KR101785955B1 - Cosmetic composition comprising upland rice extracts - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising a paddy field extract. In one embodiment, the cosmetic composition comprising the paddy rice extract comprises paddy rice bran extract, corynebacterium extract, chamomile extract, and mossy extract as an active ingredient.

Description

COSMETIC COMPOSITION COMPRISING UPLAND RICE EXTRACTS [0002]

The present invention relates to a cosmetic composition comprising a paddy field extract.

The human skin consists of three layers of epidermis, dermis and subcutis. The skin layer is composed of stratum corneum, stratum granulosum, stratum spinosum, basal layer. Melanin is produced in the melanocytes in the basal layer. These melanocytes have enzymes such as tyrosinase, which work together to oxidize an amino acid called tyrosine, which is always present in the living body, To form melanin, a dark brown pigment. The melanin thus formed migrates to the epidermal cell called keratinocyte through the dendrites of the melanocyte to show skin color of the epidermal layer.

In vivo, there is no enzymes that degrade melanin, but it is removed by falling off the skin when the keratin is separated from the epidermis. However, when melanin occurs more than necessary, or when aging of the skin occurs, hyperpigmentation such as stinging, freckles or dots is caused, resulting in poor cosmetic results.

Various factors affecting the production of melanin are known. The enhancement of melanin production caused by ultraviolet rays and the melanin deposition resulting therefrom are very important in the cosmetics field. The basic mechanisms of the active ingredients used in cosmetics for the prevention of melanin deposition include a method of inhibiting the formation of genes involved in tyrosinase formation, a method of inhibiting the activity of the formed tyrosinase, a suppression of the melanin mediator, Reduction of melanin and suppression of photooxidation, promotion of melanin excretion, and ultraviolet screening.

In Asian countries such as Korea, Japan and China, white skin has been a symbol of beauty and a measure of beauty. Recent development of technology and improvement of quality of life have increased the desire and concern for beauty, and it is necessary to develop a whitening product that prevents excessive melanin production. Accordingly, studies on the development of whitening agent and whitening cosmetic product are actively under way. Inhibitors which inhibit tyrosinase activity such as kojic acid and arbutin, hydroquinone, vitamin A, vitamin C and derivatives thereof are used in whitening products. However, they are limited in their use due to safety concerns on the skin, safety issues in the formulation and insufficient whitening effects.

On the other hand, skin cells are the most closely related to wrinkling. In general, the cause of skin aging can be classified into natural aging caused by aging and internal aging caused by external environment. Natural aging is difficult to control because it involves many genetic factors. Therefore, much research has been conducted to control environmental factors such as ultraviolet rays, oxidation, and drying.

Environmental factors mainly collagen protein, elastin protein contained in the dermis, such as destruction or denaturation causes wrinkles. In particular, ultraviolet rays of the sun are typical environmental factors for oxidizing skin and producing active oxygen. Such oxidative power and active oxygen can cause protein breakdown in skin cells, oxidation of sugar, genetic modification, and the like, which can cause skin aging and wrinkles.

Accordingly, development of a material capable of minimizing the side effects and maximizing both skin whitening and anti-wrinkle effects has been demanded. In addition, there is an increasing demand for a cosmetic composition which can be used without limitation of a content with a small risk factor such as skin irritation and cytotoxicity.

On the other hand, curcumalonga is an annual plant of ginger and is also called ganghwang (姜黄 / 薑黄). The rhizome is a massive, side cut with a yellow color and a scent. The leaf is large, 30 ~ 90cm long, 10 ~ 20cm wide, the tip of the leaf is pointed, the base is triangular, and the top is blue. Leaves are elliptical and 4-5 are collected. The rootstock is a cone-like or branched round pole, and the folded face is navy blue. The flower is an acanthum acupuncturist. It is bloomed from late spring to summer, about 30cm long, pale green oval, 4-5cm long, Yellow, 2.5cm in length, and rhizome is also called Ulgum or Cheonokgyedang. It does not use leaves or flowers but mainly roots and tubers, and it is used as a material to make spicy and bitter flavors to make curry rice. The color of the rootstock is the best when it smells like a soft orange with a curcumin, it is mostly used for cooking, but it is also used for medicine in India.

Chamomile ( Matricaria chamomilla L.) is a perennial herb that grows up to 50 ~ 100cm in height. There are few hairs, the stem is straight, round, and there are many side branches. The stem is pointed with long feathered leaves. A corolla with a width of 1 to 3 cm is pierced from the tip of the branch. The corolla is flattened and lengthens in a cone shape. Flowers bloom in May-September, and 15 tongue-shaped white border petals are bent back as the bloom grows. Flowers have an apple flavor.

Hashuo (首 首 烏, Polygonum multiflorum Thunberg ) is a perennial herbaceous perennial plant. The sewage has no hairs in its entirety, and its roots extend into the ground, sometimes forming round root roots. In the room, the roots are used as medicinal materials. The Hashuo has been known to have the efficacy of tonic, gangjeong, blood (blood), interpolation, breeze and excretion.

BACKGROUND ART [0002] The background art relating to the present invention is disclosed in Korean Patent Laid-Open Publication No. 2010-0131598 (published on Dec. 16, 2010, entitled " composition for external application for skin containing rice callus extract and method for producing the same).

It is an object of the present invention to provide a cosmetic composition comprising a paddy field extract having excellent anti-inflammatory, antioxidant and wrinkle-reducing effects.

Another object of the present invention is to provide a cosmetic composition comprising a paddy field extract having excellent skin whitening effect and skin tone improving effect.

It is still another object of the present invention to provide a cosmetic composition comprising a paddy field extract having an excellent effect of improving freckles, dullness and inflammatory hyperpigmentation.

Another object of the present invention is to provide a cosmetic composition which is free from cytotoxicity, is harmless to the human body, and contains a paddy field extract having little skin irritation.

Another object of the present invention is to provide a cosmetic composition comprising a paddy field extract having excellent productivity and economy.

One aspect of the present invention relates to a cosmetic composition comprising a paddy field extract. In one embodiment, the cosmetic composition is selected from the group consisting of rice ( Oryza sativa L.) extract, Curcumalonga extract, Matricaria chamomilla L.) extract and Polygonum multiflorum Thunberg ) extract as an active ingredient.

In one embodiment, the one or more varieties of the above varieties can be used.

In one embodiment, the paddy rice extract, corynebacterium extract, chamomile extract, and sulfoisophthalate extract may be contained in a weight ratio of 1: 0.1-5: 0.1-5: 0.1-5.

In one embodiment, the pomegranate extract, corynebacterium extract, chamomile extract and Sasa extract may be contained in an amount of 0.01 to 50% by weight based on the total weight of the cosmetic composition.

In one embodiment, the cosmetic composition may be formulated as a lotion, essence, foundation, powder, lotion, cream, pack, gel, ointment, patch, spray, mask sheet or detergent.

The cosmetic composition of the present invention is excellent in anti-inflammatory, antioxidant and anti-wrinkle effects, has no cytotoxicity, has little irritation to the skin, is harmless to human body, has excellent skin whitening effect and skin tone improving effect, And inflammatory hyperpigmentation, and can be economically superior since it has low production cost and high productivity as compared with other products having similar effects.

FIG. 1 is a graph showing the results of cell activity tests of Examples according to the present invention and Comparative Examples according to the present invention.
2 is a photograph showing the results of the antibacterial activity test of the example according to the present invention.

In the following description of the present invention, a detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to be exemplary, self-explanatory, allowing for equivalent explanations of the present invention.

Including paddy rice extract Cosmetics  Composition

One aspect of the present invention relates to a cosmetic composition comprising a paddy field extract. In one embodiment, the cosmetic composition is selected from the group consisting of Oryza sativa L. extract, Curcumalonga extract, Matricaria chamomilla L.) extract and Polygonum multiflorum Thunberg ) extract as an active ingredient.

Mulberry extract

Unlike paddy rice, upland rice ( Oryza sativa L.) means rice grown in the field. The rice paddy must be grown in paddy fields because it must supply a sufficient amount of water for almost all growing periods. On the other hand, the paddy rice is cultivated in the field because it consumes a significantly lower amount of water than the paddy rice during the growing period. It is known that the difference between the paddy rice and the paddy rice originated from a long time evolution or breeding through breeding.

In one embodiment, the paddy field and field paddy may have a phenotype or genotype difference in one or more properties. Due to these differences, the active ingredients contained in rice paddies and rice paddies may be different. For example, the paddy rice and paddy rice can be distinguished from each other through ecological characteristics, morphological characteristics or breeds.

In one embodiment, the ecological characteristics of the paddy field are that they are cultivated in the field, consume much less water than the paddy during the growth period, have durability characteristics, have a strong cold tolerance, Strong point, and strong resistance to potassium chlorate (KClO ₃ ).

In one embodiment, the morphological characteristics of paddy rice are that the roots fall deep into the ground (myocardial roots), the roots are large, the leaves are thicker, the number of pores is small, the water storage capacity is high, Thick point, large and dense spike, and well-developed point of chiken.

In one embodiment, the one or more varieties of the above varieties can be used. When the paddy rice of the above variety is applied, the content of the paddy rice active ingredient to be described later is high, so that the effective ingredient can be easily extracted.

In one embodiment, the paddy field extract can be obtained by extracting at least one of outpox, roots, leaves, stems, and middle heights.

In one embodiment, the paddy field extract may be an outpost extract of paddy field. As used herein, the term " outpost " may refer to a part of a plant to be extracted excluding parts of eggs or rice grains. For example, the paddy bark extract may be extracted from the outpost of the paddy field where the ears are removed by harvest. In this case, the extraction efficiency can be improved by preventing the impurities caused by the rice grain or the scar during the extraction process. In addition, when the outpost of the remaining rice after harvesting is used, not only the production cost of the paddy rice extract can be lowered, but also the supply of the material can be smoothly performed, thereby improving the efficiency of the production cost.

In another embodiment of the present invention, the paddy field extract can be obtained by extracting at least one of the roots, leaves, stems and the middle of the paddy field. For example, the paddy rice extract may be extracted from a mixture of roots and heavy roots of paddy rice. In another example, the paddy field extract may be extracted from a mixture of stem and leaves of the paddy field. As another example, the paddy field extract may be extracted from a mixture of roots, stem and leaves of the paddy field.

In one embodiment, the paddy rice extract may have a skin whitening effect through inhibition of tyrosinase, an antioxidative effect through elimination of active oxygen, and an anti-wrinkle effect through inhibition of elastase. In addition, even when the cosmetic composition contains a high concentration of paddy field extract having a concentration of 100 占 퐂 / ml or more, it may inhibit inflammatory factors and may not exhibit skin toxicity.

In one embodiment, the paddy rice extract may be contained in an amount of 0.003 to 20% by weight based on the total weight of the cosmetic composition. When the content is in the above range, the content of the active ingredient is high, and skin whitening, wrinkle-reducing and anti-inflammatory effects are increased, and the formulation stability of the cosmetic composition can be maintained.

Kool  extract

The koji extract of the present invention is included for the purpose of inhibiting the elastase and collagenase involved in wrinkle formation and improving wrinkles. The extract can be extracted using one or more of Curcumalonga leaf, flower, root and stem regions.

In one embodiment, the extract is from 0.003% to 20% by weight based on the total weight of the cosmetic composition. Within the above range, the wrinkle improvement, antioxidative effect and whitening effect of the present invention can be excellent.

Chamomile  extract

The chamomile of the present invention is included for the purpose of further improving the wrinkle-improving effect, the anti-inflammatory effect and the antioxidative effect. In one embodiment, the chamomile extract may be extracted using one or more of the leaves, flowers, roots, and stem regions of the chamomile.

In one embodiment, the chemomail extract may be contained in an amount of 0.003% to 20% by weight based on the total weight of the cosmetic composition. Within the above ranges, the wrinkle-improving effect, anti-inflammatory effect and antioxidative effect of the present invention can be excellent.

Sewage  extract

Hoseo (Polygonum multiflorum Thunb) Are included for the purpose of improving anti-inflammatory and antioxidant effects. In one embodiment, the Sucrose extract may be extracted using one or more of the roots and stem regions of Sucrose.

In one embodiment, the Sucrose extract may be included in an amount of 0.003 to 20% by weight based on the total weight of the cosmetic composition. Within the above ranges, the antioxidative and anti-inflammatory effects of the present invention can be excellent.

In one embodiment, the pomegranate extract, corynebacterium extract, chamomile extract and Sasa extract may be contained in an amount of 0.01 to 50% by weight based on the total weight of the cosmetic composition. When included in the above range, it is excellent in cytotoxicity and harmlessness to human body, skin whitening, antibacterial and antioxidative effect, and excellent formulation stability of cosmetic composition.

In one embodiment, the cosmetic composition of the present invention may contain the pomegranate extract, corynebacterium extract, chamomile extract, and horseradish extract at a weight ratio of 1: 0.1-5: 0.1-5: 0.1-5. When included in the above range, a synergistic action occurs between the active ingredients contained in the above components, thereby excelling in skin whitening, antibacterial and antioxidative effects, excellent wrinkle-improving effect, and excellent formulation stability of a cosmetic composition .

For example, the paddy rice extract, corynebacterium extract, chamomile extract, and sucrose extract may be contained in a weight ratio of 1: 0.1-1: 0.1-1: 0.1-1. In another example, the paddy rice extract, corynebacterium extract, chamomile extract, and sulfo extract may be contained in a weight ratio of 1: 0.2-0.4: 0.1-0.3: 0.1-0.3.

The cosmetic composition of the present invention can have a low production cost and high productivity as compared with other products having similar effects, and thus can be excellent in economy.

The cosmetic composition may further comprise a base component which can be compounded in a conventional cosmetic composition. The specific examples may further include a solubilizer, a surfactant, a moisturizer, an agitator, a pH adjuster, a preservative, an antioxidant, a sequestering agent, a bactericide, an anti-inflammatory agent, an antimicrobial agent, a solubilizing agent or a flavoring agent, .

Examples of the surfactant that can be included in the composition include anionic surfactants such as ammonium laurylsulfosuccinate and ammonium lauryl sulfate and nonionic surfactants such as polyoxyethylene alkyl ether and polyoxyethylene fatty acid ester . Also, cationic surfactants such as tertiary aliphatic amine salts, alkyltrimethylammonium chloride and the like and amphoteric surfactants such as betaine, amide betaine and the like can be used.

The moisturizing agent may be sodium hyaluronate, glycerin, propylene glycol, 1,3-butylene glycol, dipropylene glycol, sorbitol, etc. The thickener may be xanthan gum, methyl cellulose, hydroxyethyl cellulose, carrageenan, carboxy Methylcellulose, hydroxymethylcellulose, and other water-soluble polymeric compounds. The pH adjuster may specifically include citric acid, sodium hydroxide, triethanolamine, sodium citrate, phosphoric acid, sodium phosphate, lactic acid, and the like.

The preservative may be benzoic acid, paraoxybenzoic acid ester, a mixture of methylchloroisothiazolinone, phenoxyethanol, DMDM, and the like. Specific examples of the antioxidant include dibutylhydroxytoluene and ascorbic acid .

The metal ion sequestering agent may be disodium ethylenediaminetetraacetate, tetranodium ethylenediaminetetraacetate, and the like. Specifically, the disinfectant may be chlorhexidine gluconate, quaternary ammonium salt, pyrrothonol amine, zinc pyrithione suspension, urethropropylbutyl Carbamate, salicylic acid, and the like.

Examples of the anti-inflammatory component include monoammonium glycyrrhizinate, dipotassium glycyrrhizinate, allantoin, and mixtures thereof. The antibacterial (antimicrobial) component specifically includes phenoxyethanol, chlorhexidine, chlorhexidine gluconate, benzoic acid and its salts, benzyl Alcohols, salicylic acid, trichlorocarbane, zinc pyrithione suspensions, and mixtures thereof.

The solubilizing agent may specifically be a phage 60 -hydrogenated castor oil or the like, and the above-mentioned flavoring agent may be any conventional agents used in the cosmetic composition field of the present invention.

In addition, the cosmetic composition of the present invention may be prepared in any formulations conventionally produced in the art. In one embodiment, the cosmetic composition may be formulated as a lotion, essence, foundation, powder, lotion, cream, pack, gel, ointment, patch, spray, mask sheet or detergent.

Cosmetics  Method for producing a composition

Another aspect of the present invention relates to a method for producing the cosmetic composition. In one embodiment, the cosmetic composition may be prepared by preparing a mixed extract using an extracting solvent such as paddy rice, paddy rice, chamomile, and sewage, and concentrating the mixed extract.

In one embodiment, 30 parts by weight to 500 parts by weight of an extraction solvent is added to 100 parts by weight of each layer of rice paddy, wool, chomomilla, and sardine, extracted for 3 to 10 days, and then filtered to obtain an extract. When filtering under the above conditions, it is possible to easily remove the solids of the paddy rice, paddy rice, chamomile and sewage to minimize loss of the active ingredient contained therein, shorten the fractionation time of the paddy rice paddies, And can be advantageous for production and production of highly concentrated products.

In one embodiment, paddy rice, cormorant, chamomile and sewage can be used by pulverizing or sieving to a size of 0.01 mm to 10 cm by using the above-mentioned parts. Here, the " size " is defined to mean a maximum length.

Examples of the extraction solvent include water, acetone, ethyl acetate, butyl acetate, diethyl ether, benzene, chloroform, hexane, and alcohols having 1 to 10 carbon atoms. These may be used alone or in combination of two or more.

In one embodiment, the alcohols having 1 to 10 carbon atoms include lower monohydric alcohols such as methanol, ethanol, butanol, pentanol, and hexanol; alcohols such as 1,2-pentanediol, 1,5-pentanediol, hexanediol, , Intermediate dihydric alcohols such as octanediol and decanediol, lower trivalent alcohols such as propylene glycol, 1,3-butylene glycol and glycerin, and the like can be used. These may be used alone or in combination of two or more.

In one embodiment, 30 to 500 parts by weight of an extraction solvent is added to 100 parts by weight of each of the above-mentioned paddy rice, paddy rice, chomomilla and Sasa extract, and the mixture is heated at a temperature of 50 to 100 ° C for 1 to It can be extracted by heating for 72 hours. The extraction efficiency can be excellent when heated under the above conditions.

In one embodiment of the present invention, the paddy rice, paddy, chomomilla, and sulfide extracts are prepared by dipping them in the extraction solvent and then subjecting them to one or more extraction methods such as stirring, refluxing, cold extraction, ultrasonic extraction and supercritical extraction .

In one embodiment, the mixture of the above-described paddy rice, paddy rice, chomomilla, and dassauo may be separately prepared using an extraction solvent, and then mixed to prepare a mixed extract, followed by concentration. In another specific example, the paddy rice, the paddy field, the chamomile, and the sewage may be extracted using an extraction solvent, respectively, and then mixed.

In one embodiment, the concentration can be carried out using an evaporative or vacuum concentration method. More specifically, the rice paddy fields, paddy fields, chamomile, and sulfide extracts may be mixed and put in a rotary vacuum evaporator (Eyela, Japan) for concentration.

In one embodiment, the method may further comprise fractionating the concentrated rice paddy field, paddy field, chamomile, and sulfide extract (or a mixed extract thereof) to prepare a fraction extract. In a specific example, a fraction solvent may be added to the mixed extract to separate the fraction into extract fractions. More specifically, the mixed extract is introduced into a separating funnel, and a first fraction solvent is added to extract the active ingredient. Then, fraction extraction using the first fraction solvent is repeated one to three times, and then the second fraction solvent is added to extract the active component.

The first fraction solvent and the second fraction solvent may be used without limitation depending on the polarity or solubility of the desired active ingredient. For example, one or more of water, ethanol, hexane, dichloromethane, ethyl acetate and butanol may be used as the first fraction solvent and the second fraction solvent, respectively. In a specific example, fraction extraction with ethyl acetate and water as the first fraction solvent is repeated 2-3 times, and then the separated water layer is extracted again with n-butanol as a second fraction solvent have.

In one embodiment, the first fraction solvent and the second fraction solvent may be fractionated with the mixed extract contained in the separating funnel at a volume ratio of 1: 0.1 to 1:10, but are not limited thereto. Each of the fractions obtained by this method can be mixed to prepare a fraction extract. At this time, the fraction extract can be obtained by concentration. For example, the fraction extracted with the fraction solvent prepared above may be concentrated using a rotary vacuum evaporator (Eyela, Japan) to prepare a fraction extract.

In another embodiment of the present invention, the fraction extract may further comprise a chromatographic purification step.

In one embodiment, the chromatographic purification step comprises fractionating the mixed extract with a first fraction solvent using ethyl acetate and water, and separating the water layer again with n-butanol as a second fraction solvent, , The ethyl acetate layer fraction can be loaded onto silica gel column chromatography and purified using a developing solvent.

As the developing solvent, a polar solvent and a non-polar solvent may be used. As the polar solvent, one or more of methanol and acetone may be used in combination. As the nonpolar solvent, benzene, chloroform (CHCl ₃ ), hexane and ethyl acetate may be used in combination. In addition, the polarity can be controlled by adjusting the mixing ratio thereof, and purification can be carried out by collecting a plurality of times.

In one embodiment, the purification is accomplished by loading the fractionated ethyl acetate layer into silica gel column chromatography, eluting first by partitioning the polarity of the first developing solvent using hexane and ethyl acetate as the first developing solvent, And a part of the aliquots is secondarily fractionated while controlling the polarity of the second developing solvent using chloroform and methanol as the second developing solvent to obtain an aliquot.

In one embodiment, at least one aliquot obtained from the first developing solvent and the second developing solvent, respectively, can be obtained. Then, the aliquots obtained in the first fraction and the second fraction are identified by silica (SiO ₂ ) thin layer chromatography (TLC) and UV absorption patterns, and the similar portions can be collected and concentrated.

In one embodiment of the present invention, the purified extract using the above-mentioned chromatography may be used in combination with the above-obtained concentrated rice paddy field, paddy field, chamomile and Sasa extract.

Hereinafter, the structure and operation of the present invention will be described in more detail with reference to preferred embodiments of the present invention. It is to be understood, however, that the same is by way of illustration and example only and is not to be construed in a limiting sense.

The contents not described here are sufficiently technically inferior to those skilled in the art, and a description thereof will be omitted.

Example  And Comparative Example

Example  One

Sowanpyeo: The Sangnamwon variety cultivated in Jeju area was used, and the roots, leaves and stems of the paddy field were cut into 0.1 cm size.

Ulgum: The roots, leaves and stems of Ulgum were prepared by cutting into 0.1 cm size.

Chamomile: Flowers, leaves, stems and roots of chamomile were prepared by cutting into 0.1 cm size.

Sasao: Sasa stem and root were cut into 0.1 cm size.

20 kg of 50% ethanol was added to 1 kg of each of the above-mentioned pulverized rice paddies, cormorant, chamomile and sardine, and the mixture was stirred and extracted at room temperature for 3 days and filtered. The extract was repeated three times and concentrated in a rotary vacuum concentrator, Chamomile and Sasa extracts, respectively. Then, the above extracts were mixed at a weight ratio of 1: 0.6: 0.2: 0.2 to prepare a mixed extract.

Example  2

The extract was prepared in the same manner as in Example 1, except that the paddy rice extract, Ulgum extract, Chamomile extract and Sasa extract were mixed at a weight ratio of 1: 1: 1: 1.

Comparative Example  One

The extract was prepared in the same manner as in Example 1, except that the paddy bark extract, uroguan extract and the chamomile extract were mixed at a weight ratio of 1: 0.6: 0.2.

Comparative Example  2

The extract was prepared in the same manner as in Example 1, except that the paddy rice extract, the chamomile extract and the Sasa extract were mixed at a weight ratio of 1: 0.2: 0.2.

Comparative Example  3

The extract was prepared in the same manner as in Example 1, except that the paddy rice extract, paddy rice extract, and Sasa extract were mixed at a weight ratio of 1: 0.6: 0.2.

Comparative Example  4

Extracts were prepared in the same manner as in Example 1, except that Ulgum extract, Camomile extract and Sasa extract were mixed at a weight ratio of 1: 0.3: 0.3.

Experimental Example

(1) Cytotoxicity test

The cytotoxicity of each of the above Examples 1 and 2 and Comparative Example 1 was evaluated by diluting with concentrations of 0, 3.125, 6.25, 12.5, 25, 50, and 100 (占 퐂 / ml).

RAW 264.7 cells, a type of macrophage, were counted in a DMEM medium containing 1% penicillin / scrap tromycin and 10% FBS (fetal bovineserum) in the same number of 1.0 × 10 ⁴ cells / well on a 24-well plate , Followed by incubation at 37 ° C and 5% carbon dioxide for 24 hours.

The cells cultured for 24 hours were mixed with the culture medium of each of the above-mentioned Examples 1 to 2 and Comparative Example 1, and 1 ml of each was added to each well. The cells were incubated in an incubator at 37 ° C and 5% CO ₂ for 24 hours , And the supernatant of each well was separately taken, and WST-1 assay solution (ez-cytox) was added to each well. After incubation for 2 hours in an incubator, the cell viability was determined by measuring the absorbance at 450 nm using an ELISA reader Respectively.

1 is a graph showing the results of cytotoxicity tests on the mixed extracts of Examples 1 and 2 and Comparative Example 1. Fig. Referring to FIG. 1, the mixed extracts of Examples 1 and 2 showed very low cytotoxicity even at a high concentration of 100 μg / ml.

(2) Inflammatory Inducing Factor Inhibition Test

In order to measure the anti-inflammatory activity effects of the mixed extracts prepared in Examples 1 and 2 and Comparative Examples 1 to 4, the above Examples and Comparative Examples were diluted to a concentration of 50 (占 퐂 / ml) ) Was measured.

RAW 264.7 cells, a type of macrophage, were counted in a 24-well plate at a density of 1.0 × 10 ⁴ cells / well in a DMEM medium containing 1% penicillin / scrap tromycin and 10% FBS (fetal bovineserum) , And cultured at 37 ° C and 5% carbon dioxide for 24 hours.

The cells were cultured for 24 hours, and then mixed with the medium for each concentration. Then, 1 ml of each extract was added to each well, followed by incubation in an incubator at 37 ° C and 5% CO ₂ for 24 hours. At this time, LPS (lipo poly saccharide), which is an inflammatory inducer for expressing NO, was treated at a concentration of 1 μg / ml and allowed to react at 37 ° C and 5% carbon dioxide for 24 hours. Next, take the supernatant of each well separately and take 100 ml of the culture solution into each well using Nitric Oxide detection kit. Add 50 ㎕ of Griess reagent A and 50 ㎕ of Griess reagent B to each well. Min, and the absorbance was measured at 540 nm using an ELISA plate reader. The results are shown in Table 1 below.

Griess Reagent A: N-1-Naphthylethylenediamine (NEDHC)

Griess Reagent B: Sulfanilamide

Referring to the results shown in Table 1, it was found that the mixed extract of Example 1 had an effect of inhibiting the formation of nitric oxide (NO) in a concentration-dependent manner, as compared with Comparative Examples 1 to 4.

(3) Free radical removal ability test

Free radicals are active oxygen, and the oxygen that we breathe is thousands of times higher in the process of making energy and reducing it to water. It is produced by the metabolic processes of cells in plants and animals, or by stress, light stimulation, and bacterial infiltration. When excessive amount of active oxygen is present in the body, normal cells become indiscriminate attacks, diseases and aging. In addition, it acts as a pathological factor showing an immune response sensitive to external stimuli. The mixed extracts prepared in Examples 1 to 2 and Comparative Examples 1 to 4 were each in an amount of 50 (㎍ / ml). The extracts of Examples 1 and 2 and Comparative Examples 1 to 4 were tested for their antioxidant activity, ) To evaluate DPPH inhibition.

Each of the above Examples 1 to 2 and Comparative Examples 1 to 4 was mixed with a solution of 0.1M DPPH (1,1-diphenyl-2-picrylhydrazyl) in concentration and reacted at 4 ° C for 30 minutes. At the end of the reaction, each sample was placed in a 96-well plate and absorbance was measured at 520 nm using an ELISA reader.

The free radical scavenging activity (%) of the sample was calculated according to the following equation 1 using the case where the above examples and comparative examples were not used as a control group, and the results are shown in Table 2 below. At this time, ascorbic acid was used as a positive control.

[Formula 1]

Free radical scavenging ability (%) = [100- (absorbance of the group treated with the sample / absorbance of the group not treated with the sample X 100)]

Referring to Table 2, it was found that the mixed extracts of Examples 1 and 2 had better DPPH scavenging ability than Comparative Examples 1 to 4.

(4) Wrinkle-preventing efficacy Verification test

The mixed extracts prepared in Examples 1 to 2 and Comparative Examples 1 to 4 were each diluted to a concentration of 50 (占 퐂 / ml) to evaluate the degree of inhibition of Elastase.

Substrates of 1.6 mM concentration of N-succinyl- (Ala) 3-p-nitroanilide (S4760, Sigma) in Examples 1 to 2 and Comparative Examples 1 to 4 were dissolved in 10 mM Tris-HCl buffer And 0.4 U / mL of Elastase from porcine pancreas Type IV (E0258, Sigma). After incubation at 25 ° C for 5 minutes, absorbance was measured at 410 nm in a 96-well plate.

And the case where the sample was not added was used as a control, and calculated using the following formula 2, and the results are shown in Table 3 below. At this time, Oleanolic acid was used as a positive control.

[Formula 2]

Elastase inhibitory activity (%) = 100- (absorbance of the group treated with the sample / absorbance of the group not treated with the sample X 100)

Referring to Table 3, the mixed extracts of Examples 1 and 2 exhibited more excellent elastase inhibitory effect than Comparative Examples 1 to 4, indicating that they are effective in improving wrinkles.

(5) Whitening efficacy Verification test

The mixed extracts prepared in Examples 1 to 2 and Comparative Examples 1 to 4 were each diluted to a concentration of 50 (占 퐂 / ml) to measure the amount of melanin inhibition.

B16F10 melanoma cells were counted in the same manner as in the case of 2.0 × 10 ⁵ cells / well in a 60Φ Petri dish using a hemacytometer, and then cultured at 37 ° C. and 5% carbon dioxide for 24 hours .

After culturing for 24 hours, the obtained cell culture medium was removed, and each of the above-mentioned Examples and Comparative Examples were taken together with 50 nM of? -MSH to induce melanin production, mixed with the medium, and divided and reacted for 60 hours. ).

After incubating for 60 hours, 1 ml of the supernatant was taken out to measure extracellular melanin production, and absorbance was measured at 450 nm using an ELISA plate reader. The amount of extracellular melanin production was calculated using the following equation (3), and the results are shown in Table 6 below. At this time, arbutin was used as a positive control.

[Formula 3]

Melanin production (%) = (absorbance of each test substance / absorbance of control group) X100

It was found that the mixed extracts of Examples 1 and 2 had an effect of suppressing the amount of melanin formation as compared with Comparative Examples 1 to 4, and that the skin whitening effect was excellent.

(6) Antimicrobial activity Verification test

Among the mixed extracts of Examples and Comparative Examples, experiments were conducted to investigate the degree of antibacterial activity of each of Examples 1 to 2 and Comparative Example 1 as follows.

<Preparation of Evaluation Medium>

Escherichia coli (ATCC 8739), Staphylococcus aureus (ATCC 6538P) and Staphylococcus aureus were used for evaluation of the antimicrobial activity. ), Pseudomonas aeruginosa (ATCC 9027) and Candida albicans (ATCC 10231) were purchased from Korea Microorganism Resource Center (KCTC, Korea) and Korea Microorganism Conservation Center (KCCM, Korea).

Among the standard test strains, TSA medium (Tryptic Soy Agar) was used for the culture of E. coli, Staphylococcus aureus and P. aeruginosa. Yeast extract media was used for culturing Candida albicans among the standard test strains.

&Lt; Circular filter paper method &

An independent nutrient broth sterilized at 121 DEG C for 15 minutes was prepared, and each of the standard test strains prepared above was inoculated in an amount of 3 x 10 &lt; ⁵ &gt; cfu / mL.

Among the compositions of the Examples and Comparative Examples, about 15 to 20 μl of the composition of Example 1 and Comparative Example 1 was absorbed to a paper disk having a diameter of 10 mm, and then the solvent was dried and the surface of the culture medium on which the strain was inoculated And the standard test strain was cultured. Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans among the standard test strains were cultured in a 37 ° C incubator for 24 hours. After the incubation was completed, the size of the clear zone formed around the paper disk was measured to confirm the antibacterial effect. The evaluation was made on three criteria (good: good, fair: poor, X: poor) Table 5 shows the results.

2 is a photograph showing the results of the antibacterial activity test (circular filter paper method) of Example 1 of the present invention. Referring to Table 5 and FIG. 2, Example 1 showed superior antibacterial activity against Escherichia coli, Staphylococcus aureus, Candida avicans, and Pseudomonas aeruginosa, as compared with Comparative Example 1.

(7) Formulation stability evaluation

Lotions were prepared by applying the ingredients listed in Table 6 below to the mixed extracts of Examples 1 and 2 and Comparative Examples 1 to 4 using conventional methods.

To measure the stability of the cosmetic material containing the extracts of Examples 1 to 2 and Comparative Examples 1 to 4 to the degree of separation and discoloration, a thermostatic chamber kept constant at 45 ° C, 25 ° C and 4 ° C, Each of the examples 1 to 2 and the comparative examples 1 to 4 was put in a container for 5 weeks and stored for 4 weeks, and the degree of separation and discoloration was compared and the results are shown in Table 7 below. The degree of separation and discoloration of the product was classified into the following six grades and evaluated.

* Separation and discoloration evaluation criteria

4: Severely separated (discolored), 5: Severely separated (discolored), 2: slightly separated (discolored), 3: slightly separated (discolored)

Referring to the results of Table 7, it was found that the cosmetic compositions of Examples 1 and 2 hardly produced discoloration and separation phenomena.

It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

Claims (5)

( Oryza sativa L.) extract, Curcumalonga extract, Matricaria chamomilla L. extract and Polygonum multiflorum Thunberg extract at a ratio of 1: 0.6-1: 0.2-1: 0.2-1: 1 &Lt; / RTI &gt;
2. The cosmetic composition according to claim 1, wherein at least one of the above-mentioned paddy fields is selected from the group consisting of Sangnam-paddy rice and Agri-forest paddy-1.
delete [Claim 2] The cosmetic composition according to claim 1, wherein the pomace oil extract, corynebacterium extract, chamomile extract and sulfosucca extract are contained in an amount of 0.01 to 50% by weight based on the total weight of the cosmetic composition.
The cosmetic composition according to claim 1, wherein the cosmetic composition is formulated with a lotion, an essence, a foundation, a powder, a lotion, a cream, a pack, a gel, an ointment, a patch, a spray, a mask sheet or a detergent.

Images