KR20100060709A - Method for effectively extracting, purifying and processing decursin from the extract of angelica gigas nakai, and the composition for skin external application containing the decursin prepared by the method - Google Patents

Abstract

PURPOSE: A method for extracting, purifying, and processing decursin from Angelica gigas nakai extract is provided to maximize efficiency of decursin on the skin and to enhance skin absorption rate. CONSTITUTION: A method for preparing decursin from Angelica gigas Nakai extract comprises: a step of extracting Angelica gigas Nakai by supercritical extraction using carbon dioxide; a step of isolating and purifying decursin through column chromatography using hexane and ethylacetate; and a step of capturing purified decursin in a polymer nano carrier. The polymer nano carrier is polycaprolactone-block polyethylene glycol block copolymer, polyethylene glycol and poly-D,L-lactic acid-co-glycolic acid block copolymer, or linear polyethylene imine and poly-carprolactone block copolymer.

Description

A method for effectively extracting, purifying and processing decursin from the extract of Angelica gigas Nakai, and the composition for skin external application containing the decursin prepared by the method}

The present invention relates to a method for extracting, purifying and processing decursin from a extract of Angelica vulgaris and to an external composition for skin containing the decusin as an active ingredient, and more specifically, extracting Angelica pericaris with a supercritical extraction method using carbon dioxide. And then extracting, purifying and processing decancin, which is isolated and purified by a combination of specific eluting solvents and then collected in a polymer nanocarrier, thereby maximizing the efficacy in the skin of decansin, the main potent substance of Angelica extract, and enhancing skin absorption rate, and It relates to an external composition for skin containing the deckerin as an active ingredient.

The skin is the body's primary barrier, acting as a barrier to protect organs in the body from changes in temperature and humidity, stimuli from the external environment such as UV and pollutants. However, internally, as hormones age, the secretion of various hormones that regulate metabolism decreases, and the function and activity of cells of immune cells is reduced, thereby reducing the biosynthesis of immune proteins and bioconstituent proteins necessary for the living body. Externally, physical and chemical irritation and stress caused by an increase in ultraviolet rays, free radicals and free radicals due to environmental pollution such as ozone layer destruction weaken the normal function of the skin, promote aging, and cause pigmentation. In order to prevent this phenomenon, by using bioactive substances obtained from various known animals, plants and microorganisms in addition to cosmetics, efforts to effectively maintain skin's unique functions and activate skin cells to effectively suppress skin aging It has been.

However, most of the existing raw materials had various problems such as insufficient efficacy or causing skin side effects. Therefore, studies on raw materials having an anti-aging effect without causing skin side effects are being actively conducted.

In particular, various clinical and empirical prescriptions have been proven to have various efficacy and safety, reducing the risk of the approach has been attempted to find a raw material having an anti-aging effect from herbal medicines.

Angelica is the most widely used natural product among herbal medicines. Angelica is an Angelica genus belonging to the Apiaceae family, and it is originated from Angelica gigas Nakai in Korea, Angelica sinensis Diels in China, and Angelica acutiloba Kitagawa in Japan. The roots of the Angelica are collected before the flowering of the Angelica Angelica, and the root of the Angelica is used. The root of the Angelica is known to have a blood replenishment effect, which is known to be prescribed as a blood donor for anemia and gynecological diseases. Roots of Angelica gigas are coumarine derivatives such as decusin, decosinol and nodakenin, and α-pinene, limonene and β-eudesmol. And an essential oil component containing elemol and the like as a main component. The main component that shows the pharmacological action of Angelica gigas is decursin, a cumarin derivative, and active ingredients such as decursinol, umbelliferon and beta-sitosterol. It is contained. Cumarin derivatives are known to have been used during acute myocardial infarction as anticoagulants used to prevent hemorrhage in the past. In particular, decusin and its isomer decursinol angelate are not present in monodon and Chinese donkey, but are present only in the rhizome of angelica gigas nakai, native to Korea.

As a prior art, decursin and decursinol angelate, which are cumin-based substances, have anticancer activity against various cancer cell lines such as lung cancer and liver cancer cells, and have cytotoxicity against various cancer cell lines. In addition, the action of enhancing the enzyme activity of protein kinase C (protein kinase C) has been found has emerged as an anti-cancer substance. However, in the present study, studies on the biological activities of the active ingredients of decancin and decosinol angelate in the skin have been insufficient. In addition, the existing extraction and separation techniques are focused on obtaining high purity of deckerin, but research on the development of effective and economical extraction and separation purification techniques of decusin and decosinol angelate is insufficient. Therefore, the current Deckerin extraction and separation and purification technology is difficult to commercialize the use of Deckerin because it lacks economic feasibility for universal development.

Therefore, the present inventors have studied the extraction method, purification and separation method and processing method for maximizing the efficacy in the skin of the decansin, the active ingredient of Angelica, on the basis of the known oriental medicine efficacy, as a supercritical extraction method After extracting, separating and purifying with a specific eluting solvent combination and then collecting it in the polymer nanocarrier, the discovery of decosin improves the skin absorption rate and thereby maximizes the efficacy of decosin in the skin, thereby completing the present invention.

Accordingly, it is an object of the present invention to provide a method for extracting, purifying and processing Decasin from Angelica maximus to maximize efficacy in the skin.

It is also an object of the present invention to provide an external composition for skin containing decosin prepared by the above method as an active ingredient.

In order to achieve the above object, the present invention comprises the steps of: 1) extracting the Angelica gigas in a supercritical extraction method using carbon dioxide; And 2) separating and purifying deckerin from the extract of step 1) through column chromatography using a ratio of hexane and ethyl acetate in a ratio of 1.5: 1 to 4: 1. .

In another aspect, the present invention provides a method for producing a deckerin further comprises the step of collecting the deckerin purified in step 2) into the polymer nano-carrier.

In another aspect, the present invention provides a composition for external application to the skin containing decosin prepared by the above method.

In the present invention, not only to improve the production yield and skin absorption rate of the deckerin through the extraction, separation, purification and processing method of the deckerin to improve the skin absorption rate, but also protects or detoxifies skin cells by improving skin wrinkles and inhibiting reactive oxygen species. The effect of retarding skin aging, etc. was excellent, and formulation stability in the external preparation composition containing the same was also improved.

Hereinafter, the present invention will be described in more detail.

The present invention relates to a method for extracting, purifying and processing Deckerin for maximizing efficacy in the skin and enhancing skin absorption, comprising the following steps:

1) extracting the Angelica gigas by a supercritical extraction method using carbon dioxide;

2) separating and purifying the deckerin from the extract of step 1) through column chromatography using a ratio of hexane and ethyl acetate in the range of 1.5: 1 to 4: 1; And

3) optionally, collecting the decansin purified in the step 2) into the polymer nanocarrier.

First, an extraction method for obtaining deckerin from Angelica is as follows. In the present invention, eco-friendly extraction technology was introduced in order to increase the content of the desired decusin from Angelica. That is, compared with the conventional solvent extraction, the supercritical extraction method, which is an eco-friendly extraction technology, was introduced, and two improvements were achieved. First, the Angelica extract using supercritical carbon dioxide was able to significantly increase the content of dekerin compared to solvent extraction. Second, in the extraction process using supercritical carbon dioxide, the carbon dioxide at normal pressure and room temperature is changed to gas when the extract is separated after extraction. Therefore, an additional process of removing the solvent remaining in the extract is not necessary in the extraction process using supercritical carbon dioxide due to the nature of the solvent extraction, thereby improving economics and removing the risk factors for skin toxicity to the remaining solvent.

Hereinafter, the supercritical extraction method according to the present invention will be described in detail. By "supercritical carbon dioxide" is meant a state at the critical point of carbon dioxide, i.e. at a temperature of 31 DEG C, the critical temperature of carbon dioxide, and at least 74 bar, the critical pressure. Therefore, in the state above this critical point, it becomes supercritical carbon dioxide. In general, above these critical points, fluids are characterized by a sharp increase in their solubility in solutes. Therefore, in performing the extraction process using the supercritical carbon dioxide as a solvent, the maximum extraction yield can be increased by setting the extraction conditions to the critical point of the supercritical carbon dioxide. However, in setting the conditions above the critical temperature, the set temperature can be continuously increased from 35 ° C or higher, which is near the critical temperature, but is preferably maintained at 80 ° C or lower in view of the thermal instability of the solute. Even in the case of pressure, if the pressure continuously rises above the critical pressure, the melting power is also gradually increased. It is preferable to set it to 600 bar or less in terms of energy efficiency for the pressure increase compared to the increase in the melting power. Therefore, the supercritical extraction method according to the present invention is most preferably carried out at a pressure of 35 to 80 ℃, 75 to 600 bar.

In addition, since the critical condition of carbon dioxide is lower than the supercritical conditions of other solvents, the extraction method using supercritical carbon dioxide can be operated in a relatively stable condition compared to the extraction method using other extraction solvents, so it is very suitable for the extraction of natural products sensitive to temperature. Do. Indeed, the decosin obtained by the supercritical extraction method using carbon dioxide according to the present invention (see Example 1) is about two to three times higher than that obtained through the conventional method (see Comparative Example 1) from the same amount of donkey. Could be obtained in yield. Therefore, it could be confirmed that the method according to the present invention is useful for obtaining a high yield of decusin.

In addition, the method for separating and purifying decusin from the Angelica extract is as follows. In the present invention, a one-stage column separation technique has been developed in order to remove the extract, increase the content of deckerin through purification, and to remove the strong odor of the donkey. Since the extract of Angelica gigas has a strong intrinsic odor of Angelica gigas, there may be limitations on its use as a general skin external preparation. Therefore, the process of removing the donkey intrinsic odor as well as improving the content of deckerin is essential. To this end, economic and efficient separation and purification techniques must be accompanied, and in the present invention, a simple, highly efficient one-stage column separation technique using a high content of supercritical donkey extract is effective. Found out. That is, in the present invention, the ratio of hexane and ethyl acetate as the elution solvent was 1.5: 1 to 4: 1, and purified by column chromatography to remove strong odor of donkey and obtain decusin with high yield. If the proportion of the eluting solvent is out of the above range, there is a problem that formulation is difficult due to the almost no deckerin in the fraction or due to the unique smell of Angelica.

In another aspect, the present invention includes the step of encapsulating in the polymer nano-carrier for the separation, stabilization of purified decusin and enhancement of skin absorption.

The decusin concentrate prepared through the column was collected using polymer nanocarriers. Deckersin concentrate, which has been separated and purified, may be precipitated due to its low solubility due to the presence of high water phase when introduced into the external preparation for skin due to its high hydrophobicity. This phenomenon has a fatal effect on the dermal absorption of the skin. Therefore, in order to prevent this, by collecting the deckerin in the self-association type polymer nanocarrier, not only can it be stably introduced into the external external preparation, but also the absorption rate to the skin can be improved.

Briefly looking at this process, first, the ring-opening polymerization of the hydrophobic polyester polymer and the hydrophilic polymer to prepare an amphiphilic polymer having a self-association bonded to the double block form. Amphiphilic polymers having self-association include polycaprolactone-block polyethylene glycol block copolymers, polyethylene glycol and poly-D, L-lactic acid-co-glycolic acid block copolymers, and poly-D, L-lactic acid-blocks. -Polyethylene glycol block copolymers, branched polyethyleneimines and poly-caprolactone block copolymers, and linear polyethyleneimines and poly-caprolactone block copolymers, and the like, but are not limited thereto. After mixing the amphiphilic polymer having self-association and the concentrate of the decusin, the distilled water may be slowly added to the mixture, followed by stirring while warming to 50 to 60 ° C. to collect decusin in the polymer nanocarrier.

Deckerin encapsulated in the polymer nanocarrier by the above method was able to increase the efficacy of the dermisin by improving the skin absorption rate, that is, to improve the skin wrinkles, cell protection or detoxification by the removal of reactive oxygen species, and improve skin elasticity .

In addition, since the nano-carrier is a biocompatible material, it is safely biodegradable in vivo and thus has no harm.

Finally, the present invention relates to an external preparation composition for skin containing decosin prepared by the above method. The external preparation composition for skin according to the present invention contains 0.0001 to 5% by weight of the decusin, based on the total weight of the composition.

The external preparation composition for skin according to the present invention is not particularly limited in its formulation, and it is a softening longevity, astringent longevity, nourishing longevity, eye cream, nourishing cream, massage cream, cleansing cream, cleansing foam, cleansing water, powder, essence, It can be formulated into a pack, hair tonic, hair liquid, scalp treatment, hair cream, hair gel, hair spray, hair shampoo or rinse.

Hereinafter, the configuration and effects of the present invention will be described in more detail with reference to Examples and Test Examples. However, the present invention is not limited only to these examples.

Comparative Example 1 Preparation of Ethanol Extract of Angelica gigas

1 kg of dried fine Angelica root was pulverized into fine powder using a grinder and then added to 5 L of 95% ethanol and extracted with gentle stirring at room temperature. The extraction time was performed for about 6 hours, after which the extraction solution was recovered and concentrated under reduced pressure to completely remove the ethanol solvent to obtain a final donkey extract 55 g.

The high pressure liquid chromatography confirmed that the content of decusin in the Angelica extract was about 5-6% (w / w).

Example 1 Preparation of Supercritical Extract of Angelica

1 kg of dried fine donkeys were pulverized into fine powder using a grinder and then extracted using supercritical carbon dioxide. 1 kg of ground Angelica root powder was charged into the reactor and carbon dioxide was introduced into the reactor under a pressure of 400 bar and 60 ° C. At this time, the incoming carbon dioxide was stored in a preheater and a reservoir to achieve equilibrium at a desired temperature. The flow rate of carbon dioxide flowing out of the extraction reactor inlet and outlet was controlled at a rate of 100 to 120 ml / min. Supercritical carbon dioxide dissolves nonpolar substances contained in Angelica gigas, and then expands to normal pressure and room temperature in the separator, causing rapid changes in solubility, resulting in the supersaturation of the Angelica extract dissolved in supercritical carbon dioxide. Angelica extract was separated from carbon dioxide at room temperature and atmospheric pressure to obtain 65 g of the final Angelica extract.

The high pressure liquid chromatography confirmed that the content of deckerin in the Angelica extract was about 13-15% (w / w). This is an amount of about 2 to 3 times more than the content of the decusin contained in the extract of Angelica gigas of Comparative Example 1 prepared by extracting the organic solvent.

[Test Example 1] Deckerin purification conditions confirmed

2 g of the Angelica extract obtained in Example 1 was filled with 200 g of a column using silica gel as a filler. In performing purification using column chromatography, hexane and ethyl acetate were used as developing solvents. For the purpose of separation according to the degree of polarity, the fractions of hexane and ethyl acetate were obtained by varying the concentration gradient from 10: 1 to 1: 1.

As a result, in the fraction where the ratio of hexane and ethyl acetate was 4: 1, the donkey specific odor was removed, and there was almost no donkey specific odor even at 1.5: 1.

Also, no decusin was detected in 10: 1 to 5: 1, and 8 to 9 fractions of hexane to ethyl acetate ratio were 4: 1, but the amount was very small. There was almost no decusin except for the first fraction.

As a result of analysis of the decansin, an index component from the above fraction, the ratio of hexane and ethyl acetate is preferably 1.5: 1 to 4: 1, showing the highest content at 2: 1, and the second fraction of the 2: 1 fraction. [The first (1 L), middle (800 ml) and second (1 L) subdivided into the highest concentrations of deckercin, the content was 75% (w / w). The yield represents 50% (w / w), indicating that there is considerable economy.

Example 2 Decusin Tablet

2 g of the Angelica extract obtained in Example 1 was filled with 200 g of a column using silica gel as a filler. The ratio of hexane and ethyl acetate was 2: 1, and the residue was purified by column chromatography to obtain 1.1 g of decancin concentrate.

Comparative Example 2

Except that 2 g of the Angelica extract obtained in Comparative Example 1 was used, the purification was carried out in the same manner as in Example 2 to obtain 0.3 g of the decusin concentrate.

Example 3 Preparation of Deckersin Concentrated Mixture-Containing Polymer Nanoparticles Prepared Using PCL-PEG Biblock Copolymer

In a glass flask containing hexamethyldisilazine, silanized by reaction with a hydroxyl group, methoxy polyethyleneglycol and catalyst Sn (Oct) ₂ (Sigma, St. Louis, MO) were added 10 g of lactone monomers were injected and mixed uniformly. The flask containing the mixture was connected by a vacuum line, and the polymerization was carried out while leaving water at 120 ° C. after removing water and the like under vacuum. After 24 hours, the polymerized polymer was dissolved in methylene chloride and then recrystallized with excess methanol to prepare a pure polycaprolactone-block-polyethylene glycol (PCL-PEG) diblock copolymer (Korean Patent Publication No. 2004 -62379).

1.2 g of the PCL-PEG diblock copolymer (total weight average molecular weight = 10,000 Da, weight ratio of PCL: PEG = 1: 1) and 0.04 g of the decansin concentrated mixture were uniformly dissolved in an organic solvent of 50 ml of ethanol solvent, Into a 50 mL aqueous solution was formed nanoparticles by spontaneous self-association process. The ethanol organic solvent was removed under reduced pressure at about 40 ° C. by the method of evaporation. Thereafter, an aqueous solution of polymeric nanoparticles containing 4% (w / w) of the decusin concentrate was obtained.

Test Example 2 Skin Absorption Rate of Polymeric Nanoparticles Containing Deckersin Concentrated Mixture

The skin of the hairless guinea pig is cut out and fixed in a skin absorption test apparatus (Franz-diffusion cell). Then, three kinds of samples are added to the upper part, and the lower part is stirred with a buffer solution having an appropriate composition. After maintaining the 32 ℃ for 18 hours, the amount of deckerin penetrated into the skin was quantified by liquid chromatography. For effect comparison, the same test was performed on a polyethyleneglycol solution containing 4% of the decusin concentrate, such as the decusin collected on the polymer nanoparticles.

Total Decosin Absorption Deckerin's area and absorption per unit time Polyethylene glycol 2% solution 0 0 4% aqueous polymer nanoparticles solution 37.53 μg 2.68 μg / cm 2 · h 4% polyethylene glycol solution 13.68 μg 1.69 μg / cm 2 · h

As can be seen from the results of Table 1, the deckerin-containing polymer nanoparticles provided in the present invention exhibited a skin absorption rate of 159% higher than that of the polyethylene glycol solution in which decosin was dissolved.

Test Example 3 Measurement of Inhibition Effect of Reactive Oxygen Species (ROS) Formation by UV Light

The transformed human keratinocytes were cultured in a 96-well plate incubator for measurement of fluorescence at a concentration of 2 × 10 ⁴ at 37 ° C. and 5% CO ₂ for 1 day. The test materials prepared in Examples 1 to 3 were pretreated for 24 hours as shown in Table 1 below. 100 μl of DCFH-DA (2 ', 7-dichlorodihydro-fluorescein diacetate; Molecular Probes, Inc) prepared at 20 μM in HEPES-buffered control salt solution (HCSS) was incubated for 20 minutes at 37 ° C and 5% CO _2. Again washed with HCSS. After adding 100 μl of HCSS containing the same sample treated 24 hours ago, the fluorescence intensity of DCF (dichlorofluorescein), which was initially oxidized with reactive oxygen species (ROS), was changed to a fluorescent plate reader (Ex = 485 nm, Em = 530 nm). Measured. UVB (30 mJ / cm 2) was treated and the fluorescence intensity immediately after treatment and up to 3 hours was measured with a fluorescence plate reader (Ex = 485 nm, Em = 530 nm). Comparative values calculated using the UV control group as 100 are shown in Table 2 below.

Test substance content % ROS generation after 3 hours UV control - 100 Example 1 1 ppm 78 Example 2 1 ppm 55 Example 3 1 ppm 61

In the results of Table 2, it can be seen that the concentration of the decusin concentrate of the present invention and the polymer nanocarrier collected therefrom reduces the generation of reactive oxygen species (ROS) by ultraviolet rays. Therefore, it can be seen that it can effectively defend against oxidative stress and detoxify the cells from harmful environments such as air pollution, yellow dust, and ozone as well as ultraviolet rays.

Test Example 4 Measurement of the Biosynthesis Inhibitory Effect of Type I Collagenase (MMP-1) by Ultraviolet Rays

Human fibroblasts were incubated in a 48-hole plate incubator at a concentration of 10 ⁴ , and after 24 hours, UV A was irradiated at 15 J / cm 2, and the test materials prepared in Examples 1 to 3 and Comparative Example 1 were shown in Table 3 below. The medium was included together. The supernatant was harvested on day 2 of culture and the amount of type I collagenase produced was quantified using ELISA (AP biotech RPN2610) method. Comparative values calculated using the UV control group as 100 are shown in Table 3 below.

Test substance content Type I collagenase biosynthesis (%) UV control - 100 Comparative Example 1 1 ppm 81 Example 1 1 ppm 72 Example 2 1 ppm 54 Example 3 1 ppm 52

From the results of Table 3, it was found that the deckerin concentrate of the present invention and the polymer nanocarrier containing the same significantly reduced the biosynthesis of type I collagenase by ultraviolet rays compared to Comparative Example 1 using only the deckerin concentrate. Therefore, it was confirmed that collagen generated normally in the skin due to the expression of collagenase can be prevented or suppressed from wrinkles generated.

Formulation Examples 1-9 Cream Formulation

In accordance with the composition shown in Table 4 was prepared according to the conventional method to prepare an oil-in-water cream of Formulation Examples 1 to 9 containing the polymer nanoparticles prepared in Example 3 (unit: wt%).

ingredient My brother's example One 2 3 4 5 6 7 8 9 Lipophilic monostearic acid 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Glyceryl Stearate 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Cetostearyl alcohol 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 1.5 Beeswax 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Squalane 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 1.2 Polysorbate 60 0.4 0.4 0.4 0.4 0.4 0.4 0.4 0.4 0.4 Cetylethylhexanoate 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Liquid paraffin 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Methyl paraben 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Propylparaben 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 Silicone oil 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 4.0 Tocopheryl acetate 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Distilled water Balance Balance Balance Balance Balance Balance Balance Balance Balance Urea 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 glycerin 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 5.0 Butylene glycol 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 3.0 Triethanolamine 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 0.18 Carboxy Vinyl Polymer 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 Nanoparticles of Example 3 0.001 0.05 0.1 0.2 0.5 1.0 2.5 4.0 5.0

Comparative Formulation Example 1

To prepare an oil-in-water cream of Comparative Formulation Example 1 containing the decusin concentrate prepared in Comparative Example 2 according to the composition shown in Table 5 (unit: wt%).

ingredient Comparative Formulation Example 1 Lipophilic monostearic acid 3.0 Glyceryl Stearate 1.5 Cetostearyl alcohol 1.5 Beeswax 0.5 Squalane 1.2 Polysorbate 60 0.4 Cetylethylhexanoate 2.0 Liquid paraffin 2.0 Methyl paraben 0.2 Profile paraben 0.05 Silicone oil 4.0 Tocopheryl acetate 0.5 Distilled water Balance Urea 0.5 glycerin 5.0 Butylene glycol 3.0 Triethanolamine 0.18 Carboxy Vinyl Polymer 0.2 Deckerin Concentrate of Comparative Example 2 0.2

[Test Example 5] Long-term storage stability of Deckerin in the formulation

The long-term storage stability of Decasin was confirmed for Formulation Examples 4 and 9, and Comparative Formulation Example 1 above. Samples were taken at regular intervals while stored in a constant temperature bath at 25 ° C. and 45 ° C. to determine the content of deckerin, and the results are shown in Table 6 below.

Test substance Condition 7 days 15th 30 days 45 days Comparative Formulation Example 1 25 ℃ 100% 95% 92% 85% 45 ℃ 98% 96% 86% 80% Formulation Example 4 25 ℃ 100% 96% 93% 92% 45 ℃ 100% 94% 90% 89% Formulation Example 9 25 ℃ 100% 97% 92% 90% 45 ℃ 100% 95% 92% 91%

As can be seen from the results in Table 6, Formulation Examples 4 and 9 containing Deckerin prepared in the state of being collected in the polymer nanoparticles, Comparative Example 1 comprising the deconsin concentrate itself which was not collected in the nanoparticles It was confirmed that the long-term storage stability of Deckerin in the formulation is excellent. Through the above results it can be seen that by collecting the deckerin in the polymer nanoparticles of the present invention it is possible to preserve the titer of the deckerin in the formulation.

Claims (5)

1) extracting the Angelica gigas by a supercritical extraction method using carbon dioxide; And 2) separating and purifying the deckerin from the extract of step 1) through column chromatography using a ratio of hexane and ethyl acetate in the range of 1.5: 1 to 4: 1; Method for producing a decusin containing. The method of claim 1, wherein the step 1) supercritical extraction method is performed at a pressure of 35 to 80 ° C. and 75 to 600 bar. The method of claim 1, further comprising the step of capturing the deckerin purified in step 2) into the polymer nanocarrier. The method of claim 1, wherein the polymer nanocarrier is a polycaprolactone-block polyethylene glycol block copolymer, polyethylene glycol and poly-D, L-lactic acid-co-glycolic acid block copolymer, poly-D, L-lactic acid-block A polyethylene glycol block copolymer, a branched polyethyleneimine and a poly-caprolactone block copolymer, and a linear polyethyleneimine and a poly-caprolactone block copolymer. An external preparation composition for skin containing deckercin prepared by the method according to any one of claims 1 to 4 as an active ingredient.