KR20120062615A - External composition for skin using an extract or a fraction of padina arborescens - Google Patents

Abstract

PURPOSE: An external use skin composition containing Padina arborescens extract and/or fraction thereof is provided to ensure antibacterial function and to treat acne. CONSTITUTION: An antibacterial external use skin composition contains ethanol solution extract of Padina arborescens or hexane fraction, dichloromethane fraction thereof, or ethyl acetate fraction thereof as an active ingredient. The external use skin composition has antibacterial function against Staphylococcus aureus, Staphylococcus epidermidis, or Propionibacterium acne. The external use skin composition is a cosmetic, cleansing, or pharmaceutical composition.

Description

External Composition for Skin Using an Extract or a Fraction of Padina arborescens}

The present invention relates to an external preparation composition for skin using BuChatmal extract or fraction. In particular, the present invention relates to an external composition for skin having antibacterial activity or acne improving activity using a buccal extract or fraction.

Human skin is an organ that is in direct contact with the environment, and infections of bacteria, fungi, etc., which are involved in various skin diseases, occur easily.

For example, Staphylococcus aureus causing a star maturation skin Philo nose kusu aureus (Staphylococcus aureus), including the skin flora is Staphylococcus epidermidis (Staphyococcus epidermidis ), acne bacterium Propionibacterium acnes ) are present on the skin, and these bacteria break down sebum and sweat secreted on the skin to produce odors, and the breakdown products cause irritation and inflammation of the skin.

Especially Propionibacterium acnes ) breaks down sebum using fatty acid degrading enzyme lipase to produce free fatty acids. These free fatty acids not only irritate the skin but also cause inflammatory lesions such as papules, pustules and nodules, which are red rashes found in acne.

Antimicrobial compounds such as triclocarban and triclosan have been used as antimicrobial agents for these bacteria, but they are known to cause skin side effects such as skin burning, burning, and redness of the skin when used for a long time.

Therefore, recent studies on natural extracts with less skin side effects have been conducted, such as green tea catechin, eucalyptus leaf oil, lemon peel oil, stalk extract, aloe vera peel extract, and the like (Korean Patent Publication No. 2010-0088878, Domestic Publication No. 2005-0010914, National Publication No. 2010-0088296)

The genus was based on the net wealth horse chaetmal the Padina durvillaeI by Okamura`s (1982) first reported in Busan, Korea in P. arborescens Holmes, P. australis Hauck, P. boryana Thivy in Taylor, P. crassa Yamada, P. Six species, including japonica Yamada, and P. minor Yamada, are reported to be growing (Lee, YP and S. Kang. 2001. A catlogue of the seaweeds in Korea. Cheju National University Press. pp. 662).

Budina horses ( Padina arborescens ) are distributed in Japan and Korea, and Korea is known to be distributed in Hamgyeong-do, Busan, Bijin-do, Geoje-do, Namhae-do, Chuja-do, and Jeju-do. The body is flat, without middle, and fan-shaped. Forging or aging is radially crushed. The size is 6-7 cm, sometimes 25-30 cm. Lower part has brown hair. The group of germ cells generally occurs on the back side of the body, that is, on the side where the growth margin is rolled up, and is arranged in an annular alternation with the mother bar of the body. The quality is thicker and dry, especially when the lower part is hard like wood and does not stick to the ground (National Institute of Biological Resources, Ministry of Environment. Biomass of Korea, Algae). 2010. Vol. 2 No. 2, pp 46-48).

The physiological activity of BuChatmal has been reported to inhibit the oxidative stress of ECV304 cells, a vascular endothelial cell line by H ₂ O ₂ (Korean J. Oriental Physiology & Pathology 22 (6): 1431 ~ 1438, 2008 ).

The present invention discloses an external composition for skin using an antibacterial activity against skin bacteria of the buchamal extract or a fraction thereof.

It is an object of the present invention to provide an antimicrobial skin external preparation composition using BuChatmal extract or a fraction thereof.

Another object of the present invention is to provide a topical composition for improving acne using BuChatmal extract or a fraction thereof.

The specific object of the present invention will be presented below.

In one aspect, the present invention relates to an antimicrobial skin external preparation composition using BuChatmal extract or a fraction thereof.

The inventors of the present invention, as confirmed in the following Examples and Experimental Examples, in order to confirm the antibacterial activity of the BuChatmal extract or its fractions ethanol aqueous solution extract of BuChatmal (80% (v / v) ethanol as a mixed solvent of ethanol and water) Sequential fractions of hexane, dichloromethane, ethyl acetate and butanol, and extracts and sequential fractions of the skin Microorganisms such as Staphylococcus aureus ( S. aureus KCTC 1927), Staphylococcus epidermidis (including S. epidermidis SK4 and antibiotic resistant bacteria S. epidermidis SK9 and S. epidermidis SK19), And propionibacterium acnes ( P. acnes) Antimicrobial activity against ATCC 3314 and antibiotic resistant bacterium P. acnes SKA7) was evaluated by Disc diffusion assay, Minimum inhibitory concentration (MIC) and Minimal bactericidal concentration (MBC). .

As a result, in the antimicrobial activity evaluation results by the disc diffusion method with a treatment concentration of 1 mg / ml, the aqueous ethanol extract showed activity against Staphylococcus aureus and Staphylococcus epidermidis SK4, and the hexane fraction , Dichloromethane and ethyl acetate fractions showed activity against all bacteria used in the experiment, including antibiotic resistant bacteria. The remaining fractions butanol fraction and residual water layer did not show antimicrobial activity against all skin microorganisms.

In addition, in the evaluation results of the antimicrobial activity by the minimum inhibitory concentration, the ethanol aqueous solution extract showed a minimum inhibitory concentration in the range of 1 to 2 mg / ㎖ for all bacteria, hexane fraction, dichloromethane fraction and ethyl acetate fraction is 0.25 to 2 Minimal inhibitory concentrations in the mg / ml range were shown. Again, butanol fraction and residual water layer did not show antimicrobial activity against all skin microorganisms.

Finally, in the results of the evaluation of the antimicrobial activity by the minimum killing concentration, the hexane fraction showed a minimum killing concentration of 3 mg / ml only against Staphylococcus aureus and the antibiotic resistant bacterium Staphylococcus epidermidis SK9. Ethyl acetate fraction is Staphylococcus epidermidis, an antibiotic resistant bacterium. SK9 and Staphylococcus epidermidis Minimal bactericidal concentrations of 3 mg / ml and 2 mg / ml were shown for SK19, respectively.

The results of the experiment can be said that the ethanol aqueous extract and the sequential fraction of ethanol, dichloromethane and ethyl acetate fraction of Buchathal can be used as antibacterial against skin microorganisms, in particular hexane fraction and It can be said that the ethyl acetate fraction can be usefully used for antimicrobial use against skin microorganisms.

In consideration of the foregoing, the antimicrobial skin external composition of the present invention comprises an aqueous ethanol extract of BuChatmal, a hexane fraction of the ethanol aqueous extract, a dichloromethane fraction of the ethanol aqueous extract, and / or an ethyl acetate fraction of the ethanol aqueous extract. It is characterized by including as an active ingredient.

In the present specification, "buchathal ethanol aqueous solution extract" means an extract obtained by immersing buchathal in an ethanol aqueous solution. As long as the BuChat horse is obtained by immersing in an ethanol aqueous solution, the extraction method may be any method such as cooling, refluxing, heating, and ultrasonic radiation.

In addition, in the present specification, "hexane fraction of ethanol aqueous solution of chatchatal acid" is a suspension obtained by mixing the aqueous ethanol aqueous extract of ethanol aqueous extract with a mixture of hexane and the suspension of solid buchatmal ethanol aqueous solution extract from which the extraction solvent is removed. It means the fraction obtained by mixing with hexane and standing.

In addition, in the present specification, "dichloromethane fraction of the buchmalol ethanol aqueous solution extract" is a fraction obtained by mixing the buchmalar ethanol aqueous solution extract with dichloromethane, and obtained by removing the extraction solvent from the bubothermal ethanol aqueous solution extract. A fraction obtained by suspending a shape of a bubumal ethanol aqueous solution extract in water and mixing it with dichloromethane and standing still, or by distilling the solid buccal ethanol aqueous solution extract into water and dividing it sequentially into hexane and dichloromethane. It means a fraction of the dichloromethane layer obtained by. Here, "sequential dividing into hexane and dichloromethane" means that the solid butchatal ethanol aqueous solution extract is suspended in water, mixed with hexane, left to stand, and the water layer, which is the remaining layer except for the fraction of the hexane layer, is dichloromethane. This means mixing and policing with methane to fractionate.

In the present specification, "ethyl acetate fraction of the aqueous bubethanol ethanol extract" refers to a fraction obtained by mixing the aqueous bubethanol ethanol extract with ethyl acetate, the solid obtained by removing the extraction solvent from the aqueous bubethanol ethanol extract. A fraction obtained by suspending the aqueous extract of aqueous BuChatmal ethanol in water and then mixing and standing with ethyl acetate, or suspending the solid extract of BuChatmal ethanol aqueous solution in water and dividing it sequentially with hexane, dichloromethane and ethyl acetate It means a fraction of the ethyl acetate layer obtained by. Here, "sequential dividing into hexane, dichloromethane and ethyl acetate" means the solid extract of the aqueous bubethanol ethanol solution is suspended in water, and the hexane and dichloromethane fractions are obtained therefrom. It means to fractionate by mixing and politics with acetate.

In addition, the term "active ingredient" as used herein means a component that can exhibit the desired activity alone or in combination with a carrier that is not active itself.

In addition, in the present specification, "antibacterial" means killing or inhibiting the proliferation of one or more microorganisms selected from the group consisting of skin microorganisms Staphylococcus aureus , Staphylococcus epidermidis and Propionibacterium acnes It means the property or use to.

Other terms that are not specifically defined herein follow the Korean dictionary meaning or the meaning generally accepted in the art.

In the antimicrobial skin external composition of the present invention, the preferred active ingredient is a hexane fraction, a dichloromethane fraction or an ethyl acetate fraction of an aqueous bubethan ethanol extract. More preferably, it is a hexane fraction or an ethyl acetate fraction of an aqueous bubethan ethanol extract. In addition, as shown in the following experimental example for the minimum killing concentration, when the active ingredient is a hexane fraction, the antimicrobial activity is antibacterial against Staphylococcus aureus and antibiotic resistant bacteria Staphylococcus epidermidis SK9, and the active component is ethyl acetate. In the case of fractions, the antimicrobial activity is the antibiotic resistant bacterium Staphylococcus epidermidis SK9 and Staphylococcus epidermidis It is preferred to be understood as antimicrobial against SK19.

In another aspect, the present invention relates to an external preparation composition for improving acne.

The following experimental example shows that the extract of Buchchatal ethanol aqueous solution, its hexane fraction, dichloromethane fraction, and ethyl acetate fraction showed antibacterial activity against propionibacterium acnes, an acne bacterium, and especially ethyl acetate fraction. When lotion was prepared and sensory evaluation was performed, more than two-thirds of respondents indicated that acne was improved.

Therefore, the external preparation composition for improving acne of the present invention is characterized in that it contains an aqueous extract of aqueous ethanol extract, hexane fraction thereof, dichloromethane fraction and / or ethyl acetate fraction as an active ingredient.

The external preparation composition for improving acne of the present invention is an active ingredient, preferably an ethyl acetate fraction having excellent antibacterial activity and confirmed acne improving activity in sensory evaluation.

In the external composition for improving acne skin of the present invention, the meaning of the extract of the aqueous ethanol aqueous extract, the meaning of the hexane fraction, the meaning of the dichloromethane fraction, the meaning of the ethyl acetate fraction, the meaning of the active ingredient, and the like are described above. One bar is still valid.

Meanwhile, the antimicrobial skin external composition or acne skin external composition for improving acne of the present invention (hereinafter "skin external composition of the present invention") may exhibit antibacterial or acne improving activity of an extract or a fraction of ethanol aqueous solution, which is an active ingredient thereof. It may be included in any amount (effective amount) depending on the use, formulation, formulation purpose, etc. A typical effective amount will be determined within the range of 0.001% to 20.000% by weight based on the total weight of the composition. Here, the "effective amount" refers to the amount of the active ingredient that can exhibit antimicrobial activity or acne improving activity. Such effective amounts can be determined experimentally within the range of ordinary skill in the art.

The external preparation composition for skin of this invention can be grasped | ascertained as a cosmetic composition in a specific aspect.

When the composition of the present invention is understood as a cosmetic composition, the cosmetic composition can be prepared in various forms, for example, emulsions, lotions, creams (oil-in-water, water-in-oil, multiphase), solutions, suspensions (anhydrous and Aqueous), anhydrous products (oil and glycol based), gels, masks, packs, powders and the like.

The composition of the present invention may include an acceptable carrier in the cosmetic preparation in addition to including the active ingredient.

The term "acceptable carrier in cosmetic preparations" herein refers to compounds or compositions that are already known and used that can be included in cosmetic preparations or compounds or compositions that will be developed in the future and that the human body is not as toxic as applicable to contact with the skin.

The carrier may be included in the composition of the present invention from about 1% to about 99.99% by weight, preferably from about 50% to about 99% by weight of the composition.

However, it is understood that the ratios limit the scope of the present invention in any aspect because the ratios vary depending on the above-described formulation of the cosmetic and its specific application site (face or hand) or its preferred application amount. It should not be.

On the other hand, as the carrier, alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, humectants, viscosity modifiers, emulsions, stabilizers, sunscreens, coloring agents, fragrances and the like can be exemplified.

Alcohols, oils, surfactants, fatty acids, silicone oils, wetting agents, humectants, viscosity modifiers, emulsions, stabilizers, sunscreens, colorants, compounds / compositions that can be used as fragrances, etc., which can be used as the carrier are already known in the art. As a result, those skilled in the art can select and use appropriate materials / compositions.

The cosmetic composition of the present invention may further include functional substances such as skin moisturizing active substance, anti-atopic active substance, anti acne active substance, whitening active substance and the like.

The composition of this invention can be grasped | ascertained as a skin cleanser composition in another specific aspect.

The skin cleanser composition of the present invention may be prepared in a solid or liquid state, and may be commercialized as a body soap, a hand soap, a face wash, and the like.

The skin cleansing agent of the present invention may include any component used in the preparation of the cleansing agent in the art, as long as the antibacterial activity of the extract or fraction of ethanol aqueous solution of buccalmar, which is an active ingredient thereof, is not inhibited.

Examples of such optional components include anionic surfactants such as polyoxyethylene alkyl ether sulfate, nonionic surfactants such as amine oxide and polyoxyethylene alkyl ether, alcohols such as oil, silicones, lower or higher alcohols, and lanolin. Derivatives, protein derivatives, acrylic resin dispersions, cationic polymers, anionic polymers, nonionic polymers, pharmaceuticals such as vitamins, fungicides, preservatives, pH adjusters, antioxidants, metal blockers, ultraviolet absorbers, animal or plant extracts or derivatives thereof, Pigments, paints, pigments, inorganic powders, clay minerals, water-insoluble polymer powders such as nylon and polyethylene, propylene glycol, ethyl alcohol, glycerin, water and the like.

The composition of the present invention may be regarded as a pharmaceutical composition in another specific embodiment.

The pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier, excipient, and the like in addition to the active ingredient, and may be a topical formulation such as creams, lotions, ointments (semi-solid external preparations), micro-emulsions, gels, pastes, transdermal preparations. (TTS) (such as patches, bandages, etc.) and the like.

As used herein, "pharmaceutically acceptable" means that the subject of application (prescription) is not as toxic as applicable, without inhibiting the activity of the active ingredient.

Examples of pharmaceutically acceptable carriers include lactose, glucose, sucrose, starch (such as corn starch, potato starch, etc.), cellulose, derivatives thereof (such as sodium carboxymethyl cellulose, ethylcellulose, etc.), malt, gelatin, talc, Solid lubricants (such as stearic acid, magnesium stearate, etc.), calcium sulfate, vegetable oils (such as peanut oil, cottonseed oil, sesame oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, etc.), alginic acid, emulsifiers (such as TWEENS), wetting agents (Such as sodium lauryl sulfate), colorants, flavors, stabilizers, antioxidants, preservatives, water, saline, phosphate buffer solutions, and the like. The carrier may be selected from one or more of suitable pharmaceutical formulations according to the formulation of the pharmaceutical composition of the present invention.

Excipients may be selected and used according to the formulation of the pharmaceutical composition of the present invention, for example, suspending agents and dispersants such as sodium carboxymethyl cellulose, methyl cellulose, hydropropylmethyl cellulose, sodium alginate, polyvinylpyrrolidone, etc. Can be.

The daily dosage of the pharmaceutical composition of the present invention is usually 0.001 ~ 150 mg / kg body weight range, it can be administered once or divided into several times. However, since the dosage of the pharmaceutical composition of the present invention is determined in view of various related factors such as the route of administration, the age, sex, weight of the patient, and the severity of the patient, the dosage may limit the scope of the present invention in any aspect. It should not be understood as.

As described above, according to the present invention, it is possible to provide an external preparation composition for skin using BuChatmal extract or a fraction thereof. The external preparation composition for skin of the present invention may be formulated into a cosmetic composition, a cleaning composition, a pharmaceutical composition, or the like.

Figure 1 is a schematic diagram of the BuChat horse extraction and fractionation process.
Figure 2 shows the results show that the BuChatmal extract or its fraction does not show the cytotoxicity.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these examples and experimental examples.

< Example > Buchat  Preparation of Extract

  The BuChat horse collected on April 6, 2006 at Seogwipo-si, Seogwipo-si, Jeju Island was removed from adherent organisms, and the samples were dried and crushed with a crusher. The fine powder sample of Buchat was leached in 80% ethanol at room temperature for 48 hours and extracted three times. Then, the supernatant was recovered and concentrated under reduced pressure to obtain an ethanol extract. 10 g of ethanol extract was suspended in 1 L of distilled water, and then extracted sequentially with hexane (1 L × 3), dichloromethane (1 L × 3), ethyl acetate (1 L × 3) and butanol (1 L × 3). The fractions were dried in vacuo to give hexane, dichloromethane, ethyl acetate and butanol and the remaining water layer. Each fraction was dissolved in the solvent and sterilized with a 0.2 μm membrane filter (Advantec MFS Inc. USA) and stored at 4 ° C. for use in this study.

BuChat horse 59.2 g of dry powder was extracted with 80% ethanol, and the extract obtained by filtration was concentrated under reduced pressure to obtain 11.87 g of crude extract, the yield was 20.0%. Sequential fractions were suspended in 1 L of distilled water after 10 g of ethanol extract of BuChatmal, followed by hexane (1 L × 3), dichloromethane (1 L × 3), ethyl acetate (1 L × 3) and butanol (1 L × 3) sequentially divided into Fractions of 0.542 g in the hexane layer, 0.128 g in the dichloromethane layer, 0.058 g in the ethyl acetate layer, 1.470 g in the butanol layer and 4.353 g in the residue water layer were obtained. The schematic diagram of the extraction and fractionation process is shown in FIG.

< Experimental Example > Buchat To skin bacteria  Sensory evaluation experiment for antimicrobial activity test and acne improvement activity

< Experimental Example  1> Antimicrobial activity test

Experimental Example 1-1 Experiment by Disc Diffusion Method

The strains used in this experiment were three strains commonly found in skin, Staphylococcus aureus, Staphylococcus epidermidis, and Propionibacterium acnes, and some Staphylococcus epidermidis. And propionibacterium acnes were used together with antibiotic resistant bacteria.

Anaerobic bacteria, Propionibacterium acnes, were cultured in anaerobic Jar (Oxoid, USA).

The culture medium of Tryptic soy agar (TSA) was used by Difco (USA) and GAM Broth from Nissui (Japan).

Table 1 below summarizes the strains used, the medium, and the like.

Antibacterial activity was measured using the disk diffusion method (Disc diffusion assay) (Hayes, AJ, Markovic, B., 2002. Toxicity of Australian essential oil Backhousia citriodora (Lemon myrtle). Part 11. Antimicrobial activity and in vitro cytotoxicity. Food. Chem .. Toxicol 40 (4): 535-543).

Each strain was inoculated into 10 mL of liquid medium and passaged three times at 37 ° C. for 18 hours to be used as an antimicrobial test strain. Each test bacterium concentration was 0.5 ⁶ to 10 ⁸ CFU / mL at 0.5 McFarland. The inoculation plate medium was used for the pour-plate method.

Each sample was prepared at a concentration of 1 mg / ml, slowly absorbed by paper disc (Whatman No. 5, 8 mm) by concentration, and the solvent was completely evaporated. The disk was adhered to the surface of the test plate medium and incubated at the optimum temperature of 37 ℃ and the antimicrobial activity was compared by measuring the size of the growth zone (mm) generated around the disk.

The results are shown in Table 2 below.

Referring to Table 2, the ethanol extract showed activity against Staphylococcus aureus and Staphylococcus epidermidis SK4, and the hexane fraction, dichloromethane fraction and ethyl acetate fraction contained antibiotic resistant bacteria. All the bacteria used in the experiment showed activity. In particular, the hexane fraction showed a high growth inhibition of 15 mm in Staphylococcus aureus. Butanol fraction and residual water layer showed no activity against all bacteria.

<Experimental Example 1-2> The minimum inhibitory concentration (Minimum inhibitory concentration , MIC )

MIC is determined according to the micro-well dilution assay method (Amster D. 1996. Susceptibility testing of antimicrobial in liquid media, antibiotics in laboratory medicine.4th ed. Williams and Wikins, MD, USA.p 52-111). Measured. Each bacterium concentration was 10 ^{6-10 8} CFU / mL at 0.5 McFarland, and then divided into 96 well plates, and each sample was incubated with 10 μl of each sample. The growth of the bacterial culture was measured at 650 nm with a Microplate reader (Powerwave XS, BioTek). The minimum inhibitory concentration was determined by comparing the absorbance values in the wells inoculated only with bacteria.

The results are shown in Table 3 below.

Referring to [Table 3], the ethanol extract showed a minimum inhibitory concentration in the range of 1 to 2 mg / ㎖ for all the bacteria, the hexane fraction, dichloromethane fraction and ethyl acetate fraction in the range of 0.25 to 2 mg / ㎖ Minimal inhibitory concentrations were shown. In particular, the ethyl acetate fraction showed a relatively low concentration of inhibitory concentrations of 0.5 mg / ml and 0.25 mg / ml, respectively, against the antibiotic resistant bacteria Staphylococcus epidermidis SK9 and Staphylococcus epidermidis SK19.

Experimental Example 1-3 Minimum Bacterial Killing Concentration ( Minimal bactericidal concentration , MBC ) measurement

100 μl of the wells corresponding to concentrations higher than the concentration selected as MIC was plated directly onto the solid medium. Each culture was directly counted the number of colony observed visually on the plate after incubation. At this time, when it was confirmed that the colony number showed the effect of killing 99.9% of the initial inoculated bacteria inoculated from the liquid culture solution, the treatment concentration at this time was determined as the minimum bacterial kill concentration (MBC) (Cohen MA, Huband MD, Yoder). SL, Gage JW, Roland GE. , 1998, Bacterial eradication by clinafloxacin, CI-990, and ciprofloxacin employing MBC test, in-vitro time-kill and in-vivo time-kill studies. J Antimicrob Chemother . 41 (6): 605-14).

The results are shown in Table 4 below.

Staphylococcus epidermidis, a Staphylococcus aureus and antibiotic resistant bacterium For SK9, the hexane fraction showed a minimum bacterial killing concentration of 3 mg / ml, and the ethyl acetate fraction showed Staphylococcus epidermidis, an antibiotic-resistant bacterium. SK9 and Staphylococcus epidermidis Minimal bactericidal concentrations of 3 mg / ml and 2 mg / ml were shown for SK19, respectively.

Experimental Example 1-4 Cytotoxicity Evaluation by MTT Method

Cell viability was measured using MTT assay to confirm the protection and toxicity of the boochat horse ethanol extract and fractions. The MTT assay is a test that utilizes the ability of intracellular mitochondria to reduce the yellow water-soluble MTT tetrazolium to a blue-violet, water-insoluble MTT formazan crystal by dehydrogenase action.

RAW 264.7 cell lines were incubated for 12 hours in DMEM medium at a concentration of 1 × 10 ⁵ cells / well in 96 well plates, followed by incubation at 100 μg / ml of ethanol extract or fraction of BuChatmal for 24 hours. After incubation, the medium was removed and 200 μl of 3- (4,5-Dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT; SIGMA, USA) solution was added to each well and incubated at 37 ° C. for 4 hours. The supernatant was removed and 200 μl of DMSO was added to dissolve the insoluble purple formazan. The amount of dissolved formazan was measured at 570 nm using a microplate reader (BioTek, USA). The control group was cultured with only the medium without the sample added, and the cell viability was calculated based on the absorbance of the control group.

The results are shown in [FIG. 2].

Referring to FIG. 2, it can be seen that ethanol extracts and fractions of BuChat horse show no particular cytotoxicity.

< Experimental Example  2> Sensory Evaluation Experiments on Acne Improvement Activity

In <Experimental Example 1>, the sensory evaluation was performed on acne improvement activity using the nutrient lotion prepared with the ethyl acetate fraction shown to have the best antibacterial activity against the acne bacterium Propionibacterium acnes.

The nutrition lotion was prepared using the ingredients and contents shown in the following [Table 5]. In order to verify the acne improvement activity of the nutrition lotion of the preparation example, the nutrition lotion of the comparative example was prepared and used for sensory evaluation together. As a lotion, instead of the ethyl acetate fraction of Example in Table 5 below, a nutrient lotion containing purified water as much as that content was used.

number ingredient content
(weight%) One Ethyl Acetate Fraction from Buchat 2.0 2 Stearic acid 2.0 3 Cetyl alcohol 2.0 4 Glyceryl Monostearate 2.0 5 Polyoxyethylene sorbitan monostearate 0.5 6 Sorbitan sesquioleate 0.5 7 Glyceryl Monostearate / Glyceryl Stearate / Polyoxyethylene Stearate 1.0 8 Wax 1.0 9 Liquid paraffin 4.0 10 Squalane 4.0 11 Caprylic / capric triglyceride 4.0 12 Carboxyvinyl polymer 0.3 13 Butylene glycol 5.0 14 glycerin 3.0 15 Triethanolamine 0.5 16 Purified water Balance

Sensory evaluation is divided into 60 groups of men and women between 10 and 20 years of age with acne disease, each group to use the nutritional lotion of the preparation example and the comparative example of the nutritional lotion twice a day for 4 weeks twice a day The number of users (A) who answered that the results were improved, the number of users (B) who answered that they did not change, and the number of users (C) who said that they had worsened, are shown in [Table 6] below. .

division A B C Manufacturing example 22 6 2 Comparative example 6 17 7

The results in Table 6 show that when using the nutritional lotion of the preparation example, a response was obtained in more than two thirds of the total users. In the nutritional lotion of the comparative example, only 4 users answered that it was improved, supporting the ethyl acetate fraction of BuChatmal having acne improving activity.

Claims (5)

Antibacterial skin external composition comprising at least one selected from the group consisting of ethanol aqueous extract of buchamal, hexane fraction of the aqueous ethanol aqueous extract, dichloromethane fraction of the aqueous ethanol aqueous extract and ethyl acetate fraction of the aqueous ethanol aqueous extract .
The method of claim 1,
The antimicrobial composition is an antimicrobial skin external composition according to claim 1, wherein the antimicrobial agent is antimicrobial against at least one microorganism selected from the group consisting of Staphylococcus aureus , Staphylococcus epidermidis and Propionibacterium acne.
The method of claim 1,
The antimicrobial skin external composition is an antimicrobial skin external composition, characterized in that the cosmetic composition, detergent composition or pharmaceutical composition.
Skin extract for acne improvement comprising at least one selected from the group consisting of ethanol aqueous extract of buchamal, hexane fraction of the ethanol aqueous extract, dichloromethane fraction of the ethanol aqueous extract and ethyl acetate fraction of the ethanol aqueous extract External preparation composition.
The method of claim 4, wherein
The external preparation composition for acne improvement is a skin external preparation composition for improving acne, which is a cosmetic composition, a cleaning composition or a pharmaceutical composition.

Images