KR100844516B1 - Cosmetic composition containing astragalus root extract - Google Patents

Abstract

A cosmetic composition comprising an extract of Astragalus Radix is provided to be useful for anti-oxidation, skin whitening and wrinkle improving without skin side effects. A cosmetic composition for skin whitening or improving wrinkle comprises 0.001-10 wt.% of an extract of Astragalus Radix as an effective ingredient. A method for preparing the cosmetic composition comprises the steps of: (a) extracting the Astragalus Radix or crushed Astragalus Radix with C1-4 alcohol or an alcohol aqueous solution; (b) adding at least one selected from the group consisting of carbohydrolase, pectinase, and cellulase to an extract obtained from the step(a) and shake-culturing it; and (c) adding an organic solvent for fractionation to a culture material obtained from the step(b) to fractionate an extract including calycosin, formononetin or a mixture thereof.

Description

Cosmetic composition containing Astragalus extract {Cosmetic composition containing Astragalus Root extract}

Figure 1 is a high-speed liquid chromatogram of the Astragalus extract containing the isoflavone compounds calicocin and formmononetin as an active ingredient extracted and separated from the root of Astragalus.

The present invention relates to a cosmetic composition containing an extract of the increase in the content of the active ingredient and the use of such a cosmetic composition.

As you age, you may experience various physiological and physical changes. In particular, menopausal women cause significant changes mentally and physically, mainly due to changes in the female hormone estrogen (estrogen). In addition, the estrogen secretion deficiency is known to increase the onset of heart disease, osteoporosis, aging, and other cancers.

In order to improve these menopausal diseases, products such as medicines, health foods, and cosmetics containing estrogen-based substances have been developed. However, long-term use of hormone-based substances is known to cause irregular bleeding and side effects such as breast and uterine cancer. Therefore, a lot of research has been made to find a substance that can substitute for an estrogen-based substance. As a result, phytoestrogen, phytoestrogen derived from plants, has been found as a substance having an action similar to estrogen.

Vegetable-like hormone components include isoflavones such as diedzein, genistein, formmononetin, calicosin and biochanin A; Cumestan, such as coumestrol; Lignans such as matairesinol; Apigenin, luteolin, kaempferol and the like are divided into flavonoids. Since the content ratio of the components and components contained in each plant is different, the degree of bioactivity and efficacy of the extract may also vary depending on the species, the extraction site, and the extraction method of the plant.

Astragali Radix peels off the bark of perennial herbaceous Astragalus membranaceus Bunge, which belongs to the legume (Leguminosae), and its roots are dried. In addition, the 'new farming menarche' is particularly effective for the elderly and has a function of prolonging the lifespan and is called Astragalus (No Seung-hyun, Hwang-in Hwang, TV Main Herb, p332, 1999; , 2003), in Korea, it has been used as a tonic after the ginseng in the private sector since modern times (Hyundai Pharmaceutics, Research Institute of Pharmacognosy, Hakchangsa, 2000; Herbology, Culture and Culture History, 2000; Herbal Medicine, Herbal Medicine School Compilation Committee, Dongmyungsa, 2001) .

Prescriptions containing Astragalus include Huanggigunjungtang, Huanggi Jiomultang, Jeopjeondaebotang, and Banggi Hwanggitang, and their physiological activities include blood pressure lowering, diuretic, tonic, immune enhancing, and anti-inflammatory effects. (Chinese Chinese Dictionary, Sang-Hak University of Science, vol. 1, 1985; Hikino, H. et al., Planta Med., 30: 297-302, 1976; Tomoda, M. et al., Phytochemistry 31 (1): 63- 66, 1992; Zang, Y. et al., Acta. Pharm. Sin., 19: 333-337, 1994).

Astragalus also contains saponins from asragalosides and polysaccharides from the astragalalan family, camphorol, quercetin, isorhamnetin, calicosin, formmononetin, rhamnocitrin, and coumatake. Flavonoids such as kumatakenin, asparagine, glutamic acid, proline, β-aminobutyric acid, arginine and alanine are known to contain amino acids (Kitagawa, I. et al., Chem. Pharm. Bull., 31 (2)) : 709-722, 1983).

Therefore, the technical problem to be achieved in the present invention is to provide a cosmetic composition containing the extract of Astragalus and the cosmetic use of such a cosmetic composition, more preferably the Astragalus extract with improved antioxidant, skin whitening or wrinkle improvement effect of cosmetic It is to provide a cosmetic composition containing a.

In order to achieve the above technical problem, the present invention provides a cosmetic composition for antioxidant, skin whitening or anti-wrinkle, characterized in that containing the extract as an active ingredient, more preferably the extract is calicosin ( calycosin)), formononetin (formononetin) or an extract of increased content thereof provides an antioxidant, skin whitening or anti-wrinkle cosmetic composition.

More preferably, the present invention is to extract the calicosin, formonenetine or the extract thereof is increased (S1) Astragali Radix or pulverized sulfur group using an alcohol or an aqueous solution of 1 to 4 carbon atoms step; (S2) adding at least one selected from the group consisting of carbohydrate, pectin hydrolase and cellulose hydrolase to the extract of step (S1) and shaking culture; And (S3) provides a cosmetic composition for antioxidant, skin whitening or anti-wrinkle, characterized in that prepared by the manufacturing method comprising the step of adding the fractionated organic solvent to the culture of the step (S2).

Hereinafter, the cosmetic composition for antioxidant, skin whitening or wrinkle improvement containing the Astragalus extract of the present invention will be described in more detail.

The present invention provides a cosmetic composition for antioxidant, skin whitening or anti-wrinkle, characterized in that it contains Astragalus extract, more preferably calicosin, formmononetin or astragalus extract with an increased content thereof.

As the content of calicosin, formonenetine or their contents in Astragalus extract increases, antioxidant, skin whitening or anti-wrinkle effect is increased, and these effective ingredients can be increased in the following way.

Calicocin, formmononetine or the sulfur extract of which the content thereof is increased in the cosmetic composition of the present invention is extracted using an alcohol or aqueous solution having 1 to 4 carbon atoms (S1) Astragali Radix or ground sulfur. Making; (S2) adding at least one selected from the group consisting of carbohydrate, pectin hydrolase and cellulose hydrolase to the extract of step (S1) and shaking culture; And (S3) adding the organic solvent for fractionation to the culture of step (S2) and fractionating the same.

The manufacturing method for increasing the cosmetic efficacy of the Astragalus extract is extracted using (C1) alcohol or alcoholic solution containing 1 to 4 carbon atoms, such as ethanol aqueous solution, aqueous methanol solution, isopropanol aqueous solution, butanol aqueous solution (S1) or dried and crushed sulfur group It includes a step, and considering the enzymatic treatment process or toxicity of the subsequent step (S2) step is more preferable ethanol solution.

In addition, the alcohol content of the alcohol aqueous solution is preferably about 50% by weight to about 100% by weight, more preferably about 75% by weight to about 95% by weight.

Extraction of the step (S1) is preferably carried out at 60-90 ℃, if the temperature is too low extraction yield, the extraction time is long, if extracted at too high temperature may be denatured of the Astragalus extract components have.

The preparation method for enhancing the cosmetic efficacy of the Astragalus extract is also (S2) added to any one or more selected from the group consisting of carbohydrate, pectin hydrolase and cellulose hydrolase to the extract of step (S1) and shaking Culturing. Such culture is preferably carried out at 40-70 ℃.

The method also includes the step of concentrating the extract of step (S1) between step (S1) and (S2) to increase the efficiency of step (S2) using the enzyme, and dissolving the concentrate again with an ethanol aqueous solution. Preferably, the ethanol aqueous solution is preferably 5 to 30% by weight, and more preferably about 10% by weight ethanol aqueous solution, the content of ethanol is lower than the extraction step of (S1) step.

The preparation method for enhancing the cosmetic efficacy of the Astragalus extract also includes the step of fractionating (S3) by adding an organic solvent for fractionation to the culture of step (S2), and the organic solvent for fractionation is ethyl acetate, di Ethyl ether, trichloromethane, butanol and the like can be used alone or in combination.

The method also comprises the steps of (A) (S2) between the step (S2) and (S3) step (S2) fractions or fractional concentration using an organic solvent for fractionation; And (b) adding at least one selected from the group consisting of carbohydrate hydrolase, pectin hydrolase and cellulose hydrolase to the fraction or fraction concentrate in step (a) and shaking culture. The content of cosmetically useful ingredients in the extract can be further increased. When the steps (A) and (B) are added in the preparation method of the present invention, the contents of calicosin and formonenetine, which are active ingredients according to the object of the present invention, increase dramatically.

In the cosmetic composition of the present invention, the content of the Astragalus extract is preferably 0.001 to 10% by weight relative to the total weight of the cosmetic composition, and the cosmetic composition containing the Astragalus extract is used in cosmetics, essences, creams, packs, foundations, It can be prepared in various formulations of basic products such as make up bases, color products and hair products. Specifically, it can be applied in various properties such as liquid, cream, paste and solid, and can be applied to a conventional cosmetic preparation method.

The cosmetic composition of the present invention is effective in anti-oxidation, skin whitening, wrinkle improvement, and the like, and in particular, the cosmetic composition containing the calicosin, formmononetine or the increased contents of Astragalus extract usually contains an Astragalus extract. It is more effective for antioxidants, skin whitening, wrinkle improvement, etc. than the cosmetic composition.

Hereinafter, specific examples, such as examples and experimental examples, will be described to help understanding of the present invention. However, the following examples may be modified in many different forms and should not be construed as limited to the examples set forth herein.

In the present invention, the Astragalus membranaceus Bunge was used as the main origin of the Cheongbuk Jecheon known as the main production site, in the following example was used in the agricultural and forestry medicine (Korea) in the peeled and cut state. Astragalus extract was prepared as in Examples 1 to 9 below.

<Extraction Example 1-6> Preparation of Astragalus Extract

Peeled and chopped Jecheonsan Astragalus herb (50 g) was ground, 75% by weight aqueous ethanol solution (500 mL) was added and heated and extracted at 60-90 ℃ and filtered. The crude ethanol extract obtained by repeating the extraction process twice was concentrated to obtain a crude extract (10 g). This was dissolved again with aqueous ethanol solution (400 mL) and added to carbohydrate hydrolase mixture (Viscozyme), pectin hydrolase (Pectinex) and cellulose hydrolase (Celluclast) from Denmark by Novozymes. Shake culture at -70 ℃. Distilled water (100 mL) was added thereto, an extract solution and the same amount of ethyl acetate were added, and the mixture was concentrated to give Examples 1 to 6. Examples 1 to 6 depend on the type and treatment concentration of the enzyme, and the yield of the Astragalus extract according to each example is described in Table 1 below.

Extraction Example Enzyme treatment Yield (mg) One No treatment 445 2 Carbohydrate Hydrolase Mixture 186 3 Pectin hydrolase 381 4 Cellulose hydrolase 269 5 Carbohydrate Hydrolase Mixture + Pectin Hydrolase 190 6 Carbohydrate Hydrolase Mixture + Cellulose Hydrolase 162

<Example 7-8> Preparation of Astragalus Extract

Distilled water (200 mL) was added to the crude extract (10 g) obtained in the extraction examples 1 to 6, and the same amount of n-butanol or ethyl acetate and n-butanol were added twice, and concentrated. Each fraction (2.5 g, 3.5 g) was dissolved again in 10% ethanol aqueous solution (100 mL, 150 mL) and carbohydrate hydrolase mixture was added and shaken at 40-70 ° C. Distilled water (200 mL) was added thereto, an extract solution and the same amount of ethyl acetate were added, and the mixture was concentrated to obtain Examples 7 to 8. The yield of the Astragalus extract according to each Example was described in Table 2 below.

Extraction Example Fractional layer Yield (mg) 7 n-butanol fractionation layer 185 8 Ethyl acetate fractionation layer + n-butanol fractionation layer 177

Extraction Example 9 Preparation of Astragalus Extract

The crude extracts (10 g) obtained in the extraction examples 1 to 6 were subjected to one time enzymatic treatment under the conditions of extraction example 2, and then the same amount of ethyl acetate and n-butanol were sequentially fractionated and concentrated. The fractions were dissolved again in 10% ethanol aqueous solution (100 mL) and carbohydrate hydrolase mixture was added to carry out the reaction under the same conditions as above. In general, Example 9 was obtained by performing the enzyme treatment twice. Enzyme treatment was performed twice or more as necessary in the whole extraction process.

Experimental Example 1 Analysis of Calicosin and Formonenetine Content in Extraction Examples 1-9

The contents of calicosin and formonenetine of the Astragalus extract obtained in the extraction examples 1 to 9 were analyzed three times using high performance liquid chromatography (HPLC). At this time, the high-performance liquid chromatography analysis conditions are shown in Table 3 below, high-speed liquid chromatography and content of calicosin and formonenetine are described in Figure 1 and Table 4, respectively.

division Condition Chromatography Dionex Summit HPLC column Luna C ₁₈ (250mm × 4.6mm × 0.5um) Detector Ultraviolet absorption spectrophotometer (wavelength 260 nm) Flow rate 1.0 mL / min Mobile phase Gradient elution of 0.1% acetic acid solution dissolved in acetonitrile and 0.1% acetic acid solution dissolved in water

Extraction Example Calicosin content (%) Formononetine content (%) One 0.65 ± 0.02 0.61 ± 0.01 2 5.75 ± 0.05 2.95 ± 0.04 3 3.29 ± 0.04 1.96 ± 0.03 4 4.49 ± 0.04 2.27 ± 0.03 5 5.99 ± 0.06 2.73 ± 0.04 6 6.37 ± 0.05 2.76 ± 0.04 7 6.01 ± 0.07 1.75 ± 0.02 8 6.78 ± 0.06 1.27 ± 0.03 9 16.86 ± 0.15 5.44 ± 0.09

The results in Table 4 for the extraction Examples 1 to 9 showed that the content ratio of calacin and formononetin, which is an isoflavone component of the Astragalus extract, is different depending on the extraction method. In addition, it can be seen that the content of calicosin and formonenetine was increased in the enzyme treatment examples 2 to 9 compared to the extraction example 1 without the enzyme treatment.

N-butanol (Extracting Example 7) or ethyl acetate and n-butanol (Extracting Example 8) before the enzymatic treatment. Although there is no particular improvement in comparison, the extraction example 8 was measured to have a higher content of calicosin and formonenetine than the extraction example 7.

In the case of extraction Example 9, which was repeated two times of enzyme treatment, the contents of calicosin and formonenetine were drastically increased to two to three times higher than that of extraction Examples 2 to 8, which was the case of one time of enzyme treatment.

Experimental Example 1 Antioxidant Test

Free radical scavenging activity test and free radicals (superoxide anion) to determine the antioxidant effect of the skin aging improvement effect assay test for each of the extracts of Astragalus extract prepared in each of the extraction examples 1 to 8 of the present invention ) Scavenging activity test. The free radical scavenging test is a modification of the method of Kim et al. (Kor. J. Pharmacogn., 24 (4): 299-303, 1993), which is a stable free radical DPPH (1,1-diphenyl-2-picryl- hydrazyl radical, Fluka) was used.

First, each extracted sample obtained according to the extraction examples 1 to 8 in 1 mL of 0.2 mM DPPH solution was diluted in methanol at an appropriate concentration, mixed in 2 mL, left at room temperature for 10 minutes, and the absorbance was measured at 517 nm. .

The control in the free radical elimination test was set in the same manner by adding methanol instead of the sample solution, it was set to obtain the respective color correction value for the extracted sample and the control by adding methanol instead of the DPPH solution.

By using the following equation 1 to calculate the free radical removal rate as a numerical value is shown in Table 5 below. In Table 5, SC ₅₀ is the concentration of the sample required to achieve 50% of the free radical scavenging rate, which is a value used for comparison, and the smaller the value, the higher the antioxidant activity.

The active oxygen scavenging test is a modification of the method of Noro et al. (Chem. Pharm. Bull., 31: 3984-3987, 1983), and the active oxygen scavenging activity is the xanthine / xanthine oxidase (Sigma) enzyme reaction. The change in absorbance due to oxidation of nitroblue tetrazolium (NBT) by active oxygen was measured using an active oxygen generator.

2.4 mL of Na ₂ CO ₃ , 0.1 mL of xanthine (Sigma), 0.1 mL of EDTA (ethylenediamine tetraacetic acid), 0.1 mL of BSA (bovine serum albumin, Sigma), 0.1 mL of NBT and 0.1 mL of sample were mixed well. Let stand for a minute. After adding 0.1 mL of Xanthine xanthine oxidase and reacting at 25 ° C. for 20 minutes, 6 mM CuCl ₂ was added to stop the reaction, and the absorbance was measured at 560 nm.

Meanwhile, the control group in the active oxygen scavenging activity test was determined by adding purified water instead of the sample solution, and adding purified water instead of the xanthine oxidase solution to obtain respective color correction values for the extracted sample and the control group.

By using the following equation (2) to calculate the oxygen free radicals are shown in Table 5 below. In the following Table 5, IC ₅₀ is the extract sample concentration required to achieve 50% of the oxygen scavenging rate, the value used when comparing relatively, the smaller the value means that the antioxidant activity is stronger.

sample Free radical scavenging rate, SC ₅₀ (mg / mL) Free radical removal rate, IC ₅₀ (mg / mL) Extraction Example 1 0.696 1.013 Extraction Example 2 0.382 0.387 Extraction Example 3 0.400 0.587 Extraction Example 4 0.400 0.563 Extraction Example 5 0.230 0.340 Extraction Example 6 0.284 0.223 Extraction Example 7 0.270 0.103 Extraction Example 8 0.236 0.090

The results of Table 5 indicate that the free radical and free radical removal rate of 0.236 mg / mL and 0.090 mg / mL, respectively, in the extraction example 8 having high content of calicosin with respect to the antioxidant effect of Astragalus showed relatively excellent antioxidant effects. . In addition, as a whole, the content of calicosin and formmononetin was lower than that of extraction examples 3 to 4, which showed more excellent antioxidant effect. Therefore, the antioxidant effect of Astragalus extract was determined to be advantageous in the case of high content of calicosin and formonenetine as an active ingredient.

Experimental Example 2 Tyrosinase Inhibition Test

In order to test the whitening effect of the Astragalus extract samples obtained according to the respective extraction examples of the present invention, the degree of inhibition of the function of an enzyme called tyrosinase was evaluated. Tyrosinase is an enzyme that accelerates the oxidation of tyrosine in vivo and helps melanin production. This extraction example measures the degree of inhibition of the function of this enzyme to inhibit the tyrosine oxidized to form a black polymer called melanin (Pomerantz SH, J. Biochem., 24: 161-168, 1966). Application was made to determine the whitening effect.

Inhibitory activity of tyrosinase of each extract sample was measured as follows. 0.9 mL of sample, 1.0 mL of 0.1 M phosphate buffer (pH 6.8) and 1.0 mL of 1.5 mM L-tyrosine solution were added thereto, and then maintained at 37 ° C. for 10 minutes. After adding 0.1 mL of mushroom tyrosinase (1,500 units / mL) and reacting for 10 minutes at 37 ° C, absorbance was measured at 475 nm using a UV-Vis spectorphotometer (Shimadzu) to measure the inhibition rate against tyrosinase. It was.

The control group for the tyrosinase activity inhibition test was measured in the same manner by putting a buffer solution instead of the sample solution, and was set to the case where each color correction value was obtained for the Astragalus extract sample and the control group was added to the buffer solution instead of tyrosinase. To calculate the tyrosinase inhibition rate by using the following Equation 3 shown in Table 6 below. In Table 6, IC ₅₀ is the concentration of the extract sample required to achieve 50% tyrosinase inhibition rate, the value used as a comparative comparison, the smaller the value indicates that the higher the inhibition rate.

sample Tyrosinase Inhibition Rate, IC ₅₀ (mg / mL) Extraction Example 1 0.733 Extraction Example 2 0.096 Extraction Example 3 0.099 Extraction Example 4 0.066 Extraction Example 5 0.079 Extraction Example 6 0.073 Extraction Example 7 0.050 Extraction Example 8 0.046

The results of Table 6 showed a relatively excellent tyrosinase inhibitory effect in the extraction Example 8 having a high content of calicosin as in the antioxidant effect test results of Table 5. In addition, the tyrosinase activity inhibition rate was higher when the content was higher than the extraction examples 3 to 4 with less content of calicosin and formmononetin as a whole. Therefore, the effect of inhibiting tyrosinase activity as a whitening assay of the Astragalus extract was determined to be advantageous in the case where the content of calicosin and formonenetine was high as an active ingredient.

Experimental Example 3 Elastase Inhibition Test

Elastase inhibition activity test (elastase inhibition activity test) was carried out as a test for inhibiting and improving the effect of skin wrinkles on the Astragalus extract samples obtained according to each of the extraction examples 1 to 8 of the present invention. Elastin (elastin) Ella stay's substrate, N- succinyl a method of measuring the enzyme activity of ellagic stay's to decompose the carbonyl - (Ala) ₃ ρ- nitroaniline _{(N-succinyl- (Ala) 3} ρ-nitroaniline, Sigma) is a method of measuring the elastase activity by measuring the absorbance at the wavelength of 405 nm changes in color generated by the decomposition of ρ-nitroaniline. PH 8.0, 0.267 M Trizma-HCl (Sigma), substrate solution 8.8 mM N-Succinyl- (Ala) ₃ ρ-nitroaniline, enzyme solution using pig pancreatic elastase at a concentration of 10 ug / mL (Sigma) It was. After mixing 60 ul of buffer solution, 20 ul of substrate solution, and 100 ul of the diluted sample solution of the Astragalus extract samples obtained from each of the extraction examples 1 to 8 according to the concentration, 20 ul of the enzyme solution was added to a 15 ° C. water bath in a 25 ° C constant temperature water bath. After reacting for a minute, the amount of ρ-nitroaniline was measured at a wavelength of 405 nm using a microplate reader.

On the other hand, the control for measuring the elastase activity was measured by the same method to put purified water instead of the Astragalus extract sample, was set the case of obtaining a color correction value for each of the purified water instead of enzyme solution.

Using Equation 4 to calculate the elastase inhibition rate as a numerical value is shown in Table 7. In Table 7 below, IC ₅₀ is the concentration of the extract sample required to achieve 50% elastase deactivation rate, the value used as a comparative comparison, the smaller the value indicates that the higher the inhibition rate.

sample Elastase Inhibition Rate, IC ₅₀ (mg / mL) Extraction Example 1 32.570% inhibition (6.960 mg / mL) Extraction Example 2 1.810 Extraction Example 3 3.670 Extraction Example 4 2.420 Extraction Example 5 1.737 Extraction Example 6 1.399 Extraction Example 7 2.049 Extraction Example 8 2.830

The results in Table 7 are different from those of Experimental Examples 1 to 2, and relatively excellent elastase activity in the extraction examples 2, 5 and 6, all of which have a high content of formmononetin, compared to the extraction example 8 having a high content of calicosin Inhibitory effect was shown. In addition, when the content is higher than the extraction example 3 with less content of calicosin and formmononetin as a whole, it showed a better rate of inhibition of elastay activity. Therefore, the effect of inhibiting the activity of elastase as an anti-wrinkle assay of Astragalus extract was determined to be advantageous in the case where the content of calicosin and formonenetine was high as an active ingredient.

Experimental Example 4 Collagen Synthesis Test

The collagen synthesis effect was measured by culturing fibroblasts as a wrinkle improvement assay of the Astragalus extract samples obtained according to the respective extraction examples 1 to 8 of the present invention. Human normal fibroblasts were seeded in 24-well microplates (1 × 10 ⁵ cells / well) and incubated for 24 hours in a 37 ° C., 5% CO ₂ incubator. Extract samples were incubated for 24 hours in a medium containing no serum (DMEM, Jeil Biotechservices) added by concentration. Collagen synthesis amount was performed using the enzyme immunoassay (ELISA) as follows. Barries incubated for 24 hours were dispensed into 96-well microplates and coated overnight at 4 ° C. Washed three times with washing buffer (PBS-T; 0.05% Tween 20 in phosphate buffered saline) and blocking solution (5% skim milk, Fluka) was added and blocked for 1 hour at 37 ℃. The blocking solution was discarded, washed three times with a washing buffer, and a primary antibody (rabbit anti-collagen type I, Sigma) was diluted in PBS-T, added 100 ul, and reacted at 37 ° C. for 2 hours. After washing three times with the washing buffer, the secondary antibody (anti-rabbit IgG alkaline phosphatase conjugate, Sigma) was diluted in PBS-T and added to 100 ul each reaction for 2 hours at 37 ℃. After washing three times with the washing buffer, 100 ul of alkaline phosphatase substrate solution (Sigma) was added and developed at room temperature. The absorbance was measured at 405 nm using an ELISA reader (EL800, Bio-Tek Instruments, Inc.). Measured. The measured value showed the collagen synthesis effect by Equation 5 below. At this time, the reaction absorbance of the cell culture without the sample was used as a control.

At this time, in order to determine the collagen synthesis effect, ascorbic acid was set as a comparison group, and each time was performed three times to obtain an average value. Table 8 shows the experimental results.

Extraction Sample Concentration (ug / mL) Collagen Synthesis Effect (%) of Extraction Examples 1 to 8 One 2 3 4 5 6 7 8 Ascorbic acid 0 100 One 96.659 99.539 94.701 102.880 98.634 105.876 92.780 90.630 109.217 10 99.424 98.963 108.986 99.539 99.770 106.452 101.306 96.774 110.445 100 99.539 113.705 107.604 110.078 111.982 111.406 107.028 116.244 120.661

The results of Table 8 showed relatively good collagen synthesis effects in the extraction Examples 2, 5 and 6 with a high content of calicosin and formonenetine. In addition, the total amount of collagen showed a better collagen synthesis effect when the content is higher than the extraction example 1 with less content of calicosin and formonenetine as a whole. Therefore, the collagen synthesis effect as an anti-wrinkle assay of the Astragalus extract was considered to be advantageous in the case where the content of calicosin and formonenetine was high as an active ingredient.

Experimental Example 5 Collagenase (MMP-1) Expression Inhibition Test

In the ELISA method using the expression inhibitory effect of collagenase (MMP-1), the expression of which is increased by UV-A irradiation of the Astragalus extract samples obtained according to the respective extraction examples 1 to 8 of the present invention. Was carried out.

Human normal fibroblasts as in Experimental Example 4 were seeded in 96-well microplates (1 × 10 ⁴ cells / well) and incubated in a CO ₂ incubator at 37 ° C., 5% concentration for 24 hours. After irradiating UV-A in a range that does not affect the fibroblasts cultured using the UV chamber, the sample was incubated for 48 hours by replacing with the medium to which the sample was added, and then the medium was recovered and coated on a 96-well microplate. The blocking solution was treated for 1 hour to block the sites where collagenase was not bound. After reacting the secondary antibody for about 2 hours, the primary antibody (rabbit anti-MMP-1, Sigma) was treated and reacted at 37 ° C. for 2 hours, followed by color development by adding 100 ul of the substrate solution. Absorbance was measured at 405 nm using an ELISA reader (EL800, Bio-Tek Instruments, Inc.). The measured value showed the collagenase expression inhibitory effect by Equation 6 below. At this time, the reaction absorbance of the cell culture without the sample was used as a control.

At this time, in order to determine the collagenase expression inhibitory effect was set to the oleanolic acid as a comparison group, and performed three times each to obtain the average value is shown in Table 9 below.

Extraction Sample Concentration (ug / mL) Collagenase (MMP-1) expression inhibition effect (%) of the extraction examples 1 to 8 One 2 3 4 5 6 7 8 Oleanolic acid 0 0 One 4.651 8.398 2.067 1.292 10.207 12.145 6.589 4.522 5.168 10 5.297 7.752 8.010 8.656 7.493 12.403 9.689 12.174 4.910 100 7.080 13.178 14.858 16.150 16.150 13.566 18.023 16.279 12.984

The results of Table 9 also showed a relatively excellent collagenase expression inhibitory effect in the extraction example of the high content of calicosin and formmononetin, the active ingredient of the Astragalus extract as in the collagen synthesis effect. In addition, Astragalus extract containing calicosin and formonenetine as an active ingredient showed better collagenase expression inhibition effect than oleanolic acid, indicating that the wrinkle improvement effect was not affected. Therefore, the collagenase expression inhibitory effect as an anti-wrinkle assay of the Astragalus extract was determined to be advantageous in the case where the content of calicosin and formonenetine was high as an active ingredient.

<Formulation Example 1 and Comparative Example 1>

The composition of the flexible cosmetics including the Astragalus extract according to the extracting Example 9 was prepared as shown in Table 10 below. At this time, the unit of component content is weight%. Among the components identified by component numbers in the following Table 10, component 2 was first dispersed and dispersed in component 1 (purified water), and then components 3 to 5 were added, and components 7 to 9 dissolved in component 6 and fractionated component 3 It prepared by the method of mixing and adding the component 10 dissolved in order sequentially. Comparative Example 1 for Formulation Example 1, except for the component 10, except for the component composition and the manufacturing method proceeded in the same manner to set the prepared.

number ingredient Formulation Example 1 Formulation Comparative Example 1 One Purified water Remaining amount Remaining amount 2 Carbomer 0.1 0.1 3 Butylene Glycol 2.0 2.0 4 glycerin 1.0 1.0 5 Butylene Glycol 0.5 0.5 6 ethanol 4.0 4.0 7 Polysorbate 80 0.2 0.2 8 Arginine 0.1 0.1 9 antiseptic a very small amount a very small amount 10 Astragalus extract according to Extraction Example 9 0.1 -

<Formulation Example 2 and Comparative Example 2>

The composition of the nutrient cosmetics containing the Astragalus extract according to the extraction example 9 was prepared as shown in Table 11 below. At this time, the unit of component content is weight%. Among the components identified by each component number in Table 11 below, components 1 to 7 were first dissolved by heating at a temperature of 70 ° C., and then components 8 to 12 were dissolved and dispersed in component 13 and emulsified in heating to 70 ° C. Thereafter, the emulsified product was neutralized with component 14, cooled to a temperature of 56 ° C, and then, component 15 dissolved in the fractionated component 8 was added, stirred, and cooled at room temperature to prepare. Comparative Example 2 with respect to Formulation Example 2, except for the Astragalus extract according to the extraction example 9 of the component 15, the same as the ingredient composition or the manufacturing method was set to proceed in the same manner.

number ingredient Formulation Example 2 Formulation Comparative Example 2 One Cetearyl Alcohol 1.0 1.0 2 Glyceryl Stearate 0.5 0.5 3 Polysorbate 60 1.0 1.0 4 Sorbitan sesquioleate 0.3 0.3 5 Cetyloctanoate 6.0 6.0 6 Squalane 4.0 4.0 7 Flower oil 4.0 4.0 8 Butylene Glycol 4.0 4.0 9 glycerin 4.0 4.0 10 Carbomer 0.1 0.1 11 Xanthan Gum 0.03 0.03 12 antiseptic a very small amount a very small amount 13 Purified water Remaining amount Remaining amount 14 Triethanolamine 0.1 0.1 15 Astragalus extract according to Extraction Example 9 0.1 -

<Formulation Example 3 and Comparative Example 3>

Component composition of the essence containing the Astragalus extract according to the extraction example 9 was prepared as shown in Table 12. At this time, the unit of component content is weight%. Among the components identified by each component number in Table 12 below, components 2 and 3 were first dispersed and dispersed in component 1 (purified water), and then components 4 to 5 and 6 were added, and components 8 to 10 were dissolved in component 7. And component 11 dissolved in aliquot component 4 were added sequentially to prepare a mixture. Comparative Example 3 with respect to Formulation Example 3, except for the Astragalus extract according to the extraction example 9 of the component 11 was set to proceed to the same composition or manufacturing method.

number ingredient Formulation Example 3 Formulation Comparative Example 3 One Purified water Remaining amount Remaining amount 2 Carbomer 0.2 0.2 3 Hydroxyethyl cellulose 0.1 0.1 4 Butylene Glycol 8.0 8.0 5 glycerin 5.0 5.0 6 Sodium hyaluronate 10.0 10.0 7 ethanol 2.0 2.0 8 Polyoxyethylene Cured Castor Oil 0.1 0.1 9 Arginine 0.2 0.2 10 antiseptic a very small amount a very small amount 11 Astragalus extract according to Extraction Example 9 0.2 -

<Formulation Example 4 and Comparative Example 4>

The ingredient composition of the nutrient cream containing the Astragalus extract according to the extraction example 9 was prepared as shown in Table 13. At this time, the unit of component content is weight%. Among the components identified by each component number in Table 13 below, components 1 to 8 were first dissolved by heating at a temperature of 70 ° C., and then components 9 to 13 were dissolved and dispersed in component 14 and emulsified in heating to 70 ° C. Thereafter, the emulsified product was cooled to a temperature of 56 ° C., and then component 15 dissolved in the fractionated component 9 was added thereto, stirred, and cooled to room temperature to prepare. Comparative Example 4 with respect to Formulation Example 4, except for the Astragalus extract according to the extraction example 9 of the component 15, the other components of the composition or preparation method was set to proceed in the same manner.

number ingredient Formulation Example 4 Formulation Comparative Example 4 One Cetearyl Alcohol 1.5 1.5 2 Glyceryl Stearate 1.0 1.0 3 Polysorbate 60 1.0 1.0 4 Sorbitan sesquioleate 0.3 0.3 5 Cetyloctanoate 6.0 6.0 6 Squalane 8.0 8.0 7 Flower oil 4.0 4.0 8 Dimethicone 2.0 2.0 9 Butylene Glycol 4.0 4.0 10 Glyserine 4.0 4.0 11 Magnesium Aluminum Silicate 0.4 0.4 12 Xanthan Gum 0.03 0.03 13 antiseptic a very small amount a very small amount 14 Purified water Remaining amount Remaining amount 15 Astragalus extract according to Extraction Example 9 0.2 -

<Formulation Example 5 and Comparative Example 5>

The composition of the nutritional pack containing the Astragalus extract according to the extraction example 9 was prepared as shown in Table 14 below. At this time, the unit of component content is weight%. Among the components identified by each component number in Table 14 below, components 2 and 3 were first added to component 1 (purified water), and components 4 to 6 were added thereto, heated to 70 ° C. while stirring and dispersed, and then components 7 to 8 were added thereto. It was sufficiently dispersed and cooled to a temperature of 56 ° C. A substance prepared by dissolving components 10 to 11 in component 9 and component 12 dissolved in aliquoted component 2 in turn was stirred and cooled to room temperature. Comparative Example 4 with respect to Formulation Example 4, except for the Astragalus extract according to the extraction example 9 of the component 12, the same as the ingredient composition or manufacturing method was set to proceed in the same manner.

number ingredient Formulation Example 5 Formulation Comparative Example 5 One Purified water Remaining amount Remaining amount 2 Butylene Glycol 3.0 3.0 3 glycerin 1.5 1.5 4 Polyvinyl alcohol 3.0 3.0 5 Polyvinylpyrrolidone 12.0 12.0 6 Cellulose gum 0.3 0.3 7 Oxol 2.0 2.0 8 kaoline 2.0 2.0 9 ethanol 5.0 5.0 10 Polyoxyethylene Cured Castor Oil 0.5 0.5 11 antiseptic a very small amount a very small amount 12 Astragalus extract according to Extraction Example 9 0.1 -

Skin Wrinkle Improvement Test

Skin wrinkle improvement effect of the Astragalus extract sample according to the extraction Example 9 was measured as follows.

Twenty women over 35 years of age were assigned to the nutritional cream of Formulation Example 4 and the nutritional cream of Comparative Example 4 in a randomized manner to be used on a predetermined site (both eyes) for 12 weeks. At 4, 8 and 12 weeks after use of the product, the skin wrinkles were measured using a transparent silicone solution, and a R-value measurement using a skin wrinkle meter (Skin Visiometer SV600) was shown in Table 15 below.

ΔR (R before experiment-R after 12 weeks) R1 R2 R3 R4 R5 Formulation Example 4 -0.19 -0.13 -0.07 -0.03 -0.06 Formulation Comparative Example 4 -0.06 -0.05 -0.03 -0.00 -0.03

In Table 15, each R1 is the distance between the highest point and the lowest point of the wrinkle profile, R2 is an arithmetic mean of the R1 values in each region, and R3 is the equalized region in each region. R4 is the highest value of R1, R4 is the horizontal line from the top of the wrinkle profile, integrated with the area divided by the length of the midline, which means the average depth of the skin wrinkles, R5 is the profile of the midline of the wrinkle profile The area is integrated and divided by the length of the midline of the profile, which means the average roughness of the skin.

As shown in the results of Table 15, in the case of the formulation example 4 containing the extraction example 9, the R-value showing the skin wrinkle improvement effect after 12 weeks of application is significantly lower than the portion to which the comparative example 4 is applied and wrinkles It was found that this was improved.

Skin Safety Test

In order to determine the occurrence of irritation or allergic reaction through the skin safety, that is, the skin reaction of the Astragalus extract according to the extraction example 9, a human patch test was performed according to the Korea Food and Drug Administration standard skin safety test method.

Thirty healthy men and women aged twenty to fifty were selected as subjects. The patch test was carried out on the inner side of the subject's upper arm and the patch was removed after 48 hours. The first reading was performed after about 60 minutes of stability, and the second reading was taken after 96 hours of patch application. Criteria are as shown in Table 16 below.

reaction weight Criteria of Judgment - 0 No response ± 0.5 Faint erythema + One Boundary but weak erythema, edema and papules ++ 2 Pronounced erythema, papules and blisters +++ 3 Cannon

The skin readiness of the sample was calculated using Equation 7 below for 48 hours and 96 hours after patch application.

The skin irritation degree of the sample showing the positive response calculated based on the test result of the patch test is shown in Table 17 below.

sample 48 hours 96 hours Responsiveness (%) + ++ + ++ 48 hours 96 hours Average Extraction Example 8 (0.1% in 1,3-butylene glycol) - - One - 0.00 0.56 0.28

As shown in Table 17, each of the extraction Example 9 according to the skin reactivity determination criteria according to Table 18 in the human patch test was found to be used for sensitive skin irritantly sensitive. Therefore, the Astragalus extract sample according to the extraction example was found to be a safe ingredient having no skin side effects as a natural product.

Average responsiveness of the skin Criteria 0.0 to 0.9 Non-irritating 1.0 to 2.9 Light stimulation 3.0 to 4.9 Heavy stimulation 5.0 or more Strong stimulation

As described above, the present invention provides a cosmetic composition containing the Astragalus extract, more preferably Astragalus extract prepared by a specific manufacturing method, the cosmetic composition of the present invention has no skin side effects as a natural product, antioxidant, skin Very useful for whitening and wrinkle improvement.

Claims (6)

Cosmetic composition for skin whitening or anti-wrinkle, characterized in that containing the extract as an active ingredient. The cosmetic composition for skin whitening or wrinkle improvement according to claim 1, wherein the Astragalus extract is an extract containing calicosin, formonenetine or a mixture thereof. (S1) extracting Astragali Radix or crushed Astragalus using an alcohol having 1 to 4 carbon atoms or an aqueous alcohol solution; (S2) adding at least one selected from the group consisting of carbohydrate, pectin hydrolase and cellulose hydrolase to the extract of step (S1) and shaking culture; And (S3) characterized in that it comprises an extract containing calicosin, formononetin or a mixture thereof prepared by a manufacturing method comprising the step of adding a fractionated organic solvent to the culture of step (S2) Cosmetic composition for antioxidant, skin whitening or wrinkle improvement. The method according to claim 3, wherein the preparation method comprises the steps of (A) (S2) between the step (S2) and (S3) step (S2) fraction or fractional concentration using an organic solvent for fractionation; And (b) adding at least one selected from the group consisting of carbohydrate hydrolase, pectin hydrolase and cellulose hydrolase to the fraction or fraction concentrate of step (a) and shaking culture. Antioxidant, skin whitening or anti-wrinkle cosmetic composition characterized in that. The method according to claim 3, wherein the preparation method comprises the step of concentrating the extract of step (S1) between step (S1) and (S2), and dissolving the concentrate with an ethanol aqueous solution at that time, Cosmetic composition for skin whitening or wrinkle improvement. The cosmetic composition for skin whitening or wrinkle improvement according to claim 1, wherein the content of the Astragalus extract is 0.001 to 10% by weight based on the total weight of the cosmetic composition.

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