KR102002997B1 - Cosmetic composition including taxifolin glucosides and extracting method of paeonia lactiflora extracts - Google Patents

Abstract

The present invention provides a cosmetic composition containing a tachypholin glycoside as an active ingredient and a method for extracting a peony root extract. According to the extract method of extract of peony root extract of the present invention, it is possible to extract a high amount of texifolin glycosides, specifically, taxon poly-3-glucoside in an optimum yield and mass, and the peony root extract containing the peony root extract has excellent skin whitening effect, Anti-inflammatory effect, antibacterial effect, atopic improvement, and ultraviolet ray protection effect, it can be usefully used as a functional cosmetic composition and external preparation for skin.

Description

TECHNICAL FIELD [0001] The present invention relates to a cosmetic composition containing a tachypolin glycoside as an active ingredient and a method for extracting a peony extract,

The present invention relates to a cosmetic composition containing a tachypolin glycoside as an active ingredient and a method for extracting a peony root extract.

Paeonia lactiflora ) is a perennial plant of peonies, peonies, peonies, peonies, and peonies originating in Korea. It grows mainly in the foothills and valleys. The branch stretches more than 2m long on the side, has no hair but has hook like thorns, and several stems come out of a droppings and stand straight, about 60cm high, and have no hairs on leaves and stems. There are several roots, but they are thin and thick with a pointed cylindrical shape with both ends. Leaves are alternate and lower leaves are double leaves with three small leaves. The flowers bloom one at the end of the stem in May-June, and it is big and beautiful. The fruit is ovate, the end is bent like a hook, it is split along the inner line, and the seed is spherical. Flowers are mainly used for gardening, and roots are used as medicine for pain, abdominal pain, dysmenorrhea, amenorrhea, hematemesis, anemia, bruise. They are distributed in Korea, Mongolia, and East Siberia.

Patent No. 10-0804993 discloses a method for producing a cosmetic soap using a peony seed extract, and Japanese Patent Application Laid-Open No. 2005-0118819 discloses an antioxidative and moisturizing detergent composition containing a peony extract as an active ingredient. However, There have been no studies on peony root extract containing taxifolin-3-glucoside and a functional cosmetic composition containing the same.

Korean Patent No. 10-0804993 Korea Patent Publication No. 2005-0118819

In order to solve the above problems, the present invention provides a method for extracting a high amount of taxifolin glycoside from a natural plant, such as a peony root, by extracting the peony root extract, concentrating, fractionating and recrystallizing the peony root extract. It is an object of the present invention to provide a cosmetic composition using skin whitening effect, wrinkle improving effect, anti-inflammatory effect, antibacterial effect, atopic improvement and ultraviolet ray protection effect.

In order to solve the above-mentioned problems, the present invention provides a cosmetic composition comprising a tachypholin glycoside as an active ingredient.

In one embodiment of the present invention, the taxifolin glycoside may be extracted from peony root.

In one embodiment of the present invention, the taxifolin glycoside may be taxifolin-3-glucoside.

In one embodiment of the present invention, the texifolin glycoside may be contained in an amount of 0.0001 to 10% by weight based on the total weight of the composition.

In one embodiment of the present invention, the composition may be one having at least one function selected from the group consisting of skin whitening, wrinkle improvement, anti-inflammation, antibacterial, atopic improvement, and ultraviolet ray protection.

In addition, the present invention provides a method for preparing a pharmaceutical composition comprising the steps of: dissolving a taxane compound in a solvent and eluting the taxane compound;

Concentrating the eluted extract;

Placing the concentrated extract in two or more solvents and fractionating;

Passing the extract obtained in the fractioned layer through a column; And

And recrystallizing the solution by precipitating the column passing liquid.

In one embodiment of the present invention, the taxifolin glycoside may be taxifolin-3-glucoside.

In one embodiment of the present invention, the fractionation step may be to fractionate and then remove the solvent of the fractioned layer and concentrate.

In one embodiment of the present invention, the extraction method may further include drying and obtaining the recrystallized extract after recrystallization.

In one embodiment of the present invention, the solvent for extracting the solvent may be at least one selected from the group consisting of water, ethanol, methanol, butanol, ethyl acetate, ethyl ether, chloroform and hexane.

In one embodiment of the present invention, the solvent at the time of fractionation may be at least two selected from the group consisting of water, ethanol, methanol, butanol, ethyl acetate, ethyl ether, chloroform and hexane.

In one embodiment of the present invention, the resin of the column is selected from the group consisting of Amberlite XAD resin, Dow X resin, Suferite DAX resin and Diaion HP-10, 20, 30, 40, Lt; / RTI >

In one embodiment of the present invention, the recrystallization may be performed at -300 to 25 ° C and 0.5 to 800 MPa.

According to the extract method of extract of peony root extract of the present invention, it is possible to extract a high amount of texifolin glycosides, specifically, taxon poly-3-glucoside in an optimum yield and mass, and the peony root extract containing the peony root extract has excellent skin whitening effect, Anti-inflammatory effect, antibacterial effect, atopic improvement, and ultraviolet ray protection effect, it can be usefully used as a functional cosmetic composition and external preparation for skin.

1 is a flowchart illustrating a method for extracting a peony root extract according to an embodiment of the present invention.
FIG. 2 is an HPLC chromatogram of Taxin pollin-3-glucoside among peony root extracts according to one embodiment of the present invention.

Hereinafter, preferred embodiments of the present invention will be described in detail in order to facilitate the present invention by those skilled in the art.

The present invention provides a cosmetic composition comprising a tachypolin glycoside as an active ingredient.

The peony extract contains a taxifolin glycoside. Specifically, in the present invention, the taxifolin glycoside may be taxifolin-3-glucoside.

The tachesopholine glycoside may be contained in an amount of 0.0001 to 10% by weight based on the total weight of the composition. If the amount is less than 0.0001% by weight, the efficacy is not sufficiently exhibited. If the amount is more than 10% by weight, the increase in the content does not lead to a marked increase in the effect.

In one embodiment of the present invention, the composition may be one having at least one function selected from the group consisting of skin whitening, wrinkle improvement, anti-inflammation, antibacterial, atopic improvement, and ultraviolet ray protection. This can be confirmed in a test example to be described later.

The formulation of the cosmetic composition is not particularly limited and may be appropriately selected according to the purpose. For example, skin lotion, skin softener, skin toner, astringent, lotion, milky lotion, moisturizing lotion, nutrition lotion, massage cream, nutritional cream, moisturizing cream, hand cream, foundation, essence, nutrition essence, pack, soap, cleansing But the present invention is not limited thereto, and may be manufactured by any one or more formulations selected from the group consisting of a foam, a cleansing lotion, a cleansing cream, a body lotion and a body cleanser.

When the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

In the case of the solution or emulsion of the present invention, a solvent, a solvent or an emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.

The content of the active ingredient is not particularly limited, but may be in the range of 0.0001 to 10% by weight based on the total weight of the composition. When the active ingredient satisfies the above content, it can exhibit excellent efficacy without side effects.

In addition to the peony extracts, the cosmetic composition may further contain ingredients included in functional additives and general cosmetic compositions. The functional additives may include water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymeric polysaccharides, sphingolipids and seaweed extracts.

The cosmetic composition of the present invention may further contain, in addition to the above-described functional additive, components contained in a general cosmetic composition as required. Examples of the other ingredients that can be included in the composition include humectants, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbents, preservatives, bactericides, antioxidants, plant extracts, pH adjusters, alcohols, Accelerators, coolants, antiperspirants, purified water, and the like.

Further, the present invention relates to an external preparation for skin comprising the above composition, wherein the external preparation for skin is a generic term that can be applied to any external skin, and cosmetics and medicines of various formulations may be included. The external preparation for skin is not particularly limited, but may be, for example, a skin external preparation for preventing skin aging or improving skin wrinkles.

In addition, the present invention provides a method for extracting a peony root extract comprising the step of eluting a taxon purine glycoside from a peony.

The method for extracting a peony root extract of the present invention comprises the steps of: dissolving a peony bud; Concentrating the eluted extract; Placing the concentrated extract in two or more solvents and fractionating; Passing the extract obtained in the fractioned layer through a column; And recrystallizing by precipitating the column passing liquid.

First, the taxane polyphosphate glycoside, preferably taxifolin-3-glucoside, is eluted from the peony. Preferably, the solvent extraction process is carried out by adding 0.5-20.0 wt.% Peony based on the solvent to the solvent for 0.5-5 hours.

The solvent may be at least one selected from the group consisting of water, ethanol, methanol, butanol, ethyl acetate, ethyl ether, chloroform, and hexane.

Then, the crude extract of Taxus polyphenine-3-glucoside, which has been eluted through the above-mentioned extraction step, is subjected to a first concentration step for solvent removal. Concentration of the eluted extract may be carried out by any one or more selected from the group consisting of heat drying, vacuum drying, spray drying and freeze drying, as long as they are known in the conventional concentration method.

Thereafter, the concentrated extract is concentrated in two or more solvents. At this time, the two or more solvents are polar solvents having different polarities, and preferably two or more solvents selected from the group consisting of water, ethanol, methanol, butanol, ethyl acetate, ethyl ether, chloroform and hexane. Several compounds present in extracts dissolved in solvents of different polarity are polarized.

A second concentration step of removing the organic solvent of the fractionated layer after the fractionation can be performed. Concentration at this stage may be carried out by any one or more selected from the group consisting of heat drying, vacuum drying, spray drying and freeze drying, as long as they are known as conventional concentration methods.

The extract obtained in the fractioned layer is then passed through a column. At this time, the column is not limited, but it is preferably a resin column using an adsorption resin. The resin may be one or more resins selected from the group consisting of Amberlite XAD resin, Dow X resin, Suferite DAX resin, and Diaion HP-10, 20, 30, 40, and 50 resins.

By passing through the column, the pigment and chlorophyll are removed from the fraction concentrate (fraction concentrated powder).

Finally, the column passed solution is precipitated to be recrystallized. This is accomplished by adjusting the pressure and temperature of the column passed through the column to precipitate. The recrystallization may be performed at -300 to 25 ° C and 0.5 to 800 MPa. That is, the recrystallization can be performed under the conditions including the ultra-low temperature and the ultra-high pressure.

The extraction method may further comprise drying and obtaining the recrystallized extract after recrystallization. In the obtaining step, the recrystallized precipitated pellet is spray-dried or lyophilized to obtain a high-content powder of taciffiospin-3-glucoside, which is then concentrated by centrifugation, filtration and vacuum concentration, -3-glucoside can be obtained.

The method for preparing peony root extract according to the present invention is a multistage continuous process, and when the peptides are subjected to extraction, concentration, fractionation, column and recrystallization as described above, a taxonlin glycoside can be obtained with the highest yield.

The peony root extract containing the extracted taxoproline glycoside has excellent whitening effect, wrinkle improving effect, anti-inflammation, antibacterial, ultraviolet ray protecting effect and atopy improvement effect.

The present invention will be described in more detail through the following examples. However, the examples are for illustrating the present invention, and the scope of the present invention is not limited thereto.

≪ Comparative Example 1 &

The peony was added to a 70% aqueous ethanol solution in an amount of 10% by weight and extracted with a reflux condenser equipped with a condenser at a temperature of 80 캜 for 2 hours. Thereafter, the filtrate was filtered through an 80 mesh filter and a 1.2 쨉 m filter, and the obtained filtrate was concentrated under a condition of 65 h and 20 hPa using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) Concentration was carried out for 1 hour to obtain a concentrated powder. The sample thus obtained was called "sample 1" and used in the following test examples.

≪ Comparative Example 2 &

The peony was added to a 70% aqueous ethanol solution in an amount of 10% by weight and extracted with a reflux condenser equipped with a condenser at a temperature of 80 캜 for 2 hours. Thereafter, the filtrate was filtered through an 80 mesh filter and a 1.2 쨉 m filter, and the obtained filtrate was concentrated under a condition of 65 h and 20 hPa using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) Concentration was carried out for 1 hour to obtain a concentrated powder. 10 times as much weight of distilled water was added to the obtained concentrated powder to uniformly disperse it. Then, hexane of the same volume as that of distilled water was added and fractionated using a fractional distillation column. Ethyl acetate and n-butanol were sequentially fractionated and finally ethyl acetate layer . The obtained ethyl acetate fraction filtrate was concentrated for 1 hour at 65 ° C and 20 hPa using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) to obtain a concentrated powder. 2 ", which was used in the following test example.

≪ Comparative Example 3 &

The peony was added to a 70% aqueous ethanol solution in an amount of 10% by weight and extracted with a reflux condenser equipped with a condenser at a temperature of 80 캜 for 2 hours. Thereafter, the filtrate was filtered through an 80 mesh filter and a 1.2 쨉 m filter, and the obtained filtrate was concentrated under a condition of 65 h and 20 hPa using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) Concentration was carried out for 1 hour to obtain a concentrated powder. 10 times as much weight of distilled water was added to the obtained concentrated powder to uniformly disperse it. Then, hexane of the same volume as that of distilled water was added and fractionated using a fractional distillation column. Ethyl acetate and n-butanol were sequentially fractionated and finally ethyl acetate layer . The obtained ethyl acetate fraction filtrate was concentrated for 1 hour at 65 DEG C and 20 hPa using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) to obtain a concentrated powder, The mixture was re-dispersed in distilled water and added to a Diaion HP-20 column. Chromatography was performed by using a gradient method using distilled water and 99.5% ethanol. Finally, 70% -99.5% ethanol column solution And concentrated under a condition of 65 ° C and 20 hPa for 1 hour using a vacuum evaporator (Rotary Evaporator NE-series, Eyela, Japan) to obtain a concentrated powder. The sample thus obtained was referred to as "sample 3" It was used in the test example.

≪ Example 1 >

The peony was added to a 70% aqueous ethanol solution in an amount of 10% by weight and extracted with a reflux condenser equipped with a condenser at a temperature of 80 캜 for 2 hours. Thereafter, the filtrate was filtered through an 80 mesh filter and a 1.2 쨉 m filter, and the obtained filtrate was concentrated under a condition of 65 h and 20 hPa using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) Concentration was carried out for 1 hour to obtain a concentrated powder. 10 times as much weight of distilled water was added to the obtained concentrated powder to uniformly disperse it. Then, hexane as a volume as that of distilled water was added and fractionated using a fractional nozzle. Ethyl acetate and n-butanol were sequentially fractionated, . The obtained ethyl acetate fraction filtrate was concentrated for 1 hour at 65 DEG C and 20 hPa using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) to obtain a concentrated powder, The mixture was re-dispersed in distilled water and added to a Diaion HP-20 column. Chromatography was carried out using gradient water and distilled water and 99.5% ethanol. Ultrafiltration was performed using a 70% -99.5% ethanol column at a flow rate of 20 ml / (TFS-2L, TOYO KOATSU CO., LTD., Japan) at 100 MPa and 25 ° C for 1 hour. The mixture was centrifuged at 8,000 rpm and 15 ° C for 15 minutes using a centrifuge (Supra 22K, Hanil, Korea), filtered through a 1.2 μm filter, and finally dried at 60 ° C. for 1 hour to obtain a powder , And the thus obtained sample was called "sample 4" and used in the following test examples.

≪ Example 2 >

The peony was added to a 70% aqueous ethanol solution in an amount of 10% by weight and extracted with a reflux condenser equipped with a condenser at a temperature of 80 캜 for 2 hours. Thereafter, the filtrate was filtered through an 80 mesh filter and a 1.2 쨉 m filter, and the obtained filtrate was concentrated under a condition of 65 h and 20 hPa using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) Concentration was carried out for 1 hour to obtain a concentrated powder. 10 times as much weight of distilled water was added to the obtained concentrated powder to uniformly disperse it. Then, hexane of the same volume as that of distilled water was added and fractionated using a fractional distillation column. Ethyl acetate and n-butanol were sequentially fractionated and finally ethyl acetate layer . The obtained ethyl acetate fraction filtrate was concentrated for 1 hour at 65 DEG C and 20 hPa using a vacuum concentrator (Rotary Evaporator NE-series, Eyela, Japan) to obtain a concentrated powder, The mixture was re-dispersed in distilled water and loaded on a Diaion HP-20 column. Chromatography was carried out using gradient water and distilled water using 99.5% ethanol. The column was finally subjected to a 70% -99.5% ethanol column under a flow rate of 20 ml / (DF-9010, Ilshin, Korea) at -90 ° C for 2 hours. The mixture was centrifuged at 8,000 rpm and 15 ° C for 15 minutes using a centrifuge (Supra 22K, Hanil, Korea), filtered through a 1.2 μm filter, and finally dried at 60 ° C. for 1 hour to obtain a powder , And the thus obtained sample was referred to as "sample 5" and used in the following test examples.

[Test Example 1] Analysis of the content of taxifolin-3-glucoside by HPLC

The method of the present invention for preparing a crude extract containing Taxifolin-3-glucoside In order to confirm the change of the content of Taxifolin-3-glucoside according to the steps, analysis of the content of each sample by HPLC was performed as follows .

For the qualitative and quantitative analysis of the taciffin-3-glucoside of the step-by-step samples prepared in the above examples, analysis was carried out using HPLC (High Pressure Liquid Chromatography). Samples were prepared at appropriate concentrations with each solvent, filtered through a 0.2 μm filter, pretreated and injected into the instrument. The instrument was analyzed in the whole UV range using a UV detector using HPLC (Agilent 1200 series system, Agilent, USA). The C ₁₈ column (Zorbax XDB-C ₁₈ 4.6 × 250 mm, Agilent, USA) using 2% purified water and slanting method using acetic acid solvent and acetonitrile solvent. All of the solvents used in the analysis were HPLC grade reagents, and standard curves were obtained and quantitative analysis was performed. The results of the analysis are shown in Table 1 and FIG. Fig. 2 is an HPLC chromatogram of Taxifolin-3-glucoside of a peony root extract (sample 5) that has undergone five steps of preparation such as extraction, concentration, fractionation, column and cryogenic recrystallization.

Name of sample Manufacturing method steps Taxicol-3-glucoside content (%) Comparative Example 1 Extraction, concentration 2.5 Comparative Example 2 Extraction, concentration, fraction 11.4 Comparative Example 3 Extraction, concentration, fraction, column 32.9 Example 1 Extraction, concentration, fraction, column, ultrahigh pressure recrystallization 97.4 Example 2 Extraction, concentration, fractionation, column, cryogenic recrystallization 99.3

As shown in Table 1 above, it was confirmed that the content of taxifolin-3-glucoside was dramatically increased as the extraction, concentration, fractionation, column, and recrystallization were carried out from the peony root, It was confirmed that a crude extract of Panax notoginseng can be prepared which contains the content of Taxifolin-3-glucoside.

[Test Example 2] Tyrosinase activity inhibition test

In order to test the whitening effect of the peony root extract containing Taxifolin-3-glucoside of the present invention, the degree of suppression of the function of the enzyme called tyrosinase was evaluated. Tyrosinase is an enzyme that helps the production of melanin by promoting the oxidation process of tyrosine in vivo. Melanin is a black polymer, which melanin is formed in the body by oxidation as described above. When melanin is formed in the upper part of the skin, the skin turns black and produces spots and freckles. The degree of inhibition of the action of tyrosinase was measured to determine the whitening effect. The test method is as follows.

0.9 mL of the 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 the mixture was maintained at 37 DEG C for 10 minutes. 0.1 ml of Mushroom Tyrosinase (1,500 units / mL) was added and reacted at 37 ° C for 10 minutes. Absorbance was measured at 475 nm using a UV-spectrophotometer (Smartspec Plus, Biorad, USA) The inhibition rate against Nausea was measured.

As a control group for the inhibition of tyrosinase activity, a buffer solution was added in place of the sample solution, and the same method was used. A buffer solution was added instead of tyrosinase to obtain the respective color correction values for the sample and the control group .

The samples used in the test were the peony extracts containing texifolin-3-glucoside obtained in Example 2 (Sample 5) and arbutin (Arbutin synthetic, Sigma), which is known to have excellent whitening effect in various studies , The tyrosinase inhibition rate was calculated numerically using the following equation (1), and the results are shown in Table 2 below. In Table 2, IC ₅₀ is the concentration of the sample required to inhibit tyrosinase activity by 50%, and the lower the value, the higher the inhibition rate.

[Equation 1]

Inhibition rate (%) = 100 - ((absorbance of sample / absorbance of control group) x 100)

sample Inhibition rate of tyrosinase activity,
IC ₅₀ (mg / mL) Example 2 0.022 Arbutin 0.032

As shown in Table 2, it was confirmed that the peony root extract containing Taxon poly-3-glucoside of Example 2 had much better whitening effect than arbutin, which is known to have excellent whitening effect at present.

[Test Example 3] Elastase activity inhibition test

An elastase inhibition activity test was conducted to examine the skin wrinkle inhibition and improving effect of the extract of Panax notoginsenosides containing Taxifolin-3-glucoside of the present invention. Elastin is a component of the matrix layer in the dermis of the skin. As the skin ages, elastin breaks down and the matrix layer in the dermis breaks down and wrinkles occur. (Ala) 3 p-nitroaniline (N-succinyl- (Ala)), which is an elastase substrate, is used as a method for measuring the activity of Elastase, an enzyme that decomposes elastin, 3 p-nitroaniline, Sigma) to determine the change in color caused by the decomposition of p-nitroaniline by measuring the absorbance at a wavelength of 405 nm. The buffer solution was pH 8.0, 0.267 M Trizma-HCl (Sigma), the substrate solution was 8.8 mM N-succinyl- (Ala) 3 p-nitroaniline and the enzyme solution was incubated with the pancreatic islets in 10 μg / Respectively. 20 μl of the buffer solution and 20 μl of the enzyme solution were mixed with 100 μl of the sample solution diluted with the third distilled water according to the concentration of Example 2 (Sample 5), and then 20 μl of the enzyme solution was added and reacted in a constant temperature water bath at 25 ° C for 15 minutes, The amount of nitroaniline produced was measured at a wavelength of 405 nm using a microplate reader (UVT-06685, Thermo max, USA).

The control group for measuring the activity of the Elastase was prepared by adding the third distilled water instead of the sample, measuring the same method, and adding the third distilled water instead of the enzyme solution to obtain the color correction value for each.

The samples used in the test were peanut extracts containing taxol-3-glucoside obtained in Example 2 (sample 5) and retinol (> 99%, Sigma, USA) And the inhibition rate of the ELASTASE was calculated numerically using the following equation (2). In Table 3, IC ₅₀ is the concentration of the sample required to inhibit the enzyme activity by 50%, and the lower the value, the higher the inhibition rate.

&Quot; (2) "

Elastase activity inhibition rate (%) = 100 - ((absorbance of sample) / absorbance of control group) × 100)

sample Elastase activity inhibition rate,
IC ₅₀ (mg / mL) Example 2 0.20 Retinol 0.38

As shown in Table 3, the extract of Panax ginseng containing Taxifolin-3-glucoside obtained in Example 2 (Sample 5) was superior to retinol, which is known to have excellent wrinkle-reducing effect, Respectively.

[Test Example 4] Collagen synthesis effect test

Collagen aggregation performance was measured to examine the skin wrinkle-improving effect of the extract of Panax notoginsenosides containing Taxifolin-3-glucoside of the present invention. Collagen is a constituent component of the dermal matrix layer in the skin together with the elastin, and the skin gradually ages, and the components constituting the matrix layer in the dermis are decomposed to form wrinkles. The collagen is a constituent of the dermal matrix layer By confirming the synthesis effect, the effect of improving the wrinkles of the skin can be confirmed. The test method proceeds as follows.

Inoculated with normal fibroblast (human dermal fibroblast) of the person in a 24-well microplate and (1 × 10 ⁵ cells / well) 37 ℃, it was cultured for 24 hours in a CO ₂ incubator at 5% concentration. Each sample was incubated in serum-free DMEM medium for 24 hours. The amount of collagen synthesis is measured by enzyme immunoassay as follows.

The medium cultured for 24 hours was dispensed in a 96-well microplate and coated overnight at 4 占 폚. After washing three times with wash buffer (PBS-T; 0.05% Tween 20 in phosphate buffered saline), blocking solution (5% skin milk, Fluka) was added and blocked for 1 hour at 37 ° C. The blocking solution was discarded and washed three times with washing buffer. The primary antibody (rabbit anti-collagen type I, Sigma) was diluted in PBS-T, and 100 μl of each was added thereto, followed by reaction at 37 ° C for 2 hours. After washing three times with washing buffer, 100 μl of secondary antibody (anti-rabbit IgG alkaline phosphatase conjugate, Sigma) was diluted in PBS-T and reacted at 37 ° C for 2 hours. After washing three times with washing buffer, 100 μl of alkaline phosphatase substrate solution (Sigma) was added and developed at 25 ° C. The absorbance at 405 nm was measured using a microplate reader (UVT-06685, Thermo max, USA) Were measured.

The control group for measuring the collagen synthesis effect was a reaction absorbance of a cell culture solution not treated with the sample. To determine the collagen synthesis effect, ascorbic acid was set as a comparative group, and the results were averaged three times.

The collagen synthesis effect was numerically calculated using the following equation (3) and shown in Table 4 below.

&Quot; (3) "

Call
la
Gen
synthesis
castle
Hyo
and
(%) sample Concentration (/ / mL) 0 One 10 100 Example 2 100.00 111.59 122.84 133.21 Ascorbic acid (vitamin-C) 100.00 108.45 123.57 131.58

As shown in Table 4, the peony root extract containing Taxifolin-3-glucoside obtained in Example 2 (Sample 5) had almost the same collagen synthesis as that of ascorbic acid, Effect.

[Test Example 5] Evaluation of inhibition of MMP-1 expression after irradiation with ultraviolet light

Enzymatic immunoassay (ELISA) for measuring MMP-1 concentration after UV irradiation and sample addition to evaluate the inhibition of MMP-1 expression after irradiation with ultraviolet rays of the fungicide extract containing Taxifolin-3-glucoside of the present invention Respectively.

UVA is irradiated to human dermal fibroblasts at an energy of 5 J / cm < 2 > using a UV chamber. Ultraviolet irradiation dose and incubation time were established by preliminary experiment to maximize MMP expression level in fibroblasts. The negative control group was kept in UVA environment for the same time after masking with silver foil. The UVA emission was measured using a UV radiometer. The UVA-irradiated cells remained in the previously-dispensed medium, irradiated with UVA, exchanged with the medium containing the sample, cultured for 24 hours, Lt; / RTI > The primary antibody (MMP-1 (Ab-5) monoclonal antibody and MMP-2 (Ab-3) monoclonal antibody) was treated and reacted at 37 ° C for 60 minutes. The secondary antibody, anti mouse IgG conjugated) was reacted again for 60 minutes. The reaction solution was incubated with alkaline phosphatase substrate solution (1 mg / ml p-nitrophenyl phosphate in diethanolamine buffer) at room temperature for 30 minutes and absorbance was measured at 405 nm with a microplate reader. As a control group, a sample to which no sample was added was used.

As shown in Table 5, in Example 2 (Sample 5), the inhibition rate of MMP-1 inducing expression at ultraviolet irradiation was 48% or more as compared with the control not treated with the sample, Which is superior to the inhibition rate.

Test group Treatment concentration (%) MMP-1 expression inhibition rate (%) Example 2 0.1 48 Retinol 0.1 28

[Experimental Example 6] Experiment to measure melanin formation inhibitory effect using B16F1 melanocyte

In order to test the whitening effect of peony root extract containing Taxifolin-3-glucoside of the present invention, the degree of inhibition of melanin formation on B16F1 melanocyte was examined to determine whitening effect as described below.

The B16F1 melanocyte used in this test example is a cell strain derived from a mouse, and is a cell that secretes a melanin pigment called melanin. During the artificial culture of these cells, samples were treated to compare the degree of reduction of melanin pigment. The B16F1 melanocyte used in this test example was purchased from ATCC (American Type Culture Collection, Accession No. 6323).

The melanin biosynthesis inhibitory effect of B16F1 melanocyte was measured as follows. B16F1 melanocytes were dispensed into 6-well plates at a concentration of 2 × 10 ⁶ cells per well, and the cells were incubated at 72 ° C. for 72 hours at a concentration that did not induce toxicity. After incubation for 72 hours, cells were detached with trypsin-EDTA, and the number of cells was measured and centrifuged to recover the cells. Quantification of intracellular melanin was carried out using Lotan: Cancer Res . , 40: 3345-3350, 1980). Cell pellet was washed once with PBS, and 1 ml of homogenization buffer solution (50 mM sodium phosphate, pH 6.8, 1% Triton X-100, 2 mM PMSF) was added and vortexed for 5 minutes to disrupt the cells. To the cell filtrate obtained by centrifugation (3,000 rpm, 10 min), 1 N NaOH (10% DMSO) was added to dissolve the extracted melanin, and the absorbance of melanin was measured at 405 nm with a microplate reader. Melanin was quantified, (%) Of melanin production was measured. B16F1 melanogenesis inhibition rate (%) of melanoma site was calculated by the following Equation 4, IC ₅₀ value is the concentration of a substance that inhibits melanin production by 50%.

&Quot; (4) "

Inhibition rate (%) = [(A-B) / A] x 100

A: Amount of melanin in wells to which no sample was added

B: Amount of melanin in the well to which the sample was added

As shown in Table 6 below, the inhibitory effect of B16F1 melanocyte on melanin production was tested. As a result, the IC ₅₀ of Example 2 (sample 5) was 0.08%, which is similar or superior to that of conventional albutin and soluble licorice extract Respectively.

Name of sample Treatment concentration (%) Melanin synthesis inhibitory effect (IC50) Example 2 0.1 0.08% Arbutin 0.1 0.21% Usefulness Licorice extract 0.1 0.03%

[Test Example 7] Effect of mitigation of cytotoxicity by ultraviolet irradiation

The following experiment was conducted to evaluate the mitigation effect of cytotoxicity by ultraviolet irradiation on the extract of falcatum arborescens containing Taxifolin-3-glucoside of the present invention.

Fibroblasts were placed in a 24-well test plate at 1 × 10 ⁵ cells and adhered for 24 hours. Each well was washed once with PBS and 500 占 퐇 of PBS was added to each well. After irradiating the fibroblasts with 10 mJ / cm 2 of ultraviolet light using an ultraviolet B (UVB) lamp (Model: F15T8, UVB 15W, Sankyo Dennki, Japan), PBS was removed and the cell culture medium (DMEM supplemented with 10% FBS 0.0 > ml) < / RTI > was added. Here, Example 2 (Sample 5) to be evaluated was treated and cultured for 24 hours. After 24 hours, the medium was removed, and 500 μl of the cell culture medium and 60 μl of the MTT solution (2.5 mg / ml) were added to each well, followed by culturing in a 37 ° C. CO ₂ incubator for 2 hours. The medium was removed and 500 μl of isopropanol-HCl (0.04 N) was added. After shaking for 5 minutes, the cells were lysed and 100 쨉 l of the supernatant was transferred to a 96-well test plate, and absorbance at 565 nm was measured in a microplate reader. The cell viability (%) was measured by the following equation (5) and the cytotoxic relaxation rate by ultraviolet was calculated by the following equation (6).

&Quot; (5) "

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

Bo: Absorbance at 565 nm of the well of the cell culture medium alone

Bt: Absorbance at 565 nm of a well that underwent chromogenic reaction in a sample not treated with the sample

St: absorbance at 565 nm of wells treated with the sample

&Quot; (6) "

(%) = [1- (St-Bo) / (Bt-Bo)] x 100

Bo: Cell survival rate of wells not irradiated with ultraviolet light and not treated with sample

Bt: Cell viability of wells irradiated with ultraviolet light and not treated with the sample

St: Cell viability of well treated with ultraviolet light and treated with sample

Experimental Results As shown in Table 7 below, Example 2 (Sample 5) showed that cytotoxicity by ultraviolet rays was mitigated by cytotoxicity of 41% at a concentration of 0.1%, thereby effectively preventing ultraviolet cytotoxicity. It can be seen from the above experiment that the cell damage by ultraviolet rays is effectively prevented at a low concentration.

Name of sample

Treatment concentration (%)

Cytotoxic Relaxation Rate (%)
Example 2 0.1 41

[Test Example 8] Inhibitory effect of inflammatory cytokine expression by ultraviolet irradiation

The following experiment was conducted to evaluate the inhibitory effect of the extract of Panax notoginsenosides containing Taxifolin-3-glucoside of the present invention on the expression of inflammatory cytokines expressed by ultraviolet irradiation.

Fibroblasts isolated from human epidermal tissue were placed in a 24-well test plate at 5 × 10 ⁴ and adhered for 24 hours. Each well was washed once with PBS and 500 占 퐇 of PBS was added to each well. After irradiating the fibroblasts with 10 mJ / cm 2 of ultraviolet light using an ultraviolet B (UVB) lamp (Model: F15T8, UVB 15W, Sankyo Dennki, Japan), PBS was removed and the cell culture medium ≪ / RTI > medium). Here, Example 2 (Sample 5) to be evaluated was treated and incubated for 5 hours. 150 [mu] l of the culture supernatant was taken to quantitate IL-1 [alpha] to determine the effect of inhibiting the inflammatory cytokine expression of Example 2 (Sample 5). The amount of IL-1? Was quantitated using an enzyme-linked immunosorbent assay, and the production rate of IL-1? Was calculated by the following equation (7).

&Quot; (7) "

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

Bo: IL-1? Production in wells without UV irradiation

Bt: IL-1? Production in wells irradiated with ultraviolet light and not treated with the sample

St: IL-1α production in wells irradiated with ultraviolet light and treated with the sample

Experimental Results As shown in Table 8 below, Example 2 (Sample 5) inhibits the production of IL-1α, which is an inflammatory cytokine caused by ultraviolet light, by 41% at a concentration of 0.1% You can see the defense.

Name of sample
Treatment concentration (%) Inhibitory rate of inflammatory cytokine expression (%) Example 2 0.05 25 0.1 41

[Test Example 9] Antibacterial effect

The antimicrobial test was carried out by the solid culture dilution method and the paper disc method by the antimicrobial activity test for the peony extract (Example 2) containing the taxoidin-3-glucoside of the present invention. The test strains were distributed from the Korea Research Institute of Bioscience and Biotechnology. Staphylococcus aureus KCTC 6910), gram-negative bacteria such as Pseudomonas aeruginosa KCTC 1637), Escherichia coli (E. Coli KCTC 1039), a yeast Candida yeast (Candida albicans KCTC 7965), black Aspergillus (Aspergillus fungi as niger KCTC 6910) were used. The experimental methods and results are as follows.

The minimum inhibitory concentration was determined using the Agar Serial Dilution Method to determine the antimicrobial activity of the peony root extract containing Taxon poly-3-glucoside.

Tritic soy broth was used as the fungi, and the broth was inoculated on the medium and pre-cultured for 24 hours in a 37 ° C shaking incubator. Potato dextrose culture medium was used for culture of yeast, and the bacteria were inoculated on the medium and cultured for 2 days in a 25 ° C shaking incubator. Potato dextrose agar medium was used for culture of molds, and the bacteria were inoculated on the medium and pre-cultured for 7 days in a 25 ° C incubator.

More specifically, in the case of bacteria, bacteria are inoculated on a tryptic soy medium and cultured at 37 ° C for 24 hours. In the case of yeast, bacteria are inoculated on a potato dextrose medium and cultured for 2 days at 25 ° C. The filamentous fungus was inoculated on a potato dextrose agar medium and pre-cultured for 7 days in a 25 ° C. incubator. Spores of filamentous fungus formed on the surface of the medium were recovered by using a smear rod and diluted in sterilized saline.

2 mL of each extract diluted to the appropriate concentration in 5% dimethylsulfoxide (DMSO) physiological saline solution is added to the sterilized Patriedish by the sample and the experimental species, and the control is diluted to a proper concentration in 5% physiological saline solution 2 mL of each sample was added, and 2 mL of 5% DMSO physiological saline solution was added to the control. Samples were sterilized in each Patridish, and 18 mL of a tryptic soy agar medium and a potato dextrose agar medium cooled to 48 DEG C After stirring, the mixture is allowed to solidify.

Each of the pre-cultivated test bacteria was applied to each petri dish at a final concentration of about 1 × 10 ⁶ CFU / mL for bacteria, 1 × 10 ⁵ CFU / mL for yeast, about 1 X 10 < ⁴ > CFU / mL in each petri dish. Each petri dish was incubated at 37 ° C for 24 hours, the yeast was cultured at 25 ° C for 3 days, the filamentous bacteria were cultured in a 25 ° C incubator for 7 days, and then colony formation was observed in each compartment, The minimum sample concentration of the plate was defined as the minimum inhibitory concentration (MIC), and the results are shown in Table 9. At this time, the minimum inhibitory concentration means that the smaller the value, the higher the antibacterial effect.

Strain Minimum inhibitory concentration (MIC, / / mL) S. aureus 50 P. aeruginosa 21 E. Coli 12 C. albicans 110 A. niger 140

As shown in Table 9, the peony root extract containing Taxifolin-3-glucoside exhibited superior antimicrobial activity against all four bacteria, yeast and mold.

In order to test the antimicrobial activity of peony root extract containing Taxifolin-3-glucoside, the following experiment was conducted by the paper disc method. Each strain cultivated on a plate culture medium was plated in a volume of 1 pl, cultured in 10 mL of liquid culture medium for 24 hours, and then inoculated with 10 mL of the culture broth in 0.1 mL of the bacterial culture solution for 6 hours, ⁷ cfu / mL, and uniformly plated with a sterilized donor rod. Then sterilized round filter paper (6 mm, Satorius, Germany) was placed on a solid plate medium, and then sample 5 dissolved in a solvent was absorbed to 0.05 ml / disk and cultured. Staphylococcus aureus , Pseudomonas aeruginosa), Escherichia coli (E. Coli) was measured for 37 ℃, the type of fungi Candida yeast (Candida albicans), black Aspergillus (Aspergillus niger) are each 24 hours in 27 ℃, the diameter of the clear zone around the filter paper using 120 hours of incubation . As a comparative group, methyl paraben, which is known to have a strong antibacterial activity, was used, and the results are shown in Table 10 below. At this time, the transparent zone around the filter paper is an index of the degree of inhibition of proliferation of the strain, and the larger the diameter, the higher the antibacterial effect against the strain.

Strain Extraction method Transparent zone diameter (mm) S. aureus Example 2 13 Methyl paraben 11 P. aeruginosa Example 2 15 Methyl paraben 12 E. Coli Example 2 24 Methyl paraben 18 C. albicans Example 2 17 Methyl paraben 13 A. niger Example 2 25 Methyl paraben 24

As shown in Table 10, the peony root extract containing Taxifolin-3-glucoside exhibited stronger antimicrobial activity against four bacteria, yeast and mold than the methylparaben known as a strong antimicrobial agent.

[Formulation Example 1 and Formulation Comparative Example 1]

The composition of the nutritional cream containing the peony root extract containing Taxon poly-3-glucoside according to Example 2 (Sample 5) was prepared as shown in Table 11 below.

number ingredient Formulation Comparative Example 1 Formulation Comparative Example 1 One Pro-type glycerin monostearate 2.0 2.0 2 Stearic acid 1.5 1.5 3 Cetearyl alcohol 2.2 2.2 4 Wax 1.0 1.0 5 Squalane 3.0 3.0 6 Hardened vegetable oil 1.0 1.0 7 Sorbitan stearate 0.6 0.6 8 Mineral oil 5.0 5.0 9 Polysorbate 60 1.5 1.5 10 Dimethicone 1.0 1.0 11 Trioctanoin 5.0 5.0 12 Betaine 3.0 3.0 13 Triethanolamine 1.0 1.0 14 glycerin 5.0 5.0 15 Sodium hyaruronate 3.0 3.0 16 The peony extract of Example 2 1.0 - 17 Distilled water Balance Balance 18 Preservative, fragrance, pigment a very small amount a very small amount

The aqueous components of the raw materials 12 to 15 and 17 were completely dissolved by heating to 80 DEG C and then the raw materials 1 to 11 and 16 were heated to 80 DEG C to obtain the raw materials 12 to 15 and 17 (Homo Mixer Mark II, Primix, JPN) for 15 minutes at 3,000 rpm. Thereafter, the raw material 18 was added, stirred for 5 minutes, and then cooled to room temperature.

[Test Example 10] Measurement of whitening effect of formulation application

The whitening effect of the nutritional cream prepared in Formulation Example 1 and Comparative Formulation Example 1 was measured by the following method. 30 women aged 25 years or older who had spots, freckles and pigmentosis were subjected to the nutritional creams of Formulation Example 1 and Comparative Example 1 for 12 weeks. The changes in skin color were measured with a chromometer (CR-410 , Minolta, Japan) was used to measure the change in color brightness (ΔL). The average value of 30 persons was calculated. The higher the brightness change value, the higher the whitening effect. The experimental results are shown in Table 12 below.

medium Formulation Example 1 Formulation Comparative Example 1 Skin color change (ΔL) 6.19 0.22

As shown in Table 12, the nutritional cream of Formulation Example 1 to which Example 2 (Sample 5) was added had a much higher whitening effect than the nutrition cream of Formulation Comparative Example 1 to which Example 2 (Sample 5) was not added .

[Test Example 11] Measurement of wrinkle improvement effect

In order to measure the wrinkle-reducing effect of the nutritional cream prepared in Formulation Example 1 and Comparative Formulation Example 1, the following method was used. Ten women aged 30 or older undergoing skin aging were allowed to use the nutritional cream of Formulation Example 1 and Comparative Example 1 for 12 weeks on the crow feets region, and after 4 weeks, 8 weeks, 12 weeks The depth of the wrinkles (탆) was measured using a skin visiometer (SV-600, C + K, Germany) by an image analysis method using an interval wrinkle replica template, Respectively.

medium Formulation Example 1 Formulation Comparative Example 1 Before application 329 ± 13 326 ± 12 After 12 weeks of application 255 ± 11 320 ± 22 Wrinkle depth reduction rate (%) 22.5 1.8

As shown above, the nutritional cream of Formulation Example 1 to which Example 2 (Sample 5) was added had a significantly improved wrinkle-reducing effect compared with the nutritional cream of Formulation Comparative Example 1 to which Example 2 (Sample 5) .

[Test Example 12] Measurement of atopy improvement effect of application of formulation

In order to measure the atopic improvement effect of the nutritional cream prepared in Formulation Example 1 and Comparative Formulation Example 1, the following method was used. Acetone was applied as a skin barrier damaging method. When the transepidermal water loss (TEWL) reached 4.0 g / m ² / h by dispensing acetone into the aliquots of 8-12 week old hairless mice, the nutritional cream of Formulation Comparative Example 1 and Comparative Formulation Example 1 was applied Respectively. TEWL was measured with a Tewameter 210 evaporator from C + K (Cologne, Germany). After 6 hours of application, TEWL was measured and the degree of reduction of TEWL was measured to evaluate the degree of reduction of TEWL. The recovery rate used in the efficacy evaluation was calculated as shown in Equation 8 below and the results are shown in Table 14 below.

&Quot; (8) "

(Barrier recovery rate) = (1- (B _{t = 6} - B _{t = 0} ) / (B _{t = d} - B _{t = 0} )

B _{t = 6} : TEWL measurement after 6 hours after skin barrier damage

B _{t = 0} : TEWL measured before skin barrier damage

B _{t = d} : TEWL measured immediately after skin barrier damage

Recovery rate (%) medium Formulation Example 1 Formulation Comparative Example 1 6 hour recovery rate
(Based on initial TEWL) 39 12

As shown above, the nutritional cream of Formulation Example 1 to which Example 2 (Sample 5) was added had a higher recovery than the nutritional cream of Formulation Comparative Example 1 in which Example 2 (Sample 5) was not added, The effect was shown to increase.

Examples of formulations of the present invention are a soft lotion, a convergent lotion, a nutritional lotion, a massage cream, an essence and a pack, but the formulation of the cosmetic composition of the present invention should not be construed as being limited thereto. Lt; / RTI > is possible.

[Formulation Example 1] Flexible longevity

number ingredient content(%) One glycerin 5.00 2 1,3-butylene glycol 3.00 3 PEG 1500 1.00 4 Allantoin 0.10 5 DL-Panthenol 0.30 6 EDTA-2Na 0.02 7 Benzophenone-9 0.04 8 ethanol 10.00 9 Octidodeces-16 0.20 10 Polysorbate 20 0.20 11 The peony extract of Example 1 5.0 12 Preservative, fragrance, pigment a very small amount 13 Distilled water Balance

[Formulation Example 2] Convergent lotion

number ingredient content(%) One glycerin 2.00 2 1,3-butylene glycol 2.00 3 Allantoin 0.10 4 DL-Panthenol 0.30 5 EDTA-2Na 0.02 6 Benzophenone-9 0.04 7 ethanol 15.00 8 Polysorbate 20 0.20 9 The peony extract of Example 2 1.0 10 Citric acid a very small amount 11 Preservative, fragrance, pigment a very small amount 12 Distilled water Balance

[Formulation Example 3] Nutritional lotion

number ingredient content(%) One Cetearyl alcohol 1.00 2 Glyceryl stearate 0.50 3 Polysorbate 60 1.00 4 Sorbitan sesquioleate 0.30 5 Cetyl octanoate 6.00 6 Squalane 4.00 7 Shapower Oil 4.00 8 Butylene glycol 4.00 9 glycerin 4.00 10 Carbomer 0.10 11 Triethanolamine 0.10 12 The peony extract of Example 1 1.00 13 Preservative, fragrance, pigment a very small amount 14 Distilled water Balance

[Formulation Example 4] Essence

number ingredient content(%) One glycerin 10.00 2 Betaine 5.00 3 PEG 1500 2.00 4 Allantoin 0.10 5 DL-Panthenol 0.30 6 EDTA-2Na 0.02 7 Benzophenone-9 0.04 8 Hydroxyethylcellulose 0.10 9 Carboxyvinyl polymer 0.20 10 Triethanolamine 0.18 11 Octyldodecanol 0.30 12 Octyldodec-16 0.40 13 ethanol 6.00 14 The peony extract of Example 2 1.00 15 Preservative, fragrance, pigment a very small amount 16 Distilled water Balance

[Formulation Example 5] Pack

number ingredient content(%) One Polyvinyl alcohol 15.00 2 Cellulose sword 0.15 3 glycerin 3.00 4 PEG 1500 2.00 5 Sheik Destrin 0.15 6 DL-Panthenol 0.30 7 Allantoin 0.10 8 Monoammonium glycyrrhizin acid 0.20 9 Nicotinamide 0.40 10 ethanol 5.00 11 PEG 40 hardened castor oil 0.30 12 The peony extract of Example 1 1.00 13 Preservative, fragrance, pigment a very small amount 14 Distilled water Balance

Claims (17)

Containing as an active ingredient a fungicidal extract comprising a taxane polyglycoside,
The peony extract is water and ethanol extract,
The cosmetic composition for inhibiting skin water loss of atopic skin, wherein the taxifolin glycoside is taxifolin-3-glucoside. delete delete The method according to claim 1,
Wherein the crude extract is contained in an amount of 0.0001 to 10% by weight based on the total weight of the composition. delete delete delete delete Dissolving the taxane-containing glyceride in a solvent;
Concentrating the eluted extract;
Placing the concentrated extract in two or more solvents and fractionating;
Passing the extract obtained in the fractioned layer through a column; And
And recrystallizing the column passing liquid by precipitation,
Wherein said taxifolin glycoside is taxifolin-3-glucoside,
The solvent of the fractionation step is water and ethanol,
Wherein the recrystallization step is performed at -300 to 25 ° C and 0.5 to 800 MPa. 10. The method of claim 9,
Wherein the solvent of the elution step is at least one selected from the group consisting of water, ethanol, methanol, butanol, ethyl acetate, ethyl ether, chloroform and hexane. 10. The method of claim 9,
Wherein the step of concentrating the eluted extract is carried out with at least one selected from the group consisting of heat drying, vacuum drying, spray drying and freeze drying. delete 10. The method of claim 9,
Wherein the column in the column passing step is a resin column. 14. The method of claim 13,
Wherein the resin is at least one resin selected from the group consisting of Amberlite XAD resin, Dow X resin, Suferite DAX resin, and Diaion HP-10, 20, 30, 40, and 50 resins. delete 10. The method of claim 9,
Wherein the fractionation step comprises fractionating and then removing the solvent of the fractionated layer and concentrating. 10. The method of claim 9,
Wherein said extracting method further comprises the step of drying and obtaining the recrystallized extract after recrystallization.

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