particular to a hair loss prevention and blackening liquid for nourishing white hair and a preparation method thereof. Background Art

[0002] Hair not only plays an important role in human appearance, but is also one of the important indicators of human health. The color of hair depends on the number, size, distribution and nature of the pigment particles in the hair shaft. White hair refers to hair that has turned completely or partially white. White hair is caused by the reduction of melanin particles in the medulla and cortex of the hair or the filling of air.

[0003] Normally shed hair is in the catagen and telogen phases. The catagen and the newly grown hair are in a dynamic balance. Pathological gray hair loss is a common skin disease, which refers to abnormal graying or excessive hair loss. Modern people are affected by busy lives, work pressure, environmental pollution and other factors, and more and more people are experiencing gray hair.

[0004] At present, there are many cases of scalp itching, severe hair loss, and rapid increase of white hair. There are many methods to treat them. Existing hair transplant surgery has side effects such as infection, damage to superficial sensory nerves, and scarring. The actual effectiveness of current hair dyes and anti-hair loss shampoos is unstable. Some of them not only have no protective effect but also have harmful effects, which brings inconvenience to patients, so that baldness is becoming younger and younger, and white hair is becoming more and more teenage.

[0005] Although there are some drugs for treating gray hair and hair loss on the market, most of them are compound prescriptions of traditional Chinese medicine with unclear medicinal ingredients, weak specificity, and difficulty in achieving effective therapeutic effects. At the same time, the disease is prone to relapse after discontinuation of the drug, and the effect is not ideal. Summary of the invention

[0006] The purpose of the present invention is to solve the shortcomings of the prior art and to propose a hair loss prevention and black hair liquid for nourishing white hair and a preparation method.

[0007] The invention discloses a hair loss prevention and blackening liquid for nourishing white hair and black hair. The raw materials thereof include, by mass, 5-10 parts of kaempferol, 1-5 parts of geranic acid, 1-2 parts of lecithin, 1-2 parts of surfactant, 40-60 parts of vegetable oil, 1-3 parts of blue copper peptide, 1-2 parts of dopamine hydrochloride, 0.01-0.1 parts of 1-ethyl-(3-dimethylaminopropyl)carbodiimide and 1-2 parts of traditional Chinese medicine extract.

[0008] Preferably, the surfactant is at least one of polyoxyethylene sorbitan fatty acid ester, sorbitan fatty acid ester, fatty acid glyceride and cocamidopropyl betaine.

[0009] Preferably, the vegetable oil is at least one of peanut oil, rapeseed oil, soybean oil, corn oil, camellia oil, walnut oil, olive oil, palm oil, grapeseed oil and sunflower oil.

[0010] Preferably, the Chinese medicine extract is at least one of mulberry fruit extract, licorice root extract, Polygonum multiflorum root extract, Rehmannia glutinosa extract, Platycladus orientalis leaf extract, black sesame extract, Channa intestine extract, dandelion extract, mulberry leaf extract, ginseng root extract, Chuanxiong root extract, and Magnolia officinalis bark extract.

[0011] Polygonum multiflorum root extract and black sesame extract are obtained by extraction with 70% ethanol aqueous solution; while mulberry extract, licorice root extract, rehmannia root extract, arborvitae leaf extract, Channa intestine extract, dandelion extract, mulberry leaf extract, ginseng root extract, ligusticum chuanxiong root extract, and Magnolia officinalis bark extract are water extracts.

[0012] Preferably, the Chinese medicine extract includes: mulberry fruit extract, Polygonum multiflorum root extract, Rehmannia glutinosa extract, mulberry leaf extract, and ginseng root extract; the mass ratio of mulberry fruit extract, Polygonum multiflorum root extract, Rehmannia glutinosa extract, mulberry leaf extract, and ginseng root extract is 1-2:4-6:3-5:1-2:2-4.

[0013] The method for preparing the above-mentioned hair loss prevention and blackening liquid for nourishing white hair comprises the following steps: S1, adding kaempferol to ethanol aqueous solution, reflux stirring at 40-60° C. for 10-20 min, adding geranic acid thereto and continuing stirring for 1-3 h, rotary evaporation, and vacuum drying to obtain kaempferol composite material; S2, adding kaempferol compound, lecithin and surfactant to vegetable oil and stirring for 10-30 minutes, adjusting the system temperature to 40-60° C., and ultrasonically treating for 10-20 minutes to obtain an oil phase; S3. Add blue copper peptide to acetic acid aqueous solution and stir evenly, add dopamine hydrochloride and 1-ethyl-(3-dimethylaminopropyl)carbodiimide thereto, adjust the pH value of the system to 5.5-6, ultrasonically treat for 1-2 hours, dialyze for 5-10 hours, freeze-dry, and then mix with the Chinese medicine extract, add to water and stir evenly, add oil phase dropwise thereto under stirring, and stir for 5-15 minutes.

[0014] Preferably, in S2, the ultrasonic frequency is 10-15 kHz.

[0015] Preferably, in S3, the concentration of the acetic acid aqueous solution is 0.01-0.05 mol / L.

[0016] Preferably, in S3, the ultrasonic frequency is 12-16 kHz.

[0017] Preferably, in S3, dialysis is performed with an aqueous acetic acid solution having a concentration of 0.01-0.05 mol / L for 5-10 hours. Beneficial Effects

[0018] The present invention uses kaempferol as an active ingredient, which can effectively reduce the inflammation of human hair papilla cells and surrounding cells, and has a significant anti-hair loss effect. It combines with geranic acid to form a synergistic effect, comprehensively improves the microenvironment around the hair follicles, and has a significant hair darkening effect. It can be effectively penetrated into the scalp and hair follicles through interaction with lecithin, effectively activate tyrosinase, and repair damaged melanocytes. The black hair has good durability, and long-term use has almost no side effects, and the hair growth and hair darkening effects are significant.

[0019] The present invention combines dopamine with the surface of blue copper peptide, which can not only be effectively dispersed at the oil-water interface, thereby having a larger interfacial area and smaller mass transfer resistance, but also more conducive to the transdermal absorption of active ingredients, with significant anti-inflammatory effect, and can effectively reduce sebum secretion, promote blood circulation, effectively improve the body's immunity, activate hair follicles to promote hair regrowth, and can well promote scalp absorption, thereby achieving good therapeutic effects.

[0020] The inventors have found through experiments that if only blue copper peptide and dopamine are simply mixed as active ingredients, not only the hair growth effect is poor, but also the durability is poor. However, the present invention organically combines blue copper peptide and dopamine, which not only has good biocompatibility, but also can effectively help the active ingredients pass through the skin barrier, so that the active ingredients are efficiently delivered to the hair follicles, significantly promoting hair growth, not only prolonging the retention time, but also comprehensively improving the microenvironment around the hair follicles, with good effects, and is not easy to relapse, and has a good treatment effect.

[0021] The present invention simultaneously carries hydrophilic and lipophilic active components, effectively promotes the repair and development of melanocytes, and can strengthen the connection between hair papilla and blood vessels, while having a significant hair growth effect, low irritation, being able to deeply nourish hair follicles, and not causing dependence, and is suitable for large-scale promotion and use. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a comparison chart of intracellular melanin content and tyrosinase activation rate of Example 5 and Comparative Examples 1-2.

[0023] Figure 2 This is a comparison chart of the number of apoptotic hair follicle cells and the number of melanin hair follicles in the NC group, PC group, S5 group, D1 group, and D2 group.

[0024] Figure 3 It is a comparison chart of the levels of inflammatory factors TNF-α and IL-6 secreted by macrophage RAW264.7 in Example 5, Comparative Example 1, Comparative Example 2, positive control and negative control. DETAILED DESCRIPTION

[0025] The present invention will be further explained below in conjunction with specific embodiments.

[0026] The following Polygonum multiflorum root extract, Platycladus orientalis leaf extract, Ligusticum chuanxiong root extract, and mulberry fruit extract were purchased from Shaanxi Fenglingu Pharmaceutical Biotechnology Co., Ltd., and the following Rehmannia glutinosa extract, mulberry leaf extract, and ginseng root extract were purchased from Shaanxi Bolin Biotechnology Co., Ltd. Embodiment 1

[0027] A hair loss prevention and blackening liquid for nourishing white hair and black hair, the raw materials of which include: 5g of kaempferol, 1g of geranic acid, 1g of lecithin, 1g of cocamidopropyl betaine, 40g of camellia oil, 1g of blue copper peptide, 1g of dopamine hydrochloride, 0.01g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide, 0.3g of Rehmannia glutinosa extract, and 0.7g of Platycladus orientalis leaf extract.

[0028] The method for preparing the above-mentioned hair loss prevention and blackening liquid for nourishing white hair comprises the following steps: S1, adding kaempferol to 30g of 40% ethanol aqueous solution, refluxing and stirring at 40°C for 10min, adding geranic acid thereto, continuing stirring for 1h, rotary evaporation, and vacuum drying to obtain a kaempferol composite material; S2, adding kaempferol compound, lecithin and cocamidopropyl betaine to camellia oil, stirring at a speed of 100 r / min for 10 min, adjusting the system temperature to 40° C., ultrasonically treating for 10 min at an ultrasonic frequency of 10 kHz to obtain an oil phase; S3. Add blue copper peptide to 30g of 0.01mol / L acetic acid aqueous solution and stir evenly, add dopamine hydrochloride and 1-ethyl-(3-dimethylaminopropyl)carbodiimide thereto, adjust the pH value of the system to 5.5-6, ultrasonically treat for 1h, the ultrasonic frequency is 12kHz, dialyze with 0.01mol / L acetic acid aqueous solution for 5h, freeze-dry, and then mix with Rehmannia glutinosa extract and Platycladus orientalis leaf extract, then add to 100g of water and stir evenly, add oil phase dropwise thereto under stirring, and stir at a speed of 1000r / min for 5min. Embodiment 2

[0029] A hair loss prevention and blackening liquid for nourishing white hair and black hair, the raw materials of which include: 10g of kaempferol, 5g of geranic acid, 2g of lecithin, 2g of glyceryl stearate, 60g of grape seed oil, 3g of blue copper peptide, 2g of dopamine hydrochloride, 0.1g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide, 1.5g of ginseng root extract, and 0.5g of chuanxiong root extract.

[0030] The method for preparing the above-mentioned hair loss prevention and blackening liquid for nourishing white hair comprises the following steps: S1, adding kaempferol to 60g of 60% ethanol aqueous solution, refluxing and stirring at 60°C for 20min, adding geranic acid thereto, continuing stirring for 3h, rotary evaporation, and vacuum drying to obtain a kaempferol composite material; S2, adding kaempferol compound, lecithin and stearyl glyceryl to grape seed oil, stirring at a speed of 400 r / min for 30 min, adjusting the system temperature to 60° C., ultrasonically treating for 20 min at an ultrasonic frequency of 15 kHz to obtain an oil phase; S3. Add blue copper peptide to 60g of 0.05mol / L acetic acid aqueous solution and stir evenly, add dopamine hydrochloride and 1-ethyl-(3-dimethylaminopropyl)carbodiimide thereto, adjust the pH value of the system to 5.5-6, ultrasonically treat for 2h, the ultrasonic frequency is 16kHz, dialyze with 0.05mol / L acetic acid aqueous solution for 10h, freeze-dry, and then mix with ginseng root extract and Chuanxiong root extract, and then add to 200g water and stir evenly, add oil phase dropwise thereto under stirring, and stir at a speed of 2000r / min for 15min. Embodiment 3

[0031] The invention discloses a hair loss prevention and blackening liquid for nourishing white hair and black hair. The raw materials of the hair loss prevention and blackening liquid include: 7g of kaempferol, 4g of geranic acid, 1.3g of lecithin, 1.8g of Tween 60, 45g of palm oil, 2.5g of blue copper peptide, 1.3g of dopamine hydrochloride, 0.08g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide and 1.3g of traditional Chinese medicine extract.

[0032] The Chinese herbal medicine extracts include: 2g of mulberry fruit extract, 4g of Polygonum multiflorum root extract, 5g of Rehmannia glutinosa extract, 1g of mulberry leaf extract, and 4g of ginseng root extract.

[0033] The method for preparing the above-mentioned hair loss prevention and blackening liquid for nourishing white hair comprises the following steps: S1, adding kaempferol to 40g of 55% ethanol aqueous solution, refluxing and stirring at 45°C for 18min, adding geranic acid thereto, continuing stirring for 1.5h, rotary evaporation, and vacuum drying to obtain a kaempferol composite material; S2, adding kaempferol compound, lecithin and Tween 60 to palm oil, stirring at a speed of 300 r / min for 15 min, adjusting the system temperature to 55° C., ultrasonically treating for 13 min at an ultrasonic frequency of 13 kHz to obtain an oil phase; S3. Add blue copper peptide to 40g of 0.04mol / L acetic acid aqueous solution and stir evenly, add dopamine hydrochloride and 1-ethyl-(3-dimethylaminopropyl)carbodiimide thereto, adjust the pH value of the system to 5.5-6, ultrasonically treat for 80min, the ultrasonic frequency is 15.5kHz, dialyze with 0.02mol / L acetic acid aqueous solution for 9h, freeze-dry, and then mix with the Chinese medicine extract, add to 120g water and stir evenly, add oil phase dropwise thereto under stirring, and stir at a speed of 1700r / min for 8min. Embodiment 4

[0034] The invention discloses a hair loss prevention and blackening liquid for nourishing white hair and black hair. The raw materials of the hair loss prevention and blackening liquid include: 9g of kaempferol, 2g of geranic acid, 1.7g of lecithin, 1.2g of Span 80, 55g of palm oil, 1.5g of blue copper peptide, 1.7g of dopamine hydrochloride, 0.02g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide and 1.7g of traditional Chinese medicine extract.

[0035] The Chinese herbal medicine extracts include: 1g of mulberry fruit extract, 6g of Polygonum multiflorum root extract, 3g of Rehmannia glutinosa extract, 2g of mulberry leaf extract, and 2g of ginseng root extract.

[0036] The method for preparing the above-mentioned hair loss prevention and blackening liquid for nourishing white hair comprises the following steps: S1, adding kaempferol to 50g of 45% ethanol aqueous solution, refluxing and stirring at 55°C for 12min, adding geranic acid thereto, continuing stirring for 2.5h, rotary evaporation, and vacuum drying to obtain a kaempferol composite material; S2, adding kaempferol compound, lecithin and Span 80 to palm oil, stirring at a speed of 200 r / min for 25 min, adjusting the system temperature to 45° C., and ultrasonically treating for 17 min at an ultrasonic frequency of 11 kHz to obtain an oil phase; S3. Add blue copper peptide to 50g of 0.02mol / L acetic acid aqueous solution and stir evenly, add dopamine hydrochloride and 1-ethyl-(3-dimethylaminopropyl)carbodiimide thereto, adjust the pH value of the system to 5.5-6, ultrasonically treat for 100min, the ultrasonic frequency is 13.5kHz, dialyze with 0.04mol / L acetic acid aqueous solution for 7h, freeze-dry, and then mix with the traditional Chinese medicine extract, add to 180g of water and stir evenly, add oil phase dropwise thereto under stirring, and stir at a speed of 1300r / min for 12min. Embodiment 5

[0037] The invention discloses a hair loss prevention and blackening liquid for nourishing white hair and black hair. The raw materials of the hair loss prevention and blackening liquid include: 8g of kaempferol, 3g of geranic acid, 1.5g of lecithin, 1.5g of Tween 80, 50g of palm oil, 2g of blue copper peptide, 1.5g of dopamine hydrochloride, 0.05g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide and 1.5g of traditional Chinese medicine extract.

[0038] The Chinese herbal medicine extracts include: 1.5g of mulberry fruit extract, 5g of Polygonum multiflorum root extract, 4g of Rehmannia glutinosa extract, 1.5g of mulberry leaf extract, and 3g of ginseng root extract.

[0039] The method for preparing the above-mentioned hair loss prevention and blackening liquid for nourishing white hair comprises the following steps: S1, adding kaempferol to 45g of 50% ethanol aqueous solution, refluxing and stirring at 50°C for 15min, adding geranic acid thereto, continuing stirring for 2h, rotary evaporation, and vacuum drying to obtain a kaempferol composite material; S2, adding kaempferol compound, lecithin and Tween 80 to palm oil, stirring at a speed of 250 r / min for 20 min, adjusting the system temperature to 50° C., ultrasonically treating for 15 min at an ultrasonic frequency of 12 kHz to obtain an oil phase; S3. Add blue copper peptide to 45g of 0.03mol / L acetic acid aqueous solution and stir evenly, add dopamine hydrochloride and 1-ethyl-(3-dimethylaminopropyl)carbodiimide thereto, adjust the pH value of the system to 5.5-6, ultrasonically treat for 90min, the ultrasonic frequency is 15kHz, dialyze with 0.03mol / L acetic acid aqueous solution for 8h, freeze-dry, and then mix with the traditional Chinese medicine extract, add to 150g of water and stir evenly, add oil phase dropwise thereto under stirring, and stir at a speed of 1500r / min for 10min.

[0040] Comparative Example 1 The invention discloses a hair loss prevention and blackening liquid for nourishing white hair and black hair. The raw materials of the hair loss prevention and blackening liquid include: 8g of kaempferol, 3g of geranic acid, 1.5g of lecithin, 1.5g of Tween 80, 50g of palm oil, 2g of blue copper peptide, 1.5g of dopamine hydrochloride and 1.5g of traditional Chinese medicine extract.

[0041] The Chinese herbal medicine extracts include: 1.5g of mulberry fruit extract, 5g of Polygonum multiflorum root extract, 4g of Rehmannia glutinosa extract, 1.5g of mulberry leaf extract, and 3g of ginseng root extract.

[0042] The method for preparing the above-mentioned hair loss prevention and blackening liquid for nourishing white hair comprises the following steps: S1, adding kaempferol to 45g of 50% ethanol aqueous solution, refluxing and stirring at 50°C for 15min, adding geranic acid thereto, continuing stirring for 2h, rotary evaporation, and vacuum drying to obtain a kaempferol composite material; S2, adding kaempferol compound, lecithin and Tween 80 to palm oil, stirring at a speed of 250 r / min for 20 min, adjusting the system temperature to 50° C., ultrasonically treating for 15 min at an ultrasonic frequency of 12 kHz to obtain an oil phase; S3. Mix blue copper peptide, dopamine hydrochloride and Chinese herbal extract, add to 150g water and stir evenly, add oil phase dropwise therein under stirring, and stir at a speed of 1500r / min for 10min.

[0043] Comparative Example 2 The invention discloses a hair loss prevention and blackening liquid for nourishing white hair and black hair. The raw materials of the hair loss prevention and blackening liquid include: 8g of kaempferol, 3g of geranic acid, 1.5g of lecithin, 1.5g of Tween 80, 50g of palm oil, 2g of blue copper peptide, 1.5g of dopamine hydrochloride, 0.05g of 1-ethyl-(3-dimethylaminopropyl)carbodiimide and 1.5g of traditional Chinese medicine extract.

[0044] The Chinese herbal medicine extracts include: 1.5g of mulberry fruit extract, 5g of Polygonum multiflorum root extract, 4g of Rehmannia glutinosa extract, 1.5g of mulberry leaf extract, and 3g of ginseng root extract.

[0045] The method for preparing the above-mentioned hair loss prevention and blackening liquid for nourishing white hair comprises the following steps: S1. Add kaempferol, geranyl acid, lecithin and Tween 80 to palm oil, stir at a speed of 250 r / min for 20 min, adjust the system temperature to 50° C., and perform ultrasonic treatment for 15 min at an ultrasonic frequency of 12 kHz to obtain an oil phase; S2. Add blue copper peptide to 45g of 0.03mol / L acetic acid aqueous solution and stir evenly, add dopamine hydrochloride and 1-ethyl-(3-dimethylaminopropyl)carbodiimide thereto, adjust the pH value of the system to 5.5-6, ultrasonically treat for 90min, the ultrasonic frequency is 15kHz, dialyze with 0.03mol / L acetic acid aqueous solution for 8h, freeze-dry, and then mix with the traditional Chinese medicine extract, add to 150g of water and stir evenly, add oil phase dropwise thereto under stirring, and stir at a speed of 1500r / min for 10min.

[0046] The hair loss prevention and blackening liquid obtained in Example 5 and Comparative Examples 1-2 was used to culture B16 melanoma cells, and the intracellular melanin content of each group after culture was detected. The details are as follows: The density of the B16 melanoma cell suspension was adjusted to 1×10 5 1 mL was inoculated into each well of a 12-well plate to a cell density of 5 × 10 4 Each well was placed in an incubator (37°C, 5% CO 2) Incubate for 24 hours. Take out the 12-well plate, remove the culture medium, add 1 mL of culture medium containing each group of samples (concentration is 1 mg / mL), add 1 mL of blank culture medium to the blank control group and the zero adjustment group, continue to culture in the incubator for 12 hours, and then remove the culture medium; 1 mL of 1 μM α-MSH solution prepared with RPMI1640 medium was added to each well of the experimental group and the zero-adjustment group, and 1 mL of medium without α-MSH was added to the blank group, and the cells were placed in an incubator for 36 hours. The cells were washed twice with 1 mL of PBS, and 100 μL of 1M NaOH solution (containing 10% DMSO) was added to each well. After being bathed in a water bath at 80°C for 1 hour, the cell extracts of each group in the 12-well plate were transferred to a 96-well plate, and the absorbance OD value of each well was measured at 475 nm on an ELISA reader to calculate the intracellular melanin content.

[0047] Intracellular melanin content = (OD 1 -OD 3 )÷(OD 2 -OD 3 )×100% Among them, OD 1 is the absorbance value of the experimental group, OD 2 is the absorbance value of the blank control group, OD 3 is the absorbance value of the zero adjustment group.

[0048] like Figure 1 As shown, the intracellular melanin content of Example 5 group was the highest, which was better than that of Comparative Examples 1-2 groups (P < 0.05).

[0049] The anti-hair loss and blackening liquid obtained in Example 5 and Comparative Examples 1-2 were used to detect the tyrosinase activation rate, specifically as follows: using phosphate buffer as a solvent, 0.5 mg / mL L-tyrosine solution and 35 mg / mL tyrosinase solution were prepared respectively; using propylene glycol as a solvent, 1 mg / mL solution of the test sample (the anti-hair loss and blackening liquid obtained in Example 5 and Comparative Examples 1-2) was prepared.

[0050] Prepare the following 4 groups of solutions: A: 0.7 mL L-tyrosine solution + 0.7 mL pure propylene glycol + 0.6 mL phosphate buffer; B: 0.7 mL pure propylene glycol + 1.3 mL phosphate buffer; C: 0.7 mL L-tyrosine solution + 0.7 mL test sample solution + 0.6 mL phosphate buffer; D: 0.7 mL of the test sample solution + 0.6 mL of phosphate buffer.

[0051] After the above four groups of solutions were kept constant at 37°C in a water bath for 10 minutes, 0.3 mL of tyrosinase solution was added to each of them. After reacting for 5 minutes, the solutions were quickly transferred to a cuvette, and the absorbance was measured at 475 nm to calculate the tyrosinase activation rate.

[0052]

[0053] Among them, (OD 1 -OD 2 ) represents the change in OD value of the reaction solution under the condition of no sample, (OD 3 -OD 4 ) represents the change in OD value of the reaction solution under the condition of sample.

[0054] like Figure 1 As shown, the tyrosinase activation rate of Example 5 group was the highest, which was better than that of Comparative Example 1-2 groups (P < 0.05).

[0055] The hair growth and hair darkening effects of the anti-hair loss liquid obtained in Example 5 and Comparative Examples 1-2 were compared, and C57BL / 6 mice (half male and half female, clean grade, 7-8 weeks old, weighing 20-22 g) were used as experimental subjects. The details are as follows: 1. Comparative test of hair growth effect: All mice were anesthetized and their back hair was removed with paraffin-rosin, with the removal area of ​​about 2cm×2cm. They were randomly divided into 5 groups, and the corresponding drugs were applied to the depilated area from the next day, twice a day.

[0056] The NC group was smeared with distilled water, the PC group was smeared with minoxidil (2.0%, W / V), and the S5, D1, and D2 groups were smeared with the anti-hair loss liquid obtained in Example 5, the anti-hair loss liquid obtained in Comparative Example 1, and the anti-hair loss liquid obtained in Comparative Example 2, respectively.

[0057] 15 days after depilation, all mice were killed by cervical dislocation. The skin of the same part of the depilated area was fixed with 10% formaldehyde, dehydrated, embedded in paraffin, sliced, and spread on poly-lysine slides. In situ hybridization technology and terminal labeling were used to detect the growth / apoptosis of cells in the hair follicles.

[0058] 2. Comparative test of black hair effect: After all mice were anesthetized, paraffin-rosin was used to remove the hair on their backs, and the removal area was about 2cm×2cm. 12h after depilation, mice without wounds on the back were selected, and a 6% hydrogen peroxide solution was applied to the depilated area twice a day for 7 days. The successful modeling criteria were: the mice had no abnormal performance, white hair grew on the surface of the back skin, and there was no black hair on the back, no black skin and wounds.

[0059] The mice were randomly divided into 5 groups. The corresponding drugs were applied to the light-colored hair growth area from the next day after the model was successfully established, twice a day. The NC group was applied with distilled water, the PC group was applied with minoxidil (2.0%, W / V), and the S5, D1, and D2 groups were applied with the anti-hair loss liquid obtained in Example 5, the anti-hair loss liquid obtained in Comparative Example 1, and the anti-hair loss liquid obtained in Comparative Example 2, respectively.

[0060] 15 days after successful modeling, all mice were killed by cervical dislocation. The skin of the same part of the depilated area was fixed with 10% formaldehyde, dehydrated, and embedded in paraffin. Two longitudinal and cross-sections of the skin were taken for each slice, and HE staining was performed respectively. The distribution of melanin in each group in the longitudinal section was observed under a microscope.

[0061] like Figure 2 As shown, the number of apoptotic hair follicle cells in group S5 was the least, while the number of melanin hair follicles was the largest, which was better than that in groups D1 and D2 (P < 0.05) and also better than that in group PC (P < 0.05).

[0062] The anti-hair loss and blackening liquid obtained in Example 5 and Comparative Examples 1-2 were used to conduct anti-inflammatory experiments, as follows: Mouse macrophage RAW264.7 cells were cultured at 2×10 5 The cells were plated at a concentration of 100 cells / well in a 24-well plate and incubated in DMEM medium at 37°C and 5% CO. 2 The cells were cultured for 24 h under the conditions of , and then the anti-hair loss and blackening liquids obtained in Example 5 and Comparative Example 1-2 were added at a concentration of 1 mg / mL, and 100 μg / mL dexamethasone was added to the positive control group; 30 minutes after the addition of the sample, LPS was added to a final concentration of 1 μg / mL, and the cells were cultured at 37°C for 20 h. The culture medium was collected, and the secreted IL-6 and TNF-α were quantified using a mouse IL-6 enzyme-linked immunosorbent assay kit and a mouse TNF-α enzyme-linked immunosorbent assay kit, respectively.

[0063] like Figure 3 As shown, compared with the negative control, the positive control had significantly reduced secretions of inflammatory factors IL-6 and TNF-α by macrophage RAW264.7 under 100 μg / mL dexamethasone (P < 0.05), indicating that the positive control detection was effective; and the anti-hair loss and blackening liquid obtained in Example 5 made the secretions of inflammatory factors IL-6 and TNF-α by macrophage RAW264.7 similar to those of the positive control (P > 0.05), and better than those of comparative examples 1-2 (P < 0.05), confirming that the anti-hair loss and blackening liquid obtained in Example 5 can inhibit the secretion of inflammatory factors TNF-α and IL-6 by macrophage RAW264.7.

[0064] The applicant believes that the reason for the above results is that the present invention uses kaempferol as an effective ingredient, which can effectively reduce the inflammation of human hair papilla cells and surrounding cells, and has a significant anti-hair loss effect. It combines with geranyl acid to form a synergistic effect, comprehensively improves the microenvironment around the hair follicles, and has a significant hair blackening effect. It can effectively penetrate into the scalp and hair follicles through interaction with lecithin, effectively activate tyrosinase, repair damaged melanocytes, and have good durability of black hair; the present invention also combines dopamine to the surface of blue copper peptide, which can not only be effectively dispersed at the oil-water interface, thereby having a larger interfacial area and smaller mass transfer resistance, but also has good biocompatibility, can effectively help the active ingredients pass through the skin barrier, and is more conducive to the transdermal absorption of the active ingredients, so that the active ingredients are efficiently delivered to the hair follicles, with a significant anti-inflammatory effect, and can effectively reduce sebum secretion, and promote blood circulation, effectively improve the body's immunity, and can activate hair follicles to promote hair regrowth, and can well promote scalp absorption, achieving a good therapeutic effect.

[0065] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A hair loss prevention liquid for nourishing white hair and black hair, characterized in that: The raw materials include, by mass, 5-10 parts of kaempferol, 1-5 parts of geranic acid, 1-2 parts of lecithin, 1-2 parts of surfactant, 40-60 parts of vegetable oil, 1-3 parts of blue copper peptide, 1-2 parts of dopamine hydrochloride, 0.01-0.1 parts of 1-ethyl-(3-dimethylaminopropyl)carbodiimide and 1-2 parts of traditional Chinese medicine extract.

2. The hair loss prevention and darkening liquid for nourishing white hair and black hair according to claim 1, characterized in that: The surfactant is at least one of polyoxyethylene sorbitan fatty acid ester, sorbitan fatty acid ester, fatty acid glyceride and cocamidopropyl betaine.

3. The hair loss prevention and blackening liquid for nourishing white hair according to claim 1, characterized in that: The vegetable oil is at least one of peanut oil, rapeseed oil, soybean oil, corn oil, camellia oil, walnut oil, olive oil, palm oil, grapeseed oil and sunflower seed oil.

4. The hair loss prevention and blackening liquid for nourishing white hair according to claim 1, characterized in that: The Chinese medicine extract is at least one of mulberry fruit extract, licorice root extract, Polygonum multiflorum root extract, Rehmannia glutinosa extract, Platycladus orientalis leaf extract, black sesame extract, Channa intestine extract, dandelion extract, mulberry leaf extract, ginseng root extract, Chuanxiong root extract, and Magnolia officinalis bark extract.

5. The hair loss prevention and blackening liquid for nourishing white hair according to claim 1, characterized in that: Preferably, the Chinese medicine extract includes: mulberry fruit extract, Polygonum multiflorum root extract, Rehmannia glutinosa extract, mulberry leaf extract, and ginseng root extract; the mass ratio of mulberry fruit extract, Polygonum multiflorum root extract, Rehmannia glutinosa extract, mulberry leaf extract, and ginseng root extract is 1-2:4-6:3-5:1-2:2-4.

6. A method for preparing the hair loss prevention and blackening liquid for nourishing white hair as claimed in any one of claims 1 to 5, characterized in that: The steps include: S1, adding kaempferol to ethanol aqueous solution, reflux stirring at 40-60° C. for 10-20 min, adding geranic acid thereto and continuing stirring for 1-3 h, rotary evaporation, and vacuum drying to obtain kaempferol composite material; S2, adding kaempferol compound, lecithin and surfactant to vegetable oil and stirring for 10-30 minutes, adjusting the system temperature to 40-60° C., and ultrasonically treating for 10-20 minutes to obtain an oil phase; S3. Add blue copper peptide to acetic acid aqueous solution and stir evenly, add dopamine hydrochloride and 1-ethyl-(3-dimethylaminopropyl)carbodiimide thereto, adjust the pH value of the system to 5.5-6, ultrasonically treat for 1-2 hours, dialyze for 5-10 hours, freeze-dry, and then mix with the Chinese medicine extract, add to water and stir evenly, add oil phase dropwise thereto under stirring, and stir for 5-15 minutes.

7. The method for preparing the hair loss-preventing and blackening liquid for nourishing white hair according to claim 6, characterized in that: In S2, the ultrasound frequency is 10-15kHz.

8. The method for preparing the hair loss-preventing and blackening liquid for nourishing white hair according to claim 6, characterized in that: In S3, the concentration of the acetic acid aqueous solution is 0.01-0.05 mol / L.

9. The method for preparing the hair loss-preventing and blackening liquid for nourishing white hair according to claim 6, characterized in that: In S3, the ultrasound frequency is 12-16kHz.

10. The method for preparing the hair loss-preventing and blackening liquid for nourishing white hair according to claim 6, characterized in that: In S3, dialysis is performed with an aqueous acetic acid solution having a concentration of 0.01-0.05 mol / L for 5-10 hours.

Images