KR101913235B1 - Manufacturing method of cosmetic composition containing mixed extract of Tubercledfruit Sanicle, Setaria viridis, Aquilegia japonica Nakai, H. Hara and Dryopteris erythrosora - Google Patents

Abstract

The present invention relates to a cosmetic composition containing a mixed extract using a mixture of a tubercledfruit sanicle, a setaria viridis, an aquilegia japonica nakai, H. hara and a dryopteris erythrosora. More specifically, the present invention relates to a cosmetic composition having excellent skin moisturizing effect and skin irritation reducing effect. The present invention is excellent in the skin moisturizing effect and the skin irritation reducing effect, thereby improving various skin troubles caused by environmental pollution and stress and protecting a skin.

Description

[0001] The present invention relates to a method for preparing a cosmetic composition comprising a mixed extract of a plant extract, a herbaceous herb, a Japanese apricot, a herbaceous herb, and a red ginseng bracken. [0002]

The present invention relates to a cosmetic composition containing a mixture of extracts using a Korean red bean curd, a lady's bean curd, a radiant bean curd, and a red bean curd and a method for producing the same. More specifically, the present invention relates to a cosmetic composition having excellent skin moisturizing and skin irritation- .

The skin of a human being is exposed to a variety of environmental troubles and stresses, and the skin becomes more dull and sensitive as time goes by, and the skin moisture retention is reduced.

Especially, the skin is most affected by ultraviolet rays, specifically, it involves drying of the skin, deterioration of skin elasticity, wrinkle formation, etc., and thus the damaged skin takes a considerably long time to recover.

Accordingly, much efforts have been made to solve such various skin troubles, and as a representative method thereof, a method of utilizing a natural plant ingredient and a stimulant is being studied.

The following patent document 1 discloses a cosmetic composition containing a mixed extract of Quanjia, Rhododendron and Burdock for anti-inflammation and skin irritation mitigation. However, the cosmetic composition of Patent Document 1 is still unsatisfactory in skin moisturizing and skin irritation mitigation There was a limit.

Accordingly, there is a desperate need to develop a cosmetic composition in which skin moisturization enhancement and skin irritation alleviating effect are remarkably improved as compared with conventional cosmetic compositions for alleviating skin irritation of natural plant materials.

[Patent Literature]

Patent Document 1: Korean Patent Laid-Open No. 10-2008-0094388 (Oct. 23, 2008)

Therefore, in order to solve the above problem, the present invention has been made to solve the above-mentioned problems, and it is an object of the present invention to provide a method for producing It has been found that a cosmetic composition having an excellent skin moisturizing effect and skin irritation alleviating effect can be prepared using the mixed extract, and the present invention has been completed based on this finding.

Accordingly, one aspect of the present invention is to provide a cosmetic composition having a skin moisturizing effect and a skin irritation reducing effect.

A cosmetic composition according to one embodiment of the present invention is a baby chambandi (Tubercledfruit Sanicle) leaf, foxtail (Setaria viridis ) leaves, Aquilegia japonica Nakai, H. Hara Petals and Dryopteris lt; / RTI > erythrosora ) stalk.

In the cosmetic composition according to another embodiment of the invention, the mixture foxtail (Setaria based on 100 parts by weight of Arabidopsis leaf chambandi 50 to 150 parts by weight of a viridis leaf, 50 to 150 parts by weight of a flower of Aquilegia japonica Nakai , H. Hara , and Dryopteris 100 to 150 parts by weight of erythrosora stems may be mixed.

In the cosmetic composition according to another embodiment of the present invention, the mixed extract may be extracted from the mixture using a fermentation extraction method, an ultra-high pressure extraction method or an ultra high pressure extraction method after fermentation.

In the cosmetic composition according to another embodiment of the present invention, the mixed extract may be contained in an amount of 0.001 to 30% by weight based on the total weight of the cosmetic.

In the cosmetic composition according to another embodiment of the present invention, the cosmetic composition may be a cream, a softening water (skin), a nutritional lotion (milk lotion), or a pack.
The method of manufacturing a cosmetic composition according to one embodiment of the present invention is characterized in that the leaves of Tubercledfruit Sanicle , Setaria viridis , Aquilegia japonica Nakai, H. Hara , and Dryopteris erythrosora (B) a step of pulverizing the steamed mixture to obtain a pulverized product; (b) adding the pulverized product to a yeast strain culture medium to obtain a culture solution; and (c) removing the culture bacteria from the culture broth by centrifugation to obtain a fermented product; (d) heating the fermented product to 50 DEG C under a pressure of 1,000 MPa using an ultra-high pressure processor; (E) adding ethanol to the ultra-high-pressure-treated fermented product, refluxing for 3 hours each time for 5 hours, cooling and filtering Obtaining an extract solution (f) and the filtered extract was dissolved in a concentration tank freeze-dried fermented through the step (g) to obtain a lyophilized product was concentrated under reduced pressure - step 100 of obtaining a high-voltage extract; And extracting the fermentation-ultrahigh pressure extract with glyceryl stearate, stearyl alcohol, lanolin, polysorbate 60, sorbitan stearate, squalane, trioctanoin, dimethicone, tocopheryl acetate, carboxyvinyl polymer, glycerin, (200) mixing with water, methanol, ethanol, propanol, isopropanol, isopropanol, n-butanol, isopropanol,

The cosmetic composition according to the present invention is excellent in skin moisturizing and skin irritation mitigating effects by containing a mixed extract using a Korean red bean curd, And protects the skin.

Before describing the invention in more detail, it is to be understood that the words or words used in the specification and claims are not to be construed in a conventional or dictionary sense, It should be interpreted as meaning and concept consistent with the technical idea of the present invention. Therefore, the constitution of the embodiments described in the present specification is merely a preferred example of the present invention, and does not represent all the technical ideas of the present invention, so that various equivalents and variations And the like.

Hereinafter, preferred embodiments of the present invention will be described in detail so that those skilled in the art can easily carry out the present invention.

The present invention relates to a tubercledfruit Sanicle) leaves, foxtail (Setaria viridis) leaves, sky maebaltop (Aquilegia japonica Nakai, H. Hara) petals and Hong centipede fern (Dryopteris erythrosora stems. < / RTI >

The extract used in the present invention is preferably contained in an amount of 0.001 to 30% by weight, more preferably 0.001 to 20% by weight, and most preferably 0.01 to 10% by weight, based on the total weight of the cosmetic composition. . When the content of the extract is less than 0.001% by weight, the effect of improving the skin is weak. When the content of the extract is more than 30% by weight, the effect of increasing the content of the extract is insignificant and the safety and stability of the cosmetic composition are insufficient. It is not desirable.

The mixed extract to be used in the present invention foxtail based on 100 parts by weight of Arabidopsis leaf chambandi (Setaria 50-150 parts by weight of a leaf of the genus V. viridis , 50-150 parts by weight of petal of Aquilegia japonica Nakai , H. Hara , and 100-150 parts by weight of a stem of Dryopteris erythrosora .

In addition, Arabidopsis chambandi used in the production of mixed extract according to the present invention (Tubercledfruit Sanicle) leaf, foxtail (Setaria The leaves and leaves of Aquilegia japonica Nakai , H. Hara Petal and Dryopteris erythrosora stem are thoroughly steamed with high temperature steam prior to extraction to completely remove any residual micro-toxicity.

The mixed extracts used in the present invention can be extracted by a pulverization extraction method. The pulverization extraction is carried out by pulverizing a rice blossom starch leaf, a foxtail leaf, a foxtail flower petal, and a foxtail foxtail stem with a fine grinder, .

At this time, the pulverized pulverized material is preferably filtered through a sieve of 300 mesh or less.

Further, the mixed extract used in the present invention can be extracted by a solvent extraction method, and the solvent used in the solvent extraction method is, for example, water, an anhydrous or a lower alcohol having 1 to 4 carbon atoms (for example, methanol, ethanol , Propanol and butanol), propylene glycol, 1,3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane and mixtures thereof .

In addition, the mixed extract used in the present invention can be extracted by a fermentation extraction method. The fermentation extraction method used in the present invention is carried out by fermenting the rice blossom starch leaf, foxtail leaf, sky starch flower petal, and red bison fescue stem.

The fermentation extraction process is carried out at a temperature of 20 to 40 占 폚 under a temperature of 20 to 40 占 폚 under a temperature of 20 to 40 占 폚 in a strain selected from the group consisting of yeast, lactic acid bacteria, Bacillus subtilis, and mixtures thereof, 7 for 1 to 7 days, and may be carried out using yeast ( Saccharomyces cerevisisae ).

In addition, the mixed extract used in the present invention can be extracted by ultra-high pressure extraction. The ultra-high pressure extraction can be carried out at a pressure of 100 to 3,000 MPa at a temperature of 40 to 90 DEG C for 1 to 24 hours, preferably at a pressure of 500 to 1,000 MPa at a temperature of 40 to 60 DEG C 12 to 24 hours treatment, most preferably at a pressure of 1,000 MPa and at a temperature of 50 < 0 > C for 24 hours.

In addition, the mixed extract of the present invention may be extracted by mixing the above-described extraction method. For example, the mixed extract of the present invention can be obtained by pulverizing a crushed product obtained by finely pulverizing a rice stalks of Korean rice terra cotta, a leaf of a Japanese persimilis, a leaf of a Japanese persimilis, a flower of a Japanese persimilis and a stem of a Japanese red bug with a microorganism selected from the group consisting of yeast, lactic acid, And then extracting it at an ultra-high pressure for 1 to 24 hours at a temperature of 40 to 90 DEG C at a pressure of 100 to 3,000 MPa.

In addition, the mixed extract of the present invention may be subjected to a process such as filtration, organic solvent reflux extraction, concentration under reduced pressure, and freeze drying in order to adjust the concentration of the extract extracted by the above extraction method and increase the concentration of the active ingredient. The extract obtained by filtering, purifying and concentrating the extract by the above-described extraction method should also be interpreted as being included in the present invention.

According to a preferred embodiment of the present invention, the mixed extract of the present invention can be produced by the following method.

First, it is necessary to thoroughly wash the leaves of Japanese cypress lily, lily of the valley, petals of the sky, and the stem of the red flounder, crush them, steam it with high-temperature steam, and then prepare a microfiltration product having passed through a 300 mesh sieve.

The microfiltrate thus passed through the sieve is added to the yeast strain culture solution at a concentration of 100 g per liter. The culture can be performed by adding 1 to 4% (preferably 1 to 2%) of glucose and 0.1 to 1% (preferably 0.2 to 0.5%) of peptone as a nitrogen source as a carbon source. The culture is carried out using a 5-liter fermenter at a temperature of 20 to 40 ° C and for 1 to 7 days while maintaining the pH at 5-7.

After the culturing, the culture is centrifuged to remove the culture medium first, and the culture medium from which the culture medium has been removed is subjected to ultra-high pressure treatment at a temperature of 40 to 90 DEG C at a pressure of 100 to 3,000 MPa for 1 to 24 hours, At a temperature of 40 to 60 DEG C for 12 to 24 hours, most preferably at a pressure of 1,000 MPa and at a temperature of 50 DEG C for 24 hours.

Ethanol was added to the final 70% (v / v) ethanol aqueous solution, refluxed three times for 5 hours, cooled, and filtered through Whatman # 2 filter paper.

When the filtration is completed, the extract obtained in the extracting step is transferred to the concentration tank and concentrated under reduced pressure, followed by lyophilization. Concentration under reduced pressure can be carried out using a rotary vacuum concentrator. 700 ml of the extract is placed in a 1,000 ml round-bottomed flask, and the mixture is centrifuged under reduced pressure for 2 hours while rotating at a temperature of from 650 to 1,200 rpm at a temperature of from 50 to 70 ° C at a pressure of from 650 mmHg to 750 mmHg. Concentrate. The concentrate under reduced pressure was preliminarily frozen to reduce the temperature to minus 40 ° C, keep it at minus 40 ° C for 2 to 4 hours, cool the trap down to minus 80 ° C and maintain the vacuum at 10-30 mtorr The temperature is raised to minus 40 ° C over the next 120 minutes.

Thereafter, the temperature is maintained at 20 ° C for 240 minutes, at 0 ° C for 40 minutes, at 15 ° C for 340 minutes, at 30 ° C for 130 minutes, and at 1,300 ° C until completely dried.

The components contained in the cosmetic composition of the present invention may contain conventional auxiliary agents or carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances in addition to the above-mentioned effective ingredients as effective ingredients.

The cosmetic composition of the present invention can be formulated into a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray have.

More specifically, the cosmetic composition of the present invention can be manufactured in the form of a cream, a soft longevity (skin), a nutritional lotion (milk lotion) or a pack, and can be used as a nutritional cream, massage cream, essence, eye cream, cleansing cream, Cleansing water, spray, powder, and the like.

Hereinafter, the present invention will be described in more detail with reference to Examples. It should be noted, however, that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

<Preparation Example 1> Preparation of a solvent-mixed extract of Arabidopsis thaliana leaf, horsetail grass leaf,

500 g of Korean gazelle leaf, 500 g of leaf of panacea, 500 g of flower petals of 500 mg, and 500 g of red ginseng bark were selected and thoroughly washed with hot steam for 30 minutes. They were then pulverized and passed through a 300-mesh sieve to form a fine pulverized material. Ethanol was added to make a final concentration of 70% (v / v) ethanol aqueous solution, refluxed three times for 5 hours, cooled down, and filtered through a filter of whatman # 2. The filtered extract was concentrated under reduced pressure and lyophilized. The powder was dissolved in 50% glycerin to 2% (W / V) to prepare a solvent extract of the rice blossom starch leaf, haploid leaf, sky starter petal, and Honginebushari stem.

<Preparation Example 2> Preparation of fermented mixed extracts of Arabidopsis thunbergii leaf, Leptoptera frugiperum, Skywalker petal and Rhodiola balsam stem

500 g of Korean gazelle leaf, 500 g of leaf of panacea, 500 g of flower petals of 500 mg, and 500 g of red ginseng bark were selected and thoroughly washed with hot steam for 30 minutes. Thereafter, the pulverized material was made finely so as to pass through a 300 mesh sieve. The micronized pulverized material was added to the yeast strain culture at a rate of 10 g / l. To this, 2% glucose (W / V) and 0.5% (W / V) of peptone were added as a carbon source.

The cultures were incubated for 5 days at 37 ° C and pH 6 using a 5-liter fermentor. After culturing, the culture was centrifuged to remove the cultures first.

Ethanol was added to the resulting fermented product to make a final 70% (v / v) ethanol aqueous solution, refluxed three times for 5 hours, cooled, and filtered through a filter of whatman # 2. After the filtration was completed, the extract was transferred to a concentration tank, and the filtered extract was concentrated under reduced pressure and lyophilized. This powder was dissolved in 2% (w / v) of 50% glycerin to prepare a fermented extract of the rice blossom starch leaf, Japanese persimmon leaf, sky starch petal, and Honginbe bracken stalk.

<Preparation Example 3> Production of ultra-high pressure blended extracts of Arabidopsis thaliana leaf, Leaf leaf,

500 g of Korean gazelle leaf, 500 g of leaf of panacea, 500 g of flower petals of 500 mg, and 500 g of red ginseng bark were selected and thoroughly washed with hot steam for 30 minutes. Thereafter, the pulverized material finely pulverized so as to pass through a 300-mesh sieve was heated to 50 ° C under a pressure of 1,000 MPa using an ultra-high pressure processor, treated for 24 hours, and cooled to room temperature.

Ethanol was added to make the final 70% (v / v) ethanol aqueous solution, and the mixture was refluxed three times for 5 hours, cooled, and filtered through Whatman # 2 filter paper. After the filtration was completed, the extract obtained in the extraction step was transferred to a concentration tank, and the filtered extract was concentrated under reduced pressure and lyophilized. The lyophilized product was dissolved in 50% glycerin to a concentration of 2% (W / V) to prepare an ultrahigh-pressure extract of a Korean red bean leaf, a Japanese bean curd leaf,

<Preparation Example 4> Fermentation of the rice stalks of Korean rice terra cotta leaf, horsetail grass leaf, sky starch petal and Hongjin balsam stem - Production of ultra high pressure mixed extract

500 g of Korean gazelle leaf, 500 g of leaf of panacea, 500 g of flower petals of 500 mg, and 500 g of red ginseng bark were selected and thoroughly washed with hot steam for 30 minutes. Thereafter, the pulverized material was made finely so as to pass through a 300 mesh sieve. This pulverized product was added to the yeast strain culture at a rate of 10 g / l. To this, 2% glucose (W / V) and 0.5% (W / V) of peptone were added as a carbon source. The cultures were incubated for 5 days at 37 ° C and pH 6 using a 5-liter fermentor. After culturing, the culture was centrifuged to remove the cultures first.

The fermented product, which had been firstly removed from the culture, was heated to 50 ° C under a pressure of 1,000 MPa using an ultrahigh pressure processor, treated at an ultra-high pressure for 24 hours, and then cooled to room temperature. Ethanol was added to the final fermented product to make an aqueous 70% (v / v) ethanol solution, refluxed three times for 5 hours, cooled, and filtered through Whatman # 2 filter paper. After the filtration was completed, the extract obtained in the extraction step was transferred to a concentration tank, concentrated under reduced pressure at 50 ° C or below, and freeze-dried. Concentration under reduced pressure was carried out using a rotary vacuum concentrator. 700 ml of the extract was placed in a 1,000 ml round-bottomed flask, and concentrated under reduced pressure at 700 mmHg at 60 DEG C and 1,000 rpm for 2 hours. The concentrate was concentrated under reduced pressure, and the temperature was lowered to -40 ° C. The temperature was lowered to -80 ° C through trap freezing after holding at 40 ° C for 3 hours. The vacuum was maintained at 20m Torr, Lt; RTI ID = 0.0 &gt; 40 C. &lt; / RTI &gt;

Then, the temperature was maintained at minus 20 ° C for 240 minutes, at 0 ° C for 40 minutes, at 15 ° C for 340 minutes, at 30 ° C for 130 minutes, and then held for 1300 minutes until completely dried. The lyophilized product was dissolved in 2% of 50% glycerin to prepare a fermentation-ultra-high pressure extract of the rice terra cotta leaf, haploid leaf, sky starch petal, and Hongine bark stem.

&Lt; Preparation Example 5 >

2,000 g of Korean gazelle leaf was selected as a sample, and a Korean gazelle leaf extract was prepared according to the method of Example 1.

&Lt; Preparation Example 6 >

2,000 g of foxtail leaf was selected as a sample, and the foxtail leaf extract was prepared according to the method of Example 1.

&Lt; Preparation Example 7 >

2,000 g of the petals were extracted from the petals of Example 1,

<Preparation Example 8> Red ginseng stem extract

2,000 g of red ginseng bracken stems were selected as a sample, and a red ginseng bract stem extract was prepared according to the method of Example 1.

 &Lt; Preparation of cosmetics &

Cream type cosmetic preparations containing the mixed extracts of Preparation Examples 1 to 4 and the extracts of Preparation Examples 5 to 8 were prepared as follows (Formulation Example 1: Cream).

Examples 1 to 4 each show a formulation example of a cosmetic composition containing the mixed extract of Preparation Examples 1 to 4 as an active ingredient and Comparative Examples 2 to 5 show the case of using the extracts of Production Examples 5 to 8 as effective ingredients Of the composition of the present invention. Comparative Example 1 shows a formulation example of a cosmetic composition which does not contain any of the extracts of Production Examples 1 to 8.

ingredient
(Unit: wt%) Example
One Example 2 Example 3 Example 4 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Glyceryl
Stearate SE 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Stearyl alcohol 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 lanolin 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Polysorbate 60 1.3 1.3 1.3 1.3 1.3 1.3 1.3 1.3 1.3 Sorbitan stearate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Hardened vegetable oil 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Mineral oil 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Squalane 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Trioctanoin 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Dimethicone 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Tocopheryl acetate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Carboxyvinyl polymer 0.12 0.12 0.12 0.12 0.12 0.12 0.12 0.12 0.12 glycerin 6.0 6.0 6.0 6.0 6.0 6.0 6.0 6.0 6.0 1,3-butylene glycol 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Sodium hyaruronate 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Triethanolamine 0.12 0.12 0.12 0.12 0.12 0.12 0.12 0.12 0.12 Methyl paraben 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Spices 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.02 The extract of Preparation Example 1 3 0 0 0 0 0 0 0 0 The extract of Preparation Example 2 0 3 0 0 0 0 0 0 0 The extract of Preparation Example 3 0 0 3 0 0 0 0 0 0 The extract of Preparation Example 4 0 0 0 3 0 0 0 0 0 The extract of Preparation Example 5 0 0 0 0 0 3 0 0 0 The extract of Preparation Example 6 0 0 0 0 0 0 3 0 0 The extract of Preparation Example 7 0 0 0 0 0 0 0 3 0 The extract of Preparation Example 8 0 0 0 0 0 0 0 0 3 Purified water Balance Balance Balance Balance Balance Balance Balance Balance Balance

Experimental Example 1 Measurement of skin moisturizing effect

The skin moisturizing and enhancing effects of the cosmetic compositions of Examples 1 to 4 and Comparative Examples 1 to 5 were compared.

The skin moisturizing effect was performed by measuring skin conductivity proportional to the skin moisture content.

The experiment was divided into two groups of 20 persons to 40 persons and 25 persons who had no skin disease. The cosmetic creams of Examples 1 to 4 and Comparative Examples 1 to 5 were divided into two groups of 20 persons per one day. . The skin conductivity was measured in a corneometer (CM820 courage Khazaka electronic GmbH, Germany) under the constant temperature and humidity conditions (24 ° C, 40% humidity) before starting the application. The skin conductivity was measured at 1 week, 2 weeks, And the rate of increase was evaluated. The results are shown in Table 2 below.

sample After 1 week
(Skin conductivity increase rate,%) after 2 weeks
(Skin conductivity increase rate,%) After 3 weeks
(Skin conductivity increase rate,%) Example 1 33 38 41 Example 2 34 36 38 Example 3 36 38 42 Example 4 51 54 58 Comparative Example 1 10 13 15 Comparative Example 2 18 22 25 Comparative Example 3 18 23 25 Comparative Example 4 19 22 24 Comparative Example 5 20 23 25

As can be seen from the results of the above Table 2, Examples 1 to 4 containing the mixed extract of the present invention show that Comparative Example 1 containing no mixed extract of the present invention and Comparative Example 1 containing no mixed extract of the present invention, The skin conductivity was highest in Comparative Example 2 to 5, in which extracts of petals and Rhododendron ferns were individually contained. In particular, Example 4, which contained extracts of both fermentation and ultra-high pressure extract, had the highest skin conductivity .

From the results of Table 2, Examples 1 to 4 containing the mixed extract of the present invention show superior skin moisturizing effect as compared with Comparative Examples 2 to 5 containing a single extract of each ingredient and Comparative Example 1 containing no extract can confirm.

Experimental Example 2 Effect of Lactic Acid on Mitigation of Skin Stimulation

In order to examine the skin irritation mitigating effect of the mixed extract of the present invention, a skin irritation mitigation effect test with lactic acid was carried out by the following method.

Human fibroblasts (Korean Cell Line Bank, Korea) were cultured in T-75 flasks (Falcon, USA) until they grew to about 80%. It is moved to be 3 × 10 ⁴ (cells / well), again with a 96-well plate (Falcon, USA) and incubated for 24 hours. After culturing, cells were completely adhered to each other through a microscope to confirm whether they were growing well. Experiments were conducted using lactic acid as a skin cell stimulating agent. Lactic acid was diluted with 0.2% solution. That is, 200 쨉 l of DMEM (Sigma, USA) medium containing 0.2% lactic acid was added to each of the 96-wells, and the mixed extracts prepared in Preparation Examples 1 to 8 were adjusted to 1.0% (v / v) Respectively. At this time, no lactic acid or mixed extract of the present invention was added as a control group, and lactic acid only group was used as a control group. MTT (Sigma, USA) solution (3 mg / ml) was added to compare cell viability after 12 hours of addition of the test substance, and the cell viability was measured at 570 nm using an ELISA reader (Molecular Devices, And the cell viability (%) was calculated according to the following equation (1). The experimental results are shown in Table 3 below.

[Equation 1]

Cell survival rate (%) = ((control absorbance - test substance absorbance) / control absorbance) x 100

Test substance (% concentration) Cell survival rate (%) Control group 100 The extract (1%) of Preparation Example 1 99.9 The extract (1%) of Preparation Example 2 99.8 The extract (1%) of Preparation Example 3 99.9 The extract (1%) of Preparation Example 4 99.8 The extract (1%) of Preparation Example 5 99.9 The extract (1%) of Preparation Example 6 99.8 The extract (1%) of Preparation Example 7 99.8 The extract (1%) of Preparation Example 8 99.9 The comparison group (lactic acid, 0.2%) 0.0 Lactic acid (0.2%) + extract of Preparation Example 1 (1%) 77.5 Lactic acid (0.2%) + extract of Preparation Example 2 (1%) 79.2 Lactic acid (0.2%) + extract of Preparation Example 3 (1%) 80.5 Lactic acid (0.2%) + extract of Preparation Example 4 (1%) 84.5 Lactic acid (0.2%) + Extract of Preparation Example 5 (1%) 21.5 Lactic acid (0.2%) + extract of Preparation Example 6 (1%) 20.5 Lactic acid (0.2%) + Extract of Preparation Example 7 (1%) 30.5 Lactic acid (0.2%) + extract of Preparation Example 8 (1%) 22.5

From the results of Table 3, it can be confirmed that the addition of the mixed extract of the present invention remarkably alleviated cell growth inhibition by lactic acid treatment.

In addition, each of the extracts has little effect on the cell viability by itself, and thus the cell non-toxicity of these extracts can also be confirmed by the above Table 3.

<Experimental Example 3> Skin irritation test

Four New Zealand male rabbits weighing between 1.75 and 2.25 kg were used. Before application of the mixed extract of the present invention as a test substance, the dorsal part of the rabbit, which is the application site, was epilated, and only the epidermis was scratched on two sides of each side of the rabbit using a needle. Each extract was dissolved in distilled water at a concentration of 50 ㎎ / ㎖, 0.5 ml was administered to the abraded part and the intact part, 0.5 ml of distilled water was administered to the non - . After application, the fixture and non - fixture were fixed with non - magnetic tape and after 24 hours, residual material was removed. The skin reaction application time was observed after 24 hours and 72 hours by removing the patches. The results are shown in Table 4 below.

division Treatment A secretary change Erythema and scar edema Erythema and scar edema part Inspecting The Inspecting The Inspecting The Inspecting The time 24 72 24 72 24 72 24 72 24 72 24 72 24 72 24 72 Production Example 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Production Example 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Production Example 3 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Production Example 4 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 all 0 0

No experimental animals died during the experiment, and body weight was increased, but there was no apparent response to stimulation. After applying the mixed extract of the present invention to rabbits, the applied skin was pale brown but was not affected by the skin reaction test. As shown in Table 4, no irritation was observed in both the treated and non-treated skin, and erythema, edema, and eschar were not observed. According to the results of Table 4, it can be confirmed that the mixed extract of the present invention does not give any irritation to the skin.

<Experimental Example 4> Experiment of eye mucosa stimulation

Four New Zealand male rabbits weighing between 1.75 and 2.25 kg were used. Each extract was dissolved in distilled water at a concentration of 50 mg / ml. Each experimental animal was instilled 0.1 ml into the conjunctival sac of the left eye and treated with 0.1 ml of distilled water in the conjunctival sac of the right eye as an untreated control group. The test for the toxicity of eye was observed after 1, 2, 3, 4 and 7 days after the test substance administration, and the results are shown in Table 5 below.

division
group
Days after treatment (day) One 2 3 4 7
Production Example 1
cornea 0 0 0 0 0 Iris 0 0 0 0 0 conjunctiva 0 0 0 0 0
Production Example 2
cornea 0 0 0 0 0 Iris 0 0 0 0 0 conjunctiva 0 0 0 0 0
Production Example 3
cornea 0 0 0 0 0 Iris 0 0 0 0 0 conjunctiva 0 0 0 0 0
Production Example 4
cornea 0 0 0 0 0 Iris 0 0 0 0 0 conjunctiva 0 0 0 0 0 Non-treated control group 0 0 0 0 0 all 0

All rabbits were observed as normal in the mucosal irritation test for rabbits. During the 7-day test period after the instillation of the sample, there were no pathological changes such as tearing (redness), redness, swelling and pus. According to the results of Table 5, the mixed extract of the present invention showed no irritation to the mucous membranes.

<Other Formulation Examples>

&Lt; Formulation Example 2 > Flexible length (skin)

ingredient Content (% by weight) The mixed extracts of Preparation Examples 1 to 4 1.0 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxyvinyl polymer 0.1 Phage-12 nonylphenyl ether 0.2 Polysorbate 80 0.4 ethanol 10.0 Triethanolamine 0.1 Preservative, coloring, fragrance 0.1 Purified water Remainder

(Mixed Extract of Preparation Example 1: Formulation Example 2-1, Mixed Extract of Preparation Example 2: Formulation Example 2-2, Mixed Extract of Preparation Example 3: Formulation Example 2-3, Mixed Extract of Preparation Example 4: Formulation Example 2 -4)

&Lt; Formulation Example 3 > Nutritional lotion (milk lotion)

ingredient Content (% by weight) The mixed extracts of Preparation Examples 1 to 4 1.0 Squalane 5.0 Wax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 1.5 Liquid paraffin 0.5 Caprylic / capric triglyceride 3.0 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 0.1 Carboxyvinyl polymer 0.2 Triethanolamine 0.2 Preservative, coloring, fragrance 0.1 Purified water Remainder

(Mixed Extract of Preparation Example 1: Formulation Example 3-1, Mixed Extract of Preparation Example 2: Formulation Example 3-2, Mixed Extract of Preparation Example 3: Formulation Example 3-3, Mixed Extract of Preparation Example 4: Formulation Example 3 -4)

&Lt; Formulation Example 4 > Pack -

ingredient Content (% by weight) The mixed extracts of Preparation Examples 1 to 4 1.0 Polyvinyl alcohol 13.0 Sodium carboxymethylcellulose 0.2 glycerin 5.0 Allantoin 0.1 ethanol 6.0 Fingi-12 nonylphenyl ether 0.3 Polysorbate 60 0.3 Preservative, coloring, fragrance 0.1 Purified water Remainder

(Mixed Extract of Preparation Example 1: Formulation Example 4-1, Mixed Extract of Preparation Example 2: Formulation Example 4-2, Mixed Extract of Preparation Example 3: Formulation Example 4-3, Mixed Extract of Production Example 4: Formulation Example 4 -4)

[Additional test for skin toxicity]

The cosmetic compositions of Formulation Example 1 (Formulation Examples 1-1 to 1-4, respectively) and Formulation Examples 2-1 to 4-4 of Examples 1 to 4 were administered to 10 participants Skin toxicity clinical tests were performed.

Skin toxicity can be assessed by 1) checking whether or not it is pruritic to the area of application at the test site during the test, 2) by checking the test site itself after 12 hours after the test is completed and applying each cosmetic composition And the number of test participants who had an itch, rash, or other skin trouble was confirmed, and the results are shown in Table 9 below.

Skin Toxicity Test
division test item itch rash Other Skin Trouble

cream
Formulation Example 1-1 0 0 0 Formulation Example 1-2 0 0 0 Formulation Example 1-3 0 0 0 Formulation Examples 1-4 0 0 0

skin
Formulation Example 2-1 0 0 0 Formulation Example 2-2 0 0 0 Formulation Example 2-3 0 0 0 Formulation Example 2-4 0 0 0

Lotion
Formulation Example 3-1 0 0 0 Formulation Example 3-2 0 0 0 Formulation Example 3-3 0 0 0 Formulation Example 3-4 0 0 0

pack
Formulation Example 4-1 0 0 0 Formulation Example 4-2 0 0 0 Formulation Example 4-3 0 0 0 Formulation Example 4-4 0 0 0 all 0

As shown in Table 9, it was found that the cosmetic composition of the present invention did not cause any other skin troubles including itching and rashes to all of the test participants even when manufactured into various formulations. Is believed to support the safety of cosmetic compositions containing the mixed extract according to the present invention.

Claims (5)

A step of steam-plowing a mixture of Tubercledfruit Sanicle leaf, Setaria viridis leaf, Aquilegia japonica Nakai, H. Hara petal and Dryopteris erythrosora stem at the same weight ratio, respectively a),
(B) a step of pulverizing the steam-boiled mixture to obtain a pulverized product,
(C) a step of adding the pulverized product to a yeast strain culture solution and culturing to obtain a culture solution,
(D) a step of removing the culture bacteria from the culture solution through centrifugation to obtain a fermentation product,
(E) heating the fermented product to 50 DEG C under a pressure of 1,000 MPa using an ultra-high pressure processor, treating the fermented product at an ultra-high pressure for 24 hours and then cooling the fermented product to obtain an ultra-
(F) and (f) of adding ethanol to the ultra-high-pressure-treated fermentation product, refluxing for 3 hours each time for 5 hours, cooling and then filtering to obtain a filtrate extract
A step (100) of obtaining a fermentation-ultrahigh-pressure extract (100) through the step (g) of obtaining the lyophilized product by concentrating the filtered extract in a concentration tank and concentrating under reduced pressure and freeze-drying; And
Wherein the fermentation-ultra high pressure extract is selected from the group consisting of glyceryl stearate, stearyl alcohol, lanolin, polysorbate 60, sorbitan stearate, squalane, trioctanoin, dimethicone, tocopheryl acetate, carboxyvinyl polymer, glycerin, (200) by mixing with water, ethanol, glycol, sodium hypaloronate, triethanolamine and methyl paraben.











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