KR102107490B1 - Cosmetic composition containing Narcissus extract - Google Patents

Abstract

The present invention relates to a cosmetic composition, and more particularly, to a cosmetic composition having an anti-inflammatory, antioxidant and whitening effect by containing a daffodil extract.

Description

Cosmetic composition containing narcissus extract {Cosmetic composition containing Narcissus extract}

The present invention relates to a cosmetic composition, and more particularly, to a cosmetic composition having an anti-inflammatory, antioxidant and whitening effect by containing a daffodil extract.

As environmental pollution such as ozone and ultra-fine dust increases rapidly, free radicals, which are the main cause of free radicals, are concerned about skin damage and skin aging. Accordingly, in order to prepare for the threatening increase of substances that promote serious environmental pollution and skin aging, cosmetics having the function of preventing skin aging are in the spotlight, and as shown in the following patent documents, skin inflammation is relieved, antioxidant, whitening, and wrinkles Functional cosmetics are widely developed to provide an improvement effect.

<Patent Document>

Patent Document 1: Japanese Patent No. JP10-36283 (Registered on 08/06/2007) "Fibroblast Proliferation Accelerator"

Patent Document 2: Patent Publication No. 10-1077824 (announced on Oct. 28, 2011) "Cosmetic composition for improving atopy containing illite extract and method for manufacturing the same"

However, the conventional cosmetic composition has a problem in that inflammation relief and skin whitening effects are not satisfactory, and stability is not guaranteed by using minerals, retinoic acid, and the like.

The present invention is to solve the problems of the prior art described above,

An object of the present invention is to provide a cosmetic composition having an anti-inflammatory, antioxidant and whitening effect by containing a narcissus extract.

The present invention is implemented by an embodiment having the following configuration in order to achieve the above object.

According to an embodiment of the present invention, the cosmetic composition according to the present invention is characterized by containing a daffodil extract.

According to another embodiment of the present invention, the cosmetic composition according to the present invention is characterized by having anti-inflammatory, antioxidant and whitening effects.

According to another embodiment of the present invention, in the cosmetic composition according to the present invention, the narcissus extract is characterized in that it is extracted from flowers, stems and roots of narcissus.

According to another embodiment of the present invention, in the cosmetic composition according to the present invention, the narcissus extract is characterized in that 0.001 to 30% by weight based on the total weight of the cosmetic composition is used.

According to another embodiment of the present invention, in the cosmetic composition according to the present invention, the narcissus extract is extracted with hot water by adding water to flowers, stems and roots of narcissus, followed by lyophilization to obtain a powder, and ethanol is added to the powder. It is characterized by being obtained by filtration.

The present invention can obtain the following effects by the present embodiment.

The present invention has the effect of having an anti-inflammatory, antioxidant and whitening effect by containing the daffodil extract.

1 to 3 is a graph showing the evaluation of NO production inhibitory efficacy of the cosmetic composition according to an embodiment of the present invention.
4 to 12 is a graph showing the cytokine production inhibitory efficacy evaluation of the cosmetic composition according to an embodiment of the present invention.
13 to 15 is a graph showing the antioxidant efficacy evaluation of the cosmetic composition according to an embodiment of the present invention.
16 to 18 is a graph showing the activity inhibitory efficacy of mushroom tyrosinase activity of the cosmetic composition according to an embodiment of the present invention.
19 to 21 is a graph showing the cytotoxic tyrosinase activity inhibitory efficacy evaluation of the cosmetic composition according to an embodiment of the present invention.
22 to 24 are graphs showing the evaluation of melanin production inhibition efficacy of the cosmetic composition according to an embodiment of the present invention.

Hereinafter, a cosmetic composition containing a daffodil extract according to the present invention will be described in detail with reference to the accompanying drawings. Unless otherwise specified, all terms in this specification are the same as the general meaning of the term understood by a person skilled in the art to which the present invention pertains, and if there is a conflict with the meaning of the term used herein It follows the definition used in the specification. In addition, detailed descriptions of well-known functions and configurations that may unnecessarily obscure the subject matter of the present invention are omitted. Throughout the specification, when a part “includes” a certain component, it means that other components may be further included rather than excluding other components unless specifically stated otherwise.

The cosmetic composition according to an embodiment of the present invention contains a daffodil extract. The narcissus extract provides anti-inflammatory, antioxidant and whitening effects, the narcissus extract is extracted from flowers, stems and roots of narcissus, and the narcissus extract is used in an amount of 0.001 to 30% by weight based on the total weight of the cosmetic composition. The daffodil extract is obtained by adding water to flowers, stems and roots of daffodils and extracting it by hot water, followed by lyophilization to obtain a powder, and adding ethanol to the powder and filtering it. In particular, the extract obtained from the root of narcissus provides an anti-inflammatory effect, and the extract obtained from a flower of narcissus provides an antioxidant and whitening effect. The daffodils are called freshness of water, and they store a lot of water in the Alpuri, an excellent water reservoir, bloom beautiful flowers in dry land and contain Lycorin and Glukomannan on scales.

The cosmetic composition of the present invention may further include a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingo lipids, and seaweed extract.

The water-soluble vitamin may be any compound that can be blended into a cosmetic, but preferably vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinamide, folic acid, vitamin C, vitamin H, etc. Water-soluble vitamins that can be used in the present invention, such as salts (thiamine hydrochloride, sodium ascorbate salt) or derivatives (ascorbic acid-2-sodium phosphate salt, ascorbic acid-2-magnesium phosphate salt) Is included in. The water-soluble vitamin can be obtained by a conventional method such as a microorganism conversion method, a purification method from a culture of microorganisms, an enzyme method, or a chemical synthesis method.

The oil-soluble vitamin may be any compound that can be blended into a cosmetic, but preferably vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol), and the like. And their derivatives (ascorbyl palmitate, ascorbyl stearate, ascorbine dipalmitate, dl-alpha tocopherol acetate, dl-alpha tocopherol acetate nicotine E, DL-pantothenyl alcohol, D-pantothenyl alcohol, pantothenyl Ethyl ether, etc.) are also included in the useful vitamin used in the present invention. The oil-soluble vitamin can be obtained by a conventional method such as a microorganism conversion method, a purification method from a culture of microorganisms, an enzyme or a chemical synthesis method.

The polymer peptide may be any compound as long as it can be blended into a cosmetic, preferably collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, keratin, and the like. The polymer peptide may be purified by a conventional method such as a purification method from a culture medium of microorganisms, an enzyme method, or a chemical synthesis method, or may be purified and used from natural products such as dermis such as pig or cow, silkworm silkworm, and the like. .

The polymer polysaccharide may be any compound as long as it can be blended into a cosmetic. Preferably, hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate, or a salt thereof (sodium salt, etc.) may be mentioned. For example, chondroitin sulfate or a salt thereof can be used after being purified from mammals or fish.

The sphingo lipid may be any compound as long as it can be blended into a cosmetic, and preferably, ceramide, phytosphingosine, sphingoglycolipid, and the like. The sphingoli lipid can be purified by conventional methods from mammals, fish, shellfish, yeast or plants, or can be obtained by chemical synthesis.

The seaweed extract may be anything as long as it can be blended into a cosmetic, preferably, brown seaweed extract, red seaweed extract, green algae extract, and the like. In addition, these seaweed extracts are purified from these seaweed extracts, colorazine, arginic acid, and sodium arginate. , Potassium arginate and the like are also included in the seaweed extract used in the present invention. The seaweed extract can be obtained by purification from seaweed by a conventional method.

The cosmetic composition of the present invention can be blended with other ingredients that are usually blended into the cosmetic composition. For example, fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavors, blood circulation promoters, And cooling agents, limiting agents, and purified water.

Examples of the fats and oils include ester fats and fats, hydrocarbon fats and fats, silicone fats and fats, fluorine fats, animal fats and vegetable fats. Examples of the ester-based fats and oils are tri2-ethylhexanoate glyceryl, 2-ethylhexanoate cetyl, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, and stearic acid. Butyl acid, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, diadi Diisopropyl finate, isoalkyl neopentanoate, tri (capryl, capric acid) glyceryl, trimethylethylpropane tri2-ethylhexanoate, trimethylolpropane triisostearate, pentaelisley tritetraethylethylhexanoate Tol, cetyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, cetyl myristate, stearyl stearate, decyl oleate, ricinolein Cetyl acid, isostearyl laurate, isotridecyl myristate, isocetyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyldodecyl oleate, octyldodecyl oleate, isopropyl isostearate , Sethostearyl 2-ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, propylene glycol di (capryl, capric acid) , Propylene glycol dicaprylate, neopentyl glycol dicapric acid, neopentyl glycol dioctanoate, glyceryl tricaprylic acid, glyceryl triundecylate, glyceryl triisopalmitate, glyceryl triisostearate, octyl neopentanoate Dodecyl, isostearyl octanoate, octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostear isostearate Reel, octyldecyl isostearate, polyglycerin oleic acid ester, polyglycerin isostearic acid ester, triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, Triethyl citrate, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearic acid, di2-ethylhexyl succinate, diisobutyl succinate, diiso sebacinate Propyl, dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phytsteryl isostearate, phytate oleate Reel, 12-stealoylhydroxystearate isocetyl, 12-stealoylhydroxystearate stearyl, 12-s Allo one hydroxy stearic acid and the like esters such as cetearyl source. Examples of the hydrocarbon-based fats and oils include hydrocarbon-based fats and oils such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybutene, microcrystalline wax, and petrolatum. Examples of the silicone oils and fats include polymethylsilicone, methylphenylsilicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane · methylcetyloxysiloxane copolymer, dimethylsiloxane · methylstearoxylsiloxane copolymer, And alkyl-modified silicone oils, amino-modified silicone oils, and the like. Examples of the fluorine-based fats and oils include perfluoropolyethers. Examples of the animal or plant oil include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, new flower oil, soybean oil, corn oil, rapeseed oil, almond oil, palm kernel oil, palm oil, castor oil, sunflower oil, grapes Seed Oil, Cottonseed Oil, Palm Oil, Cucumin Nut Oil, Wheat Germ Oil, Rice Germ Oil, Shea Butter, Moon Seed Colostrum, Marker Demi Nut Oil, Meadow Home Oil, Egg Yolk Oil, Uji, Horse Oil, Mink Oil, Orange Rape Oil, Jojoba And animal or vegetable oils such as milk, candelary wax, carnauba wax, liquid lanolin, and hardened castor oil.

Examples of the moisturizing agent include water-soluble low molecular moisturizing agents, oil-soluble molecular moisturizing agents, water-soluble polymers, and fat-soluble polymers. Examples of the water-soluble low molecular humectant include serine, glutamine, sorbitol, mannitol, pyrrolidone-sodium carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (polymerization degree n = 2 or more), poly And propylene glycol (polymerization degree n = 2 or more), polyglycerin B (polymerization degree n = 2 or more), lactic acid, lactic acid salt, and the like. Examples of the fat-soluble low-molecular humectant include cholesterol and cholesterol esters. Examples of the water-soluble polymer include carboxyvinyl polymer, polyasparaginate, tragacanth, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitin, chitosan, dextrin, etc. Can be lifted. Examples of the fat-soluble polymer include polyvinylpyrrolidone-eicosen copolymer, polyvinylpyrrolidone-hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and polymer silicone.

Examples of the emollient agent include long-chain acyl glutamate cholesteryl ester, hydroxystearate cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid, and lanolin fatty acid cholesteryl ester.

Examples of the surfactant include nonionic surfactants, anionic surfactants, cationic surfactants, and amphoteric surfactants. As the nonionic surfactant, self-emulsifying type glycerin monostearate, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene), sorbitan fatty acid ester, POE sorbitic fatty acid ester , POE glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE cured castor oil, POE castor oil, POEPOP (polyoxyethylene polyoxypropylene) copolymer, POEPOP alkyl ether, polyether modified silicone, lauric acid alkanolamide , Alkylamine oxides, hydrogenated soybean phospholipids, and the like. Examples of the anionic surfactant include fatty acid soap, alpha-acyl sulfonate, alkyl sulfonate, alkyl allyl sulfonate, alkyl naphthalene sulfonate, alkyl sulfate, POE alkyl ether sulfate, alkylamide sulfate, alkyl phosphate, POE alkyl phosphate, alkyl Amide phosphate, alkyloyl alkyl taurine salt, N-acyl amino acid salt, POE alkyl ether carboxylate, alkyl sulfosuccinate, sodium alkyl sulfoacetate, acylated hydrolyzed collagen peptide salt, perfluoroalkyl phosphate ester, etc. Can. Examples of the cationic surfactant include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, And benzalkonium chloride, diethylaminoethyl stearate, dimethylaminopropylamide stearate, and quaternary ammonium salts of lanolin derivatives. Examples of the amphoteric surfactant include carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amidesulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, amideamine type, etc. And amphoteric surfactants.

Examples of the organic and inorganic pigments include silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, bengala, clay, bentonite, titanium film mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, and oxidation. Inorganic pigments such as aluminum, calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine, chromium oxide, chromium hydroxide, calamine, and complexes thereof; Polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluorine resin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene ㆍ styrene copolymer, And organic pigments such as silk powder, cellulose, CI pigment yellow, and CI pigment orange, and composite pigments of these inorganic and organic pigments.

As said organic powder, Metal soap, such as calcium stearate; Alkyl phosphate metal salts such as sodium cetyl acid zinc, lauryl acid zinc, and calcium lauryl acid; Acylamino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc, and N-lauroylglycine calcium; Amide sulfonic acid polyvalent metal salts such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; N such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylazine, N-alpha-paritoylolnitine, N-alpha-lauroylarginine, N-alpha-cured Uji fatty acid acyl arginine -Acyl basic amino acids; N-acylpolypeptides, such as N-lauroylglycylglycine; Alpha-amino fatty acids such as alpha-amino caprylic acid and alpha-amino lauric acid; And polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene-styrene copolymer, and ethylene tetrafluoride.

Examples of the ultraviolet absorber include paraaminobenzoic acid, ethyl paraaminobenzoate, amyl paraaminobenzoate, octyl paraaminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomentyl salicylate, cinnamic acid Benzyl, paramethoxy cinnamic acid-2-ethoxyethyl, paramethoxy cinnamic acid octyl, diparamethoxy cinnamic acid mono-2-ethylhexaneglyceryl, paramethoxy cinnamic acid isopropyl, diisopropyl-diisopropyl cinnamic acid ester mixture, Urocanoic acid, ethyl urocanoate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenonesulfonic acid and salts thereof, dihydroxymethoxybenzophenone, dihydroxymethoxybenzophenone disulfonate sodium, dihydroxybenzo Phenone, tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino-p- (carbo-2'-ethylhexyl-1'-oxy)- 1,3,5-triazine, 2- (2- And the like can be de-hydroxy-5-methylphenyl) benzotriazole.

Examples of the disinfecting agent include hinokitiol, triclosan, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, zincfilionone, benzalkonium chloride, Photoresist 301, mononitrowire sodium, undeciric acid, and the like.

Examples of the antioxidants include butyl hydroxyanisole, propyl gallic acid, and elisoric acid.

Examples of the pH adjusting agent include citric acid, sodium citrate, malic acid, sodium malate, phthalic acid, sodium phthalate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogen phosphate, and the like.

Examples of the alcohol include higher alcohols such as cetyl alcohol.

In addition, the compounding component that may be added in addition is not limited to this, and any of the above components can be blended within a range not impairing the objects and effects of the present invention, but preferably 0.01 to 5% by weight based on the total weight, More preferably, it is blended at 0.01 to 3% by weight.

The cosmetic composition of the present invention may take the form of solutions, emulsions, viscous mixtures, and the like.

The components included in the cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the above-mentioned compounds as active ingredients, for example, conventional adjuvants such as stabilizers, solubilizers, vitamins, pigments, and fragrances. And carriers.

The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, milk, cream, foundation, lotion, cosmetic liquid, hair cosmetic, and the like. Specifically, the cosmetic composition of the present invention is a skin lotion, skin softener, moisture lotion, nutrition lotion, massage cream, nutrition cream, hand cream, moisture cream, essence, pack, soap, lotion, milk lotion, gel, ointment, patch, Contains cleansing foam, body cleanser, astringent, and spray formulation. When the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. You can. When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder can be used as a carrier component, and in the case of a spray, additionally chlorofluorohydrocarbon, propane / Propellant such as butane or dimethyl ether. When the formulation of the present invention is a solution or emulsion, a solvent, solvating 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 ester, polyethylene glycol or fatty acid ester of sorbitan. When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, ethoxylated isostearyl alcohol, suspensions such as polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or trakant, etc. can be used. When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide as a carrier component Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linoline derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

Hereinafter, the present invention will be described in more detail through examples. However, these are only for explaining the present invention in more detail, and the scope of the present invention is not limited thereto.

<Example 1> Preparation of sample

(1) Cut daffodil flowers, stems, and roots into 200 g of filter cloth, add 1 L of water to extract hot water at 90 ℃, filter the extract using filter paper (0.45 μm), concentrate with a rotary concentrator, and freeze-dry To obtain a powder. After dissolving 1 g of 80% ethanol in 50 g of the powder, it was re-filtered with filter paper (0.45 μm) to prepare 80% ethanol extract having each concentration of 16 mg / mL. Each ethanol extract (16mg / mL) was vacuum concentrated at 100rpm in a 60 ℃ water bath of a rotary evaporator (HS-2005V, HAHNSHIN S & T CO., LTD) by 200mL. After concentration, each residue (80mg / mL) was filtered through a CA syringe filter (pore size: 0.45um) and stored at 4 ° C.

(2) The residue was fractionated using Prep-LC. As the Prep-LC, 1220 Infinity II LC of Agilent Technologies (Agilent, Santa Clara, CA, USA) was used, and each residue (80 mg / mL) was diluted with methanol to prepare a final concentration of 30 mg / mL. A YMC-Pack ODS-A column (250 mm × 10.0 mm id, 5 μm, 12 nm column) was used, and separated at a flow rate of 2 mL / min with two mobile phases consisting of solvent (A) distilled water and (B) acetonitrile. (0-60 min: 0: 100, 60-70 min: 100: 0). The eluate was measured at 230 nm with a VWD detector at a flow rate of 2 mL / min for 90 minutes, and the temperature of the column was maintained at 30 ° C. Through the prep-LC, 7 peaks from narcissus flower extracts, 3 from narcissus stem extracts, and 6 peaks from narcissus root extracts were obtained, and a fraction having a fourth peak of narcissus flower extract (sample 1) and narcissus The fraction having the third peak of the stem extract (sample 2) and the fraction having the third peak of the daffodil root extract (sample 3) were used in the following experiment.

<Example 2> Anti-inflammatory efficacy evaluation

1. NO (nitroc acid) measurement evaluation

(1) RAW 264.7 cells subcultured in DMEM medium were inoculated at 1 × 10 ⁶ cells / well in a 6 well plate and cultured in an incubator (5% CO _2, 37 ° C.) for 24 hours. Subsequently, samples 1 to 3 were treated for each concentration, reacted for 1 hour, and treated with LPS (final concentration 2 μg / mL) in an experimental group other than the negative control group to induce an inflammatory reaction, followed by incubation for 24 hours under the same conditions as in the previous culture. Did. The amount of NO generated was measured in the form of NO ₂ ⁻ present in the cell culture using a Griess Reagent System. 50 μL of the cell culture supernatant was collected, 50 μL of sulfanilamide solution and NED (N-1-naphthylethylenediamine) solution were sequentially added, and each was reacted in the dark for 5 minutes, and absorbance at 550 nm was measured using an ELISA reader. The amount of NO generated is compared with sodium nitrite (NaNO ₂ ) as a standard, and the results are shown in FIGS. 1 to 3.

(2) As can be seen in FIGS. 1 to 3, the production of NO was significantly increased in Raw264.7 macrophage in which inflammation was induced by LPS. In the experimental group treated with Sample 1 and Sample 3, the concentration of NO inhibition was about 60% and 57%, respectively, at a concentration of 5 μg / mL or more, and in the experimental group treated with Sample 2, at the concentration of 2 μg / mL compared to the control group. At a concentration of about 42% and 5 μg / mL, it showed NO inhibitory effect of about 66%. Therefore, it was confirmed that when the inflammation was induced, each extract of daffodils reduced the production of NO in a concentration-dependent manner.

2. Evaluation of TNF-α, IL-1β and IL-6

(1) RAW 264.7 cells passaged in DMEM medium were inoculated at 1 × 10 ⁶ cells / well in a 6 well plate and cultured in an incubator (5% CO _2, 37 ° C.) for 24 hours. After incubation, each sample 1 to 3 was treated by concentration, and after reacting for 1 hour, in order to induce an inflammatory reaction, LPS (final concentration 2 μg / mL) was treated in the experimental group except for the negative control group and cultured for 24 hours under the same conditions as in the previous culture. Did. After incubation, the TNF-α, IL-1β and IL-6 contents as the supernatant of the medium were measured using ELISA KIT. When using ELISA KIT in each experiment, after washing the plate coated with the antibody of each cytokine (TNF-a, IL-6 and IL-b), react with 100 μL of medium per well. Then, diluted detection antibody (0.25 μg / mL) and color development (1:40 dilution for TNF-a, 1:20 dilution for IL-6 and IL-b) and color development soultion for each well 100 μL Each was added sequentially and reacted for a certain period of time for each step, and the absorbance was measured at 450 nm with an ELISA reader, and the results are shown in FIGS. 4 to 12.

(2) As can be seen in FIGS. 4 to 12, the production of TNF-α, IL-1β and IL-6 was increased in the LPS-induced inflammation of the Raw 264.7 macrophage compared to the control group, and each sample was concentration-dependent. The production of inflammatory cytokines TNF-α, IL-6 and IL-1β was reduced. In the experimental group treated with Sample 1, TNF-α was about 70%, IL-1β was about 23%, and IL-6 was about 45% at a concentration of 20 μg / mL or more, which inhibited the production of each cytokine. In the experimental group treated with sample 2, TNF-α was about 79%, IL-1β was about 29%, and IL-6 was about 38% at a concentration of 5 μg / mL or more, which inhibited the production of each cytokine. In the experimental group treated with Sample 3, TNF-α was about 86%, IL-1β was about 43%, and IL-6 was about 52% at a concentration of 40 μg / mL or more, which inhibited the production of each cytokine. Therefore, it was observed that each extract of daffodils reduces the production of inflammatory cytokines TNF-α, IL-6 and IL-1β when concentration is induced.

<Example 3> antioxidant effect evaluation

(1) The free radical scavenging activity was measured by using 1,1-diphenyl-2-pycrylhydrazyl (DPPH) as a measure of the antioxidant effect. That is, 50 μL of each sample diluted by concentration (0, 100, 200, 400, 800 μg / mL) was dissolved in DMSO in 950 mL of 0.2 mM DPPH solution dissolved in ethanol, and then incubated at room temperature for 30 minutes, followed by 525 nm. The absorbance was measured at, and the results are shown in FIGS. 14 to 16.

(2) As can be seen in FIGS. 14 to 16, as a result of measuring the radical scavenging capacity of the sample, DPPH radical scavenging capacity was increased depending on the concentration, and about 93% and about 91% at a concentration of 800 μg / mL of samples 1 to 3 , It was observed that it exhibited a radical scavenging activity of about 28%.

<Example 4> Whitening effect evaluation

1. Measurement of inhibition of mushroom tyrosinase activity

(1) Samples 1 to 3 were diluted with 0.1M phosphate buffer (pH 6.5) by concentration (0, 250, 500, 1000, 2000, 4000, 8000 μg / mL), and 130 μL was added to a 96 well plate. , Mushroom tyrosinase solution (1250 U / mL) 20 μL was added to each well and reacted at 37 ° C. for 5 minutes. 120 μL of 1.5 mML-DOPA solution was added to the solution, and the mixture was reacted at 37 ° C. for 5 minutes, and absorbance was measured at 490 nm using an ELISA reader. Arbutin (10 mM), a tyrosinase inhibitor, was used as a positive control. The measurement results are shown in FIGS. 16 to 18.

(2) As can be seen in Figures 16 to 18, as a result of observing the tyrosinase inhibitory efficacy of samples 1 to 3, it was confirmed that the tyrosinase activity is decreased in a concentration-dependent manner of each sample. Each daffodil extract inhibits Mushroom Tyrosinase activity as the Mushroom Tyrosinase activity decreases to approximately 36%, Sample 2 to approximately 29%, and Sample 3 to approximately 17% at a concentration of 8000 μg / mL of each sample. It can be seen that it has a whitening effect through.

2. Measurement of cytotoxic tyrosinase activity inhibitory effect

(1) To measure the activity of tyrosinase in B16-F10 melanoma cells, B16-F10 cells were dispensed into 6 well plates at 1 × 10 ⁶ cells / well, and then cells were well in 5% CO₂, 37 ℃ incubator. The cells were cultured for 24 hours until they were attached to the bottom of about 80% or more. After incubation, the medium was removed, and each sample was replaced with a medium diluted by various concentrations, followed by incubation for 1 hour in 5% CO2, 37 ° C incubator, treatment with 20 nM α-MSH, and incubation for another 48 hours. The concentration range of the sample was determined through a toxicity test using a crystal violet assay. The cells from which the medium was removed were washed twice with PBS (phosphated buffer saline), and 500 μL of 0.1 M phosphate buffer solution containing 1% Triton X-100 and 2 mM phenylmethylsulfonyl fluoride (PMSF, sigma, USA) was added thereto. The solution was stored at -80 ° C for 30 minutes, dissolved at room temperature, transferred to a 1.5 mL tube, and centrifuged for 30 minutes at 15,000 rpm and 4 ° C. After adding 150 μL of 2 mM L-DOPA to 150 μL of the supernatant and reacting at 37 ° C. for 10 minutes, absorbance was measured at 490 nm using an ELISA reader. The measurement results are shown in FIGS. 19 to 21.

(2) As can be seen in FIGS. 19 to 21, the tyrosinase activity of the cells was increased by α-MSH, but it was confirmed that the activity of tyrosinase was decreased in a concentration-dependent manner in the experimental groups treated with samples 1 to 3. Could.

3. Measurement of melanin inhibition in cells

(1) In order to measure melanin production in B16-F10 melanoma cells, B16-F10 cells were dispensed into 6 well plates at 1 × 10 ⁶ cells / well, and then 5% CO₂, cells at 37 ℃ incubator were over 80%. Incubation was carried out for 24 hours until adhesion. After incubation, the medium was removed, and each sample was replaced with a medium diluted for various concentrations, and incubated for 1 hour in a 5% CO₂, 37 ℃ incubator. At this time, the concentration range of the sample was determined through a toxicity test using a crystal violet assay. After stimulation of the medium, to stimulate melanocytes, 20 nM α-MSH was treated in the experimental group excluding the negative control group and cultured again for 48 hours. After 48 hours, the medium was removed, washed with PBS (phosphated buffer saline), and then treated with trypsin to recover the cells. The recovered cells were centrifuged at 1200 x g for 5 minutes and dried at 60 DEG C to obtain pellets. Thereafter, 500 μL of 1 M sodium hydroxide solution containing 10% DMSO was added to obtain intracellular melanin in an 80 ° C. bath, and the melanin content was obtained by measuring absorbance at 490 nm with a microplate reader. The measurement results are shown in FIGS. 22 to 24.

(2) As can be seen in FIGS. 22 to 24, it was confirmed that the synthesis of melanin was inhibited in a concentration-dependent manner in the experimental group treated with the sample.

In the above, the applicant has described the preferred embodiments of the present invention, but these embodiments are only one embodiment for implementing the technical concept of the present invention and any modification or modification of the present invention as long as the technical concept of the present invention is implemented It should be construed as being within the scope.

Claims (5)

Contains narcissus extract, has antioxidant and whitening effects,
The daffodil extract is extracted from flowers, 0.001 to 30% by weight based on the total weight of the cosmetic composition is used,
The daffodil extract is 200 g parts by weight of finely chopped daffodil flowers into a filter cloth, 1000 parts by weight of water is added, hot water is extracted at 90 ° C to obtain an extract, filtered using 0.45 μm filter paper, concentrated with a rotary concentrator and frozen After drying, to obtain a powder, and after dissolving by adding 1000 parts by weight of 80% ethanol per 50 parts by weight of the powder, re-filtering with 0.45 μm filter paper to prepare an 80% ethanol extract having a concentration of 16 mg / mL, and then rotating the ethanol extract by 200 mL Vacuum concentrated at 100 rpm in a 60 ° C. water tank of a vacuum concentrator to obtain a residue with a concentration of 80 mg / mL, filtered with a CA syringe filter, and the filtered residue obtained by fractionation using Prep-LC Composition. delete delete delete delete

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