KR101289813B1 - Composition for anti-wrinkle comprising an extract of Sophora japonica L., Leguminosae - Google Patents

Abstract

The present invention relates to a composition for skin whitening comprising a painting flower extract or a fraction thereof as an active ingredient. Painting tree flower extract or fractions thereof according to the present invention can be usefully used as a composition for skin whitening because it exhibits a whitening effect by inhibiting the activity of tyrosinase and effectively inhibiting the production of melanin without causing toxicity to cells. . In addition, the painting flower extract or fractions thereof according to the present invention is useful as a composition for inhibiting skin aging and improving skin wrinkles because it inhibits superoxide production and has excellent free radical scavenging effect, and has an antioxidant effect and shows skin wrinkle improvement effect. Can be used.

Description

Composition for improving skin wrinkles comprising aspen active flower extract {Composition for anti-wrinkle comprising an extract of Sophora japonica L., Leguminosae}

The present invention relates to a composition for skin whitening comprising a painting flower extract as an active ingredient. The present invention also relates to a composition for inhibiting skin aging or for improving skin wrinkles, which comprises a painting flower extract as an active ingredient.

Skin is an important tissue that protects the human body while directly contacting the external environment and has biochemical and physical functions. The skin is divided into three types: epidermis, dermis, and hypodermis.

Whitening agent is a product developed to alleviate blemishes and freckles caused by UV rays, and to suppress the production of melanin pigments. It is divided into inhibition and peeling promotion.

The pigments that affect skin color include melanin, carotene, and hemoglobin, the most important of which is melanin. The production of melanin depends on race, region, sex, and age. It may cause parts such as tanning, or overall pigmentation, as well as acne and scars.

The biosynthesis of melanin consists of the conversion of tyrosine, a type of amino acid, from melanocyte melanosome to tyrosinase and conversion to dihydroxy phenylalanine. Thereafter, a series of enzymatic and non-enzymatic oxidation processes form a brown (pheomelanin) and black (eumelanin) polymer. This biosynthesis process is carried out in the organelles of a special type of brown cells called melanosomes. Melanosomes containing melanin granules move from the periphery of the nucleus to the end of dendritic projections and into the cytoplasm by the phagocytosis of keratinocytes. Accumulate around the nucleus of the latinosite.

The synthesis of melanin, the number of melanosomes, and the migration to surrounding keratinocytes are influenced by hormones and ultraviolet rays as well as genetic factors. In addition, cytokines, metal ions such as copper, zinc and iron, interferon, prostaglandin and histamine, which are involved in the expression of tyrosinase and synthesis and transfer of melanin, are known to be involved. The most important role in the production of melanin is tyrosinase, which inhibits the activity of the enzyme and can be expected to whiten the skin.

Currently, the development of whitening agents has been largely made in three directions: inhibiting melanin production in melanocytes, controlling melanocyte stimulants, and promoting melanin excretion. In particular, in order to inhibit melanogenesis, inactivation of enzymatic activity to control Cu, an essential component of the active site of tyrosinase, or to control Fe, an essential component of tyrosinase-related proteins TRP-1 and TRP-2, or to produce melanin-producing intermediates We are focusing on developing substances that inhibit

Tyrosinase inhibitors known to date include arbutin, hydroquinone, hydroxyanisole, ascorbic acid derivatives, kojic acid and retinoids, but some inhibitors cause mutations. There is an experimental result that there is a possibility of carcinogenesis, and in addition, there are problems such as safety and economics, absorption through transdermal. Because of these problems, there is a demand for the development of inhibitors having strong inhibitory activity and ensuring both safety and economy. In addition, research on the development of whitening cosmetics materials requires multi-faceted examinations such as the ability to penetrate to melanosomes, the interaction with keratinocytes, and the differentiation and regeneration of epidermal cells, and overcome the limitations and problems of existing whitening materials and provide superior raw materials. In order to find a substance, there is a need to search for an effective material among natural products for which safety has been proven.

On the other hand, dried flower buds of Sophora japonica L., Leguminosae are called Sophorae Flos, which are prescribed for the purpose of antihypertensive, bleeding, fever, convergence, hemostasis, and sedation in Chinese medicine. It contains many flavonoid compounds as well as other triterpenoids. The painting flower which contains a lot of honey is used as a yellow dye, and it is used as a tea to prevent hypertension. However, there are no studies on the whitening effect of the extract of flower of the painting.

Accordingly, the present inventors have tried to develop a natural-derived composition, as a result of painting flower extract or fractions thereof is excellent in tyrosinase inhibitory activity and melanin production inhibitory effect, excellent antioxidant activity and skin wrinkle improvement, whitening composition or skin aging inhibitory and The present invention has been confirmed that it can be usefully used as a composition for improving skin wrinkles.

An object of the present invention is to provide a composition for skin whitening that acts on the skin stably without cytotoxicity, thereby reducing melanin production.

Another object of the present invention is to provide a composition for inhibiting skin aging and improving skin wrinkles, which is excellent in antioxidant activity and skin wrinkle improvement effect.

In order to achieve the above object, the present invention provides a cosmetic composition for skin whitening comprising a painting flower extract or a fraction thereof as an active ingredient.

In another aspect, the present invention provides a pharmaceutical composition for skin whitening comprising a painting flower extract or a fraction thereof as an active ingredient.

In another aspect, the present invention provides a food composition for skin whitening comprising a painting flower extract or a fraction thereof as an active ingredient.

In another aspect, the present invention provides a cosmetic composition for inhibiting skin aging and improving skin wrinkles comprising a painting flower extract or a fraction thereof as an active ingredient.

In another aspect, the present invention provides a pharmaceutical composition for inhibiting skin aging and skin wrinkle improvement comprising a painting flower extract or a fraction thereof as an active ingredient.

Painting tree flower extract or fractions thereof according to the present invention can be usefully used as a composition for skin whitening because it exhibits a whitening effect by inhibiting the activity of tyrosinase and effectively inhibiting the production of melanin without causing toxicity to cells. have. In addition, the painting flower extract or fractions thereof according to the present invention is useful as a composition for inhibiting skin aging and improving skin wrinkles because it inhibits superoxide production and has excellent free radical scavenging effect, and has an antioxidant effect and shows skin wrinkle improvement effect. Can be used.

Figure 1 is a picture confirming the inhibitory effect of tyrosinase activity of the painting extract.
Figure 2 is a picture confirming the melanin production inhibitory effect of the cedar flower extract.
Figure 3 is a picture confirming the antioxidant activity of the painting extract.
Figure 4 is a picture confirming the superoxide generation inhibitory activity of the painting tree flower extract.
Figure 5 is a picture confirming the free radical scavenging effect of the painting tree flower extract.
Figure 6 is a picture confirming the skin wrinkle improvement effect of the painting tree flower extract
Figure 7 is a picture confirming the cytotoxicity of the painting flower extract.
8 and 9 are diagrams analyzing the fragrance component of the painting flower.

Hereinafter, the present invention will be described in detail.

Painting trees ( Sophora japonica L., Leguminosae) is a deciduous broad-leaved arborescent belonging to the legume family. The leaves are shifted and the upper right bilobal is composed of 15 to 40 leaflets. Leaflets are ovate lanceolate, with flat edges, 2.3-6 cm long. Flowers bloom in August, yellowish white, and hang on a cone-shaped inflorescence. Fruits are narrow pods, 5-8cm long, ripen in October.

Painting tree flower extract of the present invention is prepared by the following method.

Wash the painting flower with water to remove foreign materials, dry in the shade and grind. Painting tree flowers can be used without limitation, such as those grown or marketed. An appropriate amount of solvent is added to the crushed sakura flower so that it is completely immersed. A painting tree flower is extracted using a solvent selected from water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. Preferably methanol is used.

Painting tree flower fraction of the present invention is prepared by the following method.

Water was added to the methanol extract, followed by addition of methylene chloride as an organic solvent, followed by shaking to separate the organic solvent layer and the water layer first, and addition of ethyl acetate as an organic solvent to the water layer, followed by shaking to form an organic solvent layer. The water layer is secondarily separated, and butanol, which is an organic solvent, is added to the water layer, followed by shaking to obtain an organic solvent layer and an aqueous layer by extraction and fractionation.

The skin whitening composition or the composition for inhibiting skin aging and skin wrinkle improvement, comprising a painting tree flower extract or a fraction thereof according to the present invention can be used orally, parenterally, and is applied to the skin during parenteral use. It may be used, and the preferred method of use may be selected depending on the formulation.

As another aspect, the skin whitening composition or the composition for inhibiting skin aging and skin wrinkle improvement may be provided as a cosmetic composition by mixing with a dermatologically acceptable carrier.

The painting flower extract or fractions thereof in the composition of the present invention may be contained in an amount of 0.001 to 50% by weight based on the total weight of the composition.

Cosmetics prepared from the cosmetic composition containing the painting flower extract or fractions thereof of the present invention may be prepared in the form of general emulsion formulations and solubilization formulations. Cosmetics of emulsified form include nutrition lotion, cream, essence, etc., and cosmetics of solubilized form have softening longevity. In addition to cosmetics containing the extract of the flower of the present invention or fractions thereof, it may be prepared in the form of adjuvants that can be applied topically or systemically as commonly used in the field of dermatology by containing a dermatologically acceptable medium or base. .

Formulations of suitable cosmetics include, for example, emulsions, suspensions, emulsions, pastes, microemulsions, microcapsules, microgranules or ionic (liposomes) obtained by dispersing an oil phase in a solution, gel, solid or pasty anhydrous product, aqueous phase. , In the form of non-ionic vesicle dispersants, creams, skins, lotions, powders, soaps, ointments, sprays, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations or conceal sticks Can be provided. It can also be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.

“Dermatologically acceptable carriers” that can be used according to the desired formulation include purified water, oils, waxes, fatty acids, fatty alcohols, fatty acid esters, surfactants, humectants, thickening agents, antioxidants, and viscosity stabilizers. Examples include, but are not limited to, topical stabilizers, chelating agents, buffers, preservatives, lower alcohols, and the like, and their types and concentrations are varied and modifications may be made by those skilled in the art without departing from the spirit and scope of the present invention. And parts that can be changed. If necessary, a whitening agent, a moisturizer, an anti-inflammatory agent, an antibacterial agent, an antifungal agent, a vitamin, a sunscreen agent, an antibiotic, an anti-acne agent, a perfume, a dye may be included, and these are cosmetic compositions according to the present invention in amounts commonly used in the cosmetic field. Can be included. Suitable from 0.001 to 50% by weight per total weight of the composition. If the content of the painting tree flower extract or fractions thereof is less than 0.001% by weight can not expect a distinct effect, if the content exceeds 50% by weight does not see a clear increase in effect with increasing content.

Specifically, the oil may be hydrogenated vegetable oil, perm oil, cottonseed oil, olive oil, palm oil, jojoba oil, avocado oil, wax, beeswax, carnauba, candelilla, montan, ceresin, liquid paraffin Lanolin may be used. Stearic acid, linoleic acid, linolenic acid, oleic acid may be used as a fatty acid, cetyl alcohol, octyl dodecanol, oleyl alcohol, pantenol, lanolin alcohol, stearyl alcohol, hexadecanol may be used as a fatty acid. As the fatty acid ester, isopropyl myristate, isopropyl palmitate, butyl stearate and the like may be used, but is not limited thereto.

Examples of the surfactant include anionic surfactants such as sodium stearate, sodium cetyl sulfate, polyoxyethylene laurylether phosphate, sodium N-acyl glutamate; Cationic surfactants such as stearyldimethylbenzylammonium chloride and stearyltrimethylammonium chloride; Amphoteric surfactants such as alkylaminoethylglycine hydrochloride and lecithin; Glycerine monostearate, sorbitan monostearate, propylene glycol monostearate, polyoxyethylene oleyl ether, polyethylene glycol monostearate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene coconut fatty acid monoethanol arnide (monoethaolarnide ), Polyoxypropylene glycol, polyoxyethylene castor oil, nonionic surfactants such as polyoxyethylene lanolin and the like.

Glycerin, 1,3-butylene glycol, propylene glycol may be used as the moisture absorbent, and ethanol or isopropanol may be used as the lower alcohol. Examples of thickeners include sodium alginate, sodium caseate, gelatin agar, xanthan gum, starch, cellulose ethers (e.g., hydroxyethyl cellulose, methyl cellulose, carboxymethyl cellulose, hydroxy propylmethyl cellulose), polyvinyl pyrrolidone, Polyvinyl alcohol, polyethylene glycol, sodium carboxymethyl cellulose, and the like, but are not limited thereto.

As antioxidants, butylated hydroxytoluene, butylated hydroxyanisole, propyl gallate, citric acid, ethoxyquin can be used, and chelating agents include disodium edetate and ethane hydroxy diphosphate. Citric acid, sodium citrate, boric acid, borax, disodium hydrogen phosphate are available as buffers, and methyl parahydroxybenzoate, ethyl parahydroxybenzoate, dihydro as preservatives. Acetic acid, salicylic acid, benzoic acid are available, but are not limited to these.

The skin whitening composition of the present invention may further contain one or more skin whitening active ingredients exhibiting the same or similar functions in addition to the painting flower extract or fractions thereof. Skin whitening active ingredients include kojic acid and its derivatives, arbutin, ascorbic acid and its derivatives, hydroquinone and its derivatives, resorcinol, cycloalkanone, methylenedioxyphenyl alkanol, 2,7-dinitroindazole, Plant extracts such as vine tangerine extract, rice extract, licorice extract, and the like, but are not limited thereto.

As another aspect, the painting tree flower extract or fractions thereof of the present invention may be mixed with a pharmaceutically acceptable carrier to provide a pharmaceutical composition.

The pharmaceutical composition comprising the painting flower extract of the present invention or a fraction thereof as an active ingredient may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant, boron Releases, sweeteners, binders, coatings, swelling agents, lubricants, lubricants, flavoring agents, and the like can be used.

Dosage forms of the pharmaceutical compositions of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.

The pharmaceutical composition may be preferably formulated into a pharmaceutical composition including one or more pharmaceutically acceptable carriers in addition to the active ingredient described above for administration.

An effective amount of the composition refers to an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, doctor or other clinician, which is the disease or disorder being treated. Includes amounts that induce relief of symptoms. It will be apparent to those skilled in the art that the therapeutically effective dose and the number of administrations of the active ingredient of the present invention will vary depending on the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. In the case of an adult, the pharmaceutical composition of the present invention is preferably administered at a dose of 0.1 mg / kg to 250 mg / kg once or several times a day.

As another aspect, the painting flower extract or fractions thereof of the present invention may be provided as a food composition by mixing with a food acceptable carrier. When the painting flower extract or fractions thereof of the present invention is used as a food or beverage additive, the painting flower extract or fractions thereof may be added as it is, or used together with other food or food ingredients, and may be appropriately used according to a conventional method. . The mixed amount of the painting flower extract or fractions thereof may be suitably determined according to the purpose of use (prevention, health or therapeutic treatment).

There is no particular limitation on the kind of the food. Examples of the food to which the substance may be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, dairy products including gum, ice cream, various soups, beverages, teas, and drinks. Alcoholic beverages and vitamin complexes. When formulated as a beverage, the liquid component added in addition to the painting flower extract is not limited to the above, but may include various flavors or natural carbohydrates, etc. as additional ingredients, as in general beverages. Examples of the aforementioned natural carbohydrates include monosaccharides (e.g. glucose, fructose, etc.), disaccharides (e.g. maltose, sucrose, etc.) and polysaccharides (e.g. conventional sugars such as dextrins, cyclodextrins, etc.), and xylitol Sugar alcohols such as sorbitol and erythritol. The proportion of such natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention. As flavoring agents other than those described above, natural flavoring agents (taumarin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (e.g., saccharin, aspartame, etc.) can be used.

In another embodiment, the food composition of the present invention comprises various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic and natural flavors, colorants and enhancers (such as cheese, chocolate), pectic acid and salts thereof, organic acids , Protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. The food composition of the present invention may also contain pulp for the production of fruit and vegetable drinks. These ingredients may be used alone or in combination, and the proportion of such additives is generally selected in the range of 0.001 to 50% by weight, based on the total weight of the composition.

Hereinafter, the present invention will be described in more detail with reference to Examples. This embodiment is only for illustrating the present invention in more detail, it is to those skilled in the art that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention. Will be self-evident.

Example  One. Flower tree  The extract and Fraction  Produce

The flower was dried well, powdered and extracted for 3 hours with heating with methanol. After evaporating both the solvent and water, the extract was dried by a lyophilizer to obtain a powder extract, which was then used in the experiment.

In addition, water was added to the methanol extract, followed by addition of methylene chloride as an organic solvent, followed by shaking to separate the organic solvent layer and the water layer first, and then adding ethyl acetate as an organic solvent to the water layer, followed by shaking to form an organic solvent. The layer and the water layer were separated secondly, butanol which is an organic solvent was added to the water layer again, followed by shaking to obtain a fraction through an extraction and fractionation process of separating the organic solvent layer and the water layer three times, and then used in the experiment. .

Example  2. Tyrosinase  Inhibitory effect test

Tyrosinase was an enzyme that acts to form melanin from L-DOPA and measured the effect of inhibiting the production of melanin from the substrate L-DOPA. For enzymatic reaction, 1.8 ml of sodium phosphate buffer (1 / 15M pH6.8), 1.0 ml of substrate 1.5 mM L-DOPA, and 0.1 ml of a solution of 10 mg of sample dissolved in 1 ml of DMSO were used. Kojic acid (Kojic acid) was used as 0.1 ml of 200 ug / ml solution. After adding 0.1 ml of enzyme (Tyrosinase 1000U / ml) solution and incubating for 10 minutes, the enzyme activity was stopped in an ice bath and the absorbance was measured at 475 nm.

As shown in FIG. 1, ethyl acetate fraction showed tyrosinase inhibitory effect of about 25% and methylene chloride fraction showed tyrosinase inhibitory effect of about 18%. Kojic acid, a single compound as a raw material for whitening cosmetics, showed an effect of about 66%. The 25% effect of the fraction containing many compounds is a very good effect.

Example  3. Melanin production inhibitory effect assay

B16 mouse melanoma cells (murine melanoma cells) were incubated in the medium containing the extract and washed with PBS, the amount of melanin contained was calculated by measuring the absorbance at 405 nm. Arbutin was used as a positive control.

As a result, as shown in Figure 2, the melanogenesis inhibitory rate of the extract of the cedar flower shows no significant difference at 12.5-50 μg / ml, but it is clearly inhibited at 100 μg / ml to effectively produce melanin, which greatly affects skin whitening. Inhibition was confirmed.

Example  4. Lipid Peroxidation Inhibition Activity Assay

Lipid peroxide content was measured according to the method of Ohkawa et al.

The compound was dissolved in DMSO and used at a concentration of 5 mg / ml. As a positive control drug, 1.0 mg / ml of ascorbic acid dissolved in DMSO was used. 0.65 ml of 0.1 M potasium phosphate buffer (pH 7.4), 0.05 ml of sample, and 0.1 ml of homogenate were added and incubated at 37 ° C. for 1 hour. After adding 0.2 ml of 8.1% SDS, 1.5 ml of acetate buffer (pH 3.5), and 1.5 ml of 0.8% 2-thiobarbituric acid, the mixture was reacted at 95 ° C for 1 hour. After cooling to room temperature, 5 ml of n-BuOH: Pyridin (15: 1) was added, followed by centrifugation at 3,000 rpm for 10 minutes, and the supernatant was measured for absorbance at 532 nm. Lipid peroxide content was expressed in nmole as the amount of MDA (malondialdehyde) per gram of tissue.

As a result, as shown in Fig. 3, the ethyl acetate fraction was about 57% in the extract and fractions of the cedar flower, and the lipid peroxidation inhibitory effect was good, followed by the methanol extract and the methylene chloride fraction as 34-37%. Similar. Ethyl acetate fractions showed higher lipid peroxidation inhibitory activity than ascorbic acid (vitamin C), which is a positive control compound.

Example  5. Superoxide  Production suppression assay

It was measured according to the method of Gotoh and Niki (NBT assay). The compound was dissolved in DMSO and used at a concentration of 5 mg / ml. The positive control group used 1 mg / ml of ascorbic acid dissolved in DMSO. Sample 30 ul, 30 mM EDTA (pH 7.4) 100 ul, 10 mM 30 mM Hypoxanthine dissolved in 50 mM NaOH, 1.42 mM NBT (nitro blue tetrazolium) 200 ul was added and reacted at room temperature for 3 minutes. After the reaction, 100 ul of 0.5 U / ml xanthine oxidase was added, and the final volume was adjusted to 3 ml using 50 mM phosphate buffer (pH 7.4). After reacting for 20 minutes at room temperature, the absorbance was measured at 560 nm.

As shown in FIG. 4, the ethyl acetate fraction showed the best effect of about 79% among the extracts and fractions of the locust flower, and the butanol fraction (about 78%) was similar. Methanol extracts also showed more than 70% of the effects, all higher than the ascorbic acid (vitamin C) positive control compound was found to be effective in inhibiting the production of superoxide damaging skin cells.

Example  6. Free radical  Elimination effect test

Measurement was made using the Blosi method. The compound was dissolved in DMSO and used at a concentration of 10 mg / ml, and the positive control group used 1 mg / ml of ascorbic acid dissolved in DMSO. 4 ml of DPPH radical 1.5 × 10 ^-5 M ethanol solution was added to 0.1 ml of each sample, and the absorbance was measured at 520 nm after 30 minutes of reaction at room temperature. DPPH radical scavenging ability was expressed by calculating the percentage of the sample addition group and the non-addition group.

DPPH scavenging activity (%) = [(A _DPPH -A _s ) / A _DPPH ] x100

As a result, as shown in Figure 5, methanol extract and butanol fraction of the painting flower extract and fractions showed a very good free radical scavenging effect of 100%. All of them were more effective than ascorbic acid (vitamin C), a positive control compound, and it was found to be effective in eliminating free radicals that damage skin cells.

Example  7. Skin wrinkle improvement effect

Elastase is an enzyme that breaks down elastine, and when excessively secreted, elastin breaks down and loses elasticity. The enzymatic reaction was performed by dissolving a sample solution of 1300 ul and 10 mg of substrate solution in which N-succinyl-Ala-Ala-Ala-p-nitroanilide (1 mM) was added to Tris-Cl (0.12M, pH8.0) in 1 ml of DMSO. 7.5 ul of solution was used. Kojic acid (7.5 oz) was used as a 200 ug / ml solution. After addition of 92.5 ul of Tris-Cl (0.12 M, pH 8.0) solution for 10 minutes, 100 ul of enzyme (Elastase 0.0375U / ml) solution was added thereto, followed by incubation for 10 minutes, and absorbance at 410 nm. .

As a result, as shown in Figure 6, methylene chloride fraction in the extract and fractions of the painting tree flower showed about 25% skin wrinkle improvement effect, but the other fractions did not show an effect.

Example  8. Cytotoxicity

B16 BL6 cell lines were dispensed at 2.5 × 10 5 cells / ml in 6 wells of 2 ml and incubated in 37 ° C., 5% CO 2 incubator for 72 hours. Samples were treated by concentration after stabilization for 1 hour after medium replacement every 24 hours for 72 hours. After 72 hours, 100 μl of 1 mg / ml MTT (Thiazoll Blue Tetrazolium Bromide) was added and cultured for 4 hours. Formazan was dissolved in 1 ml of DMSO and absorbance was measured at 540 nm.

As a result, as shown in Figure 7, in the case of the flower extract, the cell viability was measured at 99% at 12.5 μg / ml, 25 μg / ml, 50 μg / ml, but there was no cytotoxicity at these concentrations 100 Above 85 μg / ml, it was measured as 85% -40%, and it was found that there was cytotoxicity above 100 μg / ml.

Example  9. Fragrance Analysis

The fragrance component of the painting tree flower was analyzed under the following conditions using a gas chromatograph-mass spectrometry (GC-MS) instrument, and the analysis results are shown in FIGS. 8 and 9.

Column-HP5MS (0.25 um X 0.25 mm, 30 m)

Carrier gas-He 1 ml / min

Split ratio-20: 1

Oven condition-100 ℃ 2min → 5 ℃ / min 300 ℃

Examples of formulations for the composition of the present invention are illustrated below. However, formulations comprising the compositions of the present invention are not limited in any way.

Formulation example  One. Cosmetics  Preparation of the composition

1.1 flexible lotion ( skin )

Asteraceae Flower Extract 10.00 (%)

1,3-butylene glycol 1.00 (%)

Disodium ID 0.05 (%)

Allantoin 0.10 (%)

Dipotassium glycylizate 0.05 (%)

Citrix Acid 0.01 (%)

Sodium Citrate 0.02 (%)

Glyceres-26 1.00 (%)

Arbutin 2.00 (%)

Hydrogenated Castor Oil 1.00 (%)

Ethanol 30.00 (%)

Preservative Trace

Trace amount of colorant

Trace amount of flavor

Purified water balance

1.2 Nutrition Cream

Asteraceae Flower Extract 10.00 (%)

1,3-butylene glycol 7.0 (%)

Glycerin 1.0 (%)

D-panthenol 0.1 (%)

Botanical Extracts 3.2 (%)

Magnesium Aluminum Silicate 0.3 (%)

PEG-40 Stearate 1.2 (%)

Stearic Acid 2.0 (%)

Polysorbate 60 1.5 (%)

Lipid Glyceryl Stearate 2.0 (%)

Sorbitan Sesquioleate 1.5 (%)

Cetearyl Alcohol 3.0 (%)

Mineral oil 4.0 (%)

Squalane 3.8 (%)

Carlylic / Capric Triglycerides 2.8 (%)

Vegetable Oil 1.8 (%)

Dimethicone 0.4 (%)

Dipotassium glycylizate traces

Allantoin Trace

Sodium Hyaluronate Trace

Tocopheryl acetate appropriate amount

Triethanolamine Amount

Preservative

Flavoring Agent

Purified water balance

Formulation example  2. Preparation of Pharmaceutical Composition

2.1 Sanje  Produce

20 mg of extract flower

Lactose 100 mg

Talc 10 mg

The above components are mixed and filled in airtight bags to prepare powders.

2.2 Preparation of Tablets

Dana Flower Extract 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

2.3 Preparation of capsule

Dana Flower Extract 10 mg

Crystalline cellulose 3 mg

Lactose 14.8 mg

Magnesium stearate 0.2 mg

The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.

2.4 Preparation of Injections

Dana Flower Extract 10 mg

180 mg mannitol

Sterile sterilized water for injection 2974 mg

Na ₂ HPO ₄ 2H ₂ O 26 mg

(2 ml) per 1 ampoule in accordance with the usual injection preparation method.

Formulation example  3: Preparation of food composition

3-1. Manufacture of flour food products

0.5-5.0 parts by weight of the painting extract of the present invention is added to flour, and bread, cake, cookies, crackers and noodles are prepared using the mixture to prepare foods for health promotion.

3-2. Manufacture of dairy

5 to 10 parts by weight of the painting extract of the present invention was added to milk, and various milk products such as butter and ice cream were prepared using the milk.

Claims (7)

delete delete delete delete delete A cosmetic composition for improving skin wrinkles comprising a painting tree flower methylene chloride fraction as an active ingredient.
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