A hair loss prevention essence containing kuding tea ilex leaf extract and a preparation method thereof

Abstract

The present application relates to a kind of anti-hair loss essence containing kudingcha ilex extract and its preparation method, kudingcha ilex extract is added in anti-hair loss essence, anti-hair loss essence can show the effect of promoting hair growth and inhibiting hair loss after use;Anti-hair loss essence also includes lactobacillus fermentation product and penetration enhancer;Lactobacillus fermentation product is used as conditioner, can improve the microenvironment of scalp, and cooperates with kudingcha ilex extract to improve the condition of hair loss;In addition, through the effect of penetration enhancer, the absorption degree of scalp to kudingcha ilex extract and is improved.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of hair loss prevention, and particularly relates to a hair loss prevention essence containing Ilex kudingcha and ilex hainanensis leaf extract and a preparation method thereof. BACKGROUND

[0002] Hair loss has become a global health challenge, with the incidence of hair loss rapidly rising and showing a trend of youth as mental stress and environmental pollution intensify, and thus the demand for hair loss prevention products is growing. The pathogenesis of hair loss is closely related to abnormal lipid metabolism, oxidative stress damage, microcirculation disorder of the scalp, and functional decline of the hair follicle.

[0003] The mainstream drugs approved by the FDA for treating hair loss are two: minoxidil for external use and finasteride for oral use, but long-term use of minoxidil or finasteride has a series of toxic side effects, such as drug dependence, skin irritation, cardiovascular effects, and sexual dysfunction. And compared to the complex causes of hair loss, the mechanisms of these two drugs are single and have low transdermal efficiency, and the effect of treating hair loss is limited. In non-drug treatment, plant extracts have gradually become a research hotspot in the field of hair loss prevention due to their natural and safe characteristics. For example, ginseng, radix polygoni multiflori, and cypress leaves have the effects of promoting scalp blood circulation, enhancing hair follicle nutrition, and inhibiting fungal reproduction, and can effectively improve the scalp environment and promote hair growth. However, in the prior art, there are still many problems in the application of plant extracts. For example, the extraction process is complex, the active ingredients are not clear or have poor stability, and the absorption effect of some products is not good, making it difficult to play a role in the hair follicle. In addition, some hair loss prevention products on the market lack scientific verification of their efficacy or mechanism, or are expensive, making it difficult to meet the actual needs of consumers. Therefore, it is urgent to develop safe and efficient alternative therapies with multiple mechanisms to solve the existing hair loss problems.

[0004] As an important product for improving scalp conditions and promoting hair growth, scalp essence has received widespread attention. However, existing scalp essence products have exposed many problems in actual application, making it difficult to fully meet people's demand for scalp health and strong and dense hair. From the perspective of scalp essence penetration rate, a large number of researches and actual use feedbacks show that the existing scalp essence generally has the problem of low penetration rate. The structure of the scalp is relatively complex, and the stratum corneum, the outermost layer, acts as a natural barrier to the absorption of substances. When using conventional scalp essence, the active ingredients in the essence often have difficulty in effectively penetrating the stratum corneum and reaching the hair follicles and surrounding tissues where they play a role. From the perspective of scalp environment regulation and hair growth promotion, the existing scalp essence also has obvious deficiencies. The scalp environment is a complex ecosystem, including microbial community, oil secretion, moisture content, acid-base balance, and many other aspects.

[0005] A healthy scalp environment is the basis of hair growth, however, many existing scalp serum products cannot comprehensively and effectively condition the scalp environment. Some scalp serums are designed to address only one issue of the scalp, such as oil control or moisturizing, ignoring the integrity and complexity of the scalp environment. For example, some oil control scalp serums, although can reduce sebum secretion to some extent, may at the same time disrupt the water-oil balance of the scalp, leading to excessive dryness of the scalp, and thus affecting the normal metabolism of the scalp and hair growth. Moreover, existing scalp serums have poor effects in regulating the scalp microbial community. There are a variety of microorganisms on the scalp, which are in a balanced state under normal circumstances, and together maintain the health of the scalp. However, when the scalp microbial community is imbalanced, such as excessive proliferation of Malassezia, it may cause scalp itching, increased dandruff, and other problems, and thus affect the health of hair follicles and hair growth. At present, most scalp serums lack ingredients or technologies that can effectively regulate the balance of the scalp microbial community, and thus cannot fundamentally solve the scalp problems caused by microbial imbalance, and it is difficult to create a good scalp environment conducive to hair growth, and to achieve the goal of promoting hair growth. SUMMARY

[0006] To solve the above technical problems, the present application provides a hair loss prevention serum containing Ilex kudingcha C.J. Tseng leaf extract and a preparation method thereof.

[0007] The technical solution adopted by the present application is: a hair loss prevention serum containing Ilex kudingcha C.J. Tseng leaf extract, comprising Ilex kudingcha C.J. Tseng leaf extract and lactobacillus fermentation product; Ilex kudingcha C.J. Tseng leaf is extracted by water decoction, the filtrate obtained by decoction is concentrated and dried, and the obtained solid material is Ilex kudingcha C.J. Tseng leaf extract; the mass fraction of Ilex kudingcha C.J. Tseng leaf extract in the hair loss prevention serum is 0.05%-10%; the lactobacillus fermentation product is used as a conditioner, and the mass fraction is 0.1-10%.

[0008] Preferably, the lactobacillus fermentation product is lactobacillus fermentation lysate or plant lactobacillus fermentation product.

[0009] Preferably, the lactobacillus fermentation product is replaced by Pichia pastoris fermentation lysate filtrate.

[0010] Preferably, it further comprises 2-8% of penetration enhancer, 0.8-1.35% of preservative, 2-15.5% of humectant, 0.1-1% of chelating agent, and the balance is solvent.

[0011] Preferably, the mass ratio of Ilex kudingcha C.J. Tseng leaf extract, conditioner and penetration enhancer is 1:2-7:4-10.

[0012] Preferably, the penetration enhancer is isosorbide dimethyl ether and / or ethoxydiglycol.

[0013] Preferably, the preservative is selected from one or more of nipagin, octisalol, phenoxyethanol, ethylhexylglycerin, 1,2-hexanediol, p-hydroxyacetophenone;

[0014] The humectant is selected from one or more of glycerin, D-pantenol, propylene glycol, butylene glycol, betaine;

[0015] The solvent is water or a mixture of water and ethanol.

[0016] A method for preparing a hair loss prevention serum containing Ilex hainanensis leaf extract, the solvent, Ilex hainanensis leaf extract, preservative, humectant, chelating agent are added to the emulsifying tank, and the stirring is started at a speed of 20-40 rpm; the temperature is raised to 80°C for 10-15 min;

[0017] The stirring is cooled to 40-50°C, the conditioning agent is added, and the stirring is uniform until the temperature is below 40°C, the tank is discharged, and the product is aged for 48 hours to obtain the hair loss prevention serum containing Ilex hainanensis leaf extract.

[0018] Preferably, the pH value of the hair loss prevention serum is adjusted to 5.0-7.0;

[0019] Preferably, the pH value of the hair loss prevention serum is adjusted by lactic acid, citric acid or arginine.

[0020] Preferably, the Ilex hainanensis leaf extract contains Ilex saponins active substances.

[0021] The hair loss prevention serum containing Ilex hainanensis leaf extract can promote hair growth and inhibit hair loss; the addition of the lactobacillus fermentation product as a conditioning agent can improve the microenvironment of the scalp and improve the hair loss condition in cooperation with the Ilex hainanensis leaf extract; and the absorption of the Ilex hainanensis leaf extract and the lactobacillus fermentation product by the scalp is improved through the action of the penetration enhancer. BRIEF DESCRIPTION OF DRAWINGS

[0022] Figure 1 Comparison of the activity of Ilex hainanensis leaf active substances on hair follicle stem cell proliferation;

[0023] Figure 2 Mass spectrum analysis of total saponin components in Ilex hainanensis leaf extract;

[0024] Figure 3 Determination of total saponin content in Ilex hainanensis leaf extract, Ilex latifolia leaf extract and Sabina vulgaris extract;

[0025] Figure 4 Comparison of the activity of Ilex hainanensis leaf extract, Ilex latifolia leaf extract and Sabina vulgaris total saponin on hair follicle stem cell proliferation;

[0026] Figure 5Mass spectrum of the saponin component in Ilex kudingcha, Ilex latifolia and Ilex asprella;

[0027] Figure 6 Effect diagram of Ilex kudingcha leaf extract on preventing hair loss in androgenetic alopecia mice;

[0028] Figure 7 Influence of different conditioning agents on the regulation of scalp microenvironment by the anti-hair loss serum containing Ilex kudingcha leaf extract;

[0029] Figure 8 Influence of plant lactobacillus ferment on the regulation of scalp microenvironment by the anti-hair loss serum containing Ilex kudingcha leaf extract;

[0030] Figure 9 Effect diagram of the anti-hair loss serum containing Ilex kudingcha leaf extract on preventing hair loss in androgenetic alopecia mice;

[0031] Figure 10 Hair mirror analysis diagram of the anti-hair loss serum containing Ilex kudingcha leaf extract before and after use. DETAILED DESCRIPTION

[0032] The embodiments of the present application will be described below in conjunction with the accompanying drawings.

[0033] The present application relates to an anti-hair loss serum containing Ilex kudingcha leaf extract and a preparation method thereof, which adds plant extract, Ilex kudingcha leaf extract, or a plant composition mainly containing Ilex kudingcha leaf to the hair care product serum. The anti-hair loss serum can exhibit the effects of promoting hair growth and inhibiting hair loss after use.

[0034] Ilex kudingcha is an arbor plant of Ilex in Aquifoliaceae, which is mainly produced in Hainan and Guangxi. It is mainly used as medicine and food, and has the effects of dispelling wind and heat, clearing the head and eyes, relieving thirst, etc. Ilex kudingcha has a long history in China, and the representatives are Ilex latifolia ( Ilex latifolia Thunb.), Ilex kudingcha ( Ilex kudingcha C.J. Tseng) and Ilex asprella ( Ilex cornuta Lindl. et Paxt.). The main component in Ilex kudingcha is saponin, and it also contains flavonoids, polyphenols, polysaccharides, volatile oil and other components. Through the extraction of effective components in Ilex kudingcha leaf, the obtained extract can be used for preparing anti-hair loss external preparation.

[0035] The Ilex kudingcha leaves are extracted by water decoction method to obtain the Ilex kudingcha leaf extract. The solution extracted by water decoction can be further concentrated and dried to obtain the Ilex kudingcha leaf extract which can be used as a raw material to prepare other products. In some embodiments of the present application, the Ilex kudingcha leaf extract is an effective component including total polysaccharides, total phenolic acids, total saponins and total flavonoids, etc. The total polysaccharides, total phenolic acids, total saponins and total flavonoids in the water extract of the Ilex kudingcha leaf can be separated by different concentrations of ethanol, and then the separated total polysaccharides, total phenolic acids, total saponins and total flavonoids are mixed to obtain a new type of Ilex kudingcha leaf extract. In some other embodiments of the present application, the Ilex kudingcha leaf extract specifically refers to the saponin active substance extracted.

[0036] The Ilex kudingcha leaves are boiled with water, and distilled water is added to the Ilex kudingcha leaves at a ratio of w / v = 1:10-15 to extract the active ingredients, and the extraction time is 0.5-3 h. The solution obtained by filtration is concentrated and dried to obtain the Ilex kudingcha leaf extract. Alternatively, the Ilex kudingcha leaves are boiled with water to extract the active ingredients, and the filtrate is concentrated and then 70% ethanol is added. The precipitate is the total polysaccharides. The supernatant is extracted, and the extract is loaded onto an adsorption resin column. The total phenolic acids are eluted by 30% ethanol, the total saponins are eluted by 50% ethanol, and the total flavonoids are eluted by 80% ethanol. The total polysaccharides, total phenolic acids, total saponins and total flavonoids are mixed and dried to obtain the Ilex kudingcha leaf extract. Alternatively, only the total saponins prepared by the above method are used as the Ilex kudingcha leaf extract. The water extract of the Ilex kudingcha leaves is added with 70% ethanol to remove the total polysaccharides. The separated liquid is loaded onto an adsorption resin column, and the total phenolic acids are removed by elution with 30% ethanol. The eluate obtained by elution with 50% ethanol is concentrated and dried to obtain the Ilex kudingcha total saponins.

[0037] It is found by experiments that the Ilex kudingcha leaf can significantly inhibit the levels of triglyceride, total cholesterol or 5α-reductase in an androgenic alopecia mouse model; and inhibit the overexpression of ROS or MDA or abnormal expression of inflammatory factors TNF-α, IL-1β, IL-6 and INF-γ in the hair follicle stem cells of mice induced by ultraviolet irradiation or hydrogen peroxide. The Ilex kudingcha leaf extract can be used to improve alopecia caused by various mechanisms, such as inhibition of abnormal increase in sebum synthesis and secretion of sebaceous glands in the scalp or alopecia caused by excessive androgen levels, clearance of alopecia caused by excessive ROS levels in the scalp induced by physical damage or chemical stimulation, and inhibition of alopecia caused by abnormal expression of inflammatory factors in the scalp induced by physical, chemical or biological factors.

[0038] The addition of the Ilex kudingcha C.J. Tseng and Cyclobalanopsis glaucoides extract to the scalp essence of the hair care product can play the effect of promoting hair growth and inhibiting hair loss when the scalp essence is used. Further, the synergistic effect of the conditioning agent and the penetration enhancer and other additives can better realize the beneficial effect of controlling hair loss. The mass ratio of the Ilex kudingcha C.J. Tseng and Cyclobalanopsis glaucoides extract in the scalp essence is 0.05%-10% by weight. In some embodiments of the present application, the hair care product containing the Ilex kudingcha C.J. Tseng and Cyclobalanopsis glaucoides extract further includes a penetration enhancer, a conditioning agent, a preservative, a humectant, a chelating agent and a solvent; the penetration enhancer, the conditioning agent, the preservative, the humectant, the chelating agent and the solvent are included in an amount of 2-8%, 0.1-10%, 0.8-1.35%, 2-15.5%, 0.1-1% and the balance by weight, respectively.

[0039] The penetration enhancer is one or a combination of isosorbide dimethyl ether and ethoxydiglycol; the conditioning agent is a lactobacillus fermentation product or a filter liquor of a Pichia pastoris fermentation lysate, and the lactobacillus fermentation product can be a lactobacillus fermentation lysate product or a plant lactobacillus fermentation product; the preservative is one or several of methyl-n-propyl-glycol, octoxyglycerin, phenoxyethanol, ethylhexylglycerin, 1,2-hexanediol and p-hydroxyacetophenone; the humectant is one or several of glycerin, D-panthenol, propylene glycol, butylene glycol and betaine; and the chelating agent is EDTA-2Na.

[0040] In the preparation of the anti-hair loss essence, the solvent, the Ilex kudingcha C.J. Tseng and Cyclobalanopsis glaucoides extract, the preservative, the humectant and the chelating agent are added to an emulsifying tank, and stirring is started at a speed of 20-40 rpm; the temperature is raised to 80°C and kept for 10-15 min; the stirring is started to reduce the temperature, and the temperature is reduced to 40-50°C; the conditioning agent is added, the pH value of the anti-hair loss essence is adjusted to 5.0-7.0 by lactic acid, citric acid or arginine, and the mixture is stirred uniformly until the temperature is below 40°C; the mixture is taken out of the tank, and is stored for 48 hours to obtain the anti-hair loss essence containing the Ilex kudingcha C.J. Tseng and Cyclobalanopsis glaucoides extract.

[0041] The core effects of anti-inflammatory and oil control of Ilex kudingcha C.J.Tseng depend on the active ingredients reaching the deep target points of the scalp (such as hair follicles, sebaceous glands and inflammatory cells). However, the barrier effect of the scalp stratum corneum and the opening of the hair follicle will hinder the penetration, resulting in most of the ingredients only staying on the surface and having limited effect. The penetration enhancer can help the active ingredients of Ilex kudingcha C.J.Tseng penetrate the barrier by opening the stratum corneum channel, reaching the dermis layer and the surrounding hair follicles, so that its anti-inflammatory effect is upgraded from "surface soothing" to "deep inhibition of inflammatory factors (such as IL-6 and TNF-alpha)", and the oil control effect is changed from "temporary adsorption of oil" to "inhibition of excessive secretion of sebaceous gland cells", greatly improving the improvement efficiency of scalp problems (such as dandruff, itching and greasy). The penetration enhancer promotes the penetration and absorption of active substances by changing the structure of the stratum corneum, increasing the solubility of the active substances, changing the protein structure to enhance the sebaceous gland pathway, or increasing the water content; by matching the types and proportions of conditioning agents and penetration enhancers, a scientific and effective penetration mechanism is established.

[0042] In addition, if the stratum corneum is disordered when the scalp barrier is damaged, the penetration of the penetration enhancer will be unstable; at the same time, the scalp under the damaged barrier is easy to be stimulated by the outside world, and the anti-inflammatory ingredients of Ilex kudingcha C.J.Tseng may be "tired of responding to surface stimulation" and unable to play a deep role. The conditioning agent can stabilize the physiological environment of the scalp by repairing the barrier and maintaining the water-oil balance: on the one hand, it can control the penetration efficiency of the penetration enhancer, and on the other hand, it can provide a "direct target channel" for the active ingredients of Ilex kudingcha C.J.Tseng. The penetration enhancer breaks down the penetration barrier and amplifies the anti-inflammatory and oil control effects of Ilex kudingcha C.J.Tseng and the repair and moisturizing effects of the conditioning agent; Ilex kudingcha C.J.Tseng protects other components through antioxidant and natural anti-inflammatory protection and enhances the targeted effect; the conditioning agent repairs the barrier, stabilizes the penetration environment and reduces the risk of irritation. By reasonably matching the ratio of Ilex kudingcha C.J.Tseng, penetration enhancer and conditioning agent, and by compatibility screening, the synergistic effect of the three can be maximized to achieve the effect of "deep repair + precise oil control + barrier stability" for scalp health management.

[0043] The following describes the present application in combination with the accompanying drawings, wherein the experimental methods not specifically described in the operation steps are performed according to the corresponding product instructions. The instruments, reagents and consumables used in the examples can be purchased from commercial companies unless otherwise specified.

[0044] Example 1: Preparation and characterization of Ilex kudingcha C.J.Tseng leaf extract

[0045] 1.1 Preparation of Ilex kudingcha C.J.Tseng leaf extract

[0046] Dry Ilex kudingcha C.J.Tseng leaves were crushed and then extracted by water decoction to obtain the active ingredients of Ilex kudingcha C.J.Tseng leaf extract.

[0047] Take 1000g of dried Ilex kudingcha C.J. Tseng leaves, crush them, and then extract them with distilled water at a ratio of w / v = 1:15 for 2 hours. Filter the mixture while it is hot using a 100-mesh stainless steel sieve. Concentrate the filtrate to a suspension with a density of 1.1 using a flash evaporator (JMF-320, Xi'an). Add ethanol to the Ilex kudingcha C.J. Tseng leaf extract suspension to adjust the ethanol proportion in the solution to 70%. Mix well and let it stand overnight. Wash the precipitate with anhydrous ethanol three times. The main component obtained is total polysaccharides, which amounts to 35.4 g. Further concentrate the supernatant to prepare a solution with a solid content of about 1%. Load the solution into an AB-8 macroporous adsorption resin column (diameter: 5.5 cm; column height: 35 cm; column volume: about 850 mL) at a loading speed of 2 mL / min. Equilibrate the column with 5% ethanol solution to elute water-soluble impurities. Then, perform gradient elution with different concentrations of ethanol solution at an elution speed of 1 mL / min. The components obtained from the 30% ethanol solution elution part are mainly total phenolic acid components, which amount to 18.7 g. The components obtained from the 50% ethanol solution elution part are mainly total saponin components, which amount to 21.8 g. The components obtained from the 80% ethanol solution elution part are mainly total flavonoid components, which amount to 28.4 g.

[0048] 1.2 Analysis of Ilex kudingcha C.J. Tseng leaf extract components

[0049] To further analyze the key active ingredients in Ilex kudingcha C.J. Tseng leaves, different types of active ingredients in the Ilex kudingcha C.J. Tseng leaf extract were separated and purified. Total polysaccharides, total phenolic acids, total saponins, and total flavonoids were collected, and the functions of each type of active ingredient were analyzed and verified.

[0050] Take 1000g of dried Ilex kudingcha C.J. Tseng leaves, crush them, and then extract them with distilled water at a ratio of w / v = 1:15 for 2 hours. Filter the mixture while it is hot using a 100-mesh stainless steel sieve. Concentrate the filtrate to a suspension with a density of 1.1 using a flash evaporator (JMF-320, Xi'an). Add ethanol to the Ilex kudingcha C.J. Tseng leaf extract suspension to adjust the ethanol proportion in the solution to 70%. Mix well and let it stand overnight. Wash the precipitate with anhydrous ethanol three times. The main component obtained is total polysaccharides, which amounts to 35.4 g. Further concentrate the supernatant to prepare a solution with a solid content of about 1%. Load the solution into an AB-8 macroporous adsorption resin column (diameter: 5.5 cm; column height: 35 cm; column volume: about 850 mL) at a loading speed of 2 mL / min. Equilibrate the column with 5% ethanol solution to elute water-soluble impurities. Then, perform gradient elution with different concentrations of ethanol solution at an elution speed of 1 mL / min. The components obtained from the 30% ethanol solution elution part are mainly total phenolic acid components, which amount to 18.7 g. The components obtained from the 50% ethanol solution elution part are mainly total saponin components, which amount to 21.8 g. The components obtained from the 80% ethanol solution elution part are mainly total flavonoid components, which amount to 28.4 g.

[0051] 1.3 Effect of Ilex kudingcha C.J. Tseng leaf active ingredients on the proliferation activity of hair follicle stem cells

[0052] Test the rat hair follicle stem cells (HFSCs) proliferation activity of the separated total polysaccharides, total phenolic acids, total saponins, and total flavonoids. The rat hair follicle stem cells (HFSCs) were purchased from Wuhan Punsen Life Science and Technology Co., Ltd. Place the HFSCs in a special culture medium and continuously culture them in a 37 ℃, 5% CO2 constant temperature incubator. Change the medium every other day. When the cells grow to a confluence of 70%-80%, subculture them. Take the logarithmic growth phase HFSCs and dilute them to 1×10 4100 μL inoculated in 96-well plates, set up control group, kuding tea ilex group, total polysaccharide group, total phenolic acid group, total saponin group, total flavonoid group. Among them, the control group is the basic medium, and the other groups are the basic medium solution containing 0.1% of the corresponding components. HFSCs were inoculated into the control group and each experimental group medium, and cultured in the incubator for 24 h, 10 μL of CCK-8 reagent was added, and it was returned to the incubator for 1 h, and the absorbance value was detected at 450 nm; Each group sets up 3 duplicate wells to ensure the accuracy of the experiment. Compare the growth rate of cell proliferation in each experimental group relative to the control group, calculate the cell proliferation rate, and measure the promoting effect of different experimental groups on cell proliferation;

[0053] Cell proliferation rate%= (experimental group absorbance value - control group absorbance value) / control group absorbance value x 100%.

[0054] From Figure 1 It can be seen that kuding tea ilex leaf extract has a significant promoting effect on hair follicle stem cells (HFSCs). Compared with the control group, the average cell proliferation rate increased by 37.1%. The main components (total polysaccharide, total phenolic acid, total saponin, total flavonoid) all showed certain proliferation activity, among which the total saponin group had the most prominent promoting effect (the proliferation rate increased by 58.4%). It shows that kuding tea ilex has the effect of enhancing the activity of hair follicle stem cells, and is speculated to have potential application value in preventing hair loss, and the core active component is represented by total saponin components.

[0055] 1.4 Analysis and performance comparison of total saponin components in kuding tea ilex leaf extract

[0056] Further identification and analysis of key components of total saponin, the key active component in kuding tea ilex leaf, were carried out. The components were separated and identified by mass spectrometry analysis by UPLC-Q-TOF / MS.

[0057] Chromatographic column: CORTECS T3 (2.1 mm x 150 mm, 1.6 μm); chromatographic gradient elution condition: mobile phase A is acetonitrile, B is 0.1% formic acid aqueous solution; volume flow rate 0.2 mL / min; gradient elution (0~5 min: 5% A, 95% B; 5~35 min: 25%, 75% B; 35~40 min: 40% A, 60% B; 40~45 min: 75% A, 25% B; 45~50 min: 95% A, 5% B). The results are as follows Figure 2The key saponin components are partially intercepted and presented in the figure, and the most representative saponin components are analyzed by secondary mass spectrometry fragment information as shown in Table 1. Kudinoside G, kudinoside C, kudinoside A and kudinoside D are the main components of Ilex kudingcha saponins. The four saponin components are enriched, concentrated and dried according to the elution time. The four saponin components are enriched, concentrated and dried, and the fraction from 36.5 min to 40.41 min is collected, concentrated and dried. The four kudingcha saponins are determined for promoting the proliferation activity of hair follicle stem cells according to the method described in 1.3, and the results are shown in Table 1. It can be seen that the four saponin components have the activity of promoting the proliferation of hair follicle stem cells at a concentration of 10 μg / mL, but the activity is lower than that of the fraction from 36.5 min to 40.41 min (total saponin of Ilex kudingcha) at the same concentration. Comparison shows that the mixed saponin components of different types of Ilex kudingcha (total saponin of Ilex kudingcha) have better effect.

[0058] Table 1 Total saponin components and activity analysis of Ilex kudingcha leaf extract

[0059]

[0060] Example 2: Determination of total saponin content and performance analysis of Ilex kudingcha, Ilex latifolia and Sabina

[0061] 2.1 Comparison of total saponin content in Ilex kudingcha, Ilex latifolia and Sabina

[0062] The representative Ilex kudingcha, Ilex latifolia and Sabina leaves of Ilex kudingcha were compared. The Ilex kudingcha leaves, Ilex latifolia leaves and Sabina leaves were purchased from the medicinal material market of Guangxi Zhuang Autonomous Region, Zhejiang Province and Guizhou Province, respectively, and the total saponin content in the three raw materials was detected.

[0063] The ultraviolet-visible spectrophotometry was used to determine the content of total saponin in three batches of Ilex kudingcha, Ilex latifolia and Sabina, with ginsenoside Re (purchased from Shanghai Yuan Ye Biological Company, product number A10036) as the control, and vanillin-glacial acetic acid method for color development. Specifically, 100 uL of test sample or control sample solution was precisely taken and placed in a 10 mL stoppered test tube. The solvent was dried by 60℃ water bath, 0.2 mL of 10% vanillin-glacial acetic acid solution and 0.8 mL of perchloric acid were added, and after 60 ℃ water bath for 15 min, it was taken out and placed in an ice water bath for 5 min. 5 mL of glacial acetic acid was added and mixed well, and the absorbance was measured at 550 nm.

[0064] Preparation of control solution: 10 mg of ginsenoside Re control was precisely weighed and placed in a 10 mL volumetric flask. Methanol was added to constant volume and shaken to obtain a 1 mg / mL ginsenoside Re control solution.

[0065] Precise pipetting of ginsenoside Re reference substance solution 0, 10, 20, 40, 60, 80, 100 uL (1 mg / mL) According to the above method, the absorbance A value was determined. The standard curve was drawn with A mass concentration as the abscissa (X) and A value as the ordinate (Y). The linear regression equation obtained was y = 0.0204x + 0.0017, R 2 = 0.9995.

[0066] Preparation of test sample solution: 100 g of dried Ilex kudingcha, Ilex latifolia and Sabina vulgaris leaves were weighed, 10 times the amount of water was added, soaked for 30 minutes, then heated to boiling extraction for 1 h, filtered, the filtrate was concentrated to a relative density of 1.1-1.2, dried under reduced pressure, and pulverized to obtain each extract. 10 mg of each extract powder was accurately weighed and placed in a 10 mL volumetric flask, methanol was added to constant volume, and shaken to obtain the test sample solution.

[0067] The total saponin content in the test sample was determined according to the above method, and the results are shown in Table 2. Among them, the total saponin content of Ilex kudingcha was the highest, with an average content of 7.43%; the total saponin content of Ilex latifolia was 5.18%; and the total saponin content of Sabina vulgaris was 4.96%, as shown in Figure 3 It can be seen that Ilex kudingcha has a higher content of active ingredient total saponin compared with the other two types.

[0068] Table 2 Total saponin content of different batches of Ilex kudingcha, Ilex latifolia and Sabina vulgaris extracts

[0069]

[0070] 2.2 Comparison of the effects of total saponins in Ilex kudingcha, Ilex latifolia and Sabina vulgaris on the proliferation activity of hair follicle stem cells

[0071] According to the preparation method of total saponin in Example 1, the total saponins of Ilex kudingcha, Ilex latifolia and Sabina vulgaris were separated, and the effects of the three kinds of total saponins on the proliferation activity of hair follicle stem cells were determined at a concentration of 0.1%.

[0072] HFSCs were subcultured when they reached 70%-80% confluence, and log phase HFSCs were taken, which were diluted to 1×10 4100 μL was inoculated in 96-well plates, and control group, kudingcha group, kudingcha ilex group, large leaf ilex group and sabina group were set. The control group was the basic culture medium, and the other groups were the basic culture medium solution containing 0.1% of the corresponding total saponin composition. The composition of the kudingcha group was: kudingcha ilex, large leaf ilex and sabina accounted for one third, that is, each added 0.033%. Continue to culture in the incubator for 24 h, add CCK-8 reagent 10 μL, and return to the incubator for 1 h. The absorbance value was detected at 450 nm; three duplicate wells were set in each group to ensure the accuracy of the experiment.

[0073] The cell proliferation rate was calculated according to the method in Example 1, and the test results are shown in Table 1. Figure 4 The total saponins of kudingcha group, kudingcha ilex group, large leaf ilex group and sabina group all had significant proliferation effect on HFSCs. Compared with the control group, the average cell proliferation rate of kudingcha total saponin was increased by 41.1%, the total saponin in large leaf ilex was increased by 36.8%, the total saponin in sabina was increased by 27.9%, and the total saponin in kudingcha ilex was increased by 57.9%. Therefore, at the same level, the activity of total saponin in kudingcha ilex is significantly higher than that in large leaf ilex or sabina, or the combination of the three (kudingcha), indicating that the potential anti-hair loss effect of kudingcha ilex is the best among all kinds of kudingcha.

[0074] 2.3 Analysis of kudingcha ilex, large leaf ilex or sabina ilex saponin composition

[0075] Further, the key components of the key active components of kudingcha ilex, large leaf ilex or sabina were identified and compared. The components were separated and identified by mass spectrometry analysis by UPLC-Q-TOF / MS.

[0076] According to the mass spectrometry conditions shown in Example 1, 10 mg / mL of the three samples were measured in parallel, and the test results are shown in Table 2. Figure 5 It can be seen that the three kinds of components have similarities, and the content of each component also has obvious differences. According to the relative peak area, the content of kudingcha ilex kudinoside G, kudinoside C, kudinoside A and kudinoside D is 1.57 times that of large leaf ilex, and 2.54 times that of sabina. Therefore, the content of active ilex saponin in kudingcha ilex is significantly better than that in large leaf ilex or sabina.

[0077] Meanwhile, the fraction of 36.5 min to 40.41 min was collected and concentrated, and the four kinds of Ilex kudingcha saponins were subjected to the determination of the activity of promoting the proliferation of hair follicle stem cells according to the method described in Example 1, and the results are shown in Table 3. It can be seen that the Ilex kudingcha saponin components in Ilex kudingcha, Ilex latifolia or Sabina saponin components have the activity of promoting the proliferation of hair follicle stem cells at a concentration of 10 μg / mL, but the activity is quite different. The activity of the Ilex kudingcha saponin components in Ilex kudingcha is the best, which is significantly better than that of Ilex latifolia or Sabina saponin components. It can be seen that different total saponin extracts will show different abilities to promote the proliferation of hair follicle stem cells due to the difference in the ratio of saponin components.

[0078] Table 3 Activity of Ilex kudingcha saponin components in Ilex kudingcha, Ilex latifolia or Sabina for promoting proliferation

[0079]

[0080] Example 3: Preparation of a scalp serum containing Ilex kudingcha leaf extract

[0081] The solvent, the Ilex kudingcha leaf extract, the preservative, the humectant, and the chelating agent were added to an emulsification tank, and stirring was started at a speed of 20-40 rpm. The temperature was raised to 80°C and maintained for 10-15 min. Then the temperature was lowered to 40-50°C by stirring, the conditioning agent was added, the pH value of the scalp serum was adjusted to 5.0-7.0 by lactic acid, and the mixture was stirred uniformly until the temperature was below 40°C. The scalp serum containing the Ilex kudingcha leaf extract was obtained after 48 hours of storage.

[0082] Example 4: Effect of the content of Ilex kudingcha leaf extract on the scalp serum

[0083] To explore the anti-hair loss effect of the Ilex kudingcha leaf extract, the therapeutic effect of the scalp serum containing different proportions of the Ilex kudingcha leaf extract on the androgenic alopecia model mice was compared. The scalp serum containing the Ilex kudingcha leaf extract with a weight percentage of 0.05, 1, 3.35, 5 and 10 (experimental example 1-2, experimental example 1-3, experimental example 1-4, experimental example 1-5, experimental example 1-6) was prepared according to the method of Example 3. In addition, the scalp serum without the Ilex kudingcha leaf extract was prepared as experimental example 1-1, which also contained nipagin methyl 0.2%, phenoxyethanol 0.35%, ethylhexylglycerin 0.8%, D-panthenol 0.3%, butanediol 5%, betaine 0.2% and EDTA-2Na 0.5%, and the rest was water.

[0084] Forty male C57BL / 6J mice were randomly divided into 8 groups: a blank group, a model group, experimental example 1-1, experimental example 1-2, experimental example 1-3, experimental example 1-4, experimental example 1-5, and experimental example 1-6. Except for the blank group, the rest of the groups were evenly smeared with 200 μL of testosterone (5 mg / mL) daily for modeling treatment, and after complete absorption, 200 μL of the test substance was smeared. The model group was treated with a blank base containing no active substance, and the treatment was continued for 14 days. The tape stripping test was used to evaluate hair firmness. After the experimental mice were anesthetized, the hair growth in the depilation area on the back of the mice was photographed, and then a 1 cm 2 transparent adhesive tape was used to perform the tape stripping test on the back of the test area, and the number of hairs peeled off was counted to reflect the anti-hair loss effect of each group. The hair in the shaved area was removed with a shaver, collected with weighing paper, and weighed on a 0.1 mg electronic balance.

[0085] The effects of Ilex kudingcha C.J. Tseng on the number of hair loss and hair weight of androgenetic alopecia mice are shown in Table 1. Figure 6 As can be seen from the results, experimental example 1-1 had no obvious effect on preventing hair loss, and the hair of the mice remained consistent with the model group. The Ilex kudingcha C.J. Tseng leaf extract in experimental examples 1-2 to 1-6 could significantly inhibit the occurrence of hair loss in androgenetic alopecia mice, reduce the number of hair loss, and increase the weight of the hair in the hair loss area. The effect of the treatment group containing 1% Ilex kudingcha C.J. Tseng leaf extract (experimental example 1-3) was the best, and the effects of the treatment groups containing 3.35% Ilex kudingcha C.J. Tseng leaf extract (experimental example 1-4), 5% Ilex kudingcha C.J. Tseng leaf extract (experimental example 1-5), and 10% Ilex kudingcha C.J. Tseng leaf extract (experimental example 1-6) were also the best, all of which had a significant effect on preventing hair loss.

[0086] Example 5: Evaluation of the effects of penetration enhancers on the in vitro release and transdermal properties of Ilex kudingcha scalp essence

[0087] To better exert the effects of Ilex kudingcha C.J. Tseng leaf extract, it was prepared into a scalp essence in combination with a penetration enhancer. The penetration enhancer can be selected from isosorbide dimethyl ether and / or ethoxydiglycol. To explore the effects of different types and amounts of penetration enhancers on the in vitro release and transdermal properties of the active ingredients in Ilex kudingcha scalp essence, Ilex kudingcha C.J. Tseng leaf extract scalp essence containing different types and amounts of penetration enhancers was prepared, and the composition and content (weight percentage) of the penetration enhancers are shown in Table 4.

[0088] Table 4

[0089]

[0090] The scalp essence also includes 1% Kudingcha and camptotheca acuminata leaf extract, 0.2% nipagin methyl ester, 0.35% phenoxyethanol, 0.8% ethylhexylglycerin, 0.3% D-panthenol, 5% butylene glycol, 0.2% betaine, and 0.5% EDTA-2Na, and the balance is water.

[0091] The penetration properties of the prepared scalp essence of each group were compared. The in vitro transdermal test was designed according to the guidelines of the Japan Pharmaceutical Medical Device Agency (PMDA), a pig skin / Franz diffusion cell permeability test system was constructed, and the penetration test was verified. The transdermal behavior of Kudingcha scalp essence containing different types and different amounts of penetration enhancers in pig skin was studied.

[0092] First, 7 pieces of Bama pig back skin were prepared, physiological saline was used as the receiving liquid, the pig skin was fixed on the Franz diffusion cell, the receiving liquid was added to the lower chamber, the stirring and temperature control were started, and the balance was 30 min at 350 rpm, 32±1 ℃. The samples of examples 2-1 to 2-7 were added to the pig skin, the amount of sample was 1 mL, the upper chamber was sealed with a sealing film, and the test was carried out. All samples were taken to absorb the receiving liquid for 12 h and the pig skin was removed. The receiving liquid was filtered through a 0.22 μm microporous filter membrane for subsequent detection; the transdermal area of pig skin was cut with surgical scissors, washed with physiological saline for 3 times, and the surface of pig skin was wiped with filter paper to remove the residual sample. After washing, 3 times the amount of methanol was added, cut, ultrasonicated, centrifuged, and the supernatant was taken for subsequent detection.

[0093] The content of Kudingcha and camptotheca acuminata total saponins in pig skin and receiving liquid was determined by ultraviolet-visible spectrophotometry, with ginsenoside Re (purchased from Shanghai Yuan Ye Biological Company, item number A10036) as the control, and vanillin-glacial acetic acid method for color development. The in vitro transdermal behavior was described. Specifically, 100 uL of test sample or control sample solution was precisely taken and placed in a 10 mL test tube with a stopper. The solvent was dried in a 60 ℃ water bath, 0.2 mL of 10% vanillin-glacial acetic acid solution and 0.8 mL of perchloric acid were added, and after 15 min in a 60 ℃ water bath, it was taken out and placed in an ice water bath for 5 min. 5 mL of glacial acetic acid was added and mixed well. The absorbance was measured at 550 nm.

[0094] The effects of different penetration enhancers on the in vitro release and transdermal properties of Kudingcha scalp essence are shown in Table 5. It can be seen that the penetration enhancers can have different effects on the scalp of Kudingcha and camptotheca acuminata. The transdermal absorption effect of example 2-3 is the best, and the skin permeation rate is 44.17%.

[0095] Table 5 Effects of different types and different amounts of penetration enhancers on the skin permeation rate of Kudingcha and camptotheca acuminata essence

[0096]

[0097] Example 6: Evaluation experiment of the effect of conditioning agent on scalp microenvironment

[0098] The scalp microenvironment, also known as the scalp microecology, is composed of host cells and their secretions on the scalp surface, scalp microorganisms, and the microenvironment in which they exist. When the scalp microenvironment is imbalanced, scalp microorganisms and their secretions can induce excessive keratinization of the epidermis, excessive secretion of sebum, induce apoptosis, increase chemotaxis and invasion of inflammatory cytokines, and ultimately infiltrate inflammatory cells around the hair follicle, change the normal pH of the scalp, destroy the scalp barrier, trigger symptoms such as scalp inflammation, oily scalp, scalp itching, redness, and ultimately lead to the occurrence of hair loss.

[0099] The conditioning agent can be further added in the scalp essence containing Ilex kudingcha C.J. Tseng leaf extract.

[0100] The conditioning agent is Lactobacillus ferment lysate (Shanghai Baihaobo Biological Technology Co., Ltd., batch number: LDHG-010723), Pichia pastoris ferment lysate filtrate-PFLF2 (Guangzhou Xendi Biological Technology Co., Ltd., batch number: 20240304-1), or Lactobacillus plantarum ferment product. The fermentation method of the Lactobacillus plantarum ferment product is as follows: a small amount of Lactobacillus plantarum is inoculated on a solid plate culture medium from the laboratory preserved strain. The strain is distributed on the solid plate using the streak inoculation method, and is incubated in a constant temperature incubator at 37°C for 24-36 h to allow the strain to grow and reproduce on the solid plate culture medium to form single colonies. Well-grown colonies are selected and added to 10 mL of liquid MRS medium, and incubated for 12-14 h. A 0.22 μm pore size filter membrane is used to filter out residual bacteria and impurities to obtain the Lactobacillus plantarum ferment product.

[0101] The Lactobacillus plantarum is selected from Lactobacillus plantarum 18 or 24. The Lactobacillus plantarum 18 is preserved in the China General Microbiological Culture Collection Center, with a preservation number of CGMCC No. 31277, a preservation time of July 12, 2024, a classification and naming of Lactobacillus plantarum Lactiplantibacillus plantarum , and a preservation address of No. 3, Beichen West Road, Chaoyang District, Beijing; and the Lactobacillus plantarum 24 is preserved in the China General Microbiological Culture Collection Center, with a preservation number of CGMCC No. 31278, a preservation time of July 12, 2024, a classification and naming of Lactobacillus plantarum Lactiplantibacillus plantarum , and a preservation address of No. 3, Beichen West Road, Chaoyang District, Beijing.

[0102] To investigate the effect of different conditioning agents on the scalp microenvironment, the above fermentation products containing 0.1% different types of conditioning agents were prepared, experimental example 3-1-1 (lactobacillus fermentation lysate), experimental example 3-1-2 (pichia pastoris fermentation lysate filtrate-PFLF2) and experimental example 3-1-3 (plantarum fermentation product), also including 1% kudingcha and wintergreen leaf extract, 5% isosorbide dimethyl ether, 0.2% nipagin methyl ester, 0.35% phenoxyethanol, 0.8% ethylhexylglycerin, 0.3% D-panthenol, 5% butanediol, 0.2% betaine and 0.5% EDTA-2Na, and the balance is water.

[0103] Suitable volunteers were recruited for the hair loss prevention test of kudingcha and wintergreen essence. 18 volunteers were randomly divided into 3 groups: experimental example 3-1-1, experimental example 3-1-2, experimental example 3-1-3, 6 people in each group, aged 18-50 years old, male and female (gender stratified analysis), mild to moderate hair loss, no hair growth drugs used in the past 3 months, no serious scalp diseases, etc. 1mL once a day, evenly applied to the scalp, and absorbed by the finger pulp. After the test, the scalp photos were collected using a hair mirror.

[0104] After using the test sample for 28 days, the hair pictures of the designated area were collected using a hair mirror, and the representative results of the volunteers are shown in Figure 7 It can be seen that each experimental example has an effect on the scalp microenvironment, and the scalp microenvironment of the subjects is significantly improved, and the redness and abnormal oil secretion phenomenon is significantly reduced. In addition, the dandruff of the subjects of experimental example 3-1-3 is significantly reduced.

[0105] To further investigate the effect of plantarum fermentation product on the scalp microenvironment, scalp essence containing different amounts of conditioning agents was prepared, 0.1%, 1%, 5% and 10% of plantarum fermentation product were added respectively (experimental example 3-2-1, experimental example 3-2-2, experimental example 3-2-3 and experimental example 3-2-4), also including 1% kudingcha and wintergreen leaf extract, 5% isosorbide dimethyl ether, 0.2% nipagin methyl ester, 0.35% phenoxyethanol, 0.8% ethylhexylglycerin, 0.3% D-panthenol, 5% butanediol, 0.2% betaine and 0.5% EDTA-2Na, and the balance is water.

[0106] Suitable volunteers were recruited for the hair loss prevention test of the Ilex kudingcha and Cyclobalanopsis glauca essence. 24 volunteers were randomly divided into 4 groups: experimental example 3-2-1, experimental example 3-2-2, experimental example 3-2-3, and experimental example 3-2-4, with 6 people in each group, aged 18-50 years old, both men and women (stratified analysis according to gender), mild to moderate hair loss, no use of hair growth drugs in the past 3 months, no serious scalp diseases, etc. 1 mL was applied to the scalp once a day, and the scalp was evenly coated with the fingers. After the test, the scalp was photographed using a hair mirror.

[0107] After 28 days of using the test sample, the hair in the designated area was collected and photographed using a hair mirror, and the representative results of the volunteers are shown in Figure 8 As can be seen from the table, experimental example 3-2-1, experimental example 3-2-2, experimental example 3-2-3, and experimental example 3-2-4 all had a significant effect on the scalp microenvironment, and the scalp microenvironment of the subjects was significantly improved, with a significant reduction in redness and abnormal oil secretion. In addition, the dandruff of the subjects in experimental example 3-2-3 was significantly reduced. The overall effect of experimental example 3-2-3 was the best, followed by experimental example 3-2-4, which was better than experimental example 3-2-2 and experimental example 3-2-1.

[0108] Experimental example 7: product ratio study

[0109] According to the formula design of the Ilex kudingcha and Cyclobalanopsis glauca scalp essence, the core efficacy material Ilex kudingcha and Cyclobalanopsis glauca leaf extract, conditioning agent, and penetration enhancer were comprehensively compared to evaluate the hair loss prevention effect under different ratios.

[0110] Table 6

[0111]

[0112] The Ilex kudingcha and Cyclobalanopsis glauca leaf extract in each group of scalp essence was added at a mass percentage of 1%, and the conditioning agent and penetration enhancer were added at a ratio shown in Table 6. It also included nipagin methyl 0.2%, phenoxyethanol 0.35%, ethylhexylglycerin 0.8%, D-panthenol 0.3%, butanediol 5%, betaine 0.2%, and EDTA-2Na 0.5%, with the balance being water.

[0113] According to the evaluation method of the male hair loss model mice in Example 4, 30 male C57BL / 6J mice were randomly divided into 6 groups: blank group, model group, experimental example 4-1, experimental example 4-2, experimental example 4-3, and experimental example 4-4. Except for the blank group, the remaining groups were evenly coated with 200 μL of testosterone (5 mg / mL) daily for modeling, and then 200 μL of the test substance was applied after complete absorption. The model group was treated with a blank base without active substances, and the treatment was continued for 14 days. After the experimental mice were anesthetized, the hair growth of the mouse back was photographed, and then a 1 cm 2The transparent tape is used to pull the hairs in the back test area, and the number of hairs pulled off is counted to reflect the anti-hair loss effect of each group. The hairs in the shaved area are shaved with a shaver, collected with weighing paper, and weighed with a 0.1 mg electronic balance.

[0114] The results show that the effects of the four groups of samples on the number of hairs lost and the weight of the hairs of the androgenetic alopecia mice are as shown in A, B and C of Figure 9 The results show that the effects of the four groups of samples on the number of hairs lost and the weight of the hairs of the androgenetic alopecia mice are as shown in A, B and C of

[0115] Experimental Example 8

[0116] A scalp serum including 1% Ilex kudingcha leaf extract, 5% Bacillus velezensis fermentation product, 5% isosorbide dimethyl ether, 0.2% nipagin methyl ester, 0.35% phenoxyethanol, 0.8% ethylhexylglycerin, 0.3% D-panthenol, 5% butylene glycol, 0.2% betaine and 0.5% EDTA-2Na, and the rest being water, is prepared as experimental example 5-1; in addition, a scalp serum including 45% ethanol is constructed as experimental example 5-2.

[0117] A scalp serum including 1% Ilex kudingcha leaf extract, 5% Bacillus velezensis fermentation product, 5% isosorbide dimethyl ether, 0.3% octisalate, 0.3% 1,2-hexanediol, 0.2% p-hydroxyacetophenone, 2% propylene glycol and 0.5% EDTA-2Na, and the rest being water, is prepared as experimental example 5-3.

[0118] Suitable volunteers are recruited for the anti-hair loss test of the Ilex kudingcha leaf serum, and the subjects are randomly divided into groups, with 6 people in each group, aged 18-50 years old, male and female (analysis by gender stratification), mild to moderate hair loss, no use of hair growth drugs in the past 3 months, no serious scalp diseases, etc. 1 mL is applied to the scalp once a day, and is evenly applied to the scalp and massaged until absorbed. The scalp serum is tested according to the preparation method described in experimental examples 5-1-5-3. After 28 days of use, the representative volunteer results are as shown in Figure 10As shown, it can be seen that experimental examples 5-1, 5-2 and 5-3 all have significant effects on hair loss, the scalp microenvironment of the subjects is significantly improved, the erythema and abnormal secretion of sebum are significantly reduced, a large number of new hairs grow out, the overall hair density increases, the hair roots are more robust compared with before use. In addition, the number of hair loss is also significantly reduced, and the results are shown in Table 7. Among them, the hair loss prevention effect of experimental example 5-2 is the best, the hair loss improvement rate is 78.5%; the hair loss improvement rate of experimental example 5-1 is 64.2%; and the hair loss improvement rate of experimental example 5-3 is 53.3%. In summary, the Kuding tea and wintergreen leaf essence has a significant effect on preventing hair loss.

[0119] Table 7 Hair loss quantity statistics table

[0120]

[0121] The above detailed description of the embodiments of the present application, but the content described is only the preferred embodiments of the present application, and cannot be considered as limiting the scope of the present application. Any equivalent changes and improvements made in accordance with the scope of the present application should still belong to the patent scope of the present application.

Claims

1. A hair loss prevention serum containing an extract of Ilex kudingcha C.J. Tseng leaves, characterized in that: The anti-hair loss essence liquid contains the extract of Ilex kudingcha C.J. Tseng leaves and the fermentation product of Lactobacillus, wherein the Ilex kudingcha C.J. Tseng leaves are extracted by water decoction, 70% ethanol is added to the water extract of the Ilex kudingcha C.J. Tseng leaves to remove total polysaccharides, the separated liquid part is loaded onto an adsorption resin column, and the total phenolic acid is removed by elution with 30% ethanol, and then the components eluted with 50% ethanol are collected, concentrated and dried to obtain the total saponins of the Ilex kudingcha C.J. Tseng leaves, and the obtained solid material is the extract of the Ilex kudingcha C.J. Tseng leaves, and the mass fraction of the extract of the Ilex kudingcha C.J. Tseng leaves in the anti-hair loss essence liquid is 0.05%-10%; the fermentation product of Lactobacillus is used as a conditioning agent, and the mass fraction of the fermentation product of Lactobacillus is 0.1-10%; and the pH value of the anti-hair loss essence liquid is 5.0-7.

0.

2. The anti-hair loss serum containing the Ilex kudingcha and Quercus infectoria leaf extract according to claim 1, characterized in that: The fermentation product of Lactobacillus is a fermentation lysate product of Lactobacillus or a fermentation product of Plantarum. 3.The anti-hair loss essence containing the Ilex kudingcha C.J.Tseng and Quercusini extract according to claim 1, characterized in that: The fermentation product of Lactobacillus is replaced by a fermentation lysate filtrate of Pichia pastoris.

4. The anti-hair loss serum containing the Ilex kudingcha and Quercusini leaf extract according to any one of claims 1-3, characterized in that: The anti-hair loss essence liquid further contains a penetration enhancer in a mass fraction of 2-8%, a preservative in a mass fraction of 0.8-1.35%, a humectant in a mass fraction of 2-15.5%, and a chelating agent in a mass fraction of 0.1-1%, and the rest is a solvent. 5.The hair loss prevention serum containing Ilex kudingcha C.J. Tseng and Quercusini extract according to claim 4, characterized in that: The mass ratio of the extract of Ilex kudingcha C.J. Tseng leaves, the conditioning agent and the penetration enhancer is 1:2-7:4-10. 6.The anti-hair loss essence containing the Ilex kudingcha C.J.Tseng and Quercusini extract according to claim 4, characterized in that: The penetration enhancer is isosorbide dimethyl ether and / or ethoxydiglycol. 7.The anti-hair loss essence containing the Ilex kudingcha C.J.Tseng and Quercusini extract according to claim 4, characterized in that: The preservative is selected from one or more of methylparaben, octoxinol, phenoxyethanol, ethylhexylglycerin, 1,2-hexanediol and p-hydroxyacetophenone; The humectant is selected from one or more of glycerin, D-pantenol, propylene glycol, butylene glycol and betaine; The solvent is water or a mixture of water and ethanol.

8. A method for preparing the anti-hair loss serum containing the Ilex kudingcha Camellia leaf extract according to any one of claims 1 to 7, characterized by: The solvent, the extract of Ilex kudingcha C.J. Tseng leaves, the preservative, the humectant and the chelating agent are added into an emulsifying tank, and stirring is started at a speed of 20-40 rpm; the temperature is raised to 80°C and maintained for 10-15 min. After the temperature is lowered to 40-50°C, the conditioning agent is added, and stirring is continued until the temperature is below 40°C, and then the anti-hair loss essence liquid containing the extract of Ilex kudingcha C.J. Tseng leaves is obtained.

9. The method for preparing the anti-hair loss essence containing bitter tea and holly leaf extract according to claim 8, characterized in that: The pH value of the anti-hair loss essence liquid is adjusted to 5.0-7.0.

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