KR20180040756A - Skin whitening composition comprising an extract of coixlachryma-jobi var. mayuen - Google Patents

Abstract

The present invention relates to a whitening composition, which comprises an adlay extract or a solvent fraction thereof, wherein the adlay extract of the solvent fraction thereof contains Coixol, and more particularly, to a whitening composition which exhibits an effective tyrosinase inhibitory activity. The adly extract is obtained by extracting the adlay roots from a C1 to C6 alcohol or a mixture of C1 to C6 alcohol and water.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a whitening composition comprising an extract of Yulmu mushroom extract. MAYUEN}

The present invention relates to a whitening composition comprising a yulmu extract, and more particularly, to a whitening composition comprising an extract of yulmu roots containing a cucumber sol or a solvent fraction thereof.

In general, the color of skin is affected by environment, race, sex, etc. The degree of skin whitening is determined by the content of brown melanin. Over-deposition of melanin pigment in the skin may result in hyperpigmentation, phototoxic response or other similar small-sized post-inflammatory reactions due to scarring or dermatitis, including unwanted freckles, senile spots, liver, spots, brown or black spots, It can cause fixed pigmented lesions, which can harm skin aesthetics regardless of the original color of the skin.

For example, Korean Patent Registration No. 10-0841276 discloses a composition for skin whitening containing a skin white ginseng ethyl acetate fraction having excellent whitening activity, more specifically, a skin whitening composition containing skin white ginseng ethyl acetate fraction containing cinnamic acid as an active ingredient Compositions for skin whitening are described.

Korean Patent Registration No. 10-0850686 discloses an extract of Nigella glandulifera Freyn et Sint having skin whitening activity and a whitening cosmetic composition containing the same as an active ingredient, specifically inhibiting melanin production in melanin-producing cells And a whitening cosmetic composition containing the same as an active ingredient.

Korean Patent Registration No. 10-1121736 discloses an extract of herbal medicines for skin whitening, a method for producing them, and a whitening cosmetic composition containing the same, wherein the extract is obtained by extracting from ear, wax, Is disclosed.

Korean Patent Publication No. 10-0604085 discloses a whitening cosmetic composition comprising an upper body extract and glutathione, wherein the composition is contained in an amount of 0.1 to 20.0% by weight, preferably 1.0 to 10.0% by weight based on the total weight of the composition, A whitening cosmetic composition containing 0.001 to 5.0% by weight, preferably 0.01 to 1.0% by weight, of a topical extract is described based on the total weight of the composition.

Korean Patent Registration No. 10-0345226 discloses a cosmetic composition having an excellent skin whitening effect, and more specifically, a cosmetic composition for skin whitening comprising an enamel extract as an active ingredient, wherein the enamel extract, A cosmetic composition for skin whitening, which is mixed with at least one extract selected from the group consisting of ginseng extract, motherwort extract, monascus extract, and licorice extract.

Korean Patent Publication No. 10-0515206 discloses a composition containing as an active ingredient an extract of Bacillus thuringiensis, and if necessary, ranunculin, deoxypodophyllotoxin, 3-O-α -L-laminopyranosyl (1 → 2) - [β-D-glucopyranosyl (1 → 4)] - α-L-arabinopyranoside (SB365) A whitening cosmetic composition containing at least one auxiliary component selected from the group consisting of dandruff and licorice extract is disclosed.

As described above, a whitening composition comprising various natural ingredients has been disclosed. A composition for whitening using a whitening agent has also been developed. For example, Korean Patent Registration No. 10-1579500 discloses a whitening agent comprising an extract of Ulmu root and its fractions / RTI >

In the prior art, a solvent extract and fractions thereof are prepared from a roasted roots, and as a main ingredient, a tyrosinase inhibitory activity and an inhibitory effect on the production of melanin are confirmed. However, since the prior art does not aim at obtaining a whitening effect by targeting a specific compound of Bombyx mori root, there is a limit in improving the whitening effect.

Korean Patent Publication No. 10-1579500 Korean Patent Publication No. 10-0841276 Korean Patent Publication No. 10-0850686 Korean Patent Publication No. 10-1121736 Korean Patent Publication No. 10-0604085 Korean Patent Publication No. 10-0345226 Korean Patent Publication No. 10-0515206

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and it is an object of the present invention to provide a composition exhibiting a whitening effect through a solvent fraction of a yulmu extract or a yulmu extract containing a cosolvent do.

Disclosure of Invention Technical Problem [8] The present invention provides a method for attaining the above-mentioned object, which comprises the step of extracting the extract or the solvent thereof, wherein the extract or the solvent fraction thereof contains a cosolvent.

The yulmu extract may be obtained by extracting Rhizome roots with a C1 to C6 alcohol, or a mixture of C1 to C6 alcohol and water, or a Rhizome root with a C1 to C6 alcohol or a mixture of C1 to C6 alcohol and water Methylene chloride fraction, ethyl acetate fraction and water fraction obtained by successively extracting the above-mentioned solution with hexane, methylene chloride and ethyl acetate to obtain a solution, which is then dissolved in water to obtain an extract, Or a mixture thereof.

At this time, the ethyl acetate fraction can be fractionated into 9 fractions using column chromatography, and the fraction No. 2 can be separated using high performance liquid chromatography (HPLC), and the ethyl acetate fraction can be separated into ethyl acetate and water saturated n- And extracted with butanol.

In addition, the alcohol may be ethanol.

The composition for whitening according to the present invention includes a yulmu extract or a solvent fraction thereof, and the yulmu extract or a solvent fraction thereof has an excellent whitening effect because it contains a cosolvent and can be applied to cosmetics, foods, and pharmaceuticals Effect.

FIG. 1 is a flow chart schematically illustrating the process of isolating the yulmu extract or its solvent fraction of the present invention.
2 is a ¹ H NMR analysis result of the koels obtained from the roots of the present invention.
Fig. 3 shows the ¹³ C NMR analysis results of the koexole obtained from the rhizome root of the present invention.
FIG. 4 is a high-performance liquid chromatography (HPLC) analysis result of a 70% ethanol extract (b) of koelsol (a), yuncheon acid yulmu root and a 70% ethanol extract (c) of Jecheon acid yulmu root.
FIG. 5 is a test result of inhibition of tyrosinase enzyme activity of koi sole according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail. The terms and words used in the present specification and claims should not be construed as limited to ordinary or dictionary terms and the inventor may appropriately define the concept of the term in order to best describe its invention It should be construed as meaning and concept consistent with the technical idea of the present invention.

The term yulmu, which is used in the present invention, should be understood as a term referring to yulmu root unless otherwise specified.

Yulmu is a one-year-old herbaceous plant belonging to the genus Paddy, whose flowers bloom in July, and the fruit is well-known as edible. In one room, it is called "nain" and is also used as medicine. In the present invention, yulmu can be purchased commercially, sold, or harvested or cultivated in nature.

In the present invention, the term "whitening" means a method of increasing the brightness of the skin with reduced brightness due to excessive pigment such as melanin or keeping the brightness of the skin at a certain level, And the like. For the purpose of the present invention, the whitening is carried out by means of inhibiting the synthesis of melanin formed in the skin by the yulmu extract or its fraction, but is not particularly limited thereto.

Melanin is a pigment that is widely distributed in organisms such as skin, hair, and eyes. It is synthesized in the melanosome of the melanocytes of the human epidermal layer. Tyrosinase activates tyrosine Melanin is biosynthesized by oxidation and polymerization reactions, which are converted to DOPA (3,4-dihydroxy-phenylalanine) or DOPA quinone. Biosynthesized melanin increases resistance to skin irritation such as ultraviolet rays, but because excessive melanin synthesis causes pigmentation such as spots, freckles, and black spots, tyrosinase activity measurement is considered as an important indicator of skin whitening effect have.

The whitening composition of the present invention comprises a yulmu extract or a solvent fraction thereof, wherein the yulmu extract or a solvent fraction thereof is characterized by containing a cosolvent.

The yulmu extract or its solvent fraction can be obtained through a process as described in the embodiment of Fig.

That is, in the present invention, the Yulmu extract may be obtained by extracting Yulmu fruit or Yulmu root with a C1-C6 alcohol, or a mixture of C1-C6 alcohol and water, or by extracting Yulmu fruit or Yulmu root with C1-C6 alcohol , Or a mixture of C1 to C6 alcohol and water to obtain an extract, dissolving the extract in water to form a solution, extracting the solution with hexane, methylene chloride, and ethyl acetate, Methylene chloride fraction, ethyl acetate fraction and water fraction, or a mixture thereof.

The alcohol of C1 to C6 as the extraction solvent includes methanol, ethanol, propanol, butanol, pentanol, hexanol and the like, preferably ethanol.

The yulmu extract or its solvent fraction of the present invention is characterized in that it contains a cosolvent. Coixol is a compound whose chemical name is 6-methoxybenzoxazoline-2 (3H) -one, and is a compound having the following structure (1).

The results of ¹ H NMR and ¹³ C NMR analyzes of the koi sol contained in the yuimu extract or the solvent fraction thereof of the present invention are shown in Figs. 2 and 3.

_{^{1 H NMR (700MHz, CD 3}} OD) analysis, δ 6.96 (1H, d, J = 8.4 Hz, H-4), 6.88 (1H, d, J = 2.1 Hz, H-7), 6.74 (1H, dd, J = 2.8, 8.4 Hz , H-5), 3.78 (3H, s, -OCH 3) , and the ^{observed, 13 C-NMR (175MHz,} CD 3 OD) analysis, δ 157.1 (C-2) , observed 157.0 (C-6), 145.5 (C-8), 124.4 (C-9), 110.4 (C-5), 110.1 (C-4), 97.5 (C-7), 55.9 (-OCH 3) . Further, m / z 166 [M + H] ⁺ was observed as a result of mass spectrum (ESIMS) analysis, indicating that the yulmu extract or its solvent fraction of the present invention contained coixol.

The yulmu extract or its fractions of the present invention can be applied to various applications as a whitening composition containing it as an active ingredient.

When applied to a cosmetic composition for whitening, the cosmetic composition may comprise the cosmetic composition as an amount of 0.0001 to 50% by weight based on the total weight of the cosmetic composition, and the balance thereof. At this time, the cosmetic base constituting the cosmetic composition may be a cosmetic base for forming a soft longevity (skin lotion), a nutritional lotion (nutrition lotion), an essence, a cream, a body cream or a pack.

The content of the dry powdery extract in the cosmetic composition is preferably from 0.0001 to 5% by weight, more preferably from 0.001% to 50% by weight in the case of a liquid 1.3-butylene glycol, propylene glycol, dipropylene glycol and glycerin and a mixture thereof May be added to the cosmetic in an amount of 0.01 to 1% by weight for the former, and 0.1 to 10% by weight for the latter. In addition, the composition refers to the addition of the extract to cosmetics, that is, softening longevity (skin), nutritional lotion (lotion), essence, nutrition cream, pack, body lotion, body oil and the like.

In addition, the whitening cosmetic composition of the present invention may be prepared in any formulations conventionally produced in the art, and examples thereof include solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, But are not limited to, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations, and sprays. More specifically, the whitening cosmetic composition of the present invention can be used as a cosmetic composition for softening lotion, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder .

The whitening cosmetic formulation of the present invention may be used as a carrier component in the case of a paste, a cream or a gel in which an animal oil, a vegetable oil, a wax, a paraffin, a starch, a tragacanth, a cellulose derivative, polyethylene glycol, silicone, bentonite, Zinc oxide and the like can be used. The whitening cosmetic formulation of the present invention can be used as a carrier component in the case of a powder or a spray, such as lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder. The whitening cosmetic formulation of the present invention may be a solution or emulsion, in which a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, Glycolate, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan can be used. The whitening cosmetic formulation of the present invention may contain, as a carrier component, water, a liquid diluent such as ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester Suspension, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth can be used. In the whitening cosmetic formulation of the present invention, it is preferable that, in the case of a surfactant-containing cleansing, the cosmetic composition containing as the carrier component an aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, Fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

In addition, the whitening cosmetic composition of the present invention can be used as a cosmetic preparation for skin, which contains an acute or chronic moisturizing agent (for example, a humectant, a sealant, and an agent that affects the natural moisturizing mechanism of the skin), an antioxidant, , Emollients, anti-irritants, vitamins, trace metals, antimicrobial agents, vegetable extracts, fragrances, and / or other beneficial agents and compounds such as dyes and color components.

In addition, the cosmetic composition for whitening of the present invention may contain a surfactant. The surfactant may be a compatible mixture of an anionic surfactant, a cationic surfactant, a nonionic surfactant or a surfactant. The surfactant may also be an ampholytic or amphoteric surfactant having the characteristics of anions or cations depending on the pH of the composition.

In addition, the whitening cosmetic composition of the present invention may also contain a hydrotrope. Hydrotropes are compounds that have the ability to enhance water solubility of other compounds. Specific examples of hydrotropes include, but are not limited to, sodium cumene sulfonate, ammonium cumene sulfonate, ammonium xylene sulfonate, potassium toluene sulfonate, sodium toluene sulfonate, sodium xylene sulfonate, toluene sulfonic acid and xylenesulfonic acid. Other useful hydrotropes include sodium polynaphthalene sulfonate, sodium polystyrene sulfonate, sodium methyl naphthalene sulfonate, sodium camphor sulfonate, and disodium succinate.

In addition, the whitening cosmetic composition of the present invention may contain a thickening agent or a gelling agent. The thickener or gelling agent may be, for example, a polymer or copolymer unsaturated carboxylic acid or unsaturated ester, polysaccharide derivative, gum, colloidal silicate, polyethylene glycol (PEG) and derivatives thereof, polyvinylpyrrolidone and its derivatives, Derivatives, polyacrylonitrile, hydrophilic silica gel or mixtures thereof.

Further, another aspect of the present invention is also applicable to a food composition for whitening, or a fraction thereof.

The whitening food composition may comprise the yulmu extract or a solvent fraction thereof.

The whitening food composition may be in the form of pills, powders, granules, precipitates, tablets, capsules or liquid preparations. Examples of the food to which the composition of the present invention can be added include various foods, for example, Beverages, gum, tea, vitamin complex, and health supplement foods.

In the food composition of the present invention, other ingredients that do not disturb the whitening activity may be added in addition to the yulmu extract or the solvent fraction thereof, and the kind thereof is not particularly limited. For example, various herbal medicine extracts, food-acceptable food-aid additives or natural carbohydrates, such as ordinary foods, may be added as an additional ingredient.

As used herein, the term "food-aid additive " refers to a component that can be added to foods in a supplementary manner, and is appropriately selected and used by those skilled in the art as added to produce health functional foods of each formulation. Examples of food aid additives include various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, colorants and fillers, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, and carbonating agents used in carbonated beverages. However, the types of the food-aid additives of the present invention are not limited by these examples.

Examples of such natural carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be advantageously used as flavors other than those described above .

Another aspect of the present invention provides a health functional food for preventing or alleviating a pigmentation disease comprising the food composition of the above aspect.

The term " health functional food " used in the present invention refers to a food prepared and processed in the form of tablets, capsules, powders, granules, liquids and pills using raw materials and components having useful functions in the human body. Here, the term " functional " means that the structure and function of the human body have a beneficial effect for health use such as controlling nutrients or physiological action. The health functional food of the present invention can be manufactured by a method commonly used in the art and can be prepared by adding raw materials and ingredients which are conventionally added in the art. Also, unlike general medicine, there is an advantage that there is no side effect that can occur when a medicine is used for a long time by using food as a raw material, and it is excellent in portability.

In addition, prevention or alleviation of skin pigmentation disease in the present invention can be achieved not only by preventing the deposition of melanin pigment on the skin by applying the composition containing the yulmu extract or the solvent fraction thereof to hyperpigmented skin, It can mean any action that alleviates melanin that has already been deposited on the skin.

The skin pigmentation disorder is caused by excessive pigmentation after drug use such as freckles, senile spots, liver, stain, brown or black spot, daylight pigment, cyanic melasma, calcium antagonist, sclerotherapy Post-inflammatory hyperpigmentation, phototoxic response, or other similar small fixed pigmented lesions due to scarring or dermatitis due to scarring and scarring, including burns, scabies and burns in women who have taken oral contraceptives But is not limited thereto.

The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment).

In general, in the production of food, the extract of Root extract or its solvent fraction of the present invention is added in an amount of 1 to 10% by weight, preferably 5 to 10% by weight, of the raw material composition. However, in the case of long-term ingestion intended for health and hygiene purposes or for the purpose of controlling health, the above amount can also be used below the above-mentioned range.

There is no particular limitation on the kind of the food. Examples of foods to which the above substances can be added include dairy products including meats, sausages, breads, chocolates, candies, snacks, confections, pizzas, ramen noodles, gums, ice cream, soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy foods in a conventional sense.

Another aspect of the present invention is a pharmaceutical composition for preventing or treating a pigmentation disease containing an extract of Yulmu and a fraction thereof as an active ingredient.

In the present invention, prevention or alleviation of skin pigmentation disease is not limited to prevention of deposition of melanin pigment on the skin by applying a composition comprising a rhodope root extract or a solvent fraction thereof to hyperpigmented skin, May mean all the actions of treating melanin that have already been deposited on.

The skin pigmentation disorder may include hyperpigmentation after drug use such as freckles, senile spots, liver, spots, brown or black spots, daylight pigmentation, blue blotch, calcium antagonist, aftereffects due to tissue hardening therapy, But are not limited to, hyperpigmentation, post-inflammatory hyperpigmentation due to scarring or dermatitis including scabies and burns in women taking birth control pills, phototoxic response or other similar small fixed pigmented lesions.

The pharmaceutical composition of the present invention can be used as a single preparation, and can also be manufactured and used as a composite preparation, further comprising a pharmaceutical composition known to have a recognized whitening effect.

The pharmaceutical composition of the present invention may be formulated into a pharmaceutical unit dosage form by adding a pharmaceutically acceptable carrier, excipient, or diluent.

By "pharmaceutically acceptable" as used herein is meant not significantly irritating the organism and not interfering with the biological activity and properties of the administered active substance.

The compositions of the present invention comprising a pharmaceutically acceptable carrier may be of various oral or parenteral formulations. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may contain one or more excipients such as starch, calcium carbonate, sucrose or lactose lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like are also used. Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances and preservatives may be included have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the non-aqueous solvent and the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.

The pharmaceutical composition may be in the form of tablets, pills, powders, granules, capsules, suspensions, solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, It can have one formulation.

In addition, in the composition of the present invention, the extract or its solvent fraction may be contained in a pharmaceutically effective amount. The term "pharmaceutically effective amount " as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will vary depending on the species and severity, age, sex, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts.

The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And can be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art. Preferably, the extract or the solvent fraction thereof may be contained in the composition in an amount of 0.001 to 200 μg / ml, more preferably 0.001 to 100 μg / ml.

The pharmaceutical composition comprising the extract or the solvent fraction thereof of the present invention can be administered to an individual to prevent or treat skin pigmentation diseases.

The subject is a person who is in need of prevention or treatment of skin pigmentation disease and is a person who needs treatment of skin pigmentation diseases and similar symptoms as well as human beings, horses, sheep, pigs, goats, camels, , ≪ / RTI > and the like.

The term "administering" in the present invention means introducing the composition of the present invention into a subject by any suitable method, and the administration route of the composition of the present invention may be administered through any conventional route so long as it can reach the target tissue have.

The number of times application of the composition of the present invention can be determined depending on the prescription, necessity or desire. The composition of the present invention may be prescribed in parallel with a surgical method such as a laser. It can also be incorporated into casts, bandages, dressings, gauze pads, and similar items.

The present invention includes the administration of a yulmu extract or a solvent fraction thereof in a pharmaceutically effective amount. It will be apparent to those skilled in the art that the appropriate total daily dose may be determined by the practitioner within the scope of sound medical judgment.

In addition, it can be administered once or several times in divided doses. For purposes of the present invention, however, the specific therapeutically effective amount for a particular patient will depend upon the nature and extent of the reaction to be achieved, the particular composition, including whether or not other agents are used, the age, weight, Sex and diet of the patient, the time of administration, the route of administration and the rate of administration of the composition, the duration of the treatment, the drugs used or concurrently used with the specific composition, and similar factors well known in the medical arts.

The composition for whitening according to the present invention may be prepared by obtaining the substance containing the kojols through the yulmu extract or the solvent fraction thereof. However, in order to increase the content of the kojols, as shown in Fig. 1, Ethyl acetate fractions can be fractionated into 9 fractions using column chromatography and fraction 2 fractions can be separated using high performance liquid chromatography (HPLC).

At this time, the ethyl acetate fraction can be extracted with ethyl acetate and n-butanol saturated with water to selectively separate the coixol.

In one embodiment, the process of preparing the ethanol extract of Solanum tuberosum and its solvent fraction and separating and purifying the effective ingredient, coixol, is as follows.

The extraction and fractionation process is as follows.

The dried yulmu root (1.9 kg) was cooled with ethanol twice at room temperature, and the extract was concentrated under reduced pressure to obtain 333.34 g of an ethanol extract. (3.9 L × 3 times), methylene chloride (3.9 L × 3 times), ethyl acetate (3.9 L × 3 times) and water saturated n-butanol (3.9 L × 3 times) in distilled water ) Was used to make a solvent fraction. The water saturated n-butanol corresponds to the water used to separate the ethyl acetate and water layers in FIG.

From the ethyl acetate fractions, the coixol compound was isolated as follows.

Hx fr. (14.3 g), CH ₂ Cl ₂ fr. (5.1 g), EtOAc fr. (5.4 g) and BuOH fr. (29 g) were subjected to column chromatography (solvent gradient). EtOAc fr. Was subjected to column chromatography using C18 gel and divided into 9 small fractions. Small fractions 1 to 9 obtained through C-18 column chromatography (gradient: H ₂ O? MeOH) on the EtOAc fraction were subjected to Prep HPLC to obtain two fractions to obtain a cosolvent (4.8 mg).

In order to confirm the amount contained in the ethanol extract of Clove Sol, ethanol extracts were prepared by using the same method as above for ethanol extracts of Yulchunsan yulmu root and Jecheon acid. HPLC analysis was carried out using HPLC Aglient 1200 series. The analytical column was VisionHT C18 HL-5 ㎛-C18 (4.6 x 250 mm) and analyzed under conditions of flow rate of 1 ml / min and injection volume of 20.. An ultraviolet absorption spectrophotometer (measuring wavelength 254 nm) was used as the detector.

The results of HPLC analysis are shown in Fig. Referring to FIG. 4, a specific peak originating from a Koysol sol was observed at 19.5 minutes in the ethanol extract of Rhodiola root.

In order to confirm the whitening effect of the cosmetic composition obtained, the whitening effect of the cosmetic composition was measured by measuring the in vitro tyrosinase inhibitory activity.

The mushroom tyrosinase inhibition assay was used to confirm the inhibitory effect of coixol on tyrosinase enzyme activity. 10 μl of the sample and 20 μl of the solution were added to 170 μl of a reaction mixture containing 1 mM L-tyrosine, 50 mM potassium phosphate buffer (pH 6.5), and distilled water at a ratio of 10: 10: 9 Mushroom tyrosinase aqueous solution (1000 units / ml) was added and the reaction solution was incubated at 25 ° C for 30 minutes. Then, the amount of dopachrome generated at 490 nm was measured by absorbance using a microplate reader. As a positive control, kojic acid, which is known as inhibitor of tyrosinase enzyme activity, was used.

The percentage formula of inhibition of tyrosinase enzyme activity was as follows.

 (A: sample absorbance, B: color control absorbance, C: control absorbance)

Table 1 shows the inhibitory activity of tyrosinase enzyme activity of Coixol-containing yuimu ethanol extract and its solvent fraction calculated from the above formula.

sample IC50 ([mu] M) Coax 62.4 Arbutin 618.1 Kojic acid 41.4

As shown in Table 1, the IC50 value was 62.4 μM in the case of coixol, and was similar to that of kojic acid used as a positive control. The enzyme activity was 10 times higher than that of arbutin (618.1 μM).

These results are a result of a significant improvement in comparison with Table 1 of the prior art Korean Patent Publication No. 10-1579500. In the present invention, it is possible to effectively inhibit the expression of tyrosinase by containing a large amount of coixol in the extract and its solvent fraction, and as a result, the melanin synthesis is inhibited and thereby the whitening effect as well as the diseases caused by melanin deposition can be treated .

In addition, when the results of FIG. 5 showing the inhibitory activity of tyrosinase were examined, it can be seen that the Kokes sole of the present invention still shows a significantly higher activity than kojic acid or arbutin even when the concentration of the Kokes sole is lowered to 100 μM from the stock solution.

Therefore, the yulmu extract or the solvent fraction thereof according to the present invention can be used as a whitening composition having an excellent whitening effect.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, It is possible to carry out various changes in the present invention.

Claims (6)

A composition for whitening, comprising a yulmu extract or a solvent fraction thereof, wherein the yulmu extract or a solvent fraction thereof contains a cosolvent.
The method according to claim 1,
Wherein the yulmu extract is obtained by extracting yulmu root with a C1 to C6 alcohol or a mixture of C1 to C6 alcohol and water.
The method according to claim 1,
The extract of Yulmu is extracted with a mixture of C1-C6 alcohol or C1-C6 alcohol and water to obtain an extract, which is then dissolved in water to form a dissolution liquid. The dissolution liquid is dissolved in hexane, methylene chloride , Ethyl acetate, hexane fraction, methylene chloride fraction, ethyl acetate fraction and water fraction, or a mixture thereof.
The method of claim 3,
Wherein the ethyl acetate fraction is fractionated into 9 fractions by column chromatography and fraction 2 is separated by high performance liquid chromatography (HPLC).
The method of claim 3,
Wherein said ethyl acetate fraction is obtained by extraction with ethyl acetate and n-butanol saturated with water.
The method according to claim 2 or 3,
Wherein the alcohol is ethanol.

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