KR101081918B1 - Cosemtics composition Containing the extract of the germinated Astragalus Root Seed as active ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising the extract of germinated yellow seed seed germinated from the Astragalus extract, as an active ingredient, inhibits excellent skin elasticity and wrinkle formation, as well as exerts skin improvement and skin whitening effect. In addition, by containing natural ingredients as an active ingredient, it is possible to reduce the consumer's rejection of cosmetics that contain various existing chemicals and irritate the skin.

Aging, whitening, germinating yellow seed, composition, wrinkles, elasticity, skin, cosmetics

Description

Cosmetic composition containing extracts of germinated yellow seed seed {Cosemtics composition Containing the extract of the germinated Astragalus Root Seed as active ingredient}

The present invention relates to a cosmetic composition of the present invention, and more particularly, to a cosmetic composition containing an extract of germinated yellow seed seed germinated from the yellow seed extract as an active ingredient.

Aging can result from UV rays, stress, disease conditions, environmental factors, wounds, and activation of free radicals as they age, and when these conditions intensify, antioxidant defenses in vivo are destroyed, cells and Damage to tissues promotes adult disease and aging. More specifically, skin aging is caused by oxidizing lipids, proteins, polysaccharides, and nucleic acids, which are major constituents of skin, to destroy skin cells and tissues. In particular, the oxidation of proteins is caused by severe cuts in the skin's connective tissues such as collagen, hyaluronic acid, elastin, proteoglycans, and fibronectin, resulting in severe hyperinflammatory reactions and skin elasticity. It leads to a situation of inducement and reduced immune function.

On the other hand, Astragali Radix is the root of perennial herb belonging to legumes (Leguminosae), peeled and dried. It is usually harvested in the fall 2 years after sowing, but sometimes in the fall of the current year or 3 years. Also used. From the roots of Astragalus, about 40 kinds of tritepene glycosides (saponins), polysaccharides such as astharam I and II, flavonoids like campapol and quentintin, more than 20 free amino acids, iron, magnesium and 20 kinds Trace elements were separated. Pharmacological actions include enhanced immune activity, antiviral and antibacterial actions, promotion of nucleic acid synthesis in liver and spleen, antioxidant activity, hepatoprotective action, vasodilation and blood pressure lowering action. In oriental medicine, it is used as anti-inflammatory, antipyretic, decongestion, analgesic, tonic medicine, especially weak, cold sweating.

On the other hand, cosmetics using natural products have been developed to reduce skin irritation caused by various chemicals, etc., as well as the recent increase in consumer response to cosmetics using natural materials, the development value of cosmetic raw materials has been further increased.

Cosmetics using natural products have been described in the cosmetic composition containing the Astragalus extract (Korean Patent No. 10-084451), but its use is limited due to unclear problems of efficacy or effect at the biological level.

Therefore, the present invention, as a result of studying the applicability to cosmetics in a variety of natural products, the extract of germinated yellow seed seed germinated from Astragalus extract is excellent in skin antioxidant, collagen degrading enzyme inhibition, skin wrinkle relief, whitening, skin irritation effect In addition, it was confirmed that the skin improvement effect is also superior to the existing Astragalus extract, the purpose is to provide a cosmetic composition containing the same.

In order to achieve the above object, the present invention provides a cosmetic composition comprising the extract of germinated yellow seed seed germinated from the yellow seed extract as an active ingredient.

Hereinafter will be described in detail with respect to the problem solving means of the present invention.

To maintain healthy skin, cell membranes are protected by eliminating free radicals mediated by UV radiation or inflammatory reactions that occur during metabolic processes in the body, and already damaged cells must be regenerated by active metabolism.

Aging involves not only free radicals but also an enzyme called MMP-1. In vivo, the synthesis and degradation of extracellular matrix such as collagen is properly regulated, but the synthesis decreases as aging progresses, and the substrate is an enzyme that degrades collagen. The expression of metalloproteinase (MMP-1) is promoted, thereby reducing the elasticity of the skin and forming wrinkles. In addition, these degrading enzymes are also activated by ultraviolet irradiation.

In addition, the skin may be aged by pigmentation, and pigments affecting skin color include melanin, melazoid, carotene, hemoglobin, and the like. The most important of these is melanin, and the biggest factors affecting the biosynthesis of melanin are ultraviolet rays and hormone secretion in the body. Melanin plays an important role in preventing or damaging the skin from ultraviolet rays by absorbing or scattering ultraviolet rays. There is no special maximum absorption wavelength and absorbs the light of the whole area. Sometimes melanin itself generates free radicals, other substances are reduced or oxidized by catechol or quinone in the melanin structure, and melanin itself exhibits the properties of free radicals.

The biosynthesis of melanin is brown, eumelanin through a series of oxidation processes starting with the conversion of tyrosine, an amino acid, from the melanocytes of melanocytes to tyrosinase and conversion to dihydroxy phenylalanine. It is formed of a polymer of). This biosynthesis process is carried out in a special type of brown intracellular organelles called melanosomes. Melanosomes, including melanin granules, move from the periphery of the nucleus to the dendritic end, and then into the cytoplasm by the phagocytosis of keratinocytes. Accumulate around the nucleus of the site.

On the other hand, the cosmetic composition of the present invention is a cosmetic containing an active ingredient extract of germinated yellow seed seed germinated from Astragalus extract necessarily to prevent and improve skin aging caused by the free radicals, MMP-1, melanin, etc. To provide a composition, the extract of germinated yellow seed germinated from Astragalus extract is not only excellent in the activity of metalloproteinase (MMP-1) and melanogenesis inhibitory effect on antioxidant, skin elasticity and wrinkle formation, It is because it is also excellent in alleviating skin irritation after ultraviolet irradiation. In addition, the present invention is the fact that the extract of germinated yellow seed seed germinated from the Astragalus extract is superior to the antioxidant, inhibiting skin elasticity and wrinkle formation, preventing and improving pigmentation, and alleviating the skin irritation effect than the Astragalus extract in the following experimental example Proved.

The content of the extract of germinated yellow seed seed contained in the cosmetic composition of the present invention is not limited to a specific range, and may be a range in which the effect of the extract of germinated yellow seed described above is exerted without impairing the quality aptitude of the cosmetic composition. Preferably it is preferably included 0.001 to 30.0% by weight based on the entire cosmetic composition. When the extract of germinated yellow seed is added to less than 0.001% by weight based on the entire cosmetic composition, the skin improvement effect is insignificant, and when it exceeds 30.0% by weight, the efficiency is inferior to the amount added. In addition, the cosmetic composition according to the present invention preferably contains, in addition to the extract of germinated yellow seed, other ingredients that can give a synergistic effect to the main effect within the range that does not impair the main effect of the present invention. Do.

On the other hand, the germinated yellow seed seed germinated from the Astragalus extract is not limited to that germinated by a specific method, preferably immersing the Astragalus seed for 22 to 26 hours; It was prepared from the step of spreading the soaked yellow seed on a cotton cloth and covering the yellow seed with a cotton cloth, and then soaking the cotton cloth with 0.001 to 70% of Astragalus extract for 1 to 7 days at 15 to 30 ℃ while blocking the light. good. At this time, the sprouted yellow seed germinated from the Astragalus extract is most preferably 0.1 ~ 0.4mm shoots.

On the other hand, the germinated yellow seed extract contained as an active ingredient in the present invention is not limited to that extracted by a specific method, but preferably 10 to 95% (v / v) extracting with cold ethanol and chilling; Filtering the chilled extract to obtain a filtrate; Concentrating the obtained filtrate under reduced pressure at 50 ℃ to obtain a reduced pressure concentrate; Freezing the resultant freeze-dried product to obtain a freeze-dried product; and, the obtained freeze-dried product was extracted with a solvent selected from purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, dichloromethane and hexane, and mixtures thereof. It is preferable to obtain from the step of extracting.

On the other hand, the cosmetic composition of the present invention can be used as an example for skin whitening, anti-aging, skin irritation, the formulation is most preferably a lotion, gel, water-soluble liquid, cream, essence, oil-in-water (O / W) type And W / O type of basic cosmetic formulation, ointment oil type and oil-in-water makeup base, foundation, skin cover, lipstick, lip gloss, face powder, two-way cake, eye shadow, cheek color and eyebrow pencil It is good to be any one selected from the color cosmetics formulation which consists of a kind.

It may also be applied to the skin, optionally in the form of an aerosol, or may be in the form of a solid such as a stick. It may be used as a skin care product and / or makeup product.

Compositions according to the invention may also contain conventional auxiliaries such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, fragrances, fillers, blockers, pigments, odorants and dyes in the cosmetic field. Can be. The amounts of these various adjuvants are amounts conventionally used in the art, such as from 0.01 to 20% by weight relative to the total weight of the composition. In any case, the adjuvant and its proportions will be chosen so as not to adversely affect the desirable properties of the cosmetic composition according to the invention.

As described above, the cosmetic composition of the present invention is antioxidant, inhibition of metalloproteinase (MMP-1), inhibition of expression of metalloproteinase (MMP-1) by UV irradiation, inhibition of melamine production and skin irritation after UV irradiation. By containing the extract of germinated yellow seed seed germinated from the Astragalus extract with excellent relaxation effect as an active ingredient, it inhibits excellent skin elasticity and wrinkle formation, exerts skin improvement and skin whitening effect.

In addition, the extract of germinated yellow seed seed germinated from the Astragalus extract contained as an active ingredient in the present invention has an antioxidant, inhibiting melanin production, improving skin elasticity and wrinkles, skin irritation effect due to ultraviolet rays than the conventional Astragalus extract The cosmetic composition of the present invention can exhibit an excellent skin improvement effect compared to the cosmetic composition containing the existing Astragalus extract.

In addition, the present invention can reduce the consumer's rejection of cosmetics that contain a variety of existing chemicals to stimulate the skin by containing a natural material as an active ingredient.

Hereinafter, the configuration and operation of the present invention will be described in more detail with reference to the following examples, but the scope of the present invention is not limited to the following examples, and includes modifications of equivalent technical spirit.

Comparative example  One: Hwanggi  Extract manufacturer

After dipping, the dried Astragalus root was refluxed three times for 5 hours with an aqueous 95% (V / V) ethanol solution and cooled. After cooling, the chilled extract was filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and freeze dried at 50 ° C or lower. Astragalus extract was prepared by dispersing / dissolving using purified water to contain 15 wt% of the obtained lyophilisate.

Example 1 Preparation of Extract of Germinated Astragalus Seed Germinated from Astragalus Extract

After washing the Astragalus seeds with clean water to remove impurities, etc., it was immersed in the Astragalus extract prepared in Comparative Example 1 for 24 hours and washed with water. Spread the washed yellow seed on the floor evenly over the wire mesh covered with clean cotton cloth with sufficient thickness, cover the clean cotton cloth again, and block the light at 23 ℃ for 3 days, soak the cotton cloth with 10% Astragalus extract with solvent in purified water. Germinated.

The germinated yellow seed seeds sprouting about 0.3 mm were dried and refluxed three times for 5 hours with an aqueous 95% (V / V) ethanol solution, followed by chilling. The cold extract was then filtered through Whatman # 5 filter paper. The filtered extract was concentrated under reduced pressure and freeze-dried at 50 ° C. or lower, and the extract of germinated yellow seed was prepared by dispersing / dissolving the obtained lyophilisate using purified water to contain 15% by weight.

Experimental Example  One: NBT Antioxidant Effect Measurement Experiment

To compare the antioxidant effects of the Astragalus extract and germinated Astragalus extract obtained in Comparative Example 1 and Example 1, respectively, and to compare the antioxidant level of the germinated Astragalus seed extract, the antioxidants Retinol and BHT samples were used and measured. The NBT method was used.

The active oxygen produced by xanthine and xanthine oxidase was measured by the NBT method, and the effect of removing the active oxygen by the test substance, that is, the active oxygen scavenging effect, was measured.

The NBT method is a method of generating active oxygen by xanthine and xanthine oxidase, and measuring blue at a wavelength of 560 nm by reacting the produced active oxygen with Nitro Blue Tetrazolium (NBT). Oxygen scavenging rate was measured, and the measuring method is as follows.

To a Bayer bottle were added 2.4 ml of 0.05 M Na ₂ CO ₃ , 0.1 ml of 3 mM xanthine solution, 0.1 ml of 3 mM EDTA solution, 0.1 ml of BSA solution, and 0.1 ml of 0.72 mM NBT solution to the sample solution (Comparative Example 1 , Example 1, retinol, BHT) were added, and then left at 25 ° C. for 10 minutes. After standing, 0.1 ml of xanthine oxidase solution was added and stirred rapidly, followed by incubation for 20 minutes at 25 ° C., and 0.1 ml of 6 mM CuCl ₂ solution was added thereto to stop the reaction and absorbance (St) at 560 nm was obtained. Measured. In the blank test, distilled water was used instead of the sample solution, and the absorbance (Bt) was measured in the same manner as above. Blank of the sample solution was measured for absorbance (Bo) using the same method as above using distilled water instead of xanthine oxidase solution.

The measurement result was computed by the formula (1).

Free radical scavenging rate (%) = [1- (St-So) / (Bt-Bo)] X 100

St: absorbance at 560 nm after enzyme reaction of sample solution

Bt: absorbance at 560 nm after enzymatic reaction of blank test solution

So: absorbance at 560 nm before reaction without enzyme in sample solution

Bo: absorbance at 560 nm before reaction without enzyme in blank test solution

Sample Name Treatment concentration (%) Free radical scavenging rate (%; antioxidant effect) Comparative Example 1 (Astragalus Extract) 0.1 88 Example 1 (extract of germinated yellow seed) 0.1 93 Retinol 0.1 92 BHT 0.1 89

As a result of measuring the antioxidant effect (Table 1), it was confirmed that the excellent antioxidant effect in all the samples tested at 0.1% treatment concentration, in particular the extract of germinated yellow seed not only exerts a higher antioxidant effect than the yellow seed extract, but also the antioxidant retinol And it was confirmed that there is an antioxidant effect similar to BHT.

From the above results, the cosmetic composition containing the extract of germinated yellow seed seed germinated in the Astragalus extract of the present invention as an active ingredient was able to deduce excellent antioxidant effect.

Experimental Example  2: DPPH Antioxidant Effect Measurement Experiment

To compare the antioxidant effects of the extract of Astragalus extract and germinated Astragalus seeds obtained in Comparative Example 1 and Example 1, respectively, and to determine the antioxidant effect level of the germinated Astragalus seed extract, antioxidants such as green tea extract and vitamin E were used. Antioxidant was measured by DPPH method.

DPPH method is a method of measuring the antioxidant activity by reducing power using a free group called '2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical (DPPH)'. The free radical scavenging rate is measured at a wavelength of 560 nm by comparing the degree of absorbance decrease with that of the blank test solution.

The reagent used was 0.1mM solution of '2,2-Di (4-tert-octylphenyl) -1-picrylhydrazyl free radical (Aldrich Chem. Co., MW = 618.76)', and 61.88 mg was dissolved in methanol and used as 100 ml. The measurement method is as follows.

0.15 ml of 0.1 mM DPPH solution was added to a 96-well plate, and 0.15 ml of each sample solution (Comparative Example 1, Example 1, green tea extract, and vitamin E) was added, followed by rapid stirring and incubation at 25 ° C. for 10 minutes. Next, the absorbance (St) at 560 nm was measured. In the blank test, distilled water was used instead of the sample solution, and the absorbance (Bt) was measured in the same manner as above. Blank of the sample solution was measured by using methanol instead of 0.1mM DPPH solution in the same manner as described above.

 The result of the antioxidant effect was calculated by the equation (2).

Free radical scavenging rate (%) = (1- (St-So) / (Bt-Bo)) X 100

St: absorbance at 560 nm after free radical scavenging of sample solution

Bt: absorbance at 560 nm after free radical scavenging of blank test solution

So: Absorbance at 560 nm before reaction without free radicals in sample solution

Bo: absorbance at 560 nm before reaction without free radicals in blank test solution

Sample Name Treatment concentration (%) Free radical scavenging rate (%; antioxidant effect) Comparative Example 1 (Astragalus Extract) 0.01 62 Example 1 (extract of germinated yellow seed) 0.01 78 Green tea extract 0.01 23 Vitamin E 0.01 26

As a result of measuring the antioxidant effect (Table 2), it was confirmed that Example 1 exhibits an excellent antioxidant effect than Comparative Example 1, green tea extract and vitamin E at 0.01% treatment concentration.

From the above results, the cosmetic composition containing the extract of germinated yellow seed seed germinated in the Astragalus extract of the present invention as an active ingredient was able to deduce excellent antioxidant effect.

Experimental Example  3: in in vitro in Of metalloproteinases (MMP-1)  Inhibition rate measurement

As aging progresses, the synthesis of extracellular matrix, such as collagen, is reduced in vivo, and the expression of metalloproteinase (MMP-1), an enzyme that degrades collagen, is promoted, which not only reduces skin elasticity but also wrinkles. Is formed. In Experimental Example 3, the inhibition rates of the extracts of Astragalus extract and germinated Astragalus seeds obtained in Comparative Example 1 and Example 1 for metalloproteinase (MMP-1) in vitro were measured and compared.

Inhibition rates of substrate metalloproteinases were determined in a biochemical model, based on the use of collagen and gelatin conjugated to purified collagenase and its substrate, fluorescein (EnzChek ™ gelatinase / cola). Genase kit, molarula probe). Collagenase purified from Clostridium historiescom was supplied in the EnzChek ™ gelatinase / collagenase kit.

Collagen was purified from porcine skin and the reaction buffer consisting of DQ-collagen conjugated to fluorescein and 0.05M Tris-HCl, 0.15M NaCl, 5mM CaCl ₂ and 0.2mM sodium azide (pH 7.6) was EnzChek ™. Gelatinase / collagenase kit (Molcula Probes) was used. The extract of germinated yellow seed was tested by dissolving in the reaction buffer. Dilutions of the test extracts were incubated with 25 ug / ml of DQ-collagen and 0.1 U / ml of collagenase for 15 minutes, 45 minutes, and 120 minutes at room temperature.

As a result of measuring the inhibition rate of MMP-1 at a sample concentration of 0.02% (v / v) and 0.04% (v / v), as shown in Table 3, the excellent inhibition rate of germinated yellow seed extract against MMP-1 was shown.

In each experimental condition, the control mixture corresponding to the collagenase and DQ-collagen mixture (control; the same composition as the above experiment but no sample added) was also incubated in the same manner, and blank (Blank; Containing blank) was incubated in the presence of DQ-collagen and in the absence of collagenase, and each experiment was performed three times. After 15 minutes, 45 minutes and 120 minutes, the signal corresponding to the degradation of DQ-collagen was measured by a fluorimeter (excitation: 485 nm, emission 505 nm), and based on the 'enzyme-free blank' fluorescence value of each sample, Fluorescence values were measured. Results are expressed as percent variance for fluorescence units per sample and control.

Experimental Sample % Inhibition of MMP-1 Treatment concentration 0.02% Treatment concentration 0.04% Comparative Example 1 (Astragalus Extract) 48 72 Example 1 (extract of germinated yellow seed) 53 80 Retinol 0 6 Green tea extract 51 67

As a result of measuring the inhibition rate of metalloproteinase (MMP-1) in vitro (Table 3), the tested extracts of germinated yellow seed had anticollagenase / gelatinase activity in a dose dependent manner.

The germinated yellow seed extracts inhibited the collagen degrading activity of Clostridium collagenase by 53% and 80%, respectively, when treated with 0.02% and 0.04%, which were superior to Comparative Example 1, retinol and green tea extract at the respective treatment concentrations. It shows the inhibitory effect of MMP-1.

From the above results, it can be inferred that the cosmetic composition containing the extract of germinated yellow seed seed germinated in the extract of the present invention as an active ingredient can effectively inhibit metalloproteinases and prevent the decrease in elasticity of the skin and the formation of wrinkles. .

Experimental Example  4: after UV irradiation Metalloproteinases ( MMP -1) Expression inhibition level measurement

Metalloproteinase (MMP-1), which degrades collagen and affects skin elasticity and wrinkle formation, is also activated by UV irradiation. In Experimental Example 4, germination was carried out in Astragalus extract, an active ingredient of the present invention. After the UV irradiation, the germinated yellow seed extract and the yellow seed extract were measured and compared to the extent to inhibit the expression of metalloproteinase (MMP-1).

After UV irradiation and sample addition (Comparative Example 1, Example 1, Retinol), ELISA was performed to measure metalloproteinase (MMP-1) concentration.

Human dermal fibroblasts were irradiated with UVA at an energy of 5 J / cm 2 using a UV chamber, and UV irradiation dose and incubation time were established through preliminary experiments to establish the condition of maximal MMP-1 expression in fibroblasts. The negative control was wrapped in tinfoil and kept at the same time in the environment of UVA, UVA emission was measured using a UV radiometer. Cells during UVA irradiation were intact with previously dispensed medium and irradiated with UVA, exchanged with medium containing the sample for 24 hours, followed by culture recovery and coating on 96-well plates. The primary antibody (MMP-1 (Ab-5) monoclonal antibody and MMP-2 (Ab-3) monoclonal antibody) was treated and reacted for 60 minutes at 37 ℃. After reacting the secondary antibody anti-mouse IgG (whole mouse, alkaline phosphatase conjugated) for about 60 minutes, the alkaline phosphatase substrate solution (1mg / mlρ-nitrophenyl phosphate in diethanolamine buffer solution) was reacted at room temperature for 30 minutes and then the microplate reader was reacted. Absorbance was measured at 405 nm. As a control, one without addition of a sample was used.

Test group Treatment concentration (%) MMP-1 expression inhibition rate (%) Control group - - Example 1 (extract of germinated yellow seed) 0.1 26 Comparative Example 1 (Astragalus Extract) 0.1 21 Retinol 0.1 22

As a result of measuring the degree of inhibition of the expression of metalloproteinase (MMP-1) for each sample after irradiation with ultraviolet rays (Table 4), Example 1 showed more than 20% inhibition rate compared to the control group not treated with the sample. In particular, the inhibition rate was higher than that of Comparative Example 1 and retinol.

From the above results, the cosmetic composition containing the extract of germinated yellow seed, germinated from the yellow seed extract of the present invention as an active ingredient, can inhibit the expression of metalloproteinases even after ultraviolet irradiation to prevent the decrease in elasticity of the skin and the formation of wrinkles. It could be inferred.

Experimental Example  5: B16F1 Melanosite  Measurement of melanin production inhibitory effect

In Experimental Example 5, in order to compare the whitening effect of the extract of Astragalus extract and germinated Astragalus seeds obtained in Comparative Example 1 and Example 1, respectively, the degree of inhibition of melanin production on B16F1 melanocytes was measured.

B16F1 melanocytes used in Experimental Example 5 are cell strains derived from mice, and cells that secrete a black pigment called melanin. Each sample (Comparative Example 1, Example 1, hydroquinone, arbutin, lettuce extract, oil-soluble licorice extract) was treated in artificial culture of these cells to evaluate the degree of melanin black pigment reduction. The B16F1 melanosite used in Experimental Example 5 was used for sale from ATCC (American Type Culture Collection, Accession No .: 6323).

The melanin biosynthesis inhibitory effect of B16F1 melanocytes was measured as follows. B16F1 melanocytes were dispensed in 6-well plates at a concentration of 2 × 10 ⁶ per well and attached to cells, followed by incubation for 72 hours with each sample treated at a concentration that does not cause toxicity. After 72 hours of incubation, the cells were detached with 'trypsin-EDTA', and then the number of cells was measured and centrifuged to recover the cells.

Determination of intracellular melanin was performed by Rotan: Cancer Res . , 40: 3345-3350, 1980). After washing the cell pellet with PBS once, 1 ml of homogenization buffer solution (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added to vortex for 5 minutes to disrupt the cells. 1N NaOH (10% DMSO) was added to the cell filtrate obtained by centrifuging the crushed cells (3,000 rpm, 10 minutes), the extracted melanin was dissolved, and the absorbance of the melanin was measured at 405 nm using a microplate reader. , Melanin was quantified and melanin production inhibition rate (%) of each sample was measured. Melanin inhibition rate (%) of B16F1 melanocytes was calculated by the equation (3), IC ₅₀ value is the concentration of the substance inhibits 50% melanin production.

% Melanin biosynthesis inhibition = [(A-B) / A] x 100

A: Melanin amount in wells without sample

B: Melanin amount in the wells to which the sample was added

Sample Name Treatment concentration (%) Melanin production inhibition rate (IC ₅₀ ) Comparative Example 1 (Astragalus Extract) 0.1 0.1% Example 1 (extract of germinated yellow seed) 0.1 0.06% Hydroquinone 0.1 0.04% Arbutin 0.1 0.2% Lettuce extract 0.1 42% Usefulness Licorice extract 0.1 0.03%

As a result of measuring the melanin production inhibition rate of B 16F1 melanocytes (Table 5), Example 1 showed a melanin production inhibitory effect than Comparative Example 1, in particular, showed a melanin production inhibitory effect than the extract of lettuce extract. Hydroquinone, arbutin and oil-soluble licorice extracts used as whitening agents showed similar melanogenesis inhibitory effects.

From the above results, it was possible to infer an excellent whitening effect on the cosmetic composition containing the extract of germinated yellow seed seed germinated in the extract of the present invention as an active ingredient.

Experimental Example  6: alleviation of cytotoxicity by UV irradiation

In Example 6, the alleviation effect of cytotoxicity by UV irradiation of the extracts of Astragalus extract and germinated Astragalus seeds obtained in Comparative Example 1 and Example 1 was measured and compared.

Fibroblasts (fibroblasts) were placed in a 24-well test plate 1 x 10 ⁵ and attached for 24 hours. Each well was washed once with PBS and 500 ul of PBS was added to each well. The fibroblasts were irradiated with UV 10mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15W, Sankyo Dennki, Japan), and then PBS was removed and 10% FBS was added to the cell culture medium (DMEM). 1 ml) was added. Each sample to be evaluated (Comparative Example 1, Example 1) was treated and incubated for 24 hours. After 24 hours, the medium was removed, and 500 μl of cell culture medium and 60 μl of MTT solution (2.5 mg / ml) were added to each well, followed by incubation in a 37 ° C. CO ₂ incubator for 2 hours. After incubation, the medium was removed and 500 μl of isopropanol-HCl (0.04 N) was added thereto. After shaking for 5 minutes, the cells were lysed and 100 μl of the supernatant was transferred to a 96-well test plate, and the absorbance was measured at 565 nm using a microplate reader.

Cell survival rate (%) was measured by the equation (4) and cytotoxicity relaxation by ultraviolet light was calculated by the equation (5).

Cell survival rate (%) = [(St-Bo) / (Bt-Bo)] X 100

Bo: 565 nm absorbance of wells that only reacted with cell culture media

Bt: 565 nm absorbance of the wells that developed the colorless reaction wells

St: 565 nm absorbance of the wells that reacted the sample treated wells

Reduction rate of cytotoxicity by ultraviolet ray (%) = [1- (St-Bo) / (Bt-Bo)] X 100

Bo: cell viability of wells without UV irradiation and no sample treatment

Bt: cell viability of wells that were not irradiated with UV light

St: Cell survival rate of wells treated with UV

Sample Name Treatment concentration (%) Cytotoxic Relaxation Rate (%) Comparative Example 1 (Astragalus Extract) 0.01 33 0.02 59 Example 1 (extract of germinated yellow seed) 0.01 48 0.02 73

As a result of measuring the cytotoxic mitigation effect by UV irradiation (Table 6), Comparative Example 1 showed a cytotoxic mitigation rate of 33% at 0.01% concentration, 59% at 0.02% concentration, while Example 1 was 0.01% concentration At 48%, 0.02% at a concentration of 73% showed a cytotoxic remission rate was confirmed that the excellent cytotoxic relieving effect of Example 1.

From the above results, it can be inferred that the cosmetic composition containing the extract of the germinated yellow seed seed germinated in the extract of the present invention as an active ingredient is excellent in alleviating the skin irritation effect by ultraviolet rays.

Experimental Example  7: Inhibition of Inflammatory Cytokine Expression by Ultraviolet Irradiation

Experimental Example 7 measured the degree of inhibition of inflammatory cytokine expression expressed by UV irradiation in order to compare the anti-inflammatory effects on the extract of Astragalus extract and germinated Astragalus seeds obtained in Comparative Example 1 and Example 1, respectively.

Fibroblasts isolated from human epidermal tissue (Fibroblast) were placed in a 24-well test plate ⁵ x 10 ⁴ and attached for 24 hours. Each well was washed once with PBS and 500 μl of PBS was added to each well. The fibroblasts were irradiated with UV 10 mJ / cm 2 using an ultraviolet B (UVB) lamp (Model: F15T8, UV B 15W, Sankyo Dennki, Japan), after which PBS was removed and FBS was not added to the cell culture medium (DMEM). Medium) 350 μl was added. Each sample to be evaluated (Comparative Example 1, Example 1) was treated, and then cultured for 5 hours. After incubation, 150 μl of the culture supernatant was taken to quantify IL-1α to determine the inflammatory cytokine expression inhibitory effect on each sample. The amount of IL-1α was quantified using Enzyme-linked Immunosorbent Assay (ELISA), and the production rate of IL-1α was calculated by Equation 6.

Inhibitory rate of inflammatory cytokine expression (%) = [1- (St-Bo) / (Bt-Bo)] X 100

Bo: amount of IL-1α produced in wells not irradiated with UV

Bt: amount of IL-1α produced in wells that were not irradiated with UV light

St: Production of IL-1α in Wells Treated by Ultraviolet Irradiation

Sample Name Treatment concentration (%) Inhibitory rate of inflammatory cytokine expression (%) Comparative Example 1 (Astragalus Extract) 0.01 37 0.02 62 Example 1 (extract of germinated yellow seed)
0.01 55 0.02 79

As a result of measuring the inhibitory effect of inflammatory cytokine expression by UV irradiation (Table 7), Comparative Example 1 showed the inhibition rate of 62% inflammatory cytokine expression at the concentration of 37% and 0.02% at 0.01% concentration, while Example 1 was 0.0 Inhibiting the expression of inflammatory cytokine expression of 79% at 55%, 0.02% concentration at .1% concentration was confirmed that Example 1 effectively inhibits the occurrence of inflammation even at low concentrations.

From the above results, it can be inferred that the cosmetic composition containing the extract of the germinated yellow seed seed germinated in the extract of the present invention as an active ingredient has an excellent anti-inflammatory effect.

Example  2 and Comparative example  2: Cosmetics  Produce

In Example 2 and Comparative Example 2, a cosmetic was prepared using the extract of germinated yellow seed seeds obtained in Example 1 above.

The cosmetics prepared were in the form of a cream, the composition of which is shown in Table 8.

The 'Na' phase recorded in Table 8 was heated and stored at 70 ° C. The preliminary emulsification was performed by adding the 'ga' phase to the preserved 'na' phase, and then uniformly emulsified with a homomixer, and then gradually cooled. Cream (Example 2, Comparative Example 2) was prepared.

Unit: weight% Raw material Example 2 Comparative Example 2 end Stearyl alcohol 8 8 Stearic acid 2 2 Stearic acid cholesterol 2 2 Squalane 4 4 2-octyldodecyl alcohol 6 6 Polyoxyethylene (25 mol addition) alcohol ester 3 3 Glyceryl Monostearic Acid Aster 2 2 I Example 1 (extract of germinated yellow seed) 2 - Propylene glycol 5 5 Purified water Rest about 100 Rest about 100

Experimental Example  8: Measure skin elasticity improvement

Experimental Example 8 measured the skin elasticity improvement effect on the cream prepared in Example 2 and Comparative Example 2.

The experimenters were 15 women aged 20-35 years, and the cream of Example 2 was applied to the right side of the face and the cream of Comparative Example 2 was applied to the left side of the face twice a day for three consecutive months.

After completion of the experiment, the skin elasticity improvement effect was measured using a skin elasticity measuring instrument (cutometer SEM 575, C + K Electronic Co., Germany) before and after using the product for 3 months.

The experimental results are described as 'ΔR8' values of 'Cutometer SEM 575' in Table 9 below, where 'R8' values indicate the nature of viscoelasticity of the skin.

Experimental product Skin elasticity effect (ΔR8) Example 2 0.24 Comparative Example 2 0.15

n = 15, p <0.05

As a result of measuring the skin elasticity effect (Table 9), Example 2 which is a cream containing germinated yellow seed extract was confirmed that the effect of improving the skin elasticity of the experimenter compared to Comparative Example 2.

From the above results, it was confirmed that the cosmetic composition containing the extract of germinated Astragalus seeds germinated in the Astragalus extract of the present invention as an active ingredient has an excellent skin elasticity effect.

Experimental Example  9: Measure skin whitening effect

Experimental Example 9 measured the skin lightening effect for each cream prepared in Example 2 and Comparative Example 2.

The experimenter was 15 women aged 20-35 years and applied the cream of Example 2 to the right side of the face and the cream of Comparative Example 2 to the left side of the face twice a day for 2 consecutive months. After completion of the test, the change in color brightness (ΔL) was measured by using an image analyzer and chromameter (Minolta CR300) on the left and right sides of the face. The effect was measured according to the following classification, and the whitening efficacy was evaluated by classifying it into 7 grades (-3: very worse, -2: worse, -1: slightly worse, 0: no change, 1: Slightly improved, 2: improved, 3: highly improved)

Change in brightness of skin color (ΔL) Objective evaluation of the skilled person Subjective Evaluation of the Subject Experimental product Example 2 Comparative Example 2 Example 2 Comparative Example 2 Example 2 Comparative Example 2 medium 5.0 2.2 2.7 1.8 2.8 1.6

As a result of measuring the skin whitening effect (Table 10), Example 2, which is a cream containing the extract of germinated yellow seed, showed better results than Comparative Example 2 in all of the brightness change of the skin color, the objective evaluation of the skilled person, and the subjective evaluation of the subject. It was confirmed that the effect is excellent.

From the above results, it was confirmed that the cosmetic composition containing the extract of germinated yellow seed seed germinated in the extract of the present invention as an active ingredient has an excellent skin whitening effect.

Claims (9)

Cosmetic composition characterized in that it contains an extract of germinated yellow seed seed germinated from the Astragalus extract as an active ingredient. The method of claim 1, The extract of germinated yellow seed, Cosmetic composition, characterized in that contained 0.001 to 30.0% by weight based on the entire cosmetic composition. The method of claim 1, The extract of germinated yellow seed, Sprouted with Astragalus seeds, which were soaked for 22 to 26 hours in Astragalus extract, covered with a cotton cloth again, and germinated while soaking 0.001 to 70% of Astragalus extract in cotton cloth at 1 to 7 days at 15 to 30 ° C while blocking light. Cosmetic composition, characterized in that the extract of the yellow seed. The method of claim 3, The extract of germinated yellow seed, Cosmetic composition, characterized in that the extract of the germinated yellow seed germinated so that the shoots come out about 0.1 ~ 0.4mm. The method of claim 1, The extract of germinated yellow seed, Germinated Astragalus seeds germinated from Astragalus extract were extracted with 10-95% (v / v) ethanol aqueous solution at 70-85 ° C. and cooled down, and then the extracted extract was filtered and the filtrate obtained was concentrated under reduced pressure at 50 ° C. or lower. Freeze-dried the resulting vacuum concentrate, and the obtained lyophilized extract was extracted with an extraction solvent selected from purified water, methanol, ethanol, glycerin, ethyl acetate, butylene glycol, dichloromethane and hexane and mixtures thereof. Cosmetic composition, characterized in that the extract of the yellow seed. The method of claim 1, Cosmetic composition, Cosmetic composition, characterized in that it is used for skin whitening. delete The method of claim 1, Cosmetic composition, Cosmetic composition, characterized in that it is used for skin irritation relief. The method according to any one of claims 1 to 6, Formulation of the cosmetic composition, Basic cosmetic formulation consisting of lotion, gel, water-soluble liquid, cream, essence, oil-in-water (O / W) and water-in-oil (W / O) type, ointment, oil-in-water and water-in-oil makeup base, foundation, skin cover, lipstick Cosmetic composition, characterized in that any one selected from lip gloss, face powder, two-way cake, eye shadow, teak color and eyebrow pencil formulations.