KR100763035B1 - An extraction method of a effective component improving an acne from Agrimonia pilosa Ledebour and a composition of cosmetic including the component - Google Patents

Abstract

The present invention relates to a method for extracting and isolating an effective ingredient for improving acne from sunflower seeds, and a cosmetic composition comprising the active ingredient material obtained therefrom, which comprises the step of obtaining a first extract of sunflower seeds by adding a predetermined extracting solvent to sunflower oil In the method of extracting and extracting an active ingredient material, the step of obtaining the first extract of Seok Hak can be further characterized by a step of obtaining a fraction of hexane through a step of partitioning with water and hexane. According to the present invention, it is possible to extract and isolate which maximizes the activity of the effective ingredient for acne improvement from sunflower seeds, and the cosmetic composition containing the active ingredient material extracted by the present invention can effectively inhibit the growth of acne bacteria.

Sunflower seeds, straw sandals, acne, cosmetics, supercritical fluid extraction

Description

[0001] The present invention relates to a method for extracting and extracting acne-effective active ingredient from sunflower seeds and a cosmetic composition comprising the active ingredient material obtained therefrom.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic view showing a method of preparing a cosmetic composition by extracting and isolating an active ingredient for improving acne from sunflower seeds according to the present invention. FIG.

FIG. 2 is a graph showing the growth inhibition ring size for various strains of the methanol extract (first extract) of the present invention.

FIG. 3 is a graph showing the growth curve of acne bacterium when the extract of the present invention (primary extract) is added.

The present invention relates to a method for extracting and isolating an effective ingredient for acne improvement from sunflower seeds and a cosmetic composition comprising the active ingredient material obtained therefrom. More particularly, the present invention relates to a cosmetic composition comprising a sunflower seed extract and a predetermined extracting solvent, Wherein the extraction solvent is any one selected from anhydrous methanol, hydrated methanol, anhydrous ethanol, and water ethanol, and the amount of the extraction solvent added is 5 - Wherein the extraction solvent is removed from the first extract of Sephaelexa to obtain a solid component, the solid component is dissolved in water to obtain an aqueous solution, and then hexane is added to the aqueous solution to obtain The hexane layer is subjected to a distribution process so as to elute the active ingredient material, And extracting hexane to obtain a hexane fraction. The present invention also relates to a method for extracting and separating an active ingredient for improving acne from sunflower seeds and a cosmetic composition comprising the active ingredient material obtained therefrom.

In addition to genetic and environmental factors, the onset of acne is caused by active sebum production due to changes in hormone secretion in the body, pore clogging due to abnormal keratinization of the follicular sebaceous glands (hyperkeratinization), acne bacteria ( Propionibacterium acnes ) and the four major causes of skin inflammation induced by acne and their interactions. Thus, anti-androgens that inhibit sebum production, non-steroid anti-inflammatory agents that act as anti-inflammatory agents, resorcinol with antimicrobial effect, acne by sulfur and benzoyl peroxide and tetracycline, erythromycin, clindamycin, A method of inhibiting bacterial activity and a method using a vitamin A derivative such as retinoic acid have been used as an acne treatment method.

However, these conventional methods for treating acne have various problems in terms of efficacy and skin safety. The use of hormones has been reported to cause adverse effects such as inhibition of epidermal growth and hormone abuse. Retinoic acid and benzoyl peroxide have been associated with birth defects, skin irritation, and contact dermatitis. Antibiotics such as mycine have been reported to exhibit adverse effects such as the appearance of resistant bacteria, allergic reactions, and photosensitivity reactions. On the other hand, acne treatment is not a short-term treatment but requires continuous and continuous treatment. Therefore, the acne treatment method having the above-mentioned problems can be used only for a short time or within a limited range.

In recent years, studies have been actively conducted to search for effective ingredients that can reduce the above-mentioned adverse effects from natural resources such as plants and continuously use them, and have harmless effects on the human body but have strong anti-acne bacteria effect. In Korea, it is used as a raw material for functional cosmetics and soaps related to acne treatment and extracts of mushroom, green tea and golden, willow, Saururus chinensis, Licorice and Horsetail. It is progressing. However, the effect is very weak compared with the existing acne remedy, or it is difficult to mass-produce effective substances related to acne treatment. Particularly, natural products are composed of various components in a complex manner. Therefore, the method for separating and purifying an active active substance differs depending on the physicochemical properties of the desired active substance, and effective methods for separation and purification based on various different principles Therefore, there are many difficulties in extracting and isolating a substance having anti-acne activity from natural products.

Meanwhile, Agrimonia pilosa Ledebour) is a plant with various names such as a perennial herb that grows commonly on the roads of Korea, a dragon grass, a 黄花草, and a desert grass. It has been known to have hemostatic, anti-inflammatory, anti-bacterial, analgesic, anti-cancer, hypoglycemic, hematopoietic effects, and has been used as a folk remedy and oriental medicine in ancient East and West long ago. , And it is known that there is no irritation to the skin and mucous membranes. In particular, research on the mechanism of the antiviral with seonhakcho extract (B. Min et al, Inhibitory effects of Korean plants on HIV-1 activities, Phytother Res, 15 (6), 481, (2001);.. D. Kwon et al ., Inhibition of hepatitis B virus by an aqueous extract of Agrimonia pilosa L., Phytother Res. , 19 (4), 355 (2005); Y. Li. et al., Antiviral activities of medicinal herbs traditionally used in southern mainland China, Phytother Res., 18 (9), 718, (2004)), A study on the anti-inflammatory efficacy with fraction (H. Correia et al., Polyphenolic profile characterization of Agrimonia eupatoria L. by HPLC with different detection devices, Biomed . Chromatogr . , 20 (1), 88 (2006)) and a study on the antibacterial effect of various kinds of bacteria (A. Copland et al ., Antibacterial and free radical scavenging activity of the seeds of Agrimonia eupatoria , Fitoterapia 174 , 133 (2003); J. Bae et al., Effect Agrimonia pilosa Ledeb. extract on the growth of food-borne pathogens, Korean Nutrition Society 38 (2), have been made to 112 (2005)).

Korean Patent No. 10-0439845 discloses a natural food preserving agent comprising an ethanol extract of Sunjoukcho, a food preservation method using the same, a sterilization treatment method, and a natural antimicrobial agent including an ethanol extract of Sunjogcho. Korean Patent Publication No. 2001-0053978 Discloses an anti-irritant cosmetic composition containing an extract as an active ingredient. Japanese Patent Application Laid-Open No. 6-16530 discloses a cosmetic composition having a whitening, antioxidant, and anti-inflammatory activity by combining a Korestricken liquid crystal and a sunflower seed extract, and JP-A-8-175957 Japanese Unexamined Patent Application Publication No. 10-226617 discloses an external preparation for skin which is excellent in the effect of preventing skin aging as a cosmetic preparation containing sunflower seed extract. However, Korean Patent Registration No. 10-0531503 is only used for the cosmetic composition showing the effect of improving acne.

In Korean Patent No. 10-0531503, the ethanol extract of Sung Hogakseo alone, or the crude extract, Aujar, Omega, Rhododendron and Pine bark are respectively pulverized, and the extract is obtained by ethanol extraction method. As a result, the effect of inhibiting the growth of acne bacteria was weaker than that of triclosan, an antimicrobial agent frequently used in cosmetic compositions. However, the solvent mixture of 1,3BG (Butylene Glycol) and ethanol in the same mass ratio was used as an auxiliary solvent for the release of the supercritical fluid, and the extracts obtained from the angel siege, sunghak, omega, rhubarb and pine bark were mixed at a certain ratio, The growth inhibition of acne bacterium was similar to that of triclosan. However, in order to obtain a superior acne bacterium growth inhibiting effect, only the extract of Zhaoguchi extract is insufficient. Therefore, there is a problem that an extract of other natural raw materials must be mixed and made into nano-particles. In addition, the addition ratio of the extraction solvent to sunflower seeds is low, so that not only the active ingredient is eluted as much as possible but also the activity against acne bacteria is low.

Accordingly, it is an object of the present invention to provide a method for efficiently extracting and isolating an effective ingredient for acne improvement from sunflower seeds for minimizing adverse effects associated with the existing acne treatment method and for preventing and treating acne with safety that is sustainable and safe do.

In particular, it is an object of the present invention to provide a method for effectively extracting and isolating an effective ingredient for acne improvement from sunflower seeds so that the effect of improving acne can be excellently solved with only a sunflower extract alone without mixing an extract obtained from other natural raw materials.

In addition, the acne improvement active ingredient material is extracted and separated by supercritical fluid extraction, in which the principles of solvent extraction and distillation are applied in a complex manner, And to provide optimum conditions.

It is an object of the present invention to provide a cosmetic composition comprising an acne improvement active ingredient which is extracted and separated from acorns.

In order to accomplish the above object, the present invention provides a method for extracting and extracting active ingredients for acne improvement comprising the step of adding a predetermined extracting solvent to Agrimonia pilosa Ledeb. Wherein the extraction solvent is selected from the group consisting of methanol, anhydrous methanol, hydrous methanol, anhydrous ethanol and water ethanol. The amount of the extraction solvent added is 5 to 20 ml per 1 g of the dry weight of sunflower seeds. After the step of obtaining the sunflower seeds first extract, Removing the extraction solvent from the extract to obtain a solid component, dissolving the solid component in water to obtain an aqueous solution, adding hexane to the aqueous solution, distributing the active ingredient material to elute the hexane layer, And recovering hexane to obtain a hexane fraction.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail with reference to FIG. 1 of the accompanying drawings.

First, prepare dried sunflower seeds. At this time, sunflower seeds should be dried in the shade because they have little effect when dried in the sun, and care should be taken not to grow or damage the mold when dried. It is also possible to purchase and use the dried Sunhakucho outpost, and prepare the dried Sunhakuwon as a powder using a pulverizer.

Next, in the step of obtaining the first extract of Seokhogcho, any one solvent selected from anhydrous methanol, hydrous methanol, anhydrous ethanol and hydrated ethanol is added to the seaweed extract as an extraction solvent. At this time, the amount of the extraction solvent added is 5 to 20 ml per 1 g of dry weight of sunflower seeds. The first extraction may be a conventional solvent extraction method such as immersion extraction at room temperature for 1 to 7 days or extraction at a temperature of 50 to 95 ° C for about 1 to 5 hours.

Conventionally, in extracting a natural product, the extract is prepared by filtering the solution after agitating the extract at a proper time interval while immersing the natural product in a suitable solvent for a certain period of time. Thus, the efficacy and the effect of the actual extract are difficult to be accurately shown. It is often not suitable for use as a cosmetic composition due to problems such as color, skin irritation and toxicity. Conventional extraction solvents include purified water, a pure water solvent selected from the group consisting of anhydrous or hydrous methanol having 1 to 4 carbon atoms, a solvent group including ethanol, propyl alcohol, butyl alcohol, glycerin, propylene glycol and butyleneglycol, , And the amount of the extraction solvent added to the plant material is about 1 to 50 times as much as the total volume of the plant material.

On the other hand, Korean Patent No. 10-0531503 discloses a method of pulverizing an ethanol extract or a crude angel, oak, rhubarb, and pine bark of a single sunflower seed alone, obtaining an extract by ethanol extraction method, mixing with a sunflower seed extract, And the effect of inhibiting the growth of inflammatory bacteria was very weak compared with the effect of inhibiting the growth of acne bacteria of triclosan. However, a solvent mixture of 1,3BG (Butylene Glycol) and ethanol in the same mass ratio was used as an auxiliary solvent for the release of supercritical water, and the solution was mixed at a mass ratio of about 1: 2 with gold angel, sunflower, oak, rhubarb and pine bark When the extracts obtained from the supercritical extraction method were mixed at a certain ratio, the effect of inhibiting the growth of acne bacterium was similar to that of triclosan in the case of nanosizing.

However, in the present invention, when methanol or ethanol was used as the extraction solvent, and the extraction solvent was changed from 5 to 20 ml to 1 g of the dry weight of sunflower seeds, the first extraction showed similar effect to that of triclosan. In other words, the selection of the extraction solvent and the mixing ratio of the sample and the extraction solvent showed the best degree of dissolution and activity of the acne improving active ingredient from sunflower seeds.

In order to further enhance the activity of the acne improvement active ingredient, acne improvement improving active ingredient is extracted and separated from the acute acne scarlet fever according to the present invention. In the step of obtaining the acute acne extract active ingredient, the hexane fraction is obtained through a distribution process using hexane As shown in FIG. That is, there is provided a method for preparing a solid extract, which comprises the steps of: removing a solvent from the first extract to obtain a solid component; completely dissolving the solid component in distilled water to obtain an aqueous solution; adding hexane to the aqueous solution; And recovering the hexane layer from which the active ingredient is dissolved, and then removing the hexane to obtain a hexane fraction.

In order to enhance the activity of the active ingredient material or to separate and purify only the ingredient thereof, the property of equilibrium of distribution with various solvents is generally used. In the present invention, the degree of separation or activity of the active ingredient for improving acne is increased by using hexane there was. However, when hexane is used, the step of obtaining hexane fractions through the distribution process using hexane may be omitted, because there is a possibility that the development as actual cosmetic raw materials may be obstructed due to problems such as various processes of fraction extraction and deterioration of recovery rate have.

Next, in order to further enhance the activity of the acne-improving active ingredient substance, the method for extracting and extracting acne improvement effective ingredient from acarinephrine according to the present invention comprises the step of removing the solvent from the acute acne extract first, , And further comprising a step of obtaining a second extract of Seokhocho by supercritical extraction from the solid fraction.

In order to further enhance the activity of the acne-improving active ingredient substance, the method for extracting and extracting acne-improving active ingredient from acornsia japonica according to the present invention comprises the steps of: obtaining the acute acne extract first and extracting the acute acne extract from the hexane fraction And a step of obtaining a second extract of Sanshogako by a critical extraction method.

Here, supercritical fluid extraction is an extraction method in which the principles of solvent extraction and distillation are applied in a complex manner. It is characterized by high solubility, rapid mass transfer and heat transfer, quick penetration into micropores due to low viscosity, It is a method to extract using the same advantage. Since the above-mentioned extraction method is capable of setting a high-density condition by manipulation of pressure and temperature, it is excellent in selectivity such as fractionation and separation to obtain a high-purity natural active ingredient and extracts at a relatively low temperature. There is an advantage that loss can be prevented.

Therefore, by using supercritical fluid extraction, it is possible to obtain an excellent effect for acne improvement by using the active ingredient material of sunflower seeds in a cosmetic composition without damage or loss.

In the present invention, carbon dioxide, which is a typical supercritical fluid and whose critical point is close to normal temperature and which is non-toxic, nonflammable and very inexpensive, is used. Since the critical temperature of carbon dioxide is 31 ° C and the critical pressure is 73 atmospheres, the step of obtaining the second extract of Sjogaksocho should be carried out under a temperature condition of more than 31 ° C and a pressure condition of more than 73 atm. At this time, when 3 g of the sample is placed and the flow rate of carbon dioxide is 3.0 ml / min and the extraction time is 60 min, the use of carbon dioxide as a supercritical fluid at a temperature of 38 to 44 ° C and a pressure of about 200 kg / desirable.

Generally, the supercritical fluid has a property of changing the solubility and density depending on the change of temperature and pressure, so that the activity of the extract may be changed accordingly.

On the other hand, pure supercritical fluid carbon dioxide has a large solubility and selectivity for most nonpolar materials, but it is often difficult to extract polar substances. Therefore, supercritical fluid extraction may be attempted by adding a co-solvent. As the co-solvent, alcohol, chlorinated hydrocarbon, hexane, acetone, and water are mainly used. The use of co-solvents increases the solubility and results in lower extraction pressure than pure supercritical fluid alone. Therefore, it is advantageous to separate the extract and solvent by changing the temperature only. However, the use of co- There may be a disadvantage that the problem of the residual solvent of the high-boiling solvent arises.

The present invention is characterized in that ethanol is added to the solid fraction or the hexane fraction as an auxiliary extraction solvent in the step of obtaining the second extract. When water is used as an auxiliary solvent, there is a problem in that it is difficult to dry in the sample collection process after extraction. Since ethanol is different from other polar organic solvent or methanol or propanol, there is almost no toxicity. Therefore, ethanol is used as an auxiliary solvent Is preferably used. In this case, the auxiliary extraction solvent is preferably added to the solid fraction or the hexane fraction at a flow rate of 0.2 to 0.3 ml per minute in the active diagram of the acne improvement active ingredient material.

The cosmetic composition for improving acne according to the present invention is characterized in that it contains the acne improving active ingredient substance derived from the above-mentioned second extract of Seokjeokcho or the hexane fraction, and the content of the second extract of Seokjeokcho or the hexane fraction is 0.001 to 20 By weight, more preferably 0.5 to 10% by weight. This is because, at a high concentration of 20 wt% or more, side effects such as skin irritation and toxicity can be caused, and the effect is weak at a low concentration of 0.001 wt% or less.

The cosmetic composition of the present invention may contain various components commonly used in cosmetics, for example, a water-soluble component, a powder component, and the like, as long as the effect of the present invention is not impaired, , Oil, surfactant, moisturizing agent, viscosity controlling agent, preservative, antioxidant, fragrance, pigment and the like.

The cosmetic composition containing the second extract of Seokjeokcho secondary extract or the hexane fraction of the present invention may be prepared into any formulation conventionally produced in the cosmetics industry, and may be, for example, a solution, a suspension, a paste, a gel, a cream, Powders, soaps, oils, powder foundations, emulsion foundations, wax foundations and sprays, but are not limited thereto. More specifically, it can be manufactured in the form of a flexible lotion, a convergent lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

Hereinafter, the present invention will be described in detail with reference to more specific examples, but the present invention is not limited to the following examples.

[ Example ]

One. Used strain , Medium and culture conditions

(1) Strain

The strains used in the present invention are Propionibacterium acnes KCTC3314 and Staphylococcus epidermidis KCTC1917, Staphylococcus aureus KCTC1927, Bacillus subtilis KCTC1021, and Bacillus cereus KCTC1012 were used in the Korean Gene Bank.

(2) Medium and culture conditions

The culture medium of acne bacterium was Reinforced clostridial (RC) medium (Merck, Germany) and P. acnes was stored at 4 ° C and activated 3 days before the experiment. The bacteria were inoculated in a culture medium, sealed with a gaspack system (Merck AnaerocultGaspack system, Germany) in an anaerobic jar, and anaerobically cultured in an incubator at 37 ° C for 3 days. Aerobic strains were used in Mueller-Hinton medium (Merck, Germany) and inoculated with the bacteria and incubated for 24 hours in an incubator at 37 ° C.

2. Antimicrobial activity against acne bacteria

The antimicrobial activity was evaluated using a paper disk diffusion method and a minimum inhibitory concentration (MIC).

(1) paper disk diffusion method

The acne strains were cultured in a Reinforced clostridial (RC) liquid medium for 3 days, and then the suspension of the bacteria was adjusted to a constant concentration (optical density at 620 nm, absorbance value of 0.5), which was then transferred to an RC agar plate using a sterile cotton swab And uniformly smoothed. After drying the plate, a paper disk (Φ 8 mm, Adventec. USA) was placed on the surface of the plate inoculated with the strain, and the first, second and third extracts or fractions were prepared and absorbed by concentration. The plates treated with the samples were incubated in an anaerobic jar using a gaspack system for 3 days. Antimicrobial activity was measured at the size of the inhibition zone around the disk. The antimicrobial activity of the aerobic strain was measured by using the same method as that of acne bacterium using Muller-Hinton medium and culturing in a 37 ° C incubator for 24 hours.

(2) Minimum inhibitory concentration (MIC)

MIC assay was performed by partially modifying the broth-dilution method using a 96-well plate (K. Yasunaka et al ., Antibacterial activity of crude extracts from mexican medicinal plants and purified coumarins and xanthones, J. Ethnopharmacol . , 97 , 293 (2005)). The acne bacteria cultured for 72 hours were adjusted to a McFarland standard of 0.5, and 20 μl of the strain diluted to 1.5 × ^{10 8} cells / ml was inoculated into each well containing 160 μl of the medium. The first, second and third extracts or fractions were dissolved in DMSO and prepared by RC liquid medium. 20 μl of each solution was added to a 96-well plate containing the strain and culture medium. As a control, 10% DMSO was used. The 96-well plate inoculated with the bacterial dilution and the sample was incubated for 3 days using an anaerocult gaspack system, and the minimum concentration at which no growth of the bacteria was observed was determined by MIC. The MIC of the aerobic strain was determined by using Mueller-Hinton medium in the same manner as the acne bacteria and culturing in a 37 ° C incubator for 24 hours.

3. Step to obtain first extract

(1) The step of obtaining the first extract of Seolhyeokcho

Agrimonia pilosa Ledeb. Was prepared by preparing a commercially available dry-type outpost and using a crusher to make powder.

One liter of 95% methanol was added to 50 g of prepared sunflower seeds and extracted with stirring at 300 rpm for 3 hours at room temperature. Then, the extract solution was filtered using a filter paper, and then the first extract of Seokhogcho was recovered.

(2) Antimicrobial activity of the first extract

The first extract of Seonghakhochu was diluted by DMSO (0.00625 ~ 0.05%) and absorbed by 50μl into each species. The antimicrobial activity was measured by disk diffusion method and the results were as shown in Table 1. Triclosan used as a control group was similar to the anticarcinogenic activity at a concentration of 0.05% of the methanol extract (first extract). FIG. 2 is a graph showing the size of the growth inhibitory ring as a result of treating each strain at a concentration of 0.05% in order to compare the antimicrobial activity with aerobic strains. The methanol extract (first extract) of Seongjeokcho showed the highest antimicrobial activity against acne bacteria . Even when ethanol was used as the extraction solvent, the size of the growth inhibition ring was found to be 36 mm when the concentration of the ethanol extract of sunflower seed was 0.05%.

<Table 1> Size of inhibiting growth of acne bacterium in methanol extract (primary extract)

Strain Seolhak-cho methanol extract (primary extract) Triclosan 0.00625% 0.0125% 0.025% 0.05% 0.025% P. acnes 25mm 30mm 33mm 36mm 36mm

In addition, the first extract of Seonjokcho (20 mg / ml of DMSO) was prepared by concentration using RC liquid medium and the MIC was measured to obtain the results as shown in Table 2. At this time, methyl paraben used as an antimicrobial control group and triclosan was used as a preservative. The MIC of triclosan was 0.004% (40 μg / ml) and the MIC of methylparaben was 0.2% (2 mg / ml) Respectively.

<Table 2> Minimum Inhibitory Concentration of Methanol Extract (First Extract)

Strains Seolhak-cho methanol extract (primary extract) Methyl paraben P. acnes 0.005% 0.2% B. cereus 0.0125% 0.2% B. subtilis 0.1% 0.2% S. aureus 0.05% 0.2% S. epidermidis 0.2% 0.2%

(3) Influence of primary extract of Seok Hakki on the growth of acne bacterium

The growth of the strain was measured at intervals of 12 hours while culturing for 3 days. The growth curve as shown in Fig. 3 was obtained by adding the first extract of Seonghakjecho to the RC medium with concentration (7.8 ~ 250 ㎍).

"Control" in FIG. 3 represents the acne bacterium that has not treated anything, and "DMSO" represents the growth curve of acne bacterium treated only with DMSO, and these were used as a control group. In both control groups, the bacteria started to proliferate at a very rapid rate 24 hours after the incubation and reached the growth stage for about 60 hours. 3, "7.8, 15.6, 31.3, 62.5, 125, 250" represents the growth curve of acne bacterium according to the concentration (unit: ㎍) In the test group treated with the first extract, the growth of the bacteria was inhibited until 24 hours, and gradually increased in the low concentration group (7.8 g) at 36 hours, and in the most concentration range after 60 hours It was observed to increase gradually. However, it was confirmed that the increase was not so large in the high concentration group.

(4) Evaluation of the stability of the first extract of Seolhyeokcho for temperature and pH

In order to measure the effect of temperature on the extract, the first extract of Seokhogcho was dissolved in DMSO at a concentration of 20 mg / ml, and the solution was placed in a 1.5 ml microfuge tube for 70 minutes, autoclave (121 ° C) For 15 minutes, and then the disk diffusion method was performed using these samples.

In order to measure the effect of pH change on the extract, the first extract of Seokhogcho was dissolved in DMSO at a concentration of 10 mg / ml, placed in a 15 ml conical tube, allowed to stand for 30 minutes by pH, And the disk diffusion method was performed.

As a result of confirming the effect of temperature and pH on the first extract of Seolhakucho (Table 3 and 4), it was found that the activity against acne was very stable. From these results, it can be seen that Seokhyecho extract can be used as a cosmetic raw material.

<Table 3> The stability evaluation of the first extract of Seoljeokcho

Temperature (℃) 70 80 90 100 121 Growth inhibition ring size (mm) 33 34 33 32 31.5

<Table 4> Evaluation of stability of pH-change of first extract

pH 2 3 4 5 6 7 8 9 10 11 12 Growth inhibition ring size (mm) 33.5 34 33 38 32.5 33 32.5 34 33.5 34 34

4. Hexane The fraction  Steps to Obtain

The above-mentioned methanol extract (first extract) of the above-mentioned Sunhogakcho methanol extract was concentrated under reduced pressure while recovering the solvent evaporated at 47 캜 using a distillation apparatus equipped with a cooling condenser to obtain 0.5 g of a powdery sample (solid content). This was put into a round bottom flask, filled with 200 ml of distilled water, thoroughly dissolved to make an aqueous solution, and this solution was added to a separating funnel and 200 ml of n-hexane was added. At this time, about 200 ml of a saturated saline solution was added to reduce the emulsion between the two solvents, and the hexane layer was shaken so as to sufficiently extract the substance. This was repeated several times to separate and recover the hexane layer. The solvent was completely removed using a rotary evaporator to recover 0.3 g of the hexane fraction.

At this time, fractionation was carried out using chloroform, ethyl acetate and butanol instead of hexane as a solvent, and fractions were recovered from each solvent.

As a result of measuring the inhibitory effect on growth of acne by the disk diffusion method using each fraction, it was found that the hexane fraction exhibited the most potent activity and exhibited the same efficacy at half the concentration of the first extract (Table 5). At this time, triclosan, which is widely used as an antimicrobial agent of the cosmetic composition, was treated at the same concentration as a control group.

<Table 5> Size of inhibitory growth of acne bacterium

Name of sample Treatment concentration Retaining ring (mm) Triclosan 500 μg 35 Seolhak-cho methanol extract (primary extract) 500 μg 36 Sunhogae ethanol extract (1st extract) 500 μg 36 Seolhak-cho methanol extract-hexane fraction 250 g 42 Seolhak-cho methanol extract-chloroform fraction 500 μg 27 Seolhak-cho methanol extract-ethyl acetate fraction 500 μg 8.5 Seolhak-cho methanol extract-butanol fraction 500 μg 12

5. Step of Obtaining Secondary Extract of Seolhak -Cho - Selection of Optimum Supercritical Fluid Extraction Condition

The supercritical fluid (CO2) was used as the supercritical fluid, and about 3g of the sample was filled in the extraction vessel. The supercritical fluid (SFE) system (JASCO, Japan) Extraction conditions were selected.

(1) Evaluation of the activity of secondary extract of Seokhoekcho according to addition of auxiliary solvent

3% ethanol was added as a co-solvent from 3 g of the hexane fraction of the methanol extract (primary extract) of Seonghak-cho (primary extract) under the conditions of a pressure of 200 kg / cm ² , a temperature of 40 ° C, a carbon dioxide flow rate of 3.0 ml / min, Table 6 shows the results of measuring the activity of the second extract of Seokhogcho obtained by supercritical fluid extraction (the size of the inhibitory ring of acne bacterium) when not added. In other words, the addition of ethanol to supercritical fluid showed more inhibitory effect on the growth of acne.

<Table 6> Evaluation of Activity of Secondary Extracts of Seokhogcho with Different Solvent Additives

Experimental conditions Concentration of extract (/ / disk) 12.5 25 50 One carbon dioxide 23mm 30mm 34mm 2 Carbon dioxide + 100% ethanol 29mm 34mm 38mm

(2) Evaluation of the activity of the second extract of Seokhoekcho according to the temperature change

From a 3 g portion of the hexane fraction of the methanol extract (primary extract) of Seongeukchoo, 100% ethanol was added with a pressure of 200 kg / cm ² , a carbon dioxide flow rate of 3.0 ml / min and a co-solvent at a flow rate of 0.2 ml / min and an extraction time of 60 minutes (The size of growth inhibition ring of acne bacterium) was measured by supercritical fluid extraction. The results are shown in Table 7.

<Table 7> Evaluation of Activity of Secondary Extracts of Seonjokcho by Temperature Variation

Oven temperature (℃) CO ₂ flow rate (ml / min) Mobile phase condition Flow rate of mobile phase (ml / min) Pressure (kg / cm ² ) Extract concentration (/ / disk) 3.125 6.25 12.5 25 One 38 3 100% ethanol 0.2 200 10 mm 12 mm 15 mm 17 mm 2 40 3 100% ethanol 0.2 200 12 mm 13 mm 22 mm 25 mm 3 42 3 100% ethanol 0.2 200 12 mm 13 mm 18 mm 22 mm 4 44 3 100% ethanol 0.2 200 10 mm 12 mm 17 mm 20 mm

(3) Evaluation of activity of secondary extract of Seokhoekcho according to the change of auxiliary solvent flux

100% ethanol was added as a co-solvent from 3 g of the hexane fraction of the methanol extract (first extract) of Seonghak-cho (extract) at a pressure of 200 kg / cm ² , a carbon dioxide flow rate of 3.0 ml / min, The results of measurement of the activity (size of growth inhibitory ring of acne bacterium) of the second extract of Seonjakcho obtained by supercritical fluid extraction were as shown in Table 8, and ethanol was added at 0.2 ml / min Activity was the best.

<Table 8> Evaluation of Activity of Secondary Extracts of Seohoekcho According to the Change of Auxiliary Solvent Flow Rate

Oven temperature (℃) CO ₂ flow rate (ml / min) Mobile phase condition Flow rate of mobile phase (ml / min) Pressure (kg / cm ² ) Extract concentration (/ / disk) 12.5 25 50 One 40 3 100% ethanol 0.3 200 23 mm 27 mm 30 mm 2 40 3 100% ethanol 0.2 200 24 mm 28 mm 31 mm 3 40 3 100% ethanol 0.1 200 12 mm 19 mm 26 mm

(4) Evaluation of the activity of the second extract of Seokhoekcho according to the pressure change

100% ethanol was added to 3 g of the hexane fraction of the methanol extract (primary extract) of Seonjeukcho methanol extract as an auxiliary solvent at a flow rate of carbon dioxide of 3.0 ml / min, a temperature of 40 ° C and an extraction time of 60 minutes, Table 9 shows the results of the activity of the second extract of Seonjakcho extract obtained by supercritical fluid extraction (the size of growth inhibition circle of acne bacterium) by varying the pressure, and the activity was the best when the pressure was 200 kg / .

<Table 9> Evaluation of Activity of Secondary Extracts of Seokhogcho with Pressure Changes

Oven temperature (℃) CO ₂ flow rate (ml / min) Mobile phase condition Flow rate of mobile phase (ml / min) Pressure (kg / cm ² ) Extract concentration (/ / disk) 12.5 25 50 One 40 3 100% ethanol 0.2 150 9 mm 10 mm 12 mm 2 40 3 100% ethanol 0.2 200 20 mm 24 mm 27 mm 3 40 3 100% ethanol 0.2 250 11 mm 16 mm 18 mm 4 40 3 100% ethanol 0.2 300 - 8.5 mm 10 mm

(5) Activity of Seohoekcho Secondary Extracts under Optimum Supercritical Fluid Extraction Conditions

From the 3 g of the hexane fraction of the methanol extract (first extract) obtained from Seongeukchecho, 100% ethanol was added with a carbon dioxide flow rate of 3.0 ml / min, a temperature of 40 캜, an extraction time of 60 minutes, and an auxiliary solvent at a flow rate of 0.2 ml / The activity of the second extract of Seok Hak - Cho obtained by supercritical fluid extraction at 200kg / ㎠ was the best. The activity of this extract was found to be 27 ~ 38mm at the concentration of 50μg / disk. It was found that the stronger antimicrobial activity against the stronger phlebotrophic bacteria than the hexane fractions of Seongjeokcho methanol extract (primary extract) Indicating strong activity.

As shown in the above results, the first extract of Seokhogcho showed excellent antimicrobial activities. The hexane fractions obtained from the first extract of Seokhogcho and the second extracts obtained by supercritical fluid extraction were found to be stronger Activity. &Lt; / RTI &gt;

The cosmetic composition according to the present invention, which comprises the acne improvement effective ingredient material derived from the acantholyticus extract of the present invention or the hexane fraction thereof, may contain 0.001 to 20% by weight of the acantholyticus extract or the hexane fraction. At this time, it is more preferable that the content of Seongseogae extract or hexane fraction is 0.5 to 10 wt%.

The cosmetic composition containing the extract of the present invention of the present invention can be used variously in cosmetics and cleanser for improving the acne skin, and the formulations can be arbitrarily selected. Examples of products to which the present composition can be added include cosmetics such as creams, milky lotions, foundations, skins, and the like, and cleansing, cleansing agents, soaps, treatments, and essences.

Hereinafter, specific formulation examples of the cosmetic composition containing the sunflower seed extract or the hexane fraction of the present invention are shown, and the formulations containing the cosmetic composition of the present invention are not limited thereto.

&Lt; Formulation Example 1 > Soft skin length (skin toner)

Ingredients Content (% by weight) Seokhyeok extract or hexane fraction 2.5 glycerin 5.0 1,3-butylene glycol 3.0 ethanol 5.0 Polyoxyethylene nonylphenyl ether 0.5 Spices 0.2 antiseptic Proper amount Purified water to 100

&Lt; Formulation Example 2 >

Ingredients Content (% by weight) Seokhyeok extract or hexane fraction 5.0 glycerin 3.0 Citric acid 0.1 ethanol 10.0 Polyoxyethylene oleyl ether 1.0 Spices 0.1 Sorbitol 2.0 antiseptic Proper amount Purified water to 100

Formulation Example 3 Lotion (emulsion)

Ingredients Content (% by weight) Seokhyeok extract or hexane fraction 4.0 glycerin 5.0 1,3-butylene glycol 8.0 Squalane 10.0 Monooleic acid polyoxyethylene sorbitan (6E.0) 2.0 Triethanolamine 1.5 Glyceryl stearate 0.5 Stearyl glycyrrhetinate 0.2 Carboxyvinyl polymer 0.1 Arginine 0.1 Spices 0.2 antiseptic Proper amount Purified water to 100

&Lt; Formulation Example 4 >

Ingredients Content (% by weight) Seokhyeok extract or hexane fraction 1.0 glycerin 5.0 Stearic acid 8.0 Squalane 5.0 Self emulsifying glycerin monostearate 2.5 Polyoxyethylene sorbitan monostearate 1.5 Propylene glycol 4.0 Stearyl glycyrrhetinate 0.2 vaseline 2.0 Antioxidant Proper amount Spices 0.2 antiseptic Proper amount Purified water to 100

&Lt; Formulation Example 5 > A liquid foundation

Ingredients Content (% by weight) Seokhyeok extract or hexane fraction 0.5 Cetostearyl alcohol 0.5 Liquid lanolin 2.0 Liquid paraffin 3.0 Isopropyl myristate 5.0 Carboxymethylcellulose sodium 0.2 Bentonite 0.5 glycerin 3.0 Titanium dioxide 8.0 Talc 4.0 Colored pigment Proper amount Triethanolamine 1.0 Stearic acid 2.0 Antioxidant Proper amount Spices 0.2 antiseptic Proper amount Purified water to 100

&Lt; Formulation Example 6 >

Ingredients Content (% by weight) Seokhyeok extract or hexane fraction 1.0 Palm oil fatty acid 1.0 Titanium dioxide 0.3 Ethylenamine Sodium tetraacetate 0.1 Spices 0.5 Pigment Proper amount glycerin 1.0 Antioxidant Proper amount Soap chip (base) to 100

According to the present invention, it is possible to extract and isolate which maximizes the activity of the effective ingredient for acne improvement from sunflower seeds, and the cosmetic composition containing the active ingredient material extracted by the present invention can effectively inhibit the growth of acne bacteria. According to the present invention, there can be obtained a cosmetic composition for improving acne which has no side effects as compared with the conventional acne treatment method, and which can be used continuously and has excellent safety.

Claims (9)

delete A method for extracting and extracting an active ingredient for acne improvement comprising the step of adding a predetermined extracting solvent to an acne extract to obtain a primary acne extract, Wherein the extraction solvent is any one selected from anhydrous methanol, hydrated methanol, anhydrous ethanol and hydrated ethanol, The amount of the extraction solvent added is 5 to 20 ml per 1 g of dried syrup- After the step of obtaining the first extract, the extraction solvent is removed from the first extract, and the solid content is dissolved in water to obtain an aqueous solution. Then, hexane is added to the aqueous solution to make the hexane layer effective And extracting the hexane layer to remove hexane to obtain a hexane fraction. The method of extracting and extracting acne-improving active ingredient according to claim 1, further comprising the step of: A method for extracting and extracting an active ingredient for acne improvement comprising the step of adding a predetermined extracting solvent to an acne extract to obtain a primary acne extract, Wherein the extraction solvent is any one selected from anhydrous methanol, hydrated methanol, anhydrous ethanol and hydrated ethanol, The amount of the extraction solvent added is 5 to 20 ml per 1 g of dried syrup- The step of obtaining the first extract of Seolhyeokcho further includes a step of removing the extraction solvent from the first extract of Seolhyeokcho to obtain a solid fraction and obtaining a second extract of Seolhyeokcheol by supercritical fluid extraction from the solid fraction using a supercritical fluid Wherein the acne-improving effective ingredient is selected from the group consisting of: 3. The method of claim 2, Further comprising the step of obtaining a second extract of Seongjeok first by the supercritical extraction method using the supercritical fluid from the hexane fraction. The method according to claim 3 or 4, Wherein the step of obtaining the second extract of Seok Hak can use carbon dioxide as a supercritical fluid under a temperature condition of 38 to 44 캜 and a pressure of about 200 kg / cm 2. 6. The method of claim 5, Wherein the step of obtaining the second extract comprises extracting ethanol as an auxiliary extraction solvent in the solid fraction or the hexane fraction. The method according to claim 6, Wherein the ethanol added as the auxiliary extraction solvent is added to the solid fraction or the hexane fraction at a flow rate of 0.2 to 0.3 ml per minute. A cosmetic composition for improving acne characterized by comprising a fraction of hexane according to claim 2 or a second extract of Seongjeokcho according to claim 3. 9. The method of claim 8, Wherein the content of the hexane fraction or the second extract of Seok Hak cane is 0.001 ~ 20 wt%.

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