KR20230036757A - Steam ultrasonic extract for induction of black hair and preventing of white hair - Google Patents

Abstract

The present invention relates to a steamed ultrasonic extract for inducing black hair and preventing white hair, and a composition containing the same as an active ingredient. The composition containing a Nardostachys jatamansi extract as an active ingredient of the present invention exhibits radical scavenging activity, superoxide dismutase (SOD)-like activity, xanthine oxidase inhibitory activity, and an effect of increasing hair melamine synthesis by oxidation of L-DOPA converted from tyrosine.

Description

Steamed ultrasonic extract for inducing black hair and preventing white hair {STEAM ULTRASONIC EXTRACT FOR INDUCTION OF BLACK HAIR AND PREVENTING OF WHITE HAIR}

The present invention relates to a steamed ultrasonic extract for inducing black hair and preventing gray hair, and a composition containing the same as an active ingredient, and more specifically, an extract containing spikerad ( Nardostachys jatamansi ) as an active ingredient for prevention of gray hair, induction of black hair, and hair growth It relates to a composition expressing simultaneously.

The main component of hair is protein, and it is largely composed of three parts: the epidermis, the cortex, and the medulla. The cortex surrounding the medulla is the thickest part of the hair and is a granule called melanosome composed of melanin pigment, lipid membrane, TRP-1 (Tyrosinase Related Protein-1), TRP-2 (Tyrosinase Related Protein-2) and other proteins. contains

Melanin pigment is synthesized in melanosomes, which are organelles within melanocytes, and migrates to surrounding keratinocytes through the resinous projections of melanocytes.

Looking at the synthesis pathway of melanin, tyrosinase is involved in the step in which tyrosine, an amino acid, is first oxidized into dopa and then oxidized again to produce dopaquinone. It is known that the same enzyme is involved and accelerates the reaction.

The color of hair is determined by factors such as the type and distribution of eumelanin and pheomelanin, which are formed from the amino acid tyrosine as a raw material, and the degree of distribution, thereby revealing a unique hair color for each person. Eumelanin is the most common and darkest pigment, producing the color of brown and black hair, while pheomelanin, a lighter shade, produces blonde hair.

Gray hair is a phenomenon of physiological aging and is caused by a decrease in the number of hair melanocytes and a decrease in melanin production due to a decrease in the function of melanocytes.

Dyeing agents for hiding gray hair have been studied a lot and are widely marketed, but black hair dyeing is a temporary method that requires re-dyeing by the growth of gray hair, and currently widely used oxidative black hair dyeing damages the skin by using an oxidizing agent. give Therefore, it can be confirmed through Korean Publication No. 2002-0059906 and the like that various studies are being conducted on an anti-gray hair agent that prevents the occurrence of gray hair.

However, it is common for conventional anti-gray hair agents to contain large amounts of chemicals harmful to the human body, and in the case of anti-gray hair agents using natural substances, sufficient effects in preventing white hair and causing black hair have not been confirmed.

On the other hand, Spikerad ( Nardostachys jatamansi ) is a perennial plant of the Apiaceae family, and its roots contain 1-alristolene-2-one, nardostachone, and 1,8,9,10-tetrades. It contains ingredients such as tetrad-hydroaristolan-2-one.

Spikerad root decoction water has been used for mental disorders, insomnia, blood disorders and circulatory disorders (Uniyal, MR., et al., J. Res. Indian Med. 4, 83 (1969)), and is the main therapeutic component of Spikerad. is nardosinone, which enhances neurotrophic activity and central nervous system (CNS) effects (Li, p. et al., Journal of Pharmacology Science, 93, 122-125 (2003)).

Additionally, an alcoholic extract of spikerad protects rats from thioacetamide-induced liver damage (Bagchi, A., et al.). However, there has been no disclosure or disclosure of Spike Rad ( Nardostachys jatamansi ) in relation to Spike Rad's black hair-inducing and gray hair-preventing performance.

Accordingly, while the present inventors were researching a composition that provides a hair growth effect in addition to preventing gray hair and inducing black hair, spikerad ( Nardostachys jatamansi ), basil ( Ocimum basilicum ) and saffron ( Crocus sativus ) were used to prevent gray hair and black hair from a mixture. The present invention was completed by confirming that the hair growth effect could be expressed with induction.

An object of the present invention is to provide a composition containing a powder of a steamed ultrasonic extract containing spikerad ( Nardostachys jatamansi ) or an extract thereof as an active ingredient, which simultaneously expresses gray hair prevention, black hair induction, and hair growth.

The above and other objects of the present invention can all be achieved by the present invention described below.

In order to achieve the above object, the present invention

Spikerad ( Nardostachys jatamansi ), basil ( Ocimum basilicum ), and saffron ( Crocus sativus ) Provided is a composition for inducing black hair and preventing white hair, comprising a powder of a steamed ultrasonic extract or an extract thereof as an active ingredient.

Spike lard ( Nardostachys jatamansi ), basil ( Ocimum basilicum ), and saffron ( Crocus sativus ) may satisfy a weight ratio of 1:0.25 to 2:0.5 to 1.0 of spiked lard (a): basil (b): saffron (c) .

The steamed ultrasonic extract may be an ultrasonic extract of a steamed dried product having a water content of 10% or less by drying the steamed product steamed at 90 to 110 ° C. for 1 to 48 hours at 15 to 35 ° C. for 1 to 24 hours.

The ultrasonic extraction may be performed for 10 to 120 minutes at a frequency of 25 to 75 kHz.

The solvent of the ultrasonic extraction may include at least one selected from chloroform, ethanol, methanol, water, ethyl acetate, nucleic acid, and diethyl ether in an amount of 5 to 40 times the dry weight of the solid phase.

The steamed ultrasonic extract may be extracted for 2 to 5 hours at 60 to 90 ° C. under reflux.

According to the present invention, Spikerad simultaneously expresses antioxidant effects by radical scavenging activity, SOD (superoxide dismutase)-like activity, xanthine oxidase inhibitory activity, and hair melamine synthesis increasing effect by oxidation of L-DOPA converted from tyrosine It is possible to provide a composition for preventing gray hair, inducing black hair, and treating hair growth, using the steamed ultrasonic extract as an active ingredient.

1 is a graph comparing DPPH and ABTS radical scavenging activities for four types of samples (NOC-4) obtained in Example 1 and samples (NOC-1 to NOC-3) obtained in Comparative Examples 1 to 3 .
2 shows the degree of melanin production by in vitro oxidation of L-DOPA for four types of samples (NOC-4) obtained in Example 1 and samples (NOC-1 to NOC-3) obtained in Comparative Examples 1 to 3 It is a graph for
Figure 3 is a graph comparing the degree of melanin production in melanoma cells for four types of samples (NOC-4) obtained in Example 1 and samples (NOC-1 to NOC-3) obtained in Comparative Examples 1 to 3 am.
Figure 4 is a graph comparing the hair growth effect for four types of samples (NOC-4) obtained in Example 1 and samples (NOC-1 to NOC-3) obtained in Comparative Examples 1 to 3.
5 is a graph comparing the cytotoxicity of four types of samples (NOC-4) obtained in Example 1 and samples (NOC-1 to NOC-3) obtained in Comparative Examples 1 to 3.

Hereinafter, the present invention will be described in more detail to aid understanding of the present invention.

Terms or words used in this specification and claims should not be construed as being limited to their usual or dictionary meanings, and given that the inventors may appropriately define the concept of terms in order to best describe the invention. Therefore, it should be interpreted as meaning and concept consistent with the technical spirit of the present invention.

As confirmed in the following Examples and Experimental Examples, the present invention, when preparing a powder or extract of a steamed ultrasonic extract using spikerad ( Nardostachys jatamansi ), basil ( Ocimum basilicum ) and saffron ( Crocus sativus ), the powder or The extract simultaneously expresses radical scavenging activity, SOD (superoxide dismutase)-like activity, antioxidant activity by xanthine oxidase inhibitory activity, and hair melamine synthesis activity by oxidation of L-DOPA converted from tyrosine, thereby preventing gray hair and black hair A composition for inducing and inducing hair growth may be provided.

Ethanol stirring extract, ethanol reflux extract, or ethanol steamed extract have antioxidant effects by radical scavenging activity, SOD (superoxide dismutase)-like activity, and xanthine oxidase inhibitory activity, and hair by oxidation of L-DOPA converted from tyrosine. Although the melamine synthesis effect was not shown at the same time and is not shown in the experimental example below, the total amount of spiked rad ( Nardostachys jatamansi ), basil (Ocimum Ocimum basilicum basilicum), and saffron ( Crocus sativus ) was 1: Even if the weight ratio of 0.25~2:0.5~1.0 is not satisfied, antioxidant effect by radical scavenging activity, SOD (superoxide dismutase) similar activity, xanthine oxidase inhibitory activity, and oxidation of L-DOPA converted from tyrosine The hair melamine synthesis effect was not shown at the same time or the effect was reduced.

The composition of the present invention, which simultaneously realizes the antioxidant effect by radical scavenging activity, SOD (superoxide dismutase)-like activity, xanthine oxidase inhibitory activity, and the hair melamine synthesis effect by oxidation of L-DOPA converted from tyrosine, To be provided based on the results, the composition for inducing black hair and preventing white hair of the present invention is spiked ( Nardostachys jatamansi ) mixed with basil ( Ocimum basilicum ) and saffron ( Crocus sativus ) at a weight ratio of 1:0.25 to 2:0.5 to 1.0. It is characterized in that it contains, as an active ingredient, a steamed ultrasonic extract obtained by treating the steamed and dried product of the obtained solid phase product with ultrasonic waves using an extraction solvent.

In the present description, unless otherwise specified, an active ingredient means a component that exhibits the desired activity alone or that can exhibit activity in combination with a carrier that is not active by itself.

In this description, unless otherwise specified, the extract refers to an extract obtained by using an extraction solvent for the solid substance itself to be extracted, or a fraction obtained by fractionating the extract, and the extraction method refers to the polarity of the active substance, the degree of extraction, and the degree of preservation. In consideration of this, methods such as cooling, reflux, heating, supercritical extraction, and ultra-high pressure extraction can be applied. In the case of a fractionated extract, the fraction or crude extract obtained by suspending the extract in a specific solvent, mixing with a solvent of different polarity, and standing still is adsorbed on a column filled with silica gel, etc. It means including the fraction obtained by.

The solid material may be obtained in the form of a solid material after thoroughly washing spikerad ( Nardostachys jatamansi ), basil ( Ocimum basilicum ), and saffron ( Crocus sativus ). After cleaning, it can be dried by any method such as natural drying (shade drying), hot air drying, etc. to remove some of the moisture, and then crush it and then use it for steaming.

In the present invention, unless otherwise specified, spikerad, basil and saffron mean obtained by extraction from various organs or parts (eg leaves, branches, flowers, roots, stems, fruits, etc.), and preferably each leaf Or an extract obtained from a fruit, and most preferably an extract obtained from each leaf.

Bilberry ( Vaccinium vitis-idaea ) is a small evergreen shrub of the angiosperm family Rhododendron and bears fruit. Both the leaves and fruits of basil are used as medicine, especially for rheumatism or gout. In the past, it was used as a substitute for antibiotics. Recently, it is called lingonberry and is known to be rich in vitamins.

In addition, saffron ( Crocus sativus ) is a plant belonging to the iris family, and is known to be effective in preventing vascular disease, anti-cancer activity, stabilizing the brain, secreting hormones, and promoting learning due to its content of components such as crocin, carotenoids, and anthocyanins. . In addition, it is known to be effective in skin beauty because it has a good effect of discharging waste materials accumulated in the body to the outside.

In addition, it can be used in the form of pulverized material or in a dried state after steaming in stages, and when used in a dried state after steaming in stages, it is possible to provide the effect of increasing the usefulness of storage and active ingredients.

Unless otherwise specified, the steamed extract in the present description is a steamed product steamed at 90 to 110 ° C, or 95 to 105 ° C for 1 to 48 hours, or 10 to 30 hours at 15 to 35 ° C, or 20 to 30 ° C. for 1 to 24 hours, or 6 to 15 hours It may be dried and steamed and dried with a moisture content of 10% or less. In this case, it is preferable because it can provide the effect of increasing the active ingredient.

At this time, a normal steaming device can be used for the steaming, a normal drying device such as a hot air dryer can be used for drying, and a normal drying device such as a freeze dryer can be used for drying to adjust the moisture content. .

The steamed extract of the present invention is preferably obtained by adjusting the moisture content to 20% or less through pre-drying treatment and using the steaming and drying.

In the steaming and drying, if necessary, the steaming time is 6 to 15 hours in the first steaming and drying, the steaming time is 8 to 15 hours in the second steaming and drying, and the steaming time is 8 to 12 hours in the third steaming and drying. This can be done in multiple steps over time.

The steamed dried product is subjected to ultrasonic treatment.

The ultrasonic treatment is a treatment method using energy generated by ultrasonic vibration. The ultrasonic wave can destroy the insoluble solvent contained in the sample in the extraction solvent, and due to the high local temperature generated at this time, the reactant particles located around the Since the kinetic energy is increased, sufficient energy required for the reaction is obtained, and the impact effect of the ultrasonic energy induces high pressure to increase the mixing effect of the substance and the solvent contained in the sample, thereby increasing the extraction efficiency.

Extraction solvent that can be used in the ultrasonic extraction method may be used, for example, one or a mixture of two or more selected from chloroform, ethanol, methanol, water, ethyl acetate, nucleic acid, and diethyl ether.

The extraction solvent is preferably added in an amount of, for example, 5 to 40 times, specifically, 8 to 25 times the weight of the solid dried material, and ultrasonic treatment is performed to maximize reaction efficiency.

It is preferable to use alcohol as a solvent during sonication because it can provide improved performance compared to the case of alcohol extraction after sonication.

In addition, it is preferable to use 50 to 90% (v / v) ethanol as the alcohol because it can provide high antibacterial activity and polarity, and in particular, 70% (v / v) ethanol has a higher concentration. The sterilization power can be improved than using.

That is, the ultrasonic extract may be a 70% ethanol ultrasonic extract of dried solids using spikerad ( Nardostachys jatamansi ), basil ( Ocimum basilicum ) and saffron ( Crocus sativus ).

The ultrasonic extraction may be performed for 10 to 120 minutes at a frequency of 25 to 75 kHz, for example. In this case, it is possible to provide an increase in effective material extraction efficiency. Specifically, the ultrasonic extraction may be performed for 30 to 90 minutes at a frequency of 35 to 60 kHz.

In addition, it is possible to change the properties of the active ingredient according to the temperature and pressure conditions of extraction. For example, when the ultrasonic extract is extracted under low-temperature conditions, the concentration of sugar in the extract decreases due to the low solubility of the active ingredient. The concentration of the active ingredient is higher than that of the extract in the second half. In this extraction method, it is possible to adjust the content of free fatty acids and the content of active ingredients by adjusting the fractionation time. There is a way to lower the degree of concentration by mixing the sections.

The free fatty acid content of raw materials varies depending on the country of origin or cropping, and since raw materials with low acid values are usually formed at high prices, they are less competitive as raw materials. In addition, since there is no raw material without free fatty acids, free fatty acids are concentrated in the extract concentration section, so even if a product is made by setting a standard for using raw materials with a low acid value, it is easy to meet the acid value standard while satisfying the active ingredient content at a certain concentration or higher. It's not work. Therefore, in order to have industrial stability in the process of producing a high-concentration fraction using an ultrasonic extract, a technique for removing free fatty acids by repeated extraction under reflux (also referred to as reflux extraction) is required.

The reflux extraction may be repeated at 20 to 75°C, or at 50 to 70°C for 12 hours or less, or 1 to 6 hours. The number of repetitions of extraction may be 5 times or less, for example, 2 to 3 times.

A purification process that removes the above-mentioned free fatty acids without affecting the efficacy without loss of active ingredients is required. For this purpose, a method of filtering using a fine filtration membrane and then concentrating under reduced pressure to remove free fatty acids is preferred, and more excellent efficacy It is more preferable to use a filtration membrane of 1.0 μm or less, preferably 0.45 μm or less, for providing the effect.

An effective extract can be obtained through the above-described purification process.

In the present description, the effective extract includes a concentrated liquid extract or solid extract from which the ultrasonic extraction solvent has been removed by freeze drying, vacuum drying, hot air drying, spray drying, vacuum concentration, or the like.

In the present disclosure, the form of the resulting extract is not particularly limited and may be in the form of a powder or other extract, including pills, granules, and powders.

The method for preparing an extract according to the present disclosure includes, for example, spiked rad ( Nardostachys jatamansi ), basil ( Ocimum basilicum ) and saffron ( Crocus sativus ), and then steaming and drying the obtained solid or dry product; It may include; and ultrasonically extracting the steamed dried product.

In the present disclosure, the spikerad ( Nardostachys jatamansi ), basil ( Ocimum basilicum ), and saffron ( Crocus sativus ) can be obtained in the form of solids after being cleaned. After cleaning, some of the moisture is removed by drying in an arbitrary method such as natural drying (shade drying) or hot air drying (in the form of pulverized material so that the moisture content of the solid material is 10% (w / w) or less) 10 to 10 It may be crushed to 50 mesh, specifically 30 to 50 mesh to increase the surface area and then used for ultrasonic pretreatment.

If necessary, it may be pulverized after drying at 95° C. or less, or 50 to 90° C. for 36 hours or more, or 48 hours or more.

The solid may be subjected to a pre-extraction step prior to ultra-high pressure extraction.

There is no particular limitation on the solids used in this description, and the pre-extraction step is for removing harmful substances such as microorganisms, residual pesticides, preservatives, etc. treatment, treatment to increase the contact area of ultrasonic waves, and the like.

The pre-extraction method is not particularly limited, but one or more of UV irradiation, heat treatment, peeling, grinding, and roasting may be used.

It is then subjected to sonication, which allows grinding to high purity while preventing the formation of additional by-products.

In the case of the ultrasonic treatment, it is preferable to increase the treatment efficiency by supplying an extraction solvent to the solid dried material. At this time, the usable extraction solvent is at least one selected from chloroform, ethanol, methanol, water, ethyl acetate, nucleic acid and diethyl ether can be used. At this time, the extraction solvent may be included in an amount of 5 to 40 times the weight of the solid dry matter.

The ultrasonic treatment is preferably carried out for 10 to 120 minutes at a frequency of 25 to 75 kHz, for example, to increase treatment efficiency.

Subsequently, repeated reflux extraction may be performed under reflux of the ultrasonic treatment liquid (also referred to as ultrasonic extraction liquid). At this time, extraction may be repeated at 20 to 75° C., specifically at 50 to 70° C. for 12 hours or less, and specifically for 1 to 6 hours. The number of repetitions of extraction may be 5 times or less, for example, 2 to 3 times.

The steamed ultrasonic extract can be purified and concentrated under reduced pressure as described above to remove the extraction solvent provided during ultrasonic treatment and separate the effective extract.

The composition of the present invention may be prepared by including a hair protection component, a hair elasticity component, a UV protection component, a hair moisturizing component, etc. known in the art in order to reinforce or add hair-related activities in addition to the active ingredient. One or more of these components may be included in the composition of the present invention together with the active ingredient.

The composition of the present invention may contain the active ingredient in any amount (effective amount) as long as it can exhibit gray hair prevention, black hair induction, hair growth effect, etc. depending on product type, product formulation, etc., and the effective amount is based on the total weight of the composition It may be determined within the range of 0.001 to 15 wt% as a standard.

In the present invention, the effective amount, unless otherwise specified, when the composition of the present invention is administered to a mammal, preferably a human, during the administration period according to the advice of medical experts, etc. It refers to the amount of the active ingredient included in the composition of the present invention that can exhibit the intended medical/cosmetic effect. Such an effective amount can be determined empirically within the ordinary skill of the skilled artisan.

In a specific aspect, the composition of the present invention can be identified as a cosmetic composition.

Even when the composition of the present invention is identified as a cosmetic composition, the cosmetic composition may be classified as an arbitrary product in terms of its use and law, and specifically, it may be a functional cosmetic having a purpose such as a hair growth effect, a non-functional general cosmetic, etc. .

In product form, it may take any product form, specifically, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax It can be formulated as a foundation, spray, and the like. In a specific product form, it may be a softening lotion, nutrient lotion, nutrient cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder formulation.

The cosmetic composition of the present invention may include, in addition to the active ingredient, ingredients commonly used in cosmetic compositions, for example, stabilizers, solubilizers, surfactants, vitamins, auxiliary agents such as pigments and fragrances, and carriers.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane / May contain a propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol, fatty acid esters of sorbitan, and the like can be used.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, or polyoxyethylene sorbitan ester, microcrystals Star cellulose, aluminum metahydroxide, bentonite, agar, and the like can be used.

When the formulation of the present invention is surfactant-containing cleansing, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

The cosmetic composition of the present invention is a cosmetic composition commonly performed in the art, except for containing active ingredients exhibiting radical scavenging activity, SOD (superoxide dismutase)-like activity, xanthine oxidase inhibitory activity, and hair melamine synthesis. It can be prepared according to the manufacturing method of.

In another specific aspect, the composition of the present invention can be regarded as a food composition.

The food composition of the present invention can be prepared in any form, for example, beverages such as tea, juice, carbonated beverages, and ionic beverages, processed oils such as milk and yogurt, chewing gum, rice cakes, traditional Korean snacks, bread, snacks, noodles, etc. It can be manufactured into health functional food preparations such as food, tablet, capsule, pill, granule, liquid, powder, flake, paste, syrup, gel, jelly, bar, etc.

In addition, the food composition of the present invention may take any product classification as long as it conforms to the enforcement regulations at the time of manufacture / distribution in functional classification. For example, it may be a health functional food according to the Act on Health Functional Food, or a special purpose food according to each food type in the Food Codex (MFDS notice, food standards and specifications) of the Food Sanitation Act.

The food composition of the present invention may include food additives in addition to the active ingredient. Food additives are generally understood as substances that are added to, mixed with, or infiltrated into food in manufacturing, processing, or preserving food, and since they are taken daily and for a long period of time with food, their safety must be guaranteed. According to the Food Additive Code (Ministry of Food and Drug Safety Notice, Food Additive Standards and Specifications) under the Food Sanitation Act, safety-guaranteed food additives are classified into chemical synthetic products, natural additives, and mixed formulations and are limitedly regulated.

These food additives can be classified into sweeteners, flavors, preservatives, emulsifiers, acidulants, thickeners, etc. in terms of functionality.

Sweeteners can be used in amounts that give the food a moderately sweet taste, and can be natural or synthetic. Preferably, a natural sweetener is used, and examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavors may be used to improve taste or aroma, and both natural and synthetic flavors may be used. Preferably, it is the case of using a natural one. In case of using natural ones, in addition to flavor, the purpose of enhancing nutrition can also be combined. As a natural flavoring agent, it may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, etc., or obtained from green tea leaves, roundworms, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo, etc. can also be used. Natural flavors can be liquid concentrates or solid extracts.

Synthetic flavors may also be used, and synthetic flavors may include esters, alcohols, aldehydes, terpenes, and the like.

As preservatives, sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. may be used, and as emulsifiers, acacia gum, carboxymethylcellulose, xanthan gum , pectin, and the like.

As the acidulant, acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, and the like may be used. Acidulants may be added to the food composition to have an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to improving taste.

A suspending agent, a sedimentation agent, a gel forming agent, a swelling agent, and the like may be used as the thickening agent.

The food composition of the present invention, in addition to the above-mentioned food additives, may include physiologically active substances or minerals known in the art for the purpose of supplementing or reinforcing functionality and nutrition and ensuring stability as food additives.

Examples of such physiologically active substances include catechins contained in green tea, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoylthiamine, and the like. Examples of minerals include calcium preparations such as calcium citrate and magnesium stearate. magnesium preparations such as the like, iron preparations such as iron citrate, chromium chloride, potassium iodine, selenium, germanium, vanadium, zinc, and the like.

The food composition of the present invention may include the above-mentioned food additives in an appropriate amount to achieve the purpose of addition according to the type of product. Regarding other food additives that may be included in the food composition of the present invention, reference may be made to the Food Codex or Food Additives Codex.

In a specific embodiment, the composition of the present invention can be identified as a pharmaceutical composition.

The pharmaceutical composition of the present invention may be formulated into an oral formulation or a parenteral formulation according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient.

In the present invention, pharmaceutically acceptable means that, unless otherwise specified, it does not inhibit the activity of the active ingredient and does not have toxicity more than is adaptable to the subject of application (prescription).

When the pharmaceutical composition of the present invention is prepared as an oral dosage form, powder, granule, tablet, pill, dragee, capsule, liquid, gel, syrup, suspension, wafer according to a method known in the art together with a suitable carrier. It can be prepared in formulations such as

Suitable pharmaceutically acceptable carriers include, for example, sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, and wheat starch, cellulose, methylcellulose, and ethyl Cellulose, sodium carboxymethylcellulose, cellulose such as hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol , vegetable oil, and the like. If necessary, diluents and/or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants may be included in the formulation.

When the pharmaceutical composition of the present invention is prepared as a parenteral formulation, it may be formulated in the form of an injection, transdermal administration, nasal inhalation, and suppository along with a suitable carrier according to a method known in the art.

When formulated as an injection, suitable carriers include sterile water, ethanol, polyols such as glycerol or propylene glycol, or mixtures thereof, preferably Ringer's solution, PBS (phosphate buffered saline) containing triethanolamine or sterilization for injection water, isotonic solutions such as 5% dextrose, and the like can be used.

When formulated as a transdermal formulation, it may be formulated in the form of an ointment, cream, lotion, gel, external agent, pasta, liniment, or air roll.

In the case of nasal inhalation, it may be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, or the like.

Bases for suppositories include witepsol, tween 61, polyethylene glycols, cacao fat, laurin fat, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, and sorbitan fatty acid esters. can be used

Formulation of pharmaceutical compositions is known in the art, and may be specifically referred to in Remington's Pharmaceutical Sciences (19th ed., 1995). These documents are considered as part of this specification.

The preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 10 g/kg per day, depending on the patient's condition, body weight, sex, age, severity of the patient, and route of administration. It may be in the range of 1 g/kg. Administration can be done once a day or divided into several times. These dosages should not be construed as limiting the scope of the present invention in any respect.

The cosmetic composition, food composition, and pharmaceutical composition of the present invention may be prepared according to the preparation method of the composition commonly performed in the art, except for including active ingredients known in the art.

In describing the composition for inducing black hair and preventing white hair of the present description, other conditions or equipment not explicitly described may be appropriately selected within the range commonly practiced in the art, and are not particularly limited.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily practice the present invention. However, the present invention may be embodied in many different forms and is not limited to the embodiments described herein.

[Example]

<Example 1> Preparation of steamed ultrasonic spikerad extract

(1) Spikerad ( Nardostachys jatamansi ) leaves, basil ( Ocimum basilicum ) leaves, and saffron ( Crocus sativus ) leaves were thoroughly washed, dried at 70 ° C. for 48 hours, and pulverized to a size of 40 mesh or less. The pulverized raw materials were mixed at a constant weight (100 g: 100 g: 100 g).

(2) The mixed raw materials were steamed at 100 ℃ for 12 hours and then naturally dried for 12 hours.

(3) 70% ethanol 10 times the total weight of the steamed dry matter was added and treated for 60 minutes at a frequency of 50 kHz using an ultrasonic extractor.

(4) Then, reflux extraction was performed by repeating the ultrasonic extract at 70° C. for 3 hours 3 times.

(5) The obtained extract was filtered through a 0.45 μm membrane filter and then ethanol was removed using a vacuum concentrator to prepare a steamed ultrasonic spikerad extract (sample NOC-4).

<Comparative Example>

<Comparative Example 1> Preparation of Stirred Spikerad Extract

(1) Spikerad ( Nardostachys jatamansi ) leaves, basil ( Ocimum basilicum ) leaves, and saffron ( Crocus sativus ) leaves were thoroughly washed, dried at 70 ° C. for 48 hours, and pulverized to a size of 40 mesh or less. The pulverized raw materials were mixed at a constant weight (100 g: 100 g: 100 g).

(3') 70% ethanol 10 times the total weight of the solids was added, and extraction at room temperature at 200 rpm for 12 hours was repeated three times.

(5) The obtained extract was filtered through a 0.45 μm membrane filter and then ethanol was removed using a vacuum concentrator to prepare a stirred Spikerad extract (sample NOC-1).

<Comparative Example 2> Preparation of Reflux Spikerad Extract

(1) Spikerad ( Nardostachys jatamansi ) leaves, basil ( Ocimum basilicum ) leaves, and saffron ( Crocus sativus ) leaves were thoroughly washed, dried at 70 ° C. for 48 hours, and pulverized to a size of 40 mesh or less. The pulverized raw materials were mixed at a constant weight (100 g: 100 g: 100 g).

(3") 70% ethanol in an amount of 10 times the total weight of the solids was added.

(4') Then, reflux extraction was performed three times at 70 °C for 3 hours.

(5) The obtained extract was filtered through a 0.45 μm membrane filter, and ethanol was removed using a vacuum concentrator to prepare a reflux Spikerad extract (sample NOC-2).

<Comparative Example 3> Preparation of Steamed Spikerad Extract

(1) Spikerad ( Nardostachys jatamansi ) leaves, basil ( Ocimum basilicum ) leaves, and saffron ( Crocus sativus ) leaves were thoroughly washed, dried at 70 ° C. for 48 hours, and pulverized to a size of 40 mesh or less. The pulverized raw materials were mixed at a constant weight (100 g: 100 g: 100 g).

(2) The mixed raw materials were steamed at 100 ℃ for 12 hours and then naturally dried for 12 hours.

(3") 70% ethanol in an amount of 10 times the total weight of the solids was added.

(4') Then, reflux extraction was performed three times at 70 °C for 3 hours.

(5) The obtained extract was filtered through a 0.45 μm membrane filter and then ethanol was removed using a vacuum concentrator to prepare a steamed Spikerad extract (sample NOC-3).

<Experimental example>

<Experimental Example 1> Antioxidant activity test 1

Antioxidant tests on the samples obtained in Example 1 and the samples (NOC-1 to 4) obtained in Comparative Examples 1 to 3 were performed to evaluate the radical scavenging ability of the sample by ABTS method and DPPH method.

(1-1) ABTS radical scavenging activity

Each of the samples (NOC-1 to NOC-4) is diluted in water to prepare test solutions with concentrations of 100 μg/mL, 250 μg/mL, and 500 μg/mL. 7 mM ABTS and 2.45 mM potassium persulfate are mixed and reacted at room temperature for 12 hours in a dark room to form ABTS cations. Thereafter, ethanol was added to adjust the absorbance value at 734 nm to be 0.70 ± 0.02.

100 μl of the test solution and 100 μl of the prepared ABTS solution were added to a 96-well plate, reacted at room temperature for 7 minutes, and measured at 734 nm. Compared with the blank test solution, the ABTS clearance rate was obtained as a percentage (%) as follows and is shown in FIG. 1 below.

ABTS radical scavenging ability (%) = [Control - (Sample - Blank)]/Control Х 100

(However, Control: absorbance of ABTS reagent, Sample: absorbance of Sample + ABTS reagent, Blank: absorbance of Sample + Blank)

(1-2) DPPH radical scavenging activity

Each of the samples (NOC-1 to NOC-4) is diluted in water to prepare test solutions with concentrations of 100 μg/mL, 250 μg/mL, and 500 μg/mL. 100 μl of the test solution and 100 μl of 0.2 mM DPPH were put in a 96-well plate, and after 30 minutes, the absorbance was measured at 517 nm using a microplate reader. Compared to the blank test solution, the DPPH radical scavenging rate was obtained in percentage (%) as follows and is shown in FIG. 1 below.

DPPH radical scavenging ability (%) = [Control - (Sample - Blank)]/Control Х 100

(Control: Absorbance of DPPH reagent, Sample: Absorbance of Sample + DPPH reagent, Blank: Absorbance of Sample + Blank)

As shown in FIG. 1, as a result of ABTS radical analysis, ascorbic acid as a control group was found to be the highest at 99.3% at 500 μg/mL, followed by 64.9% at 500 μg/mL in the sample (NOC-4) according to Example 1. was confirmed as high, and then the sample according to Comparative Example 3 (NOC-3), the sample according to Comparative Example 2 (NOC-2), and the sample according to Comparative Example 1 (NOC-1) in the order of 500 μg / mL 59.8%, 54.7%, and 51.1%, respectively.

In addition, as a result of the DPPH radical analysis, the same tendency as the above-mentioned ABTS analysis result was shown. Specifically, the sample according to Example 1 (NOC-4), the sample according to Comparative Example 3 (NOC-3), the sample according to Comparative Example 2 (NOC-2), and the sample according to Comparative Example 1 (NOC-1) ) showed 65.4%, 63.8%, 60.7%, and 57.5% at 500 μg/mL, respectively. At this time, ascorbic acid as a control was analyzed at 98.3% at 500 μg/mL.

<Experimental Example 2> Antioxidant activity test 2

As an antioxidant test on the samples obtained in Example 1 and the samples (NOC-1 to 4) obtained in Comparative Examples 1 to 3, SOS-like activity and xanthine oxidase inhibitory activity were tested.

(2-1) SOS-like activity

Each of the samples (NOC-1 to NOC-4) is diluted in water to prepare test solutions with concentrations of 100 μg/mL, 250 μg/mL, and 500 μg/mL. To 0.2 mL of the test solution, add 2.6 mL of tris-HCl buffer corrected to pH 8.5 and 0.2 mL of 7.2 mM pyrogallol, and react at 25 °C for 10 minutes. Add 0.1 mL of 1 N HCl to the reacted solution to stop it. The amount of oxidized pyrogallol is determined by measuring the absorbance at 420 nm. Compared to the blank test solution, the SOD-like activity was obtained in percentage (%) and is shown in Table 1 below.

(2-2) Xanthine oxidase inhibitory activity

Samples (NOC-1 to NOC-4) were diluted in water, respectively, to prepare assays with concentrations of 100 μg/mL, 250 μg/mL, and 500 μg/mL. Add 0.6 mL of 0.1 M potassium phosphate buffer (pH 7.5) and 0.2 mL of 1 mM xanthine to 1.0 mL of the test solution. Then, 0.1 mL of 0.2 U/mL xanthine oxidase was added to stop the reaction. The resulting uric acid is obtained by measuring the absorbance at 292 nm. Compared with the blank test solution, the Xanthine oxidase inhibitory activity was obtained in percentage (%) and is shown together in Table 1 below.

Extract SOD-like activity (%) Xanthin oxidase Inhibition (%) 100 µg/mL 250 µg/mL 500 µg/mL 100 µg/mL 250 µg/mL 500 µg/mL Control 94.0±3.7 96.1±4.2 97.9±4.2 95.2±4.8 97.2±3.5 98.2±4.2 NOC-1 11.2±1.4 19.7±2.8 30.1±1.2 13.4±3.1 19.1±1.0 24.4±3.3 NOC-2 13.2±2.1 20.8±1.6 31.2±2.3 15.4±0.8 20.7±2.3 26.7±2.5 NOC-3 18.2±1.8 21.8±2.5 32.4±3.9 17.5±2.8 21.7±1.5 28.6±2.1 NOC-4 19.2±2.4 23.9±1.8 34.3±2.4 19.6±1.6 23.1±2.5 30.8±3.1

*Control: ascorbic acid

As shown in Table 1, as a result of analyzing the SOD-like activity, the sample according to Example 1 (NOC-4) was found to be the highest at 34.3% at 500 μg / mL, followed by the sample according to Comparative Example 3 (NOC-4). 3), the sample according to Comparative Example 2 (NOC-2), and the sample according to Comparative Example 1 (NOC-1) showed 32.4%, 31.2%, and 30.1% at 500 μg/mL in that order. All test materials were analyzed for scavenging activity in a concentration-dependent manner, and all were analyzed to be lower than the control ascorbic acid.

In addition, as a result of the analysis of xanthine oxidase inhibitory activity, the sample according to Example 1 (NOC-4) was found to be the highest at 500 μg / mL at 30.8%, followed by the sample according to Comparative Example 3 (NOC-3) , the sample according to Comparative Example 2 (NOC-2), and the sample according to Comparative Example 1 (NOC-1) were analyzed to show 28.6%, 26.7%, and 24.4% at 500 μg / mL in that order, and the above-mentioned SOD The same tendency as the analysis result of similar activity was confirmed.

<Experimental Example 3> Hair growth melamine synthesis test 1

An L-DOPA oxidation experiment was performed as a hair growth melamine synthesis test for the samples obtained in Example 1 and the samples (NOC-1 to 4) obtained in Comparative Examples 1 to 3. L-DOPA oxidation converts L-DOPA from tyrosine, and then these L-DOPAs are oxidized to synthesize melamine.

Specifically, samples (NOC-1 to NOC-4) are diluted in water to prepare test solutions having concentrations of 100 μg/mL, 250 μg/mL, and 500 μg/mL. 480 μg/ml of 0.1 M phosphate buffer (pH6.5) and 20 μg/ml of L-3,4-dihydroxyphenyl alanine (L-DOPA), which is involved in melanin synthesis, and 2,000 U/ml mushroom tyrosinase 40 After adding μg/ml in order, it was reacted at 37℃ for 1 hour. In addition, the amount of melanin production was confirmed by measuring the absorbance at 490 nm using a microplate reader, and then shown in FIG. 2 below.

As shown in FIG. 2, as a result of in vitro melanin production analysis, the sample (NOC-4) according to Example 1 was found to be the highest at 500 μg / mL at 162.6%, followed by the sample according to Comparative Example 3 ( NOC-3), the sample according to Comparative Example 2 (NOC-2), and the sample according to Comparative Example 1 (NOC-1) showed 156.5%, 149.4%, and 147.1% at 500 μg/mL in that order. All test substances were analyzed to have increased melanin production compared to the control group in vitro .

<Experimental Example 4> Whitening activity test 2

An experiment was performed to determine the degree of hair growth melanin production for the samples obtained in Example 1 and the samples (NOC-1 to 4) obtained in Comparative Examples 1 to 3. By stimulating melanin production in melanoma cells (B16F1) using L-DOPA, the relative amount of melanin production of the samples (NOC-1-4) and the control group (water) was compared.

Specifically, 1Х10 ⁴ cell/ml of melanoma cells (B16F1) are dispensed in DMEM medium and pre-incubated for 24 h under 37 °C and 5% CO ₂ conditions. Thereafter, samples (NOC-1 to NOC-4) at a concentration of 100 to 500 μg/mL were added and L-DOPA was added to stimulate melanin production, followed by incubation for another 48 hours. After incubation, the cells are collected, centrifuged at 1,200 rpm for 5 minutes to precipitate, and then dissolved in 1 ml homogenization buffer (50 mM sodium phosphate pH 6.5, 1% Triton X-100, 2 mM phenyl methyl sulfonyl fluoride) to obtain a pellet. . After adding and mixing 200 μl of 1N NaOH (10% DMSO) to the pellet, the amount of melanin produced was confirmed by measuring the absorbance at 405 nm, and then shown in FIG. 3 below.

As shown in FIG. 3, as a result of analyzing the amount of melanin produced in mouse melanoma cells, in the case of the sample (NOC-4) according to Example 1, it was found to be the lowest at 500 μg / mL at 162.6% compared to the control group, and according to Comparative Example 3 Sample (NOC-3), sample according to Comparative Example 2 (NOC-2), and sample according to Comparative Example 1 (NOC-1) showed 156.3%, 149.4%, and 147.1% at 500 μg/mL in that order. .

It was found that all samples showed an increase in melanin production compared to the control group, and the analysis result was confirmed to be consistent with the above-described in vitro experiment trend.

<Experimental Example 5> Hair growth effect test

In order to evaluate the hair growth efficacy of the samples obtained in Example 1 and the samples (NOC-1 to 4) obtained in Comparative Examples 1 to 3, the experimental group (NOC-1 to 4) for 4 weeks was 250, 500 mg / kg It was administered orally, and the control group was administered with the same amount of water to evaluate the hair growth effect.

Specifically, 6-week-old CL57BL/6 mice (Orient Bio Co., Ltd.) were purchased and sufficiently supplied with water and solid feed to adapt to the laboratory environment (temperature 22±2°C, humidity 55±5%, 12-hour light/dark cycle). and managed. They were divided into 5 groups, 5 animals per group, and the experiment was conducted after first depilating the hair on the back of the mice. For 4 weeks, the experimental groups (NOC-1~4) were orally administered 250 and 500 mg/kg each, and the control group was administered the same amount of water.

In order to confirm the degree of hair growth, after measuring the growth period hair growth value to 0.1 mm with vernier calipers, experimental animal tissues were collected and fixed in 10% formalin for 24 hours. After cutting the fixed tissues and embedding them in paraffin, paraffin tissue sections were prepared with a thickness of 4 μm using a microtome. Tissues were stained with Hematolxylin & Eosin, and histological changes in hair follicle tissue were observed with an optical microscope (DP71, Olympus).

As shown in FIG. 4, as a result of analyzing the hair growth efficacy, it was confirmed that the sample according to Example 1 (NOC-4) had the most excellent hair growth at 500 mg/kg at 0.94±0.10 mm.

In addition, in the sample (NOC-3) according to Comparative Example 3, it was 0.91 ± 0.09 mm at 500 mg / kg, followed by the sample according to Comparative Example 2 (NOC-2) and the sample according to Comparative Example 1 (NOC- 1) were analyzed in order, and were analyzed at 0.85±0.10 mm and 0.81±0.09 mm at 500 mg/kg, respectively. The control group was confirmed to have the slowest hair growth at 0.65±0.10 mm.

Furthermore, the hair follicle count analysis showed the same trend as the hair growth efficacy analysis result. Specifically, the sample according to Example 1 (NOC-4), the sample according to Comparative Example 3 (NOC-3), the sample according to Comparative Example 2 (NOC-2), the sample according to Comparative Example 1 (NOC-1) 11.9 ± 1.4, 10.5 ± 1.2, 9.5 ± 1.4, and 9.1 ± 1.1 at 500 mg/kg in the order of . The control group was 4.5±1.8 pieces, and it was confirmed that the test substance induces an increase in the number of hair follicles.

<Experimental Example 6> safety test

In order to evaluate the safety of the samples obtained in Example 1 and the samples (NOC-1 to 4) obtained in Comparative Examples 1 to 3, the cytotoxicity of the test substance (NOC-1 to 4) was tested on mouse melanoma cells. To confirm, MTS analysis was performed. The concentration of the test substance was set to 100 to 500 μg/mL, and the cytotoxicity of the test substance was confirmed by culturing mouse melanoma cells (B16F10, ATCC) cells.

Specifically, mouse melanoma cells (B16F10, ATCC) were dispensed at 1Х10 ⁴ cell/ml and pre-incubated for 24 h at 37 °C and 5% CO ₂ , and samples at a concentration of 100 to 500 μg/mL (NOC-1 ~4) and control (water) were added and cultured for 24 hours. Thereafter, 20 μl of MTS reagent was added and incubated for 2 hours, and the absorbance was measured with an ELISA reader (Epoch ^TM 2, BioTek, USA) at 570 nm using a microplate reader.

Cell viability was calculated using Equation 1 below, and the results are shown in FIG. 4 below.

[Equation 1]

Cell viability (%) = [(Exp. - Blank)/Control] Х 100

(In Equation 1, Exp: absorbance of extract containing cells, Blank: absorbance of extract without cells, Control: absorbance of distilled water containing cells.)

As shown in FIG. 5, as a result of cytotoxicity analysis in mouse melanoma cells, it was confirmed that the cell viability was 95% or more at all concentrations.

Therefore, in the sample (NOC-4) according to Example 1, the sample (NOC-3) according to Comparative Example 3, the sample (NOC-2) according to Comparative Example 2, and the sample (NOC-1) according to Comparative Example 1 It can be seen that there is no cytotoxicity at the concentration of 100 to 500 μg/mL.

In conclusion, when the powder or the extract of steamed ultrasonic extract using spikerad ( Nardostachys jatamansi ), basil ( Ocimum basilicum ) and saffron ( Crocus sativus ) is included as an active ingredient, the obtained composition has ABTS, DPPH free radical scavenging activity It can be confirmed that it is suitable for applications requiring gray hair prevention, black hair inducing effect, and hair growth effect, such as the increase in hair melamine synthesis due to the oxidation of L-DOPA converted from tyrosine. there was.

Claims (6)

Spikerad ( Nardostachys jatamansi ), basil ( Ocimum basilicum ) and saffron ( Crocus sativus ) A composition for inducing black hair and preventing white hair, comprising a powder of a steamed ultrasonic extract or an extract thereof as an active ingredient. According to claim 1,
The spiked lard ( Nardostachys jatamansi ), basil ( Ocimum basilicum ) and saffron ( Crocus sativus ) are spiked lard (a): basil (b): saffron (c) satisfying a weight ratio of 1: 0.25 to 2: 0.5 to 1.0 A composition for inducing black hair and preventing white hair. According to claim 1,
The steamed ultrasonic extract is 1 to 48 hours at 90 to 110 ℃ The steamed product is boiled for 1 to 24 hours at 15 to 35 ° C. It may be an ultrasonic extract of a steamed dried product having a water content of 10% or less after being dried. According to claim 1,
The ultrasonic extraction is a composition for inducing black hair and preventing white hair, which is extracted for 10 to 120 minutes at a frequency of 25 to 75 kHz. According to claim 1,
The solvent of the ultrasonic extraction contains at least one selected from chloroform, ethanol, methanol, water, ethyl acetate, nucleic acid and diethyl ether in an amount of 5 to 40 times the dry weight of the solid phase. Composition. According to claim 1,
The steamed ultrasonic extract is extracted for 2 to 5 hours at 60 to 90 ° C. under reflux, a composition for inducing black hair and preventing white hair.

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