KR101909225B1 - A composition comprising the Isoscopoletin isolated from the extract of Artemisia Capillaris for anti-wrinkle - Google Patents

Abstract

The present invention relates to the use of Artemisia The present invention relates to a composition for improving wrinkles containing isoscopoletin isolated from a Capillaris extract as an active ingredient, which comprises (1) Artemisia Capillaris ) with water, an alcohol or a mixed solvent thereof; And (2) fractionating the mugwort-mugwort extract of step (1) with an organic solvent to prepare a mugwort fraction; . According to the present invention, there is provided a method for producing an extract of Artemisia syrup extract having wrinkle improving activity or an extract of Artemisia syrup extract having enhanced wrinkle improving activity, thereby preventing or improving skin aging and wrinkles.

Description

[0001] The present invention relates to a composition for improving wrinkles comprising isoskopoletin isolated from Artemisia sp.

The present invention relates to the use of Artemisia The present invention relates to a wrinkle-improving composition containing isoscopoletin isolated from an extract of Capillaris as an active ingredient. More particularly, the present invention relates to a waxy extract having enhanced wrinkle-improving activity or a method for producing the fraction.

In human skin, various physical and chemical changes occur during the aging process. The cause is largely classified into intrinsic aging and photo-aging, and research has been conducted actively. Specifically, free radicals may be activated by ultraviolet rays, stress, disease states, environmental factors, wounds, and aging. If such conditions are deepened, the antioxidant defense network existing in the living body may be destroyed, Thereby promoting adult diseases and aging. Therefore, it is necessary to protect the cell membrane by destroying the free radicals mediated by free radicals, ultraviolet rays, and inflammation that occur during the metabolism of the body and to proliferate cells by regenerating already damaged cells by active metabolism. Lt; / RTI >

Artemisia is a perennial herbaceous plant of Compositae which is distributed in Korea and Asia and Europe. It is known that there are about 400 species. About 300 species are native to Korea, 10-0401472 (Functional beverage containing Artemisia sp. Extract and method for producing the same) describes Artemisia sp. Extract for use in blood vessel relaxation, stroke prevention, or anticancer beverage. A method for obtaining fractions using solvent polarity for natural products such as mugwort is to sequentially treat hexane, chloroform, dichloromethane, ethyl acetate, and butanol to fractionate the fractions according to the polarity. However, this method has an advantage of being capable of subdividing the substance according to the polarity, but in the experiment for separating the active ingredient, the active ingredient is dispersed.

Accordingly, the inventors of the present invention have devised a method of preventing the dispersion of the active ingredient and obtaining a larger amount of the active fraction as compared with the known solvent fractionation method, and by applying it to the extract of Artemisia capillaris, (Isoscopoletin) isolated from Artemisia sp. Extract or Artemisia sp. Extract, which can be easily expressed, as an active ingredient.

An object of the present invention is to provide a cosmetic composition, a soap composition, a pharmaceutical composition and a food composition which are harmless to the human body and are effective in improving wrinkles by providing an extract of Artemisia mugwort and its fractions which can best exhibit wrinkle- .

In order to achieve the above object, the present invention provides a composition for improving wrinkles containing isoscopoletin isolated from Artemisia sp. Extract as an active ingredient.

The composition is characterized by being a cosmetic composition, a soap composition, a pharmaceutical composition, or a food composition.

The present invention relates to a method for producing a composition for improving wrinkles containing isoscopoletin isolated from an extract of Artemisia capillaris as an active ingredient, which comprises (1) Artemisia Capillaris ) with water, ethanol or a mixed solvent thereof; And (2) fractionating the mugwort-mugwort extract with an organic solvent to prepare an mugwort fraction; The present invention is characterized in that it includes another feature.

The organic solvent in step (2) is ethyl acetate, methanol, hexane or a mixture thereof.

In the above production method, it may further comprise the step of separating and purifying the produced Artemisia spp. Fraction.

According to the present invention, there is provided a method for producing an extract of Artemisia syrup extract having wrinkle improving activity or an extract of Artemisia syrup extract having enhanced wrinkle improving activity, thereby preventing or improving skin aging and wrinkles.

FIG. 1 is a schematic view showing a process for producing an extract of Artemisia princeps or Artemisia sp., According to an embodiment of the present invention.
FIG. 2 is a result of a medium pressure liquid chromatography (MPLC) fraction according to an embodiment of the present invention.
FIG. 3 is a result of HPLC analysis of Isoscopoletin isolated and purified from Artemisia sp. Fractions (MPLC fraction 5) according to an embodiment of the present invention.
Figure 4 shows the results of analysis of hydrogen nuclear magnetic resonance ( ¹ H NMR, ¹ H Nuclear Magnetic Resonance) of isoscopoletin according to one embodiment of the present invention.
Figure 5 shows the results of carbon nuclear magnetic resonance ( ¹³ C NMR, ¹³ C nuclear magnetic resonance) of isoscopoletin according to one embodiment of the present invention.
FIG. 6 is a graph illustrating cytotoxicity of the extract of Artemisia princeps, which is extracted with ethanol according to an embodiment of the present invention.
FIG. 7 is a graph showing Elastase activity of the extract of Artemisia princeps, extracted with ethanol according to an embodiment of the present invention.
8 is a graph showing cytotoxicity of isoscopoletin according to an embodiment of the present invention.
9 is a graph showing the results of measuring the elastase activity of isoscopoletin according to an embodiment of the present invention.
10 is a photograph of a result of Western blotting of isoscopoletin according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail.

There is provided a composition for improving wrinkles containing isoscopoletin isolated from Artemisia sp. Extract as an active ingredient according to an embodiment of the present invention.

Artemisia spp., Which is a member of the mugwort, capillaris ; AC), and its young stem and leaves are known as in Jin Chen (阴 Chen 蒿), and it is known that dandruff, asthma suppression, anthelmintic effect, diuretic effect and the like include scoparone, chlorogenic acid, Capillin, etc. are known (Information Supervision and Shin Min-gyo, Encyclopedia of Enlightenment, pp 1016-1017, Young Lim, 1998). A perennial herb belonging to Asteraceae Mugwort is several species as follows. Even if they belong to the same genus, they exhibit different effects due to differences in their characteristics, contained components and active substances.

The composition according to an embodiment of the present invention may be contained in any amount within a range that the extract of Artemisia princeps or its fraction is not toxic to the human body, and may be contained in an amount of 0.001 to 99.99% by weight based on the total weight of the composition.

In a composition for improving wrinkles containing isoscopoletin isolated from Artemisia sp. Extract according to the present invention as an active ingredient, isoscopoletin may be represented by the following formula (1).

[Chemical Formula 1]

The composition according to this embodiment can be used as a cosmetic composition. Forms such as emulsions, lotions, creams (water-in-oil type, water-in-water type, multiphase), solutions, suspensions (anhydrous and aqueous), anhydrous compounds (oil and glycol), gels, masks, packs or powders Oils, surfactants, fatty acids, silicone oils, wetting agents, moisturizers, viscosifying agents, emulsifiers, stabilizers, sunscreen agents, and the like, which can be suitably selected and used by those skilled in the art in cosmetic formulations , A coloring agent, a perfume, and the like.

The composition according to this embodiment can be used as a soap composition. Emulsifying agent, water softening agent, etc. as an additive to a soap base such as palm oil, palm oil, soybean oil, pharmacopoeial, olive oil and palm kernel oil, or animal fats such as tallow, lard, have. As the above-mentioned skin moisturizing agent, glycerin, erythritol, polyethylene glycol, propylene glycol, butylene glycol, pentylene glycol, hexyl glycol, isopropyl myristate, silicone derivatives, aloe vera, sorbitol, Wax fatty alcohols, hydrocarbons, natural plant extracts and the like can be used. As the water softening agent, tetrasodium edetate and the like can be used. The additive may further include an antimicrobial agent, a dispersant, a foam inhibitor, a solvent, a scouring inhibitor, a corrosion inhibitor, a perfume, a dye, a sequestering agent, an antioxidant, an antiseptic and the like.

The composition according to this embodiment can be used as a pharmaceutical composition. Specifically, it is a skin irritant caused by acne, atopy, psoriasis, dermatitis and the like caused by inflammation reaction, skin aging caused by increase of active oxygen, wrinkles, and stain, freckles, blotch, It can be used as a pharmaceutical composition having a preventive or therapeutic effect on a skin pigmentation disease such as a pigment, a pigment, a drug-induced pigmentation, and an inflammation-induced pigmentation. The pharmaceutical composition can be administered at various times during clinical administration, including oral, transdermal, subcutaneous, intravenous, or muscular, and can be applied as an external preparation for skin. In addition, formulations can be prepared in various formulations. In the case of formulation, formulations can be prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants that are usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose and gelatin, . Examples of liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances and preservatives . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations and suppositories. Examples of the non-aqueous solvent and the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Witepsol, macrogol, tween 61, cacao paper, laurin, glycerol and gelatin may be used as a base for suppositories.

In addition, oral administration or parenteral administration may be carried out according to the intended method, and parenteral administration may be selected by external or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, . The dosage varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity. The dosage varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of disease, and the daily dosage is based on the amount of the extract or fraction thereof 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg, and can be administered 1 to 6 times a day. The composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

The composition according to this embodiment can be used as a food composition. The health food can be used as it is or directly with other food or food ingredients, and can be suitably used according to conventional methods. There is no particular restriction on the type of health food. Examples include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, soups, , An alcoholic beverage and a vitamin complex, and includes all the health foods in a conventional sense. The health beverage composition may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 g of the composition of the present invention. In addition to the above, the health food of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, , A carbonating agent used in carbonated drinks, and the like. It may also contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

In the production method for the iso Scoring retinoic pole (Isoscopoletin) injinssuk separated from the extract according to one embodiment of the present invention as an active ingredient, improving wrinkles containing composition, (1) injinssuk (Artemisia Capillaris ) with water, an alcohol or a mixed solvent thereof, and (2) fractionating the Artemisia sp. Extract with an organic solvent to produce Artemisia sp. Fractions.

Step (1) is to produce an extract of Artemisia princeps, using water, alcohol or a mixture thereof, and the extracted Artemisia extract may be filtered or concentrated under reduced pressure and dried. As the alcohol, C ₁ to C ₂ lower alcohol is preferably used, and as the lower alcohol, ethanol or methanol is preferably used. As the extraction method, it is preferable to use shaking extraction, Soxhlet extraction or reflux extraction, but it is not limited thereto. The extraction solvent is preferably added 1 to 5 times, more preferably 1 to 2 times, more preferably 1.5 times, more preferably 1 to 5 times. The extraction temperature is preferably 20 to 100 ° C, more preferably 20 to 40 ° C, most preferably room temperature, but is not limited thereto. Further, the extraction time is preferably 10 to 48 hours, more preferably 12 to 36 hours, but is not limited thereto. The number of times of extraction is preferably 1 to 5 times, more preferably 1 to 2 times, more preferably 1 time, but is not limited thereto. Artemisia Capillaris ) may be used without restrictions, such as cultivated or marketed. It is most preferable that all organs such as leaves, flowers, roots, stems, and seeds are available, but it is not limited thereto.

In the drying of the extract, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the concentration under reduced pressure, but the concentration is not limited thereto, and it is preferable that the concentration is about 2 to 10 Brix as measured by a sugar content meter , And 3 to 6 bricks, but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.

Step (2) is a step of extracting the extract of Artemisia japonica extract obtained in step (1) with an organic solvent to prepare an extract of Artemisia japonica, wherein Isoscopoletin is extracted from water extract, alcohol, For example. The organic solvent may be ethyl acetate, methanol, hexane, or a mixed solution thereof. Preferably, the organic solvent is a mixed solution of water and ethyl acetate, or a mixed solution of methanol and hexane. And fractionating the extract of Artemisia mugwort with a mixture of water and ethyl acetate as a solvent, whereby the ethyl acetate fraction of the present invention can be obtained. At this time, the concentration of ethyl acetate in the mixed solution of water and ethyl acetate is preferably 40 to 70% by volume, more preferably 50 to 60%, most preferably 50% by volume.

And then fractionating the ethyl acetate fraction using a mixture of methanol and hexane as a solvent, whereby the methanol fraction of the present invention can be obtained. At this time, the concentration of methanol in the mixed solution of methanol and hexane is preferably 70 to 95% by volume, more preferably 80 to 90%, most preferably 90% It is possible.

Further, the methanol fraction may be separated by using a mixed solvent of ethyl acetate and hexane, ethyl acetate or methyl alcohol as a solvent, and the fraction (Fr.1 to Fr.8) of the present invention can be obtained have. At this time, the concentration of ethyl acetate in the mixed solution of ethyl acetate and hexane is preferably 10 to 70%, more preferably 20 to 50% or 55 to 70%, more preferably 20 to 50%, or 55 to 66% Most preferably, but not exclusively. Separation can be performed using normal silica gel or reversed phase ODS (octadecylsiyl silica gel), but it is not limited to such a method as long as the concentration gradient is possible. In the method for preparing the fraction, the extraction temperature is preferably 20 to 100 ° C, more preferably 20 to 40 ° C, most preferably room temperature, but is not limited thereto. Further, the extraction time is preferably 10 to 48 hours, more preferably 12 to 36 hours, but is not limited thereto. Further, the fractionation can be obtained by repeating the fractionation process 1 to 5 times, preferably 3 times. It is preferable, but not limited, to fractionate and concentrate under reduced pressure.

In addition, the method according to an embodiment of the present invention may further include a step of purifying the fraction from Artemisia sp. This is a step for separating and purifying the fractions (Fr.1 to Fr.8) prepared in the step (2) so that a large amount of isoscopoletin can be contained, And may be a step of separation through recrystallization, thereby obtaining isoscopoletin isolated from Artemisia sp. Extract of the present invention. When the extract of Artemisia princeps or its fractions are produced through this example, the active ingredient can be prevented from being dispersed and a larger amount of the active fraction can be obtained. Thus, the extract or fraction which can best exhibit the wrinkle- Can be obtained.

When the extract of Artemisia officinalis is extracted with a mixture of water and ethyl acetate, the aqueous layer contains water-soluble components and the ethyl acetate layer contains a lipid-soluble component. At this time, only the ethyl acetate layer shows activity Soluble component. In addition, when only the ethyl acetate layer is separated and extracted with a mixed solution of methanol and hexane, hexane will contain lipid components, and the methanol layer will contain components with slightly higher polarity such as flavonoid and lignan, Means that the component exhibiting activity is a component having a somewhat higher polarity among fat-soluble components.

On the contrary, when a conventional solvent fractionation method is used, the active ingredients are dispersed in various solvents depending on the polarity, so that when the active ingredients are mostly polarized as in the present invention, a large amount of active ingredients It will be disadvantageous to obtain. Therefore, the production method according to one embodiment of the present invention is an effective method for easily obtaining a larger amount of active ingredient by overcoming the limitations of the existing method when the active ingredient is separated using Artemisia japonica.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will now be described in detail with reference to the accompanying drawings. It will be apparent to those skilled in the art that this embodiment is for illustrative purposes only and that the scope of the present invention is not construed as being limited by these embodiments.

EXAMPLE 1 Preparation of Artemisia cinerea Extract According to an Example of the Invention

The mushroom collected from the Hwasun area in Jeollanam-do was dried and stored at 4 ℃. The mixture was extracted with 100% ethanol (71.343 g) as a solvent at room temperature for 3 days. The mixture was filtered under reduced pressure using a filter paper (Advantec No.2, Toyo, Japan) and then subjected to vacuum filtration with a rotary vacuum evaporator (EYELA, Japan) The concentrate was lyophilized and used as a sample. (Fig. 1)

Example 2: Preparation of the Artemisia sp. Fractions according to one embodiment of the present invention

 For the solvent fraction of the ethanolic Artemisia monocytogenes extract prepared according to Example 1, the ethanol extract of Artemisia monocytogenes was mixed with water and ethyl acetate at a ratio of 1: 3, followed by solvent fractionation. Then, an ethyl acetate layer 38.56 g) was further subjected to solvent fractionation with 1: 3 of 90% methanol (24.19 g) and? -Hexane. 5 g of the prepared 90% methanol fraction of artificial wormwood was dissolved in 20 ml of ethyl acetate, and then silica gel 40 (SIL-25,, YMC) and MPLC were carried out. The experiment was developed under the conditions of ethyl acetate and a-hexane (2: 8 to 10: 0, 60 minutes), ethyl acetate and methyl alcohol to separate into eight fractions (FR) .

 Operating condition Time (min) EtOAc (%) Hexane (%) FR. No 0 20 80
FR.1 - 6 60 100 0 70 100 0 FR. 7 MeOH MeOH washing FR. 8

Example  3. In the extract Isoskopoletin ( Isoscopoletin ) Separation purification (recrystallization) is carried out

In order to increase the purity of MPLC fraction 5 in the MPLC fraction obtained in Example 2, fractions were dissolved in ethyl acetate and methanol, and the product was kept in a frozen state for recrystallization. At this time, the supernatant was removed and only the recrystallized material was separated and purified.

Example  4. Work of the invention In the embodiment  Refined eggplant The fraction (FR.5) HPLC  Through analysis Isoskopoletin (Isoscopoletin) analysis.

(FR.5) was analyzed by high performance liquid chromatography (Waters 2000 series, Japan). The column was a YMC C18 column (5 占 퐉, 250 mm 占 4.6 mm) (254 nm, 285 nm, 366 nm) was used. The mobile phase was water and methanol (MeOH), and the gradient was set through the pump program. The flow rate was 1 ml / min. The composition and slope conditions of the mobile phase solvent were set at various conditions as shown in Table 3 below, and the analysis experiments were performed. Samples used for analysis were dissolved in methanol at a concentration of 1 mg / ml, and 10 μl of the prepared solution was injected into an autosampler. (Fig. 3)

 Operating condition Time
(min) MeOH (%) Water (%) 0 40 60 40 40 60 50 100 0 55 100 0 60 40 60 70 40 60

Example  5. Work of the invention In the embodiment  Isolated from Artemisia sp. Of Isoscopoletin  Nuclear Magnetic Resona nce , NMR) analysis

¹ H-NMR and ¹⁴ C-NMR were measured using a 400 MHz nuclear magnetic resonance spectrometer (Varian, 400-MR) from Myongji University Instrumental Analysis and Information Support Center to determine the structure of the separated compounds. Chloroform was used as the medium (Figs. 4 and 5).

Experimental Example  One. MTT  assay to determine wrinkle-reducing efficacy

MTT assay was performed to confirm the wrinkle-reducing effect of this example using fibroblasts. Inoculate 24 well plates with 24 well plates at 1.5 x 10 ^ 4/500 μl per well. After 24 hours, change the culture medium to 500 μl of DMEM (HIGH GLUCOSE FBS, + Antibiotics) medium. After 24 hours, treat each extract concentration. After 24 hours, 50 μl of MTT reagent dissolved in PBS at 5 mg / ml is dispensed. And incubated for 3 hours in an incubator. After the medium is removed, 300 μl of dimethyl sulfoxide (DMSO) is added and the absorbance is measured at 570 nm (FIGS. 6 and 8).

Experimental Example  2. Elastinolytic enzyme assay ( Elastase  assay to determine wrinkle-reducing efficacy

It is inoculated to a 6 well plate at a density of 1.5 × 10 5/2 ml per well. After 24 hours, the medium is changed to 2 ml of DMEM (HIGH GLUCOSE FBS, + Antibiotics) medium. After 24 hours, treat each extract concentration, and after 48 hours, remove the medium. Place the cold PBS (phosphate buffer saline) on the plate on ice and stop the reaction. After removing the PBS, add 150 μl of lysis buffer. Scrape the cells in the wells using a scraper on ice and put them in a tube (E-tube) and lightly mix the tube (E-tube). Respond to ice for 15 minutes (repeat twice). Centrifuge at 13000 rpm, 15 min, 4 캜. Add 90ul of the supernatant to a 96-well plate and dispense into each well. STANA (N-succinyl-Ala-Ala-Ala-p-nitroanilide), which reacts with elastinase, is made to 50 mM and added with 1.8 ul (total volume 2%). React for 2 hours in the incubator. Absorbance is measured at 405 nm. The value is calculated as sample / control x 100 (Figs. 7 and 9).

Experimental Example  3. Western Western blot  Check wrinkle improving efficacy through

Inoculate with 2 × 10 ^ 5/2 ml per well on a 6-well plate. After 24 hours of culture, the medium is removed and the medium is replaced with 2 ml of DMEM (HIGH GLUCOSE FBS, + Antibiotics) medium. After 24 hours, the extract is treated with 1 ml per well (concentration) per concentration. After culturing for 24 hours, the medium is collected and stored frozen. (RIPA (Radioimmunoprecipitation Assay) buffer (10 ml) is added to 10 μl of Protease Inhibitor, the cells are lysed with 120 μl, scraped with a scraper and placed in tube And centrifuged at 13,000 rpm for 15 min at 4 ° C. The supernatant was quantitated using BCA Protein Assay kit. The protein was separated by electrophoresis on 10% SDS-PAGE and analyzed by using a Trans-Blot Turbo (BIORAD) Transfer to a poly-vinyl difluoride (PVDF) membrane 9 to 10 ml of blocking buffer is added to the membrane and allowed to react for 30 minutes at room temperature Tris-buffered saline and Tween 20 buffer (MMP-1 sb137332 Abcam 1: 3000) Slowly react for one day at 4 ° C Wash rapidly 3 times for 10 min with TBS-T . (Goat Anti-Rabbit IgG (H + L) BIORAD 1: 1000) at room temperature for 1 hour. After washing three times, the fluorescence is detected to detect the band (Chemiscope 3000 Fluorescence / Chemiluminescence Imaging System) (Figure 10).

Having described specific portions of the present invention in detail, it will be apparent to those skilled in the art that this specific description is only a preferred embodiment and that the scope of the present invention is not limited thereby. It will be obvious. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (8)

(1) preparing Artemisia sp. Extract with Artemisia Capillaris as a water, an alcohol or a mixed solvent thereof;
(2) filtrating, purifying and concentrating the mugwort extract;
(3) fractionating the concentrated Artemisia sp. Extract of step (2) with an organic solvent to prepare an Artemisia sp. Fraction; And
(4) recrystallization of the above artichoke fraction,
In step 1), the temperature for extracting the mugwort extract is 20 to 40 ° C,
The method for preparing a wrinkle-improving composition according to claim 2, wherein, in step (2), the concentration of the extract is from 3 to 6 Brix, wherein the concentrate is isoscopoletin isolated from an extract of Artemisia princeps.
The method according to claim 1,
In the fractionation with an organic solvent in the step (3)
(a) using an ethyl acetate mixed solution and fractionating to produce an ethyl acetate fraction;
(b) preparing a fraction of the methanol fraction by using and fractionating hexane and methanol mixed solution into the ethyl acetate fraction; And
(c) dissolving the methanol fraction in ethyl acetate and then fractionating the extract. The method for producing a wrinkle-improving composition according to claim 1, wherein the isocyanate is isopropylbenzene.
The method according to claim 1,
The recrystallization of step (4) may be performed by adding a composition for improving wrinkles containing isoscopoletin as an active ingredient, which comprises separating and purifying only the recrystallized material by maintaining the frozen state to enhance the purity of the artichoke spp. Gt;
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