KR20140086747A - Health Functional Food Composition Comprising Red Ginseng Extract, Benincasa Hispida Extract, and Apios Americana Extract for Improving Skin Beauty - Google Patents

Abstract

The present invention relates to a functional health food composition having a use for improving skin beauty, and more specifically, to a functional health food composition containing a fermented red ginseng extract, a Benincasa hispida extract, and an Apios americana extract as active ingredients for improving skin beauty, and to a functional health food manufactured by using the same. A mixture of the fermented red ginseng extract, the Benincasa hispida extract, and the Apios americana extract of the present invention has an effect of ameliorating skin wrinkles by inhibiting MMP-1 activity, an effect of whitening the skin by inhibiting tyrosinase activity and inhibiting melanine generation, and an effect of preventing the skin aging through anti-oxidative activity. Therefore, the composition of the present invention containing the fermented red ginseng extract, the Benincasa hispida extract, and the Apios americana extract as active ingredients can exhibit an excellent effect of improving the skin beauty by ameliorating skin wrinkles, whitening the skin, and preventing the skin aging, and thus can be useful as materials for functional health food or functional health assistant food. In particular, the mixture of the fermented red ginseng extract, the Benincasa hispida extract, and the Apios americana extract of the present invention is an extract derived from natural products, and very safe to the human body.

Description

(Health Functional Food Composition Comprising Red Ginseng Extract, Benincasa Hispida Extract, and Apios Americana Extract for Improving Skin Beauty), which comprises fermented red ginseng,

The present invention relates to a health functional food composition having skin beauty enhancement use, and more particularly, to a skin health improving functional food composition comprising fermented red ginseng, frogs and apiose extract as an active ingredient, and a health functional food .

Today, as the quality of life improves, unlike the past, interest in appearance has increased greatly. This leads to a demand for a new type of cosmetics beyond the satisfaction of existing skin cosmetics. Recently, there has been a tendency to prevent skin aging through food, and to increase skin whitening and skin moisturizing.

Skin is the surrounding tissue that protects the body, regulates body temperature, excretion and skin respiration. The skin is divided into three layers from the outer layer to the epidermis, dermis and subcutaneous fat and consists of epidermal appendages such as hair, sweat glands, nails, and sebaceous glands. The epidermis is divided into the horny layer, the granular layer, the visible layer and the basal layer. The epidermis consists mainly of keratinocyte and melanocyte. The dermis, a fibrous connective tissue below the epidermal layer, is composed of a papillary dermis protruding from the upper part and a net dermis distributed in the lower part of the net. The dermis consists of collagenous fibers and some elastic fibers, which are vaginal and elastic, and have capillaries and nerves. The subcutaneous fat layer is composed of adipocytes and is known to protect internal organs from external shocks, block heat, maintain body temperature, and store energy.

Skin aging can be divided into intrinsic aging and photo aging. Endogenous aging is an inevitable aging phenomenon that naturally occurs over time, and photoaging is observed in areas exposed to sunlight It is caused by a combination of endogenous aging and ultraviolet rays. The characteristics of endogenous aging are slight wrinkles, dryness of skin, reduction of elasticity, etc. The characteristics of photoaging are coarse and deep wrinkles and many fine wrinkles, irregular pigmentation on skin, increase of pigment diseases, decrease of elasticity due to rough and dry skin And the like.

The aging process of the skin progresses differently from the epidermis to the dermis. First, the epidermis decreases in the thickness and the water content of the stratum corneum, resulting in a decrease in elasticity and a change in cell shape such as a sunburn. In addition, the pigment cells are reduced, the decolorizing part appears at a specific site, and the number of Langerhans cells responsible for skin immunity is decreased, resulting in diseases such as skin cancer. In the dermis, collagen fibers and elastic fibers that maintain skin elasticity decrease, skin elasticity decreases, and fine lines increase. In particular, collagen in the dermis is a major component of skin aging, accounting for 77% of the dry weight of the entire skin, excluding fat, and 90% of the dermis fiber component. Since collagen plays a role in maintaining the strength, elasticity and flexibility of the skin, inhibition of synthesis and inhibition of collagen synthesis is considered to be very important in relation to skin cosmetic and skin aging inhibition.

Clean skin is another major concern of skin care, as it is one of the strong desires of modern people to have white, clear and transparent skin. The skin color of a person is determined by the concentration and distribution of melanin present in the epidermis of the skin. It is affected not only by genetic factors but also by environmental or physiological conditions such as ultraviolet rays, fatigue, and stress. Melanin is produced by a nonenzymatic oxidation reaction after tyrosine, an amino acid, is converted to DOPA and Dopaquinine by an enzyme called tyrosinase. Melanin protects the body from the sun's ultraviolet rays, but excessive melanin synthesis in the skin can darken skin tone and cause spots or freckles. Therefore, inhibiting the synthesis of melanin pigment in the skin is a major approach for skin whitening.

On the other hand, it is well known that active oxygen having a high mobility in an incomplete ion state attacks an unsaturated fatty acid to cause a peroxidation reaction to accumulate lipid peroxidation in the body, resulting in deterioration of biological functions and aging. Therefore, intake of food having antioxidant effect is considered to prevent aging by removing active oxygen in the body, and also to be very helpful for skin beauty.

As described above, in order to improve skin beauty, it is necessary to approach various directions such as skin wrinkle improvement, skin whitening and anti-aging, and excavation of materials having such functions simultaneously is useful for functional foods for skin care enhancement It can be used.

Accordingly, the present inventors have screened about 400 kinds of natural products from natural materials in order to find a material which is safe to humans and exhibits excellent skin-beauty-enhancing effect. As a result, it was confirmed that the fermented red ginseng, frog and apiose extract were effective for antioxidation, wrinkle improvement and whitening, and after mixing them at a certain ratio, the antioxidative effect, the effect of improving skin wrinkles by inhibiting MMP-1 expression, Inhibiting and inhibiting melanin synthesis, thereby completing the present invention.

Korean Patent Publication No. 10-2008-0094499

Accordingly, an object of the present invention is to provide a health functional food composition for enhancing skin beauty which is effective for improving skin wrinkles, skin whitening, and prevention of aging.

Another object of the present invention is to provide a health functional food for enhancing skin beauty, which is effective for improving skin wrinkles, skin whitening and preventing aging.

In order to accomplish the above object, the present invention provides a health functional food composition for promoting skin beauty comprising fermented red ginseng, frogs and apiose extract as an active ingredient.

In one embodiment of the present invention, the fermented red ginseng, frog and apiose extract may be water, an organic solvent or a mixed solvent extract thereof.

In one embodiment of the present invention, the organic solvent may be selected from the group consisting of methanol, ethanol, propanol, isopropanol, and butanol.

In one embodiment of the present invention, the fermented red ginseng, frog and apiose extract may be 70% ethanol extracts of fermented red ginseng, frozen ginseng, and apiose, respectively.

In one embodiment of the present invention, the mixture is prepared by mixing a fermented red ginseng extract, a frog extract and an apiose extract in a weight ratio of 1: 1: 1, 2: 1: 1, 1: 2: 1 or 1: w: w).

In one embodiment of the present invention, the mixture may be included in the composition at a concentration of from 0.1 to 10,000 μg / ml.

In one embodiment of the present invention, the skin cosmetic enhancer may be used for improving skin wrinkles, for whitening skin or for preventing skin aging.

In one embodiment of the present invention, the skin wrinkle improvement can be achieved through inhibition of MMP-1 expression.

In one embodiment of the present invention, the skin whitening can be achieved through inhibition of tyrosinase activity and inhibition of melanin formation.

In one embodiment of the present invention, the anti-aging of the skin can be achieved through antioxidant activity.

In addition, the present invention provides a health functional food for enhancing skin beauty prepared from the composition.

In one embodiment of the present invention, the food is selected from the group consisting of beverage, meat, chocolate, foods, confectionery, pizza, ram noodles, gums, candy, ice cream, alcoholic beverages, .

The mixture of fermented red ginseng, frost and apiose extract of the present invention not only has a skin whitening effect by inhibiting MMP-1 expression, an inhibition of tyrosinase activity and a skin whitening effect by inhibiting melanin formation, It has anti-aging effect at the same time. Therefore, the composition of the present invention containing it as an active ingredient can exhibit an excellent skin cosmetic enhancement effect by improving skin wrinkles, skin whitening and anti-aging, so that it can be usefully used as a material for functional health food or health supplement have. Further, the mixture of the fermented red ginseng, frog and apiose extract of the present invention is a natural extract and has an advantage of being safe for human body.

FIG. 1A is a graph showing cytotoxicity of a mixture of the present invention to human fibroblasts. FIG.
FIG. 1B is a graph showing cytotoxicity of the mixture of the present invention on mouse melanoma cell line. FIG.
Fig. 2 is a graph showing MMP-1 production inhibitory activity of the mixture of the present invention.

The present invention is characterized by providing a health functional food composition for enhancing skin beauty comprising a mixture of fermented red ginseng, frogs and apiose extract as an active ingredient.

The "red ginseng (Panax ginseng used in the fermentation of the present invention ginseng CA Meyer 'has been widely used in the private sector, especially in the Korean peninsula, since it has been steamed or steamed by ginseng or other methods. Red ginseng has been reported to have immune enhancement, cardiovascular control and anticancer effects like ginseng, which is a raw material. It is well known that ginsenoside, which is steroidal saponin, is present as a major physiologically active substance of red ginseng.

Of the present invention, donggwaja (Benincasa hispida Cogniaux) is an annual plant belonging to the genus Bacillus, and refers to the mature seed of the amber pumpkin. It is well known that frogs contain a large amount of urease and are effective for diuretic relief, drainage and skin care.

The term " Apios " americana Medikus) is a vine plant in soybean, called 'Indian potato'. Apiosis is known to be effective for hypertension and high cholesterol, restoring fatigue, enhancing stamina, and also for treating dermatitis such as atopy. In addition, Apiose has excellent nutritional value compared to potatoes such as 4 times for iron, 5 times for fiber, 6 times for protein, and 30 times for calcium.

The 'mixture of fermented red ginseng, frost and apiose extract' of the present invention means a mixture of fermented red ginseng, frozen ginseng, and apiose, respectively.

The extracts of fermented red ginseng, frozen ginseng, and apiose according to the present invention can be obtained by extracting and isolating from natural substances by using extraction and separation methods known in the art, and the 'extract' For example, crude extracts of fermented red ginseng, frog and apiose, polar solvent-soluble extracts, or nonpolar solvent-soluble extracts, all of which are extracted from fermented red ginseng, frostbite and apiose using an appropriate solvent.

As an appropriate solvent for extracting the extract from each of the fermented red ginseng, frozen ginseng, and apiose, any pharmaceutically acceptable organic solvent may be used, and water or an organic solvent may be used. Examples thereof include, but are not limited to, For example, alcohols having 1 to 4 carbon atoms, acetone, ether, etc., including purified water, methanol, ethanol, propanol, isopropanol, butanol, Various solvents such as benzene, chloroform, ethyl acetate, methylene chloride, hexane and cyclohexane may be used alone or in combination.

In one embodiment of the present invention, the extracts of fermented red ginseng, frozen fruits and apiose of the present invention may be 70% ethanol extract of fermented red ginseng, 70% ethanol extract of frogs and 70% ethanol extract of apiose.

As the extraction method, any one of the methods such as hot water extraction method, cold extraction method, reflux cooling extraction method, solvent extraction method, steam distillation method, ultrasonic extraction method, elution method and compression method can be selected and used. In addition, the desired extract may be further subjected to a conventional fractionation process or may be purified using a conventional purification method. There is no limitation to the method for producing the fermented red ginseng, frosty and apiose extract of the present invention, and any known method may be used.

For example, each of the extracts of fermented red ginseng, frozen ginseng and apiose contained in the composition of the present invention is subjected to an additional step such as vacuum distillation, freeze drying, spray drying, or the like, And can be produced in a powder state. Further, the primary extract can be further purified by using various chromatographies such as silica gel column chromatography, thin layer chromatography, high performance liquid chromatography and the like, You can get it.

Therefore, in the present invention, each of the extracts of fermented red ginseng, frozen ginseng, and apiose is a concept including all the extract, fraction and purified product obtained in each step of extraction, fractionation or purification, and diluted solution, concentrate or dried product thereof.

The method for preparing the extracts of fermented red ginseng, frozen persimmon, and apiose according to an embodiment of the present invention will be described in more detail as follows.

First, 100 g of each of fermented red ginseng, frogs, and apiose was added to 1 L of 70% ethanol, followed by heating and extracting the mixture. Each of the filtrates thus obtained was concentrated under reduced pressure, Followed by pulverization.

The health functional food composition for enhancing skin beauty of the present invention contains as an active ingredient, a mixture of the fermented red ginseng, the frog and the apiose, respectively.

In one embodiment of the present invention, the mixture comprises a mixture of fermented red ginseng extract, frog extract and apiose extract in a weight ratio of w: 1: 1: 1, 2: 1: 1, 1: 2: : w).

The present inventors have found that a mixture of fermented red ginseng, frogs, and apiose extract has an effect of improving skin wrinkle through inhibition of MMP-1 activity, inhibiting tyrosinase activity and improving skin whitening through inhibition of melanin production, It has been confirmed that the mixture of fermented red ginseng, frozen ginseng and apiose extract has a good skin cosmetic enhancing effect and is useful as a material for functional health food or health supplement food It was the first time that I could do that.

In the following Experimental Example 1 of the present invention, the free radical scavenging activity of the mixture of the present invention was measured and it was confirmed that the DPPH free radical scavenging activity was increased depending on the treatment concentration of the mixture of the present invention (see Table 1) .

In the following Experimental Example 2 of the present invention, the cytotoxicity of the mixture of the present invention to human fibroblast and mouse melanoma cell line was confirmed, and it was confirmed that cytotoxicity was not induced in both of the two cells (FIGS. 1A and 1B) 1b)

In Experiment 3 of the present invention, the inhibitory rate of MMP-1 expression of the mixture of the present invention was measured. As a result, it was confirmed that the expression of human fibroblast MMP-1 was inhibited in all the mixtures of the present invention depending on the treatment concentration 2).

In the following Experimental Examples 4 and 5 of the present invention, the skin whitening activity of the mixture of the present invention was measured. As a result, tyrosinase activity was inhibited depending on the treatment concentration in all of the inventive mixtures (see Table 2) (See Table 3).

The present inventors have found that a mixture of fermented red ginseng, frogs and apiose extract is very effective in preventing skin wrinkles and skin whitening as well as preventing skin aging, and thus can exert an excellent effect on skin beauty enhancement I have experimentally proved the facts.

The composition of the present invention may be a health functional food composition. In addition to containing the above fermented red ginseng, frog and apiose extract as active ingredients, the health functional food composition may contain various flavors or natural Carbohydrates and the like as additional components.

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

The health functional food composition may further comprise one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients, and may be suitably formulated into a health functional food composition.

The form of the health functional food composition may be a tablet, a capsule, a powder, a granule, a liquid, a ring, or the like. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like. Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile water and sterile water suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added.

In one embodiment of the present invention, the mixture of fermented red ginseng, corn steep liquor and apiose extract of the present invention may be contained in the composition at a concentration of 0.1 to 10,000 μg / ml, and may be 0.1 to 95% ≪ / RTI >

The health functional food composition of the present invention can be formulated as described above and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolates, foods, confectionery, pizza, ram noodles, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes, .

In addition, the health functional food composition may further comprise a flavoring agent such as various nutrients, vitamins, minerals (electrolytes), a synthetic flavor and a natural flavor, a coloring agent and an aging agent Cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols and carbonating agents used in carbonated beverages. In addition, the health functional food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks.

The present invention also provides a health functional food for enhancing skin beauty comprising a mixture of fermented red ginseng, frozen persimmon and apiose extract as an active ingredient.

The skin cosmetic enhancement may be skin cosmetic enhancement through skin wrinkle improvement, skin whitening or skin aging prevention.

The health functional food of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and circles for the purpose of enhancing skin beauty.

In the present invention, the term " health functional food " refers to a food prepared or processed using raw materials or ingredients having useful functions in accordance with the Act on Health Functional Foods, and the nutrients are controlled Physiological action, etc., for the purpose of obtaining a beneficial effect.

The health functional foods of the present invention may contain conventional food additives and, unless otherwise specified, whether or not they are suitable as food additives are classified according to the General Rules for Food Additives approved by the Food and Drug Administration, Standards and standards.

Examples of the items listed in the above-mentioned "food additives" include chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as persimmon extract, licorice extract, crystalline cellulose, high color pigment and guar gum; L-glutamic acid sodium preparations, noodle-added alkalis, preservative preparations, tar coloring preparations and the like.

For example, a health functional food in the form of tablets may be prepared by granulating a mixture of the fermented red ginseng, frog and apiose extract, which is an effective ingredient of the present invention, with an excipient, a binder, a disintegrant and other additives in a usual manner Next, a lubricant or the like may be put in a compression mold, or the mixture may be directly compression molded. In addition, the health functional food of the tablet form may contain a mating agent or the like if necessary.

The hard capsule of the capsule-type health functional food can be prepared by filling a mixture of the fermented red ginseng, the frog and the apiose extract, which are the active ingredients of the present invention, with an additive such as an excipient into a normal hard capsule, The capsule can be prepared by filling a mixture of the active ingredient with an excipient such as an excipient into a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative and the like, if necessary.

The ring-shaped health functional food can be prepared by molding a mixture of the fermented red ginseng, frozen duck, and apiose extract, which is an effective ingredient of the present invention, together with excipients, binders, disintegrants, etc. by a known method, If necessary, it may be coated with white sugar or other skin care agent, or the surface may be coated with a substance such as starch or talc.

The granular health functional food may be prepared by granulating a mixture of the above fermented red ginseng, frozen gruel and apiose extract, excipient, binder, disintegrant, etc., which are effective ingredients of the present invention, by a conventionally known method, If necessary, a flavoring agent, a mating agent, and the like may be contained.

The health functional food may be a beverage, a meat, a chocolate, a food, a confectionery, a pizza, a ramen, a noodle, a gum, a candy, an ice cream, an alcoholic beverage, a vitamin complex and a health supplement food.

The present invention also provides a cosmetic composition for enhancing skin beauty comprising a mixture of fermented red ginseng, frogs and apiose extract as an active ingredient.

The cosmetic composition of the present invention may contain not only the above-mentioned mixture of fermented red ginseng, frogs and apiose extract, but also components commonly used in cosmetic compositions such as antioxidants, stabilizers, dissolving agents, vitamins, , ≪ / RTI > and carriers.

In addition to the above-mentioned mixture of fermented red ginseng, frog and apiose extract, the composition of the present invention may be mixed with an organic ultraviolet screening agent which has been used in the past so long as it does not impair the skin protecting effect by reacting with the above-mentioned mixture.

Examples of the organic ultraviolet screening agent include glyceryl paraben, drometrizol trisiloxane, drometrizol, dicaloyl triolate, disodium phenyldibenzimidazole tetrasulfonate, diethylhexylbutamidotriazone, Hydroxybenzoylhexyl benzoate, di-methoxy cinnamate, a mixture of Rawson and dihydroxyacetone, methylenebis-benzotriazolyltetramethylbutylphenol, 4-methylbenzylidene camphor, menthyl anthranylate, benzophenone Benzophenone-4, benzophenone-8 (dioxyphenyl) benzene, butylmethoxydibenzoylmethane, bishexyloxyphenol methoxyphenyltriazine, synoxate, , Octocrylene, ethylhexylmethylmethylphenyl, ethylhexylmethoxycinnamate, ethylhexylsalicylate, ethylhexyltriazone, isoamyl-p-methoxy cinnamate, polysilicon-15 (dimethicone ethyl benzene A carbonate), terephthalic ylidene dikaem peoseol sulphonic Acid and their salts, tea yieyi - may be used salicylate and at least one member selected from the group consisting of amino-benzo ripening Acid (paba).

Examples of products to which the cosmetic composition of the present invention can be added include cosmetics such as astringent lotion, softening longevity lotion, nutrition lotion, various creams, essences, packs, foundation and the like, cleansing, cleanser, soap, .

As a specific formulation of the cosmetic composition of the present invention, there may be mentioned a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, It includes formulations such as soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, latex, lipstick, makeup base, foundation, press powder, loose powder, eye shadow.

In one embodiment of the present invention, the content of the mixture of the fermented red ginseng, frozen fruits and apiose extract of the present invention is 0.0001-20% by weight, preferably 0.5-20%, based on the total weight of the composition, 1.0 to 20% by weight.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited to these examples.

< Example >

Comparative Example  One: Fermented red ginseng  Preparation of extract

About 10 times of water or alcohol was added to 100 g of fermented red ginseng (Sejong Korean ginseng, dried ginseng, Hyanggun, Chungbuk province) and refluxed at 90-100 ℃ for 4 hours. The extract was filtered with a filter paper (Whatmann), and then concentrated under reduced pressure using a rotary evaporator at 60 ° C. Thereafter, the concentrate was lyophilized to obtain 16.4 g of fermented red ginseng extract as a dry weight, and this was used as the experimental sample described below.

Comparative Example  2: Frog  Preparation of extract

About 10 times as much water or alcohol was added to 100 g of frozen fish (Asen Korea, Bupyung-gu, Incheon city) and the mixture was refluxed at 90-100 ℃ for 4 hours. The extract was filtered with a filter paper (Whatmann), and then concentrated under reduced pressure using a rotary evaporator at 60 ° C. Thereafter, the concentrate was lyophilized to obtain 18.1 g of a frog extract as a dry weight, and this was used as the following experimental sample.

Comparative Example  3: Apios  Preparation of extract

About 10 times water or alcohol was added to 100 g of Apiose (Asen Korea, Bupyung-gu, Incheon city) and the mixture was refluxed at 90-100 ℃ for 4 hours. The extract was filtered with a filter paper (Whatmann), and then concentrated under reduced pressure using a rotary evaporator at 60 ° C. Thereafter, the concentrate was lyophilized to obtain 21.7 g of Apiose extract as a dry weight, and this was used as the experimental sample described below.

Example  One

Fermented red ginseng extracts, frog extracts and apiose extracts of Comparative Examples 1, 2 and 3 were mixed at a weight ratio of 1: 1: 1 and used as the experimental sample described below.

Example  2

Fermented red ginseng extracts, frog extracts, and apiose extracts of Comparative Examples 1, 2 and 3 were mixed at a weight ratio of 2: 1: 1 and used as the following experimental sample.

Example  3

Fermented red ginseng extracts, frog extracts, and apiose extracts of Comparative Examples 1, 2 and 3 were mixed at a weight ratio of 1: 2: 1 and used as the following experimental sample.

Example  4

Fermented red ginseng extracts, frog extracts, and apiose extracts of Comparative Examples 1, 2 and 3 were mixed at a weight ratio of 1: 1: 2 and used as the following experimental sample.

Experimental Example  One: DPPH Radical Scatters  Effect of inhibiting skin aging by measuring

To investigate the antioxidative effect of the mixture of the present invention, a method of Kilani et al. (Kilani et al al ., Environ. Toxicol. Phar. 1,1-diphenyl-2-picrylhydrazyl (DPPH; Sigma Co., USA) was assayed for its radical scavenging activity. DPPH is widely used to measure the hydrogen donating ability of antioxidants through the measurement of absorbance because it reacts with a substance having an antioxidant activity to decolorize with a dark purple color due to radical scavenging. Vitamin C (ascorbic acid), a well-known antioxidant, was used as a positive control in this experiment.

100 μl of the test substance diluted with methanol (100, 500 and 1,000 μg / ml) was added to each well of a 96-well plate, 100 μl of DPPH solution (0.2 mM DPPH in Methanol) was added, Absorbance was measured at 517 nm using a reader (BioTEK, USA). The radical scavenging ability of each test substance was calculated as the percentage of electron donating ability by the following formula.

[Equation 1]

Electron donating ability (%) = (1 - absorbance of test substance addition group / absorbance of no addition group) x 100

Test substance Electron donating ability (%) 100 [mu] g / ml 500 / / ml 1,000 ㎍ / ml Positive control group 82.4 91.7 95.2 Comparative Example 1 10.7 33.4 41.8 Comparative Example 2 50.0 69.8 77.4 Comparative Example 3 67.9 83.3 89.5 Example 1 48.5 63.1 71.9 Example 2 28.7 47.5 63.4 Example 3 52.6 65.3 76.0 Example 4 61.9 78.0 83.8

As shown in the results of the above Table 1, the group treated with fermented red ginseng (Comparative Example 1), frog (Comparative Example 2), Apiose (Comparative Example 3) or various mixtures (Examples 1, 2, 3 and 4) It was confirmed that the radical scavenging ability was lower than that of vitamin C, which is a control group. However, it was confirmed that the group treated with the test substances except fermented red ginseng generally had excellent radical scavenging ability. In particular, the group treated with the apiose-treated group and the apiose-treated group (Example 4) showed excellent radical scavenging ability .

Experimental Example  2: Measurement of cell survival rate

Prior to evaluating the skin wrinkle improvement and skin whitening efficacy, human fibroblasts (CCD-985sk cell; Korea Cell Line Bank, Seoul, Korea) and mouse melanoma cell line MTT assay was performed using B16F10 cell; Korean Cell Line Bank).

Human fibroblasts and mouse melanoma cell lines were inoculated into 96-well plates at a density of 1.5 × 10 ⁴ cells / well. DMEM containing 10% (v / v) fetal bovine serum (FBS; Gibco, USA) USA) medium at 37 ° C in a 5% CO ₂ incubator for 18 hours. Each of the test substances was diluted with DMEM medium containing no fetal bovine serum and treated for 24 hours at various concentrations (0, 25, 50, 100 and 200 / / ml). Then, 5 mg / ml of MTT (Sigma, USA) was added to a final concentration of 1 mg / ml and further cultured for 4 hours. Finally, the supernatant was discarded and 200 μl of dimethylsulfoxide (DMSO; Sigma, USA) was added to each well. The crystal violet was dissolved in a shaking incubator (TW3, Fine PCR, Korea) for 30 min. Absorbance was measured. The survival rate of human fibroblasts and mouse melanoma cells was expressed as a percentage of the survival rate of the control group not treated with the test substance as 100%.

As shown in FIGS. 1A and 1B, the cytotoxicity of each test substance was measured, and no statistically significant cytotoxicity was observed even at the highest concentration of 200 μg / ml in both human fibroblast and mouse melanoma cell lines. Based on these results, it was confirmed that the mixture of the present invention has a certain degree of safety against toxicity.

Experimental Example  3: MMP -1 wrinkle improvement effect by inhibition of expression

MMP (matrix metalloproteinase) is a group of enzymes involved in the degradation of extracellular matrix and basement membrane. It has been shown to be an interstitial collagenase, stromelysin, gelatinase gelatinase), membrane-type MMP (MT-MMP) and membrane-type MMP (membrane-type MMP) can be divided into four subfamily. It is known that MMP-1, MMP-2 and MMP-9 are mainly present in the skin, and MMP-1 is known to be most important for collagen degradation. Therefore, the present inventors tried to confirm the wrinkle-reducing effect of the test materials through inhibition of MMP-1 protein expression.

The human fibroblasts (CCD-986sk cell) in ^{24 well plate 1 × 10 5 cell} / well inoculated and, 10% (v / v) fetal bovine serum by using a DMEM medium 37 ℃, 5% CO ₂ And cultured in an incubator for 24 hours. The cells were washed once with PBS and replaced with DMEM medium lacking fetal bovine serum. Ultraviolet A was irradiated to the extent of not affecting the cells using an ultraviolet irradiator (ELC-500, Lightning Enterprisis, USA). Each test substance was treated with concentration (0, 25, 50 and 100 / / ml) and then incubated for 2 days. The amount of MMP-1 was measured using an ELISA kit (Amersham, USA). The degree of inhibition of MMP-1 expression by treatment with test substances was expressed as% of the control group in which only UV light was not irradiated and 100% of the control group was treated, and TFG-β (transforming growth factor-β) was used as a positive control.

As shown in Fig. 2, expression of MMP-1 was inhibited depending on the treatment concentration of each test substance. Therefore, it was found that the mixture of the present invention inhibits biosynthesis of MMP-1, a skin tissue degrading enzyme generated by endogenous or photoaging, thereby suppressing skin aging and improving skin wrinkles.

Experimental Example  4: Tyrosinase  Skin whitening effect through inhibition analysis

In the continuous enzymatic oxidation reaction of tyrosine to form melanin, tyrosinase is known to be the enzyme that firstly oxidizes tyrosine to dopa and the next step, dopa, is a two step reaction that is oxidized to dopaquinone. Thus, in order to confirm that the mixture of the present invention effectively inhibits tyrosinase involved in skin whitening, it was tested for tyrosinase inhibitory activity in vitro using mushroom tyrosinase assay.

200 μl of a substrate solution containing 10 mM L-DOPA dissolved in 500 μl of 67 mM sodium phosphate buffer (pH 6.8) and 100 μl of a sample solution were added to 200 μl of 110 Unit / ml mushroom tyrosinase, followed by reaction at 25 ° C for 2 minutes The absorbance of DOPA chrome produced in the reaction solution was measured at 475 nm using a microplate reader. The tyrosinase inhibitory activity of each test substance was expressed by the inhibition rate of tyrosinase by the following equation.

&Quot; (2) &quot;

Tyrosinase inhibition rate (%) = 100- (reaction absorbance of test substance / reaction absorbance of control group x 100)

Test substance Inhibition rate of tyrosinase (%) 25 [mu] g / ml 50 ug / ml 100 [mu] g / ml Control group 0 0 0 Comparative Example 1 10.2 28.9 43.0 Comparative Example 2 24.6 41.5 76.3 Comparative Example 3 20.9 43.7 54.2 Example 1 19.1 37.2 74.0 Example 2 16.5 32.4 48.8 Example 3 27.8 44.9 82.3 Example 4 26.0 42.6 83.8

As shown in the results of Table 2, tyrosinase activity was inhibited depending on the treatment concentration of the test substance, and it was confirmed that the inhibition rate of tyrosinase in Comparative Example 2, Comparative Example 3, Example 3 and Example 4 was relatively high . It was found that the extract of Copper and Apis extract in the composition of the present invention is more effective for whitening, and when they are used in combination, the whitening effect is more excellent due to the synergy effect thereof.

Experimental Example  5: Skin whitening effect through inhibition of melanin formation

The mouse melanoma cell line was inoculated in a 24-well plate at 1 × 10 ⁵ cells / well and cultured in DMEM medium containing 10% (v / v) fetal bovine serum at a concentration of 80% or more in a 5% CO ₂ incubator at 37 ° C. Lt; / RTI &gt; After washing once with PBS, each test substance was treated with concentrations (0, 25, 50 and 100 ㎍ / ml) using DMEM medium without fetal bovine serum, and then cultured for 2 days. After removing the culture medium and washing with PBS, the cells were dissolved with 1 N sodium hydroxide and absorbance was measured at 400 nm using a microplate reader.

&Quot; (3) &quot;

Melanin production inhibition rate (%) = 100- (absorbance of test substance / absorbance of control x 100)

Test substance Melanin production inhibition rate (%) 25 [mu] g / ml 50 ug / ml 100 [mu] g / ml Control group 0 0 0 Comparative Example 1 9.6 16.0 22.7 Comparative Example 2 28.1 43.7 49.3 Comparative Example 3 17.4 30.5 41.2 Example 1 22.1 31.6 39.4 Example 2 15.6 26.0 33.8 Example 3 27.4 39.8 53.5 Example 4 31.0 41.7 48.3

As shown in the results of Table 3, melanin production was inhibited depending on the treatment concentration of all the test substances, and it was confirmed that the melanin production was greatly suppressed in the group treated with copper extract and apiose extract as in the result of the previous tyrosinase inhibition experiment there was. It was also found that the inhibitory effect on melanogenesis is better when the extracts of Copper or Apiose are mixed with each other than when they are treated alone.

The present invention has been described with reference to the preferred embodiments. 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. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (12)

Fermented red ginseng, frogs and apiose extract as active ingredients. The method according to claim 1,
Wherein the fermented red ginseng, frog and apiose extract are extracts obtained by extracting each of fermented red ginseng, frozen ginseng and apiose with water, an organic solvent or a mixed solvent thereof. 3. The method of claim 2,
Wherein the organic solvent is selected from the group consisting of methanol, ethanol, propanol, isopropanol, and butanol. The method according to claim 1,
Wherein the fermented red ginseng, frog and apiose extract are 70% ethanol extracts of fermented red ginseng, frog ginseng, and apiose, respectively. The method according to claim 1,
The mixture is characterized in that the fermented red ginseng extract, frog extract and apiose extract are mixed in a weight ratio (w: w) of 1: 1: 1, 2: 1: 1, 1: 2: 1 or 1: Wherein the health functional food composition for enhancing skin beauty. The method according to claim 1,
Wherein the composition is contained in the composition at a concentration of 0.1 to 10000 g / ml. The method according to claim 1,
Wherein said skin beauty enhancing agent is for skin wrinkle improvement, skin whitening or skin aging prevention. 8. The method of claim 7,
Wherein said improvement of skin wrinkles is achieved through inhibition of MMP-1 expression. 8. The method of claim 7,
Wherein said skin whitening is achieved through inhibition of tyrosinase activity and inhibition of melanin formation. 8. The method of claim 7,
Wherein said skin aging prevention is achieved through antioxidant activity. A health functional food for enhancing skin beauty produced by the composition of any one of claims 1 to 10. 12. The method of claim 11,
Wherein said food is selected from the group consisting of beverage, meat, chocolates, foods, confectionery, pizza, ramen, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes and health supplement foods.

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