Preparation method of ginger root extract and anti-hair loss composition

Abstract

The invention relates to the technical field of plant extraction, and discloses a preparation method of a ginger root extract and an anti-hair loss composition. Firstly, the ginger root extract is obtained by adopting a liquid phase pulse discharge-supercritical CO2 extraction process for the first time, so that the extraction rate can be greatly improved; secondly, the ginger root extract, the cacumen biotae extract, the tea seed extract and the hydrolyzed keratin are combined into the anti-alopecia composition for the first time, the composition comprehensively considers multiple aspects of environmental factors, endocrine factors, nutritional factors and the like, the alopecia problem is remarkably improved through the synergistic effect of multiple active substances, and the anti-alopecia composition has a good anti-alopecia effect. The shampoo has the advantages of high absorptivity, no scalp irritation, no wound, quick response and the like, and can achieve better anti-hair loss and hair care effects.

Description

Technical Field

[0001] The invention relates to the technical field of plant extraction, and in particular to a preparation method of a ginger root extract and an anti-hair loss composition. Background Art

[0002] In recent years, the problem of hair loss has become more and more common among younger people, becoming a problem for more and more people. It not only affects the beauty of personal image, but also easily brings huge mental burden and social pressure to patients. Common factors of hair loss are as follows: (1) Environmental factors: excessive Malassezia and Propionibacterium in the scalp will affect the scalp microecology, resulting in dandruff, scalp itching, hair follicle inflammation, etc., which ultimately lead to hair loss; (2) Endocrine factors: the increase of substances such as androgen leads to excessive sebum secretion, and the hair follicles gradually shrink and destroy, causing permanent hair loss; (3) Nutritional factors: The metabolism of hair is inseparable from the supply of nutrition. Lack of nutrition will affect the growth of hair, and severe malnutrition will lead to hair loss. In summary, under the influence of various factors, the phenomenon of hair loss is becoming more and more serious and the living standards are generally improved, which has changed people's requirements for shampoo and shampoo itself. In addition to cleaning hair and scalp, it is also required to have functions such as dandruff removal, itching relief, and hair loss prevention. Nowadays, plant-based shampoo has been expected and accepted by more and more consumers. Various types of Chinese herbal shampoos on the market have become common products.

[0003] Ginger is the fresh rhizome of the perennial herb Zingiber offcinale Roscoe of the Zingiberaceae family, and is widely grown in central, southeastern, and southwestern my country. Due to the composition of ginger root extract, researchers have begun to develop the use of ginger in anti-hair loss shampoo. For example, the patent with application number 202310512666.4 discloses a shampoo containing ginger root extract and a preparation method thereof: ginger extract, arginine and other active ingredients are penetrated into the hair follicles through the skin to effectively exert their active effects, and a shampoo that can cure hair loss is prepared; the patent with application number 202311312081.4 discloses a ginger anti-hair loss shampoo: ginger extract, Polygonum multiflorum extract, and lecithin are combined to prepare a liposome suspension to prepare an anti-hair loss shampoo that has the effects of significantly activating hair follicle tissue, nutrient absorption of hair roots, strengthening hair roots, inducing and promoting hair growth; the patent with application number 202311704161.4 discloses a plant composition and shampoo with anti-hair loss effect: through the compounding of various raw materials, a hair conditioner containing multiple plant extracts such as Polygonum multiflorum root extract, Artemisia argyi extract, ginseng root extract, ginger root extract, etc., the prepared plant composition has excellent anti-hair loss and hair solidification effects.

[0004] In the above patents, some of them are about the active ingredients of ginger extract acting on hair follicle tissue, reflecting the effect of the active ingredients of ginger root extract on hair follicles and exerting active anti-hair loss effects; some are about compounding ginger root extract with a variety of raw materials to obtain hair conditioners. However, these solutions only prepare anti-hair loss shampoo products around ginger root extract from endocrine factors or nutritional factors. Therefore, there is still a lack of anti-hair loss and hair solidification shampoo products that comprehensively consider environmental factors, endocrine factors, and nutritional factors, so that the products are suitable for people with hair loss in different age groups. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides a preparation method of ginger root extract and an anti-hair loss composition. First, the present invention innovatively adopts liquid pulse discharge-supercritical CO2 extraction process to obtain ginger root extract, which can significantly break through the upper limit of the extraction rate of a single extraction technology. At the same time, this method can also significantly shorten the extraction time, reduce energy consumption, and save organic solvents. Secondly, the present invention combines ginger root extract, Platycladus orientalis leaf extract, tea seed extract and hydrolyzed keratin for the first time. The obtained anti-hair loss composition comprehensively considers environmental factors, endocrine factors, nutritional factors and other aspects, and the synergistic cooperation of multiple active substances can achieve better anti-hair loss effects.

[0006] The specific technical scheme of the present invention is: In a first aspect, the present invention provides a method for preparing a ginger root extract, comprising the following steps: 1) Pretreatment: Grind the ginger root.

[0007] 2) Crude extraction: The obtained ginger root residue was added to an ethanol aqueous solution and subjected to liquid phase pulse discharge extraction.

[0008] 3) Concentration: Filter the obtained ginger root crude alcohol extract, and concentrate the filtrate to obtain an extract paste.

[0009] 4) Refined extraction: Using ethanol aqueous solution as the extraction solvent, the extract paste is subjected to supercritical CO2 extraction to obtain a refined extract.

[0010] 5) Secondary concentration: Filter the refined extract, and concentrate the filtrate to dryness to obtain the ginger root extract.

[0011] In the applicant's previous experiments, different extraction processes were tried to obtain ginger root extract, but it was found that a high extraction rate could not be obtained. Finally, the present invention innovatively adopted a liquid phase pulse discharge-supercritical CO2 extraction process to obtain ginger root extract, and found that compared with a single liquid phase pulse discharge extraction and supercritical CO2 extraction technology, it can significantly break through the upper limit of the extraction rate of a single extraction technology (that is, even if the extraction time of a single extraction process is extended, the extraction rate cannot be further improved). In addition, the method of the present invention can also significantly shorten the extraction time, reduce energy consumption, and save the amount of organic solvents.

[0012] During the liquid phase pulse discharge process, strong liquid turbulence and cavitation will occur. These violent physical phenomena will lead to the destruction of the cell structure, enhance the mass transfer of solutes, and increase the mass transfer rate, thereby greatly shortening the extraction time, while improving the yield and purity of the extract. The unique advantage of supercritical CO2 fluid extraction technology is that it is extracted at room temperature, which ensures the original nature of the extract and can effectively extract volatile substances without oxidation and deterioration. It is particularly suitable for the separation of unstable natural products and physiologically active substances. Liquid phase pulse discharge combined with supercritical CO2 extraction can firstly release the active ingredients of ginger root through liquid phase pulse discharge, and further purify the active substances through supercritical CO2 extraction, so that the active ingredients in the ginger root are fully extracted to obtain higher yields and purer substances.

[0013] It should be emphasized that the extraction order of the two is crucial. If supercritical CO2 extraction is used first and then liquid phase pulse discharge technology is used, on the one hand, the ginger root plant tissue has not been pretreated, some active ingredients cannot be fully released, and the extraction effect cannot be improved well; on the other hand, the same extraction effect cannot better reduce time costs, energy costs, etc.

[0014] Preferably, step 1) specifically comprises: using ginger root as raw material, washing, drying, crushing and passing through a 20-100 mesh sieve to obtain ginger root residue for later use.

[0015] Preferably, in step 2), the conditions for the liquid phase pulse discharge extraction are: the concentration of the ethanol aqueous solution is 15-35wt%, the solid-liquid ratio is 1g:(40-10)mL, the discharge voltage is 0-7KV, the temperature is 10-50°C, and the extraction time is 1-5min.

[0016] The present invention found that the above factors have a certain influence on the extraction rate of ginger root extract, specifically: ① ethanol aqueous solution concentration: the extraction rate of ginger root extract shows a trend of first rising and then falling with the increase of ethanol concentration. The reason is that when the ethanol concentration is at a high level, only a part of the total incident energy of the liquid phase pulse discharge is used to form an underwater plasma channel, and the remaining energy is deposited in the solution. At this time, the discharge intensity decreases with the increase of ethanol concentration; ② extraction time: at the same extraction solvent concentration, the extraction rate also shows a trend of first rising and then falling with the extension of extraction time. This is because liquid phase pulse discharge extraction will greatly damage plant tissues and cells in the initial stage, so that a large amount of target compounds are quickly dissolved in the solvent. Sufficient extraction time can ensure the completion of the extraction process and the diffusion process of the target compound. However, as the extraction time is further extended, mass transfer has reached equilibrium, and the yield of ginger root extract remains unchanged or even decreases; ③ solid-liquid ratio: a higher solid-liquid ratio will lead to insufficient extraction of the target compound, a lower solid-liquid ratio can ensure that the target compound will not reach saturation before complete dissolution, and a higher intracellular and extracellular concentration gradient will promote mass transfer. However, excessive solvent will increase the energy absorbed by the solvent, reduce the effective energy of the system, increase the resistance in the extraction process, and thus reduce the yield of the target compound; ④Discharge voltage: The discharge voltage determines the intensity of secondary phenomena such as shock waves, cavitation, and chemical reaction substances during the extraction process, thereby affecting the destruction efficiency of plant tissues and the mass transfer rate of active ingredients. The yield increased significantly with the increase of voltage, however, after exceeding this voltage, the extraction yield decreased slightly. This is due to the occurrence of corona discharge instead of arc discharge at a lower voltage. Corona discharge is a local discharge with a weaker shock wave intensity compared to arc discharge. Therefore, the energy generated by the discharge is not enough to completely extract the active substances contained in the ginger root extract.

[0017] Preferably, in step 2), the liquid phase pulse discharge extraction adopts a multi-stage differentiation procedure, which is as follows: (a) The first stage: discharge voltage 0-1KV, extraction time 0-2min; (b) The second stage: discharge voltage 1-4KV, extraction time 2-3min; (c) The third stage: discharge voltage 4-6KV, extraction time 3-4min; (d) Stage 4: discharge voltage 6-7KV, extraction time 4-5min; The concentration of the ethanol aqueous solution in each stage is 15-35wt%, the solid-liquid ratio is 1g:(40-10)mL, and the temperature is 10-50°C.

[0018] In order to further improve the extraction rate of ginger root extract, the present invention has designed multi-stage differentiated parameters according to the characteristics of different stages in the liquid phase pulse discharge extraction process. The principle is: in the liquid phase pulse discharge extraction process, the discharge voltage determines the intensity of secondary phenomena such as shock waves, cavitation and chemical reaction substances in the extraction process, thereby affecting the destruction efficiency of plant tissues and the mass transfer rate of active ingredients. According to the inventor's research, liquid phase pulse discharge extraction can be roughly divided into the following four stages: pre-breakdown stage (streamer) → breakdown stage (arc) → disintegration of cell structure → enhanced mass transfer process.

[0019] In the first stage, the discharge voltage and extraction time are set within the above range because this stage is the formation and propagation of streamers; it injects energy directly into the aqueous solution through the plasma channel formed by the high current / high voltage discharge between the two immersed electrodes, and the ionized gas channel generated by the streamer constitutes a conductive path. In the second stage, the discharge voltage is further increased and the extraction time is extended. When the streamer reaches the ground surface electrode, an arc is generated in the streamer channel (breakdown phase). This stage is the formation and propagation of the arc; in the third stage, with the increase of the discharge voltage and the extension of the extraction time, the product is broken, the cells are broken, and the cell structure is disintegrated; in the fourth stage, the mass transfer process is enhanced. Energy input is one of the most important parameters to be considered in the optimization of discharge extraction. Discharge pretreatment can obtain a higher solute yield in a shorter diffusion time.

[0020] Preferably, in step 4), the conditions for supercritical CO2 extraction are: the concentration of ethanol aqueous solution is 50-80wt%, the solid-liquid ratio is 1g:(30-10)mL, the extraction pressure is 4500psi, the extraction temperature is 45-55°C, the CO2 flow rate is 1-3mL / min, and the extraction time is 10-30min.

[0021] The present invention found that the above factors have a certain influence on the extraction rate of ginger root extract, specifically: ① Concentration of ethanol aqueous solution: ethanol concentration has a significant influence on the extraction rate of ginger root. Since ethanol is a highly polar solvent, it can enhance the solubility and selectivity of supercritical CO2, making it easy to extract substances with strong polarity and large relative molecular mass, and plays a reinforcing role in extraction. Therefore, the ethanol concentration in supercritical CO2 extraction is greater than that in liquid phase pulse discharge extraction; ② CO2 flow rate: With the increase of CO2 flow rate, the extraction rate shows a trend of first rapidly increasing and then slowly decreasing. In the initial stage of extraction, as the CO2 flow rate increases, the contact opportunity between the ginger root extract and the supercritical CO2 fluid increases, which can improve the intermolecular mass transfer efficiency and eventually reach equilibrium. Therefore, a larger CO2 intake within an appropriate range can promote the extraction reaction. In the process of decreasing extraction rate, the increase in CO2 flow rate causes CO2 to flow out of the device before it has time to fully contact with the ginger root extract, thereby reducing the extraction rate. In addition, a higher CO2 flow rate will increase the difficulty of the extraction operation. ③ Extraction temperature: The extraction rate of ginger root extract gradually increases with increasing temperature. The reasons for this are as follows: within a certain range, a higher temperature can reduce the viscosity of the supercritical CO2 fluid, increase the vapor pressure of the ginger root extract, and promote the diffusion of the ginger root extract molecules. It is manifested in that the ginger root extract can be well dissolved in the CO2 solvent; when the extraction temperature rises to a certain temperature, the extraction rate of the ginger root extract decreases. The reasons are analyzed as follows: the molecular activity of the supercritical CO2 fluid increases, which increases the average distance between its molecules, which is manifested as an increase in the fluid volume. When a certain fluid mass is reached, the density drops rapidly, resulting in a decrease in the solubility of the ginger root extract in CO2, that is, the extraction rate of the ginger root extract decreases; ④ Extraction time: The extraction rate of the ginger root extract first increases gradually with time, and the ginger root extract is fully freed with the extension of time, and can penetrate and diffuse well in the supercritical CO2 fluid, and gradually reach equilibrium and saturation; if the time continues to be extended, the extraction rate will no longer increase or even decrease, and the time cost and energy consumption will be higher.

[0022] Preferably, the concentration in step 3) and / or the secondary concentration in step 5) is vacuum concentration: the temperature is 40-55° C. and the pressure is 0.09-0.10 MPa.

[0023] In a second aspect, the present invention provides a use of the ginger root extract obtained by the above preparation method in cosmetics.

[0024] Preferably, the cosmetic comprises a shampoo product, a hair care product, or a scalp care product.

[0025] In a third aspect, the present invention provides an anti-hair loss composition, comprising the following components in mass percentage: 0.01-0.25% ginger root extract (further preferably 0.01-0.2%), 0.01-0.25% orientalis leaf extract, 0.01-0.3% hydrolyzed keratin, 0.01-0.25% tea seed extract, and a matrix.

[0026] Most of the anti-hair loss products in the prior art are designed based on endocrine factors or nutritional factors. At present, there is still a lack of anti-hair loss and hair-fixing shampoo products that comprehensively consider environmental factors, endocrine factors, and nutritional factors. The present invention combines active ingredients such as ginger root extract, Platycladus orientalis leaf extract, tea seed extract, and hydrolyzed keratin for the first time. The obtained anti-hair loss composition takes environmental factors, endocrine factors, and nutritional factors into consideration. The above active substances can achieve better anti-hair loss effects under synergistic cooperation.

[0027] Specifically: Ginger root extract can achieve the effect of antibacterial and antimicrobial scalp protection within the appropriate concentration range, considering the impact of environmental colonies. Platycladus orientalis leaf extract, in collaboration with ginger root extract, has a significant inhibitory effect on 5α-reductase from the perspective of endocrine factors, with mild efficacy, reducing the production of androgen receptors, and regulating the growth of hair follicles. Tea seed extract, from the perspective of nutritional factors, acts on the roots of the scalp to promote blood circulation, nutrient supply, promote hair follicle health, and nourish the scalp. The above points focus more on scalp care, hair follicle health, etc. On this basis, hydrolyzed keratin, hair is keratin that grows from scalp follicles. Hydrolyzed keratin focuses more on hair care, which can smooth out roughness and make hair smooth and shiny.

[0028] Preferably, the tea seed extract includes at least one of camellia oleifera seed extract, tea seed extract and camellia seed extract.

[0029] Preferably, the matrix is ​​a shampoo matrix, a conditioner matrix, or an essence matrix.

[0030] Preferably, the shampoo base comprises the following components in mass percentage: 8-20% surfactant, 0-2.5% hair conditioner, 0-0.5% antidandruff agent, 0-0.5% preservative, 0-0.7% sodium chloride, 0-0.2% citric acid, 0-0.5% rheology modifier, 0-0.3% fragrance and water.

[0031] Further preferably, the matrix comprises the following components in mass percentage: 8-15% surfactant, 1.5-2.5% hair conditioner, 0.2-0.5% antidandruff agent, 0.1-0.3% preservative, 0.5-0.7% sodium chloride, 0.1-0.2% citric acid, 0.1-0.3% rheology modifier, 0.1-0.3% fragrance and water.

[0032] Compared with the prior art, the beneficial effects of the present invention are: (1) The present invention innovatively adopts a liquid phase pulse discharge-supercritical CO2 extraction process to obtain ginger root extract, which can significantly break through the upper limit of the extraction rate of a single extraction technology. In addition, the method of the present invention can also significantly shorten the extraction time, reduce energy consumption, and save the amount of organic solvent used.

[0033] (2) The present invention combines active ingredients such as ginger root extract, Platycladus orientalis leaf extract, tea seed extract and hydrolyzed keratin for the first time. The obtained anti-hair loss composition takes environmental factors, endocrine factors and nutritional factors into consideration. The above active substances cooperate synergistically to achieve a better anti-hair loss effect. DETAILED DESCRIPTION

[0034] The present invention will be further described below in conjunction with the embodiments.

[0035] Overall embodiment The present invention provides a method for preparing a ginger root extract, comprising the following steps: 1) Pretreatment: Grind the ginger root.

[0036] In some specific implementation cases, the pretreatment specifically includes: using ginger root as raw material, washing, drying, crushing and passing through a 20-100 mesh sieve to obtain ginger root residue for later use.

[0037] 2) Crude extraction: The obtained ginger root residue was added to an ethanol aqueous solution and subjected to liquid phase pulse discharge extraction.

[0038] In some specific implementation cases, the conditions for the liquid phase pulse discharge extraction are: the concentration of the ethanol aqueous solution is 15-35wt%, the solid-liquid ratio is 1g:(40-10)mL, the discharge voltage is 0-7KV, the temperature is 10-50°C, and the extraction time is 1-5min.

[0039] In some other specific implementation cases, the liquid phase pulse discharge extraction adopts a multi-stage differentiation procedure: (a) The first stage: discharge voltage 0-1KV, extraction time 0-2min; (b) The second stage: discharge voltage 1-4KV, extraction time 2-3min; (c) The third stage: discharge voltage 4-6KV, extraction time 3-4min; (d) Stage 4: discharge voltage 6-7KV, extraction time 4-5min; The concentration of the ethanol aqueous solution in each stage is 15-35wt%, the solid-liquid ratio is 1g:(40-10)mL, and the temperature is 10-50°C.

[0040] 3) Concentration: Filter the obtained ginger root crude alcohol extract, and concentrate the filtrate to obtain an extract paste.

[0041] In some specific implementation cases, the concentration is vacuum concentration: the temperature is 40-55° C., and the pressure is 0.09-0.10 MPa.

[0042] 4) Refined extraction: Using ethanol aqueous solution as the extraction solvent, the extract paste is subjected to supercritical CI2 extraction to obtain a refined extract.

[0043] In some specific implementation cases, the conditions of the supercritical CI2 extraction are: the concentration of ethanol aqueous solution is 50-80wt%, the solid-liquid ratio is 1g:(30-10)mL, the extraction pressure is 4500psi, the extraction temperature is 45-55°C, the CO2 flow rate is 1-3mL / min, and the extraction time is 10-30min.

[0044] 5) Secondary concentration: Filter the refined extract, and concentrate the filtrate to dryness to obtain the ginger root extract.

[0045] In some specific implementation cases, the secondary concentration is vacuum concentration: the temperature is 40-55° C., and the pressure is 0.09-0.10 MPa.

[0046] The present invention provides an application of the ginger root extract obtained by the above preparation method in cosmetics. In some specific implementation cases, the cosmetics are shampoo products, hair care products, or scalp care products.

[0047] The invention provides an anti-hair loss composition, comprising the following components in mass percentage: 0.01-0.25% (more preferably 0.01-0.2%) of ginger root extract, 0.01-0.25% of orientalis leaf extract, 0.01-0.3% of hydrolyzed keratin, 0.01-0.25% of tea seed extract and a matrix.

[0048] In some specific implementation cases, the tea seed extract includes at least one of camellia oleifera seed extract, tea seed extract and camellia seed extract.

[0049] In some specific implementation cases, the matrix is ​​a shampoo matrix, a conditioner matrix, or an essence matrix.

[0050] In some specific implementation cases, the shampoo base includes the following components in percentage by mass: 8-20% surfactant, 0-2.5% hair conditioner, 0-0.5% antidandruff agent, 0-0.5% preservative, 0-0.7% sodium chloride, 0-0.2% citric acid, 0-0.5% rheology modifier, 0-0.3% fragrance and water.

[0051] Further preferably, the matrix comprises the following components in mass percentage: 8-15% surfactant, 1.5-2.5% hair conditioner, 0.2-0.5% antidandruff agent, 0.1-0.3% preservative, 0.5-0.7% sodium chloride, 0.1-0.2% citric acid, 0.1-0.3% rheology modifier, 0.1-0.3% fragrance and water.

[0052] Specific Examples and Comparative Examples The technical solution of the present invention is further described below by specific implementation methods, so as to better understand the purpose and technical solution of the present invention and make the advantages clearer. The embodiments are only to help understand the present invention and should not be regarded as specific limitations of the present invention.

[0053] (I) Preparation of ginger root extract 1) Pretreatment: Use ginger root as raw material, wash, dry, grind with a universal grinder, and then pass through a 60-mesh sieve to obtain ginger root residue for later use.

[0054] 2) Crude extraction: The ginger root residue was subjected to liquid pulse discharge assisted extraction using ethanol aqueous solution (15-25 wt%, see Table 1) as the extraction solvent. Liquid pulse discharge assisted extraction was performed under the conditions of a discharge voltage of 5 KV, a treatment time of (see Table 1) and a solid-liquid ratio of 1:40 g / mL, all at room temperature and pressure.

[0055] 3) Concentration: Filter the crude alcohol extract of ginger root, and concentrate the filtrate under vacuum at 50° C. and a pressure of 0.10 MPa to obtain an extract paste.

[0056] 4) Refined extraction: The extract paste was subjected to supercritical CO2 extraction, with CO2 and ethanol aqueous solution (50-70wt%, see Table 1 for details) as solvent, the solid-liquid ratio was 1g:20mL, the extraction pressure was 4500psi, the extraction temperature was 50°C, the CO2 flow rate was 2mL / min, the extraction time was 20-30min (see Table 1 for details), and a refined extract was obtained (see Table 1 for details).

[0057] 5) Secondary concentration: Filter the extract, and concentrate the filtrate to dryness under reduced pressure at 50° C. and a pressure of 0.10 MPa to obtain a concentrated extract, i.e., ginger root extract.

[0058] Table 1: Effects of different extraction times and extraction solvents on the extraction rate of ginger root extract From the test results in Table 1, we can see that: Comparative Examples 1-4 and Example 1 only used the liquid phase pulse discharge extraction process, and the extraction rate of Comparative Examples 1-3 showed a trend of first increasing and then decreasing with the increase of the extraction solvent concentration under the same extraction time condition (5min). When the ethanol concentration reaches a certain level, the reason why the yield of ginger root extract gradually decreases with the further increase of its concentration is that at a higher ethanol concentration, only a part of the total incident energy of the discharge is used to form an underwater plasma channel, and the rest of the energy is deposited in the solution. Therefore, the discharge intensity decreases with the increase of the ethanol concentration, which leads to a decrease in the extraction rate.

[0059] Comparing Comparative Example 2 with Example 1 and Comparative Example 4, it can be found that at the same extraction solvent concentration, the extraction rate also shows a trend of first increasing and then decreasing with the extension of extraction time. This is because liquid phase pulse discharge extraction will greatly damage plant tissues and cells in the initial stage, causing a large amount of target compounds to quickly dissolve in the solvent. Sufficient extraction time can ensure the completion of the extraction process and the diffusion process of the target compound. However, as the extraction time is further extended, the yield of ginger root extract remains basically unchanged, because the mass transfer has reached equilibrium when the extraction time is 4 minutes.

[0060] Comparative Examples 5-6 and Example 2 both used only supercritical CO2 extraction technology. As the extraction time changed, the extraction rate of Example 2 obtained a slightly higher value under the corresponding conditions.

[0061] Comparative Example 7, Example 3 and Comparative Example 8 all adopted the "liquid phase pulse discharge + supercritical CO2 extraction" process. By comparing the data with single liquid phase pulse discharge extraction and single supercritical CO2 extraction, it can be seen that the "liquid phase pulse discharge + supercritical CO2 extraction" process can further significantly improve the extraction rate on the basis of the aforementioned single process. It shows that the combination of the two above-mentioned extraction processes can break through the upper limit of the extraction rate of a single extraction process (even if the extraction time of a single extraction process is extended, the extraction rate cannot be further improved). Considering the extraction time and the amount of organic solvent used in the extraction solvent, it is finally found that Example 3 is the optimal process selection. The extraction yield under this extraction process is higher and the energy consumption is lower, which shows that the combination of liquid phase pulse discharge and supercritical CO2 extraction is an efficient green extraction technology, which has important research significance for the development and utilization of ginger root extract.

[0062] Example 4 On the basis of Example 3, the present invention further studies the effect of using the "multi-stage liquid phase pulse discharge + supercritical CO2 extraction" process. The difference between Example 4 and Example 3 is only that the conditions of liquid phase pulse discharge extraction in step 2) are different, as follows: 2) Crude extraction: For ginger root residue, multi-stage liquid phase pulse discharge extraction was performed at room temperature and pressure using 20 wt% ethanol aqueous solution as the extraction solvent and a solid-liquid ratio of 1:40 g / mL: (a) The first stage: discharge voltage 1 kV, extraction time 1 min; (b) Second stage: discharge voltage 3 KV, extraction time 2 min; (c) The third stage: discharge voltage 5 KV, extraction time 3 min; (d) The fourth stage: discharge voltage 6 KV, extraction time 4 min.

[0063] Table 2: Effect of multi-stage liquid pulse discharge extraction combined with supercritical CO2 extraction on the extraction rate of ginger root extract Experimental Group Extraction rate (%) Example 3 15.8 Example 4 18.6 From the test results in Table 2, it can be seen that in Example 4, compared with Example 3, the extraction rate of ginger root extract is further improved after the conventional liquid phase pulse discharge extraction method is replaced by the multi-stage liquid phase pulse discharge extraction method. This shows that the extraction rate of ginger root extract can be further improved after the discharge voltage and extraction time are adjusted in a targeted manner at different stages of the liquid phase pulse discharge extraction process.

[0064] (II) Preparation of anti-hair loss composition (a) Environmental factors A daily chemical cleaning and anti-hair loss composition shampoo is composed of the following components in percentage: 0-0.25% of ginger root extract obtained in Example 4, 0.15% of Platycladus orientalis leaf extract, 0.05% of hydrolyzed keratin, 0.2% of camellia seed extract and the remainder of shampoo base. Other conditions remain unchanged, and only the percentage of ginger root extract is changed (see Table 3 for details).

[0065] The composition of the shampoo base is as follows: 8% sodium laureth sulfate, 1.2% cocamide MEA, 0.2% polyquaternium-7, 0.6% dimethiconol, 0.15% dimethicone, 0.32% cocamide DEA, 0.3% guar hydroxypropyltrimonium chloride, 0.24% piroctone olamine (OCT), 0.25% sodium salicylate, 0.05% benzyl alcohol, 0.05% propylene glycol, 0.7% sodium chloride, 0.15% citric acid, 0.2% styrene / acrylic acid (ester) copolymer, 0.3% fragrance and the balance water.

[0066] Preparation of culture medium Solid culture medium: Weigh 76g of modified Dixon agar (MD), 10g of Tween 40, 50mg of chloramphenicol, 1000mL of pure water, and sterilize at 121℃ for 30 minutes. Liquid culture medium: Weigh 20g of malt extract, 2g of yeast extract, 20g of glucose, 20g of peptone, 20mL of olive oil, 1000mL of pure water, and sterilize at 121℃ for 30 minutes.

[0067] The activated strain of Malassezia for testing was purchased as a lyophilized powder in an ampoule tube. Use a small grinding wheel to draw three circles in the middle of the ampoule tube, wipe the outer wall of the ampoule tube with 75% ethanol, wrap the scratched part of the outer wall of the ampoule tube with dry cotton, and then break it by hand. Inject about 400μL of sterile water into the ampoule tube and blow gently to dissolve it completely into a bacterial suspension, then put the bacterial suspension into the surface of the solidified solid culture medium (200μL / piece), apply it evenly with a sterile coating rod, and invert and culture it in a 37℃ constant temperature incubator for 60h. When the strain grows, relevant tests can be carried out. Preparation of bacterial suspension Under sterile conditions, take the activated colony and dilute it to 10 with sterile saline. 7 ~10 8 CFU / mL bacterial solution for later use.

[0068] The diameter of the inhibition zone was determined by the culture medium punching method: add about 15 mL of solid culture medium cooled to about 50 ° C to the culture medium plate, rotate the plate to spread it evenly, add 200 μL of bacterial solution after complete solidification, and spread it evenly with a sterile coating rod to make a bacterial plate. Use a sterile puncher to punch holes (d = 6 mm) on the bacterial plate, add 10 μL of Chinese medicine volatile oil to each hole, and culture it in a 37 ° C constant temperature incubator for 72 hours, observe and record the results. Each Chinese medicine volatile oil was tested in parallel for 2 times, and the diameter of the inhibition zone (mm) was determined by the cross-cross method. The experimental data of the inhibition zone diameter were processed using Excel, and the Duncan multiple comparison test method was used to analyze its significance using SPSS21.0 data statistical analysis software. The criteria for determining the antibacterial effect are detailed in Table 3.

[0069] Table 3: Criteria for determining antibacterial effect Diameter of inhibition zone / mm ＞20 15～20 10～15 8～10 ＜8 Sensitivity Extremely sensitive Highly sensitive Moderately sensitive Low sensitivity Insensitive Table 4: Determination of the diameter of the inhibition zone ( mm)(n=3) As can be seen from the test results in Table 4, the difference between Comparative Examples 9-11 and Examples 5-8 is the different content of ginger root extract. When the concentration of ginger root extract is 0 (Comparative Example 9) or the concentration is too low (Comparative Example 10), the antibacterial effect is at a relatively low sensitivity and does not reach the effective antibacterial concentration, that is, the antibacterial effect is not obvious. On the contrary, when the concentration of ginger root extract is too high (Example 7 and Example 8), the present invention team found that its main active ingredient can cause apoptosis of hair follicle dermal cells, thereby inhibiting hair growth. Ginger root extract is the main component for inhibiting hair loss. The 6-gingerol content in the ginger root extract with too high a concentration is too high, which inhibits the growth of dermal papilla cells, that is, it is easy to cause the extension of the resting period in the body, causing hair growth to slow down. Therefore, the ginger root extract must be adjusted to an appropriate concentration to ensure the antibacterial effect while avoiding its side effects.

[0070] (b) Endocrine factors A daily chemical cleaning and anti-hair loss composition shampoo is composed of the following components in percentage: 0-0.25% of ginger root extract obtained in Example 4, 0-0.25% of Platycladus orientalis leaf extract, 0.05% of hydrolyzed keratin, 0.2% of camellia seed extract and the balance of shampoo base (same as the formula in (a) environmental factors). Other conditions remain unchanged, and the percentage of ginger root extract and Platycladus orientalis leaf extract is changed (see Table 4 for details).

[0071] The 5α-reductase (5AR) enzyme-linked immunosorbent assay (ELISA) was used to analyze and test the 5α-reductase (5AR) level of the sample. This kit uses the double antibody sandwich method to determine the 5α-reductase (5AR) level in the sample. The microplate is coated with purified 5α-reductase (5AR) antibody to prepare a solid-phase antibody. 5α-reductase (5AR) is added to the microwells coated with the monoclonal antibody in sequence, and then combined with the HRP-labeled 5α-reductase (5AR) antibody to form an antibody-antigen-enzyme-labeled antibody complex. After thorough washing, the substrate TMB is added for color development. TMB is converted into blue under the catalysis of HRP enzyme, and converted into the final yellow under the action of acid. The depth of color is positively correlated with the 5α-reductase (5AR) in the sample. The absorbance (OD value) is measured at a wavelength of 450nm using an enzyme reader, and the activity concentration of 5α-reductase (5AR) in the sample is calculated using the standard curve.

[0072] Steps: (1) Dilution of standard: This kit provides one original standard (320U / L). Users can dilute it in a small test tube by stepwise dilution. For example, 150μl of original standard is added to 150μl of standard diluent, and the final concentration is (160U / L, 80U / L, 40U / L, 20U / L, 10U / L).

[0073] (2) Sample addition: Set up blank wells (blank control wells do not add samples and enzyme-labeled reagents, and the rest of the steps are the same), standard wells, and test sample wells. Accurately add 50 μl of the standard sample to the enzyme-labeled plate, add 40 μl of sample diluent to the test sample wells, and then add 10 μl of the test sample (the final sample dilution is 5 times). Add the sample to the bottom of the well of the enzyme-labeled plate, try not to touch the well wall, and gently shake to mix.

[0074] (3) Incubation: Seal the plate with a sealing film and incubate at 37°C for 30 min.

[0075] (4) Liquid preparation: Dilute the 30-fold concentrated washing solution with 30-fold distilled water and set aside.

[0076] (5) Washing: Carefully peel off the sealing film, discard the liquid, spin dry, fill each well with washing solution, let it stand for 30 seconds and then discard. Repeat this process 5 times and pat dry.

[0077] (6) Add enzyme: Add 50 μl of enzyme-labeled reagent to each well, except for the blank well.

[0078] (7) Incubation: Same operation as (3).

[0079] (8) Washing: The operation is the same as (5).

[0080] (9) Color development: First add 50 μl of color developer A to each well, then add 50 μl of color developer B, gently shake to mix, and develop the color at 37°C in the dark for 10 min.

[0081] (10) Termination: Add 50 μl of stop solution to each well to terminate the reaction (the blue color immediately turns yellow).

[0082] (11) Measurement: Use the blank well as the zero setting and measure the absorbance (OD value) of each well in sequence at a wavelength of 450 nm. The measurement should be performed within 15 minutes after adding the stop solution.

[0083] Calculate the standard curve on the coordinate paper with the concentration of the standard as the horizontal axis and the OD value as the vertical axis. Find the corresponding concentration from the standard curve based on the OD value of the sample; then multiply it by the dilution factor; or use the concentration of the standard and the OD value to calculate the linear regression equation of the standard curve, substitute the OD value of the sample into the equation, calculate the sample concentration, and then multiply it by the dilution factor to get the actual concentration of the sample.

[0084] Table 5: Inhibition rate of 5α-reductase From the test results in Table 5, it can be seen that in Comparative Examples 12-14 and Examples 9-14, the other ingredients remain unchanged, and the addition of ginger root extract (Comparative Example 14) or Platycladus orientalis leaf extract (Comparative Example 13) alone has a certain inhibitory effect on 5α-reductase, but it is not obvious. However, as the concentration of ginger root extract and Platycladus orientalis leaf extract added at the same time increases, the inhibitory effect of 5α-reductase gradually becomes obvious, and even doubles. Therefore, it can be found that in terms of inhibiting 5α-reductase, ginger root extract and Platycladus orientalis leaf extract have a mutually promoting effect, thereby achieving a better effect.

[0085] Platycladus orientalis leaf extract has mild efficacy and is a natural plant extract. It contains anti-hair loss and hair growth ingredients, rich volatile oils, flavonoids, phenols and other ingredients. Hair follicle growth regulation can improve the blood circulation of hair follicles, increase the nutrient supply of hair follicles, promote hair growth, promote the proliferation of dermal papilla cells, reduce the production of androgen receptors, reduce the damage of free radicals to hair follicles, protect hair health, and inhibit the activity of 5α-reductase and the level of dihydrotestosterone in hair follicles, prevent the influence of androgen, and fully extend the growth period of hair follicles, thereby slowing down the individual's hair loss process. On this basis, it is shown that the combination of ginger root extract and Platycladus orientalis leaf extract will further increase the efficacy.

[0086] (c) Nutritional factors A daily chemical cleaning and anti-hair loss composition shampoo is composed of the following components in percentage: 0.15% of ginger root extract obtained in Example 3, 0.15% of Platycladus orientalis leaf extract, 0.05% of hydrolyzed keratin, 0-0.25% of camellia seed extract and the balance of shampoo base (same as the formula in (a) environmental factors). Other conditions remain unchanged, and only the percentage of camellia seed extract is changed (see Table 5 for details).

[0087] Growth factor determination: Three mice were randomly selected from each group (Comparative Examples 15-17, Examples 15-16), and a wax / resin mixture (1:1) was used to depilate a 2×3 cm area on the back of the mice. Starting from the second day, the test was continued every day. After the 28th day of the test, the skin on the back of the mice was taken, cut into pieces, weighed, and the procedure was performed according to the operating instructions of the one-step animal tissue active protein extraction kit. Add 9 times the amount of tissue extract, incubate on ice for 20 minutes, centrifuge at 4°C, take the supernatant, and use the kit to determine the vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) in the skin. The operating steps were carried out according to the instructions of the kit. The absorbance value of vascular endothelial growth factor in the sample was determined at a wavelength of 450nm using an enzyme marker, and the absorbance value of epidermal growth factor in the sample was determined at a wavelength of 560nm.

[0088] Table 6: Expression of growth factors in the skin of mice treated with different methods From the test results in Table 6, it can be seen that in Comparative Examples 15-17 and Examples 15-16, other ingredients remain unchanged. As the concentration of camellia seed extract increases, it has a certain promoting effect on the expression of vascular endothelial growth factor and epidermal growth factor. Vascular endothelial growth factor and epidermal growth factor are key indicators of hair follicle nutrition supply. More secretion of vascular endothelial growth factor provides sufficient nutrition for hair follicles, and the generated hair shaft diameter is thicker and the hair is longer. Epidermal growth factor is the switch for the transformation of hair follicle growth phase and regression phase. The present invention has previously found that the effect of epidermal growth factor in promoting hair growth is closely related to its dosage. Its expression level is only at 2×10 3 ~2×10 4 pg / mL range can promote hair growth. Therefore, controlling the expression of epidermal growth factor within an appropriate range can also effectively prevent hair loss and achieve the purpose of treating hair loss. Similarly, as shown in Table 6, in Examples 15-16, the epidermal growth factor is within the normal expression range. It can be seen that the addition of an appropriate amount of camellia seed extract has a significant effect in promoting blood circulation, nutrient supply, promoting hair follicle health, and nourishing the scalp.

[0089] The raw materials and equipment used in the present invention, unless otherwise specified, are all commonly used raw materials and equipment in the art; the methods used in the present invention, unless otherwise specified, are all conventional methods in the art.

[0090] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any way. Any simple modification, change and equivalent transformation made to the above embodiment based on the technical essence of the present invention still falls within the protection scope of the technical solution of the present invention.

Claims

1. A method for preparing ginger root extract, characterized in that The following steps are involved: 1) Pretreatment: crush the ginger root; 2) Crude extraction: Add the obtained ginger root residue into ethanol aqueous solution and perform liquid phase pulse discharge extraction; 3) Concentration: Filter the obtained ginger root crude alcohol extract, and concentrate the filtrate to obtain the extract paste; 4) Extraction: Using ethanol aqueous solution as the extraction solvent, the extract paste is subjected to supercritical CO2 extraction to obtain the extract solution; 5) Secondary concentration: Filter the refined extract, concentrate the filtrate to dryness to obtain ginger root extract.

2. The preparation method according to claim 1, characterized in that: Step 1) specifically comprises: taking ginger root as raw material, washing, drying, crushing and passing through a 20-100 mesh sieve to obtain ginger root crumbs for later use.

3. The preparation method according to claim 1, characterized in that: In step 2), the conditions of the liquid phase pulse discharge extraction are: the concentration of the ethanol aqueous solution is 15-35wt%, the solid-liquid ratio is 1g: (40-10)mL, the discharge voltage is 0-7KV, the temperature is 10-50°C, and the extraction time is 1-5min.

4. The preparation method according to claim 1, characterized in that: In step 2), the liquid phase pulse discharge extraction adopts a multi-stage differentiation procedure: (a) Discharge voltage 0-1KV, time 0-2min; (b) Discharge voltage 1-4KV, time 2-3min; (c) Discharge voltage 4-6KV, time 3-4min; (d) Discharge voltage 6-7KV, time 4-5min; The concentration of the ethanol aqueous solution in each stage is 15-35wt%, the solid-liquid ratio is 1g: (40-10)mL, and the temperature is 10-50°C.

5. The preparation method according to claim 1, 3 or 4, characterized in that: In step 4), the conditions of supercritical CO2 extraction are: the concentration of ethanol aqueous solution is 50-80wt%, the solid-liquid ratio is 1g:(30-10) mL, the extraction pressure is 4500psi, the extraction temperature is 45-55°C, the CO2 flow rate is 1-3mL / min, and the extraction time is 10-30min.

6. The preparation method according to claim 1, characterized in that: The concentration in step 3) and / or the secondary concentration in step 5) is vacuum concentration: the temperature is 40-55° C. and the pressure is 0.09-0.10 MPa.

7. Use of the ginger root extract obtained according to the preparation method according to any one of claims 1 to 6 in cosmetics.

8. An anti-hair loss composition, characterized in that: The following components are included in mass percentage: 0.01-0.25% of the ginger root extract obtained by the preparation method according to any one of claims 1-6, Tea seed extract 0.01-0.25%, Platycladus orientalis leaf extract 0.01-0.25%, Hydrolyzed keratin 0.01-0.3%, Matrix.

9. The anti-hair loss composition according to claim 8, characterized in that: The tea seed extract includes at least one of camellia oleifera seed extract, tea seed extract and camellia seed extract.

10. The anti-hair loss composition according to claim 8 or 9, characterized in that: The matrix is ​​a shampoo matrix, a conditioner matrix, or an essence matrix.

Images