CN109010112B - Use of composition for external use for skin containing ginsenoside Y as main active ingredient - Google Patents

Abstract

The present invention relates to the use of a composition for external application to the skin containing ginsenoside Y as a main active ingredient, and more particularly, to a composition for external application to the skin containing ginsenoside Y having excellent antioxidant ability and capable of providing skin problems such as improvement of acne, antibacterial, inhibition of lipid production and allergy.

Description

Use of composition for external use for skin containing ginsenoside Y as main active ingredient

The present application is a divisional application of chinese patent application having application number 201480025030.6, application date 5/2/2014, and entitled "topical skin composition containing ginsenoside Y".

Technical Field

The present invention relates to a composition for external application to skin containing ginsenoside Y, and more particularly, to a composition for external application to skin containing ginsenoside Y, which has excellent antioxidant activity, and can provide effects of improving the skin moisturizing ability, anti-inflammatory effect, improving skin problems such as acne and allergy, whitening effect, sebum-regulating effect, pore-shrinking effect, improving the general state of skin such as complexion by improving blood circulation, and improving the state of scalp and hair such as dandruff-preventing effect, hair-growing effect, and white hair-preventing effect.

Background

Human skin, as a first protective membrane of the human body, has a function of protecting internal organs from changes in temperature and humidity, and external environmental stimuli such as ultraviolet rays and harmful substances, and as it ages, the skin undergoes changes due to various intrinsic and extrinsic factors. That is, there are inherent problems that secretion of various hormones regulating metabolism is decreased, function and cell activity of immune cells are decreased, biosynthesis of immune proteins and structural proteins of living body required by living body is decreased, and also, there are inherent problems that ultraviolet ray content reaching the ground surface in sunlight is increased due to ozone layer destruction, and as environmental pollution is more serious, free radicals and active harmful oxygen are increased, so that skin causes various changes, that is, not only thickness is thinned, wrinkles are increased, elasticity is decreased, but also skin color is darkened, skin problems are frequently occurred, spots, freckles and senile plaques are increased, color is deteriorated and skin color is darkened.

In order to prevent such skin state changes due to intrinsic and extrinsic factors and maintain healthy skin state, efforts have been made to improve skin state by adding physiologically active substances derived from various animals, plants, microorganisms, etc. known in the art to cosmetics, and in particular, many studies and developments have been made on using ginseng (Panax ginseng, Panax ginseng c.a.meyer) as a component of cosmetics. Ginseng has been already well established for its function on the skin and is widely used in cosmetic compositions, and extracts of ginseng roots or leaves, ginsenosides (aglycons), ginseng polysaccharides, and the like are commonly used. However, these cosmetics containing ginseng extract, essence, polysaccharide, etc. contain an extremely small amount of active substances, and have a disadvantage that the effect is not very remarkable as compared with other raw materials.

Disclosure of Invention

Technical subject

Accordingly, the present inventors have found that ginsenoside Y contained in ginseng provides not only whitening and moisturizing effects, but also skin condition improving effects such as a complexion improving effect, sebum regulating effect, and pore shrinking effect, and also scalp and hair condition improving effects such as dandruff prevention, hair growth prevention, and white hair prevention, and have completed the present invention.

Accordingly, an object of the present invention is to provide a composition for external application to skin, which contains ginsenoside Y and is capable of improving the general state of skin.

Means for solving the problems

In order to achieve the above object, the present invention provides a skin external composition containing ginsenoside Y as an active ingredient.

In addition, the present invention provides a skin external composition for whitening skin containing ginsenoside Y as an active ingredient.

In addition, the present invention provides a skin external composition for moisturizing containing ginsenoside Y as an active ingredient.

In addition, the present invention provides a skin external composition for improving acne containing ginsenoside Y as an active ingredient.

In addition, the present invention provides a composition for external use for skin comprising ginsenoside Y as an active ingredient for improving allergy.

In addition, the present invention provides a skin external composition for improving complexion and complexion comprising ginsenoside Y as an active ingredient.

In addition, the present invention provides a skin external composition for shrinking pores, comprising ginsenoside Y as an active ingredient.

The present invention also provides a skin external composition for regulating sebum, which contains ginsenoside Y as an active ingredient.

In addition, the present invention provides a composition for external use for skin comprising ginsenoside Y as an active ingredient for preventing dandruff.

In addition, the present invention provides a skin external composition for hair growth comprising ginsenoside Y as an active ingredient.

In addition, the present invention provides a skin external composition for preventing canities, which contains ginsenoside Y as an active ingredient.

In addition, the present invention provides a skin external composition using ginsenoside Y as a natural preservative.

Advantageous effects

The composition of the present invention has excellent antioxidant ability due to the inclusion of ginsenoside Y, and can provide not only an effect of improving skin moisturizing ability, an anti-inflammatory effect, an effect of improving skin problems such as acne and allergy, a whitening effect, an effect of regulating sebum, an effect of shrinking pores, an effect of improving the overall state of skin such as facial complexion due to improvement of blood circulation, but also an effect of improving the state of scalp and hair such as an effect of preventing dandruff, an effect of growing hair, and an effect of preventing white hair.

Detailed Description

The composition for external use for skin according to the present invention contains ginsenoside Y as an effective ingredient, and particularly, the ginsenoside Y can be extracted from ginseng.

Ginsenoside Y used in the present invention has the structure of the following chemical formula 1

Chemical formula 1

Ginseng belongs to the genus Panax of Araliaceae (Araliace), and is a perennial root dicotyledonous herbaceous plant. Production is carried out in many countries such as korea, china, japan and the united states, and the main cultivation areas are: in east Asia, the east longitude is 85-140 degrees, the north latitude is 22-49 degrees, in North America, the west longitude is 70-97 degrees, and the north latitude is 34-47 degrees. Ginseng thus produced all over the world is different in composition, and most important when it shows various physiological and pharmacological effects of ginseng is ginseng saponin (saponin), which is called ginsenoside, and ginseng produced in korea has the highest content of ginseng saponin. About 34 kinds of ginsenosides have been isolated so far, and their pharmacological effects are different depending on the kind of saccharide bound to aglycone, the number of bound saccharide, or the binding site. Ginsenosides are classified into Protopanaxadiol (Protopanaxadiol) type, Protopanaxatriol (Protopanaxatriol) type, and oleanane (oleana) type according to structural characteristics. In addition, it also contains ginseng alkene (panacen), polyacetylene compound, polyphenol compound, flavonoid and vitamins as the natural aroma components of ginseng.

The traditional Chinese medicine effect of ginseng has been recognized since ancient times, and thus, it is used for treating various diseases. The effects of ginseng on traditional Chinese medicines are known to be physical strength enhancement, metabolism improvement, stress relieving, diabetes treatment, respiratory disease improvement, digestive organ improvement and anticancer effects, and recent studies have disclosed that ginseng has anticomplementary activity, antiulcer effect, immunity enhancement, anticancer effect, antihyperglycemic effect and the like. The effects related to skin in the research publications include anti-inflammatory effects (Korea J. Dermatol.18 (1): 39-42 (1980)), Korea J. Dermatol.14 (4): 335-, elasticity and hydration enhancement (Fitotterapia 57 (1): 15-28 (1986)), Fitotterapia 57 (4): 217-.

The ginsenoside Y of the present invention may be extracted from a plant, synthesized by a method known in the art, or used as a commercially available product. In addition, in consideration of the technical level in the art, it is clear to those skilled in the art that among the inducers obtained by subjecting the ginsenoside Y to substitution and substitution reactions of substituents generally practiced in the art, inducers showing an effect of improving skin and hair and a preservative effect are also included in the scope of the present invention.

Ginsenoside Y used in the present invention may be extracted from a ginseng extract in particular. The kind of ginseng used is not particularly limited, and ginseng, red ginseng, white ginseng, taiji ginseng, and ginseng radix can be used. The ginseng extract includes a leachate obtained by leaching and transferring ginseng, a concentrate obtained by concentrating the leachate again partially or entirely, or a chemical substance which is produced by drying the concentrate again and exerts a main effect by being contained in ginseng, a precipitate, a decoction, a tincture, a fluid essence, and the like, and it may include a plant itself, and an extract of all parts of ginseng such as a stem, a root, a leaf, a flower, a fruit, and the like, and is not limited to an extract of a specific part. In addition, known methods can be used to extract ginsenoside Y from the ginseng extract.

Specifically, after a ginseng extract is produced from ginseng using water or an organic solvent by a method commonly used in the art, the ginsenoside Y is separated therefrom. The organic solvent used in the present invention may be selected from the group consisting of ethanol, methanol, butanol, ether, ethyl acetate, chloroform and a mixed solution of these organic solvents and water, and 80% ethanol is preferably used. In this case, the extraction temperature is preferably 10 to 80 ℃ and the extraction time may be 3 to 24 hours. When the extraction temperature and the extraction time are exceeded, the extraction efficiency may be reduced or the composition may be changed.

The composition according to the present invention contains ginsenoside Y in an amount of 0.001 to 50 wt%, preferably 0.01 to 30 wt%, and more preferably 0.1 to 10 wt% based on the total weight of the composition. If the content of ginsenoside Y is less than 0.001 wt%, the effect or effect is weak depending on the components, and if it exceeds 50 wt%, there is a problem in skin safety or formulation.

Ginsenoside Y is a component having excellent antioxidant ability, and thus, the composition of the present invention containing ginsenoside Y provides excellent antioxidant ability to the skin.

The composition of the present invention can be used as a whitening composition, and can provide excellent whitening effects because it can inhibit tyrosinase activity and inhibit melanin production.

The composition of the present invention can be used as a moisturizing skin external composition, which can strengthen the skin barrier function and induce the differentiation of skin keratinocytes. Therefore, the composition can be effectively used as a composition for external skin preparations for preventing or improving xeroderma, allergic dermatitis, contact dermatitis, psoriasis, and the like, which are caused by incomplete epidermal differentiation.

The composition of the present invention can be used as a skin external composition for acne improvement, which provides an antibacterial effect, particularly an excellent antibacterial effect against acne-causing bacteria, and in addition, provides an anti-inflammatory effect.

The composition of the present invention can be used as a skin external composition for improving complexion and complexion, which can dilate capillaries and promote blood circulation when applied to the skin, thereby smoothly supplying nutrients to the skin and inhibiting skin aging, and thus, has a significant effect of improving complexion and complexion.

The composition of the present invention can be used as a skin external composition for shrinking pores, regulating sebum, and improving skin problems, which, when applied to the skin, suppresses excessive sebum secretion, and has an excellent effect of shrinking pores by promoting the removal of active oxygen and the synthesis of collagen, and suppressing skin problems due to the reduction in the expression of inflammatory factors. In addition, the skin irritation can be prevented due to the excellent antioxidant ability.

The combination of the present invention is a composition for external use for skin capable of being used as dandruff prevention, which can effectively discharge toxins accumulated on hair and scalp to cleanse scalp, can inhibit the proliferation and growth of dandruff bacteria to prevent scalp inflammatory reaction, and in addition, can provide the effects of stabilizing scalp and strengthening scalp, and strengthening original defense ability, since the antioxidant effect of inhibiting the generation and action of active oxygen is significant.

The composition of the present invention can be used as a hair-fostering skin external composition which promotes the growth of hair by promoting the transition from the resting-period hair cycle to the growing-period hair cycle and provides an effect of preventing hair loss.

The composition of the present invention can be used as a skin external composition for preventing white hair by significantly increasing the expression of melanocytes to MITF, thereby inhibiting white hair and inducing long-lasting black hair.

In addition, ginsenoside Y used in the composition for external preparation for skin of the present invention can provide the effect of natural preservatives.

The composition for external application to the skin of the present invention can be formulated into a dosage form as a cosmetic composition, and can be formulated into a dosage form including a cosmetically or dermatologically acceptable vehicle or base. It may take all forms suitable for topical use, for example, in the form of a solution, a gel, a solid, a stirred anhydrous product, an emulsion obtained by dispersing an oil phase in a liquid phase, a suspension, a microemulsion, microcapsules, microspheres or vesicular dispersions of ionic (liposomes) and nonionic type, or in the form of a cream, lotion, skin cream, foundation, ointment, spray or concealer, etc. In addition, it is also possible to use the form of an aerosol composition in the form of a foam (foam) or further containing a compressed propellant. These compositions can be manufactured by methods common in the art.

In particular, when the composition for external application to the skin of the present invention is used for preventing dandruff, hair growth, or white hair, the composition can be formulated into a composition for scalp and hair, and the formulation is not particularly limited, and for example, the composition can be formulated into a hair tonic, a scalp care solution, a hair conditioner, a shampoo, a conditioner, a hair cream, or a scalp and hair care solution.

In addition, the composition according to the present invention may contain a fatty substance, an organic solvent, a dissolving agent, a concentrating agent, a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, water, an ionic or nonionic softening agent, a filler, a metal ion-blocking agent, a chelating agent, a preservative, a vitamin, a blocking agent, a humectant, an essential oil, a dye, a pigment, a hydrophilic or lipophilic active agent, a lipid vesicle, or an auxiliary agent generally used in the field of cosmetics or dermatology, which is the same as any other ingredient generally used in cosmetics. The amount of the adjuvant to be added is an amount generally used in the field of cosmetics or skin science.

In addition, the composition of the present invention may contain a substance that promotes skin absorption in order to increase the skin improvement effect.

The structure and effects of the present invention will be described in more detail below with reference to experimental examples and dosage forms. However, these test examples and dosage forms are merely illustrative for facilitating understanding of the present invention, and the scope of the present invention are not limited by the following examples.

Reference example 1

Ginsenoside Y used to test the effect of the composition of the present invention was purchased and used from AMBO institute.

Test example 1

Inhibiting Reactive Oxygen Species (ROS) production

Keratinocytes (keratinocytes) were removed from human epidermal tissue in 24-well plates, each plate containing 5X 10 cells⁴Then, the adhesion was performed for 24 hours. After 16 hours, the ginsenoside Y was processed to 1%. At this time, ginsenoside Y was not treated in the control group for comparison. After two hours, the culture medium was removed, and then 100. mu.l of Phosphate Buffered Saline (PBS) was added to each well. The keratinocytes were irradiated with 30mJ/cm of radiation using an ultraviolet B (UVB) lamp (Model: K5T8, UV B15W, Sankyo Dennki Co., Ltd., Japan)²After UV, PBS was separated and 200. mu.l of the solution was added to each wellKeratinocyte culture solution. Here, ginsenoside Y was retreated, and the content of active oxygen increased by ultraviolet stimulation was measured at regular time intervals. The ROS content was measured by referring to the Tan method (Tan et al, 1998, j.cell biol.vol.141, pp1423-1432), that is, by measuring the ROS content by measuring the fluorescence spectrum of dichlorodiacetate (DCFDA) oxidized by ROS, the ratio of ROS to the control group treated with only vehicle was calculated, and the results thereof are shown in table 1 below.

TABLE 1

As is clear from the results of table 1 above, ginsenoside Y according to the present invention can effectively inhibit the generation of ROS, which is known to cause skin cell damage, and the amount of ROS after uv stimulation can inhibit the generation of ROS more than when uv is not irradiated, and thus it is understood that the antioxidant effect is very prominent. It was thus confirmed that ginsenoside Y according to the present invention can inhibit oxidation, prevent aging, thus preventing pore widening, and improve skin problems by preventing the generation of skin irritation.

Test example 2

Tyrosinase inhibitory effect

Tyrosinase was extracted from mushrooms (Mushroom) and produced by Sigma (Sigma) corporation. First, tyrosine as a substrate was dissolved in distilled water to prepare a solution of 0.3mg/ml, and 1.0ml of the above solution was added to each test tube, followed by addition of 1.0ml of potassium-phosphate buffer (0.1 molar concentration, pH 6.8) and 0.7ml of distilled water thereto.

Ginsenoside Y of the present invention was mixed in an ethanol solution at an appropriate concentration, and 0.2ml of each prepared sample solution was added, followed by reaction in a thermostatic bath at 37 ℃ for 10 minutes. In this case, the control group was prepared by adding only 0.2ml of solvent instead of each sample solution, and the positive control group used ascorbic acid. To the reaction solution, 0.1ml of a 2500 unit/ml tyrosinase solution was added, and the reaction was carried out again for 10 minutes in a 37 ℃ thermostat. The reaction solution was rapidly cooled in ice water to terminate the reaction, and the absorbance at a wavelength of 475nm was measured by a photoelectron spectroscopic analyzer, and the results are shown in Table 2 below. The respective tyrosinase inhibitory effects were calculated by the following mathematical formula 1.

Mathematical formula 1

TABLE 2

Test substance	Tyrosinase inhibition (%)
		Control group (not added)	0
Ascorbic acid	52
		Ginsenoside Y	59

In the results of table 2 above, it is understood that the tyrosinase inhibition rate of ginsenoside Y according to the present invention is much higher than that of ascorbic acid, which is a known tyrosinase inhibitor, and thus the whitening effect is very excellent.

Test example 3

Inhibitory Effect on melanogenesis by Using B16/F10 melanoma cells

Test materials samples each containing 0.1 wt% of ginsenoside Y and kojic acid were added to the culture solution of B16/F10 melanoma cells (korean cell line bank) at a certain concentration, and after 3 days of culture, the culture solution was removed, washed with PBS, the cells were lysed with 1N NaOH, and the absorbance was measured at 405 nm. The test substance to which no cell was added was used as a control group, and the melanin content in the control group was compared to determine the degree of inhibition of melanin production by each test substance. The melanin production inhibition rate was calculated according to the formula 2, and the results are shown in table 3 below.

Mathematical formula 2

TABLE 3

Test substance	Melanin production inhibition (%)
		Control group (not added)	0
Kojic acid	53
		Ginsenoside Y	65

In the results of table 3 above, it is understood that the ginsenoside Y according to the present invention has a higher melanin production inhibition rate than kojic acid, which is a known melanin production inhibitor, and thus has very excellent whitening effects.

Dosage form example 1 and comparative dosage form example 1

A nourishing cream (unit: wt%) was manufactured according to the composition of Table 4 below by a usual method.

TABLE 4

Test example 4

Determining the effect of enhancing skin moisturization

In order to determine the effect of ginsenoside Y on enhancing skin moisturizing ability, the above formulation example 1 and comparative formulation example 1 were used, and the following evaluation was made.

20 adults of 40-50 years old, male and female, classified as dry skin, were divided into two groups of 10 persons each, and the face was applied with the nourishing cream for each of the two groups of dosage form example 1 and comparative dosage form example 1 twice a day for 4 weeks. The skin moisture content was measured using a skin moisture content meter (Corneometer CM825, C + K Electronic co., germany) under constant temperature and humidity conditions (24 ℃, 40% relative humidity) before the start of application, after 1 week, 2 weeks, 4 weeks, and after 2 weeks after the termination of application (total 6 weeks). The results are shown in table 5 below, and the results in table 5 are expressed as a percentage of the increase in the measured value after a certain period of time based on the measured value of the skin moisture content measured immediately before the start of the test.

TABLE 5

As seen from the results of table 5 above, it was confirmed that the comparative formulation example 1 showed a moisture increase rate of about 30% at 4 weeks of application, but the skin moisture content sharply decreased after the application was terminated, whereas the formulation example 1 containing ginsenoside Y showed a skin moisture increase rate of 30% or more in most cases after the application was terminated. From this, it is found that the composition of the present invention containing ginsenoside Y has an excellent skin moisturizing effect.

Test example 5

Measurement of the Effect of promoting keratinocyte differentiation

In order to understand the effect of ginsenoside Y on the promotion of keratinocyte differentiation, the content of Cornified Envelope (CE) produced during keratinocyte differentiation was measured using absorbance.

First, human keratinocytes isolated from the epidermis of a newborn and subjected to primary culture were placed in a culture flask, the keratinocytes were allowed to adhere to the bottom of the flask, ginsenoside Y was treated in a culture medium to a concentration of 5ppm, and the cells were cultured for 5 days until the cells grew to a level of 70 to 80% of the flat bottom area. At this time, a low calcium (0.03mM) treated group and a high calcium (1.2mM) treated group were used as a negative control group and a positive control group, respectively. Thereafter, the cultured cells were extracted, washed with Phosphate Buffered Saline (PBS), 1ml of 10mM Tris-HCl buffer (Tris-HCl, pH 7.4) containing 2% Sodium Dodecyl Sulfate (SDS) and 20mM Dithiothreitol (DTT), and subjected to sonication, boiling (foaming), and centrifugation, and the precipitate was again precipitated in 1ml of PBS, thereby measuring the absorbance at 340 nm. Separately from the above-described operation, a part of the solution after the ultrasonic treatment was taken, and the protein content was measured and used as a standard for evaluating the degree of cell differentiation. The results are shown in Table 6 below.

TABLE 6

Test substance	Differentiation ability in keratinocytes (%)
		Low calcium (0.03mM) solution (negative control group)	100
High calcium (1.2mM) solution (positive control group)	210
		Ginsenoside Y	290

As shown in table 6 above, it can be confirmed that when ginsenoside Y is treated, the effect of promoting keratinocyte differentiation is excellent.

Test example 6

The effect of restoring the skin barrier was determined

In order to determine the effect of ginsenoside Y in restoring the barrier function of the skin damaged by skin injury, the following experiment was performed. The upper arms of 10 adult men and women were injured on the skin barrier by Tape Stripping (Tape striping), two groups of formulation example 2 and comparative formulation example 2, which were manufactured with the compositions of table 7, were applied, respectively, and the degree of recovery of skin moisture loss (TWEL) was measured and compared once a day over 7 days using a Vapometer (Delfin, finland). Among them, comparative formulation example 2 as a negative control group was excipient (vehicle). The results of the experiments are shown in Table 8 below. The results in table 8 are compared with the difference between the results before barrier damage and the results after barrier damage before treatment as a 100% standard.

TABLE 7

Compounding ingredients	Dosage form example 2	Comparative formulation example 2
			Purified water	69	70
Propylene glycol	30	30
			Ginsenoside Y	1	-

TABLE 8

As can be confirmed from table 8 above, when comparative formulation example 2 containing no ginsenoside Y was treated, the amount of skin moisture loss increased more and more with the passage of time, and on the contrary, when formulation example 2 containing ginsenoside Y was treated, the amount of skin moisture loss rapidly recovered to normal and barrier damage recovered.

Test example 7

Improving the effect of the gas color

In order to evaluate the effect of the cosmetic composition according to the present invention on promoting blood circulation of the skin, the degree of blood circulation in the skin was measured using a Laser Doppler Perfusion Imager (LDPI). LDPI is a well-known device for measuring blood circulation in the skin and, as a device commonly used today, is a very sensitive device that is capable of measuring not only the flow rate and volume of blood in the skin capillaries, but also the flow in arterioles and venules.

In a constant temperature and humidity chamber, after washing face with soap, the solution is adapted for 30 minutes, and an initial value is measured by LDPI. First, the initial blood flow volume of the lower forehead of 30 women with cold hands and feet was measured by LDPI. After that, the blood flow rate was measured after the subject used the two groups of formulation example 1 and comparative formulation example 1 for 1 week, and the result of comparison between the measured blood flow rate and the initial value (change in skin blood flow rate) is shown in table 9 below.

TABLE 9

LDPI results before and after cosmetic application-skin blood flow

Test substance	Change rate of skin blood flow after 1 week of use (%)
		Dosage form example 1	11
Comparative formulation example 1	5

As can be confirmed from the results of table 9 above, the cosmetic composition according to the present invention has significantly increased skin blood flow compared to comparative formulation example 1 containing no ginsenoside Y, and thus the improvement of complexion by promoting blood circulation. This finally indicates that the cosmetic composition containing ginsenoside Y of the present invention can effectively deliver nutrients in the skin and can inhibit and delay skin aging.

Test example 8

Improving skin color

In order to understand the skin color improving effect of the above formulation example 1 and comparative formulation example 1, 30 subjects were allowed to use the skin color improving composition (applied in the evening, 1 time/day, 1 week in total), and then the skin color improving degree was evaluated by Facial Stage DM-3(Moritex, japan). The skin color improvement rate was judged using the measured values of skin brightness and color and the values of skin brightness and color changes, and the results are shown in table 10 below. Greater values of intensity and color change indicate improved skin tone.

Watch 10

As shown in the results in table 10, comparative formulation example 1 containing no ginsenoside Y of the present invention did not exhibit a significant skin color improvement effect, whereas formulation example 1 containing ginsenoside Y as an active ingredient exhibited a much improved skin color after use compared to before use.

Test example 9

Effect of shrinking pores

1. Pore-shrinking effect by promoting collagen synthesis

The effect of promoting collagen synthesis was determined by comparing ginsenoside Y according to the present invention with TGF- β. First, seeding 10 in each of 24 wells (wells)⁵Individual fibroblasts (fibroplasts) were cultured to the extent that the cells grew to 90%. After 24 hours of culture in serum-free DMEM medium, ginsenoside Y and TGF-beta of the present invention dissolved in serum-free medium were treated to 10. mu.g/ml, respectively, and cultured in CO₂The culture was carried out in an incubator for 24 hours. The supernatant of these solutions was taken out, and the increase or decrease of procollagen (procollagen) was observed using a procollagen type I ELISA kit (procollagen type (I)). The results are shown in Table 11, where the untreated group was set to 100, and the performance of collagen synthesis was compared with that of the 100 untreated group.

TABLE 11

Test substance	Collagen Synthesis Performance (%)
		Untreated group	100
TGF-β	183.5
		Ginsenoside Y	189.9

As can be determined from the results of table 11 above, ginsenoside Y according to the present invention showed higher and superior collagen synthesis performance than TGF- β of the positive control group. Therefore, it can be confirmed that ginsenoside Y according to the present invention can shrink enlarged pores by increasing the amount of collagen production around the pores.

2. Effect of shrinking pores

In order to understand the effect of shrinking pores of the dosage form example 1 and the comparative dosage form example 1, the following evaluations were performed. The male and female testees with large pores are selected to be divided into two groups, 10 testees are selected in each group, and the nourishing cream of the preparation example 1 and the nutrition cream of the comparative preparation example 1 are applied to the face of each group every day for 4 weeks. Photographs were taken before and after 4 weeks of the experiment, respectively, and evaluated by the naked eye of an expert to judge the effect of shrinking pores. The results are shown in Table 12 below (evaluation scale: 0-no shrinkage at all; 5-much shrinkage).

TABLE 12

Test substance	Rating of evaluation
		Dosage form example 1	4
Comparative formulation example 1	0

As is clear from table 12 above, although comparative formulation example 1 has no effect of shrinking pores, formulation example 1 shows a pore-shrinking effect to a degree that can be determined with the naked eye, and thus ginsenoside Y according to the present invention has an excellent effect of reducing the size of pores.

Test example 10

Inhibiting effect on sebum secretion

1. Effect of inhibiting skin hypersecretion by inhibiting 5 alpha-reductase activity

To determine the effect of inhibiting 5. alpha. -reductase activity, the value of [ alpha ] -R2 in HEK 293-5. alpha. -R2 cells¹⁴C]Conversion of testosterone into [ alpha ], [ alpha ]¹⁴C]The ratio of dihydrotestosterone. P3 XFLAG-CMV-5. alpha. R2 was transfected into HEK293 cells and cultured in 24-well plates, 2.5X 10 per well⁵Individual cells (Park et al, 2003, jds.vol.31, pp.191-98). The next day, the medium was changed to a new medium supplemented with enzyme substrate and inhibitors. The substrate of the culture medium is 0.05. mu. Ci 2¹⁴C]Testosterone (Amersham Pharmacia biotech, UK).

To determine the extent of inhibition of 5 α -reductase activity, ginsenoside Y was added and 5% CO was added at 37 deg.C₂Was cultured in the incubator of (1) for 2 hours. At this time, a culture solution to which ginsenoside Y was not added was used as a negative control group, and a culture solution to which finasteride (finasteride) was added was used as a positive control group. Thereafter, the culture solution was extracted, the steroid was extracted with 800. mu.l of ethyl acetate, the upper organic solvent layer was separated and dried, and then the remaining residue was dissolved again with 50. mu.l of ethyl acetate and applied with acetic acid on Silica gel 60F254(Silica plastic sheet kit gel 60F254)Ethyl ester-nucleic acid (1:1) was developed as a solvent.

The plastic sample was dried in the air, and in order to measure the isotope content, a bas system was used in which the dried plastic sample and X-ray film were put together in a bas box, the isotope content of testosterone and dihydrotestosterone remaining on the film was measured after one week, and then the conversion rate and the inhibition rate were calculated according to the following mathematical formulas 3 and 4, respectively, and the results thereof are shown in table 13 below.

Mathematical formula 3

Mathematical formula 4

Watch 13

Test substance	Conversion (%)	Inhibition ratio (%)
			Negative control group	48.0	-
Positive control group	27.6	42.5
			Ginsenoside Y	13.9	62.8

As can be seen from the results shown in table 13, the 5 α -reductase converted testosterone into dihydrotestosterone, bound to cytoplasmic receptor proteins, entered into the nucleus to activate sebaceous gland cells and promote differentiation, and thus the 5 α -reductase had the effect of excessively secreting sebum in the sebaceous glands, while ginsenoside Y effectively inhibited the activity of 5 α -reductase, thereby blocking the conversion of testosterone into dihydrotestosterone, and showed more excellent inhibitory effects than the known finasteride inhibiting the activity of 5 α -reductase. Therefore, ginsenoside Y can inhibit sebum hypersecretion by effectively inhibiting the activity of 5 α -reductase.

2. Inhibiting effect on sebum secretion

In order to understand the effect of suppressing sebum secretion in the above formulation example 1 and comparative formulation example 1, the following evaluation was performed. The test subjects who considered to be hyperseborrhea were selected as male and female 30 persons, and the nutrition creams of the formulation example 1 and the comparative formulation example 1 were applied to the designated sites every day for 4 weeks. The effect of sebum reduction was judged by measuring the average sebum reduction (%) after 2 weeks and 4 weeks, respectively, using a sebum amount meter (Sebumeter SM810, C + K Electronic co., germany), and the results are shown in table 14 below.

TABLE 14

As is apparent from the results of table 14 above, dosage form example 1 containing ginsenoside Y according to the present invention as an active ingredient can more effectively suppress excessive sebum secretion than comparative dosage form example 1 containing no ginsenoside Y.

Dosage form example 3 and comparative dosage forms examples 3 to 4

Dosage form example 3, comparative dosage form example 3, and comparative dosage form example 4 were prepared according to the ingredients and contents (wt%) shown in table 15 below. Specifically, ginsenoside Y was added to formulation example 3, comparative formulation example 3 did not contain any effective ingredient for improving acne skin at all, and comparative formulation example 4 contained erythromycin (erythromycin), which is a standard substance for judging antibacterial performance and is commonly used as an acne therapeutic agent.

The manufacturing methods of dosage form example 3, comparative dosage form example 3, and comparative dosage form example 4 are as follows. The ingredients in a of table 15 below were completely dissolved, and after the ingredients in B were completely dissolved in an additional dissolution tank, B was added to a to perform mixing and dissolution. At this time, the present composition was prepared by adding the components in C in the formulation ratios shown in table 15, mixing them uniformly, and then filtering.

Watch 15

Test example 11

Antibacterial ability test against acne bacteria

Using each of the cosmetic compositions prepared with the compositions of formulation example 3, comparative formulation example 3 and comparative formulation example 4, an antibacterial ability test was performed on Propionibacterium acnes (ATCC 6919: Medium-BHI liquid Medium (broth)) as a pathogenic bacterium for acne.

The test method for the antibacterial ability against acne bacteria is as follows.

(1) Preparation of test bacterial solution

Propionibacterium acnes was cultured in a medium inoculated into BH1 liquid medium and anaerobically cultured.

(2) Preparing a dilute solution

To 15ml of BH1 broth (pH 6.8) or LB broth (pH 4.5), 0.15ml of the above-mentioned test bacterial solution was added, and the resulting mixture was used as a diluted solution.

(3) Preparing the sample

The raw solutions of the cosmetic compositions manufactured by formulation example 3, comparative formulation example 3, and comparative formulation example 4 were used as samples.

(4) Test for antibacterial ability

1) Samples were added in line with the initial concentration in the first row of a 96well cell culture tube (96well plate) and dilution solution was added until the total amount was 200 μ l.

2) After the mixed solution in the first line was mixed uniformly, 100. mu.l of the mixed solution was added to the second line and mixed uniformly, and then 100. mu.l of the mixed solution was added to the third line, and double dilution (double dilution) was performed in the above-described manner.

3) After the culture was allowed to stand at 32 ℃ for 24 hours and 48 hours, the degree of growth of the bacteria was judged to be suspended, and the Minimum Concentration at which the bacteria were not grown was determined as the MIC (Minimum Inhibitory Concentration) value. If the mixture is opaque and it is difficult to determine whether or not the bacteria have proliferated, the proliferation is confirmed by observing the mixture under a microscope.

The results of the antimicrobial efficacy test against acne bacteria are shown in Table 16 below. The MIC is converted into the concentration of the active ingredient contained in the dosage form and is labeled.

TABLE 16

Item	pH	Propionibacterium acnes
			Dosage form example 3	5.7	>50ppm
Comparative formulation example 3	5.7	Maximum concentration (no antibacterial ability)
			Comparative formulation example 4	5.7	>100ppm

The lower the MIC, the more effective the antibacterial activity against acne bacteria, and the ppm concentration in formulation example 3 was significantly lower than that in comparative formulation example 4 using erythromycin, which is known as an acne therapeutic agent, and therefore, it could be confirmed that the composition containing ginsenoside Y had more excellent antibacterial activity against the test bacteria.

Test example 12

Test for inhibition of lipid Synthesis (Lipogenesis)

3T3-L1 cells as a fibroblast line (fibroblast cell line) of live mice were cultured at 1X 10⁵The amount of (2) was attached to a 6-well plate (culture plate) to which a DMEM (Dulbecco's modified eagle's medium, GIBCO BRL, Life technologies Co.) medium containing Fetal Bovine Serum (FBS) was added. After 2 days, the medium was replaced with fresh DMEM (containing 10% FBS) and cultured for another 2 days. Then, the above cultured cells were induced again to differentiate by DMEM (containing 10% FBS) containing 1. mu.g/mL of insulin (insulin), 0.5mM of IBMX, and 0.25. mu.M of dexamethasone (dexamethasone), and 50. mu.M of ginsenoside Y and caffeine were treated, and after 2 days, they were again changed to DMEM containing insulin, and cultured for 5 days. After 5 days, the medium was changed to normal medium (DMEM, containing 10% FBS) again, and the culture was observed until the cells became adipocytes in shape.

In order to evaluate the effect of ginsenoside Y on inhibiting fat accumulation in adipocytes, Sudan III staining was performed using the differentiated 3T3-L1 adipocytes described above (S4136, sigma-aldrich). At normal temperature, adipocytes were fixed with paraformaldehyde (pH 7.2) in phosphate buffer, followed by washing with PBS (phosphate buffered saline), followed by staining with Sudan III, followed by photography, and comparison by the naked eye. The control group used only the medium to which the test substance or the comparative substance was not added, and the other control group was treated with 50. mu.M of caffeine. The degree of inhibition of fat accumulation was given a grade by dividing the degree of staining into +++, ++, -. Wherein the more toward +++ the greater the degree of staining. The results are shown in Table 17 below.

TABLE 17

Sample (I)	Inhibition ratio (%)
		Control group	+++
Comparison group	+
		Ginsenoside Y	-

As shown in table 17 above, it is understood that ginsenoside Y used in the present invention not only reduces the amount of fat accumulated in adipocytes, but also has a superior lipid synthesis inhibitory effect over caffeine, which is a known lipid synthesis inhibitory substance. Thus, by inhibiting lipid synthesis, sebum is reduced, thereby enabling the inhibition of acne production.

Test example 13

Test for improving acne, reducing sebum secretion and having no irritation

30 persons with acne were divided into 3 groups of 10 persons, and the corresponding subjects in each group were allowed to use the above-mentioned cosmetic compositions manufactured by dosage form example 3, comparative dosage form example 3 and comparative dosage form example 4, respectively, for 1 month. The scale for acne improvement was set at 1 to 5 points, with 1 point indicating "none", 3 points indicating "normal", and 5 points indicating "very good". The results of the experiments are shown in Table 18 below, where the labeling was done on an average of 10 people.

The disappearance period of acne was based on the date of disappearance judgment, and the recurrence of acne was based on the result after 1 month. The sebum secretion reduction was set at 1 to 5 points, 1 point for "none", 3 points for "normal", and 5 points for "excellent". The results are reported in Table 22, below, where the 10-person average score is used. The presence or absence of skin irritation was observed by (number of persons with irritation reaction)/(total number of subjects).

Watch 18

As shown in table 18 above, in the formulation example 3, compared with the comparative formulation example 3, acne did not recur, and the effect of improving acne was excellent as a whole. In addition, the acne improvement effect was shown in comparative formulation example 4 containing an antibacterial ability standard substance, but the irritation was large at the time of use and thus it was not suitable for long-term use, whereas the composition according to the present invention was shown to be non-irritating and suitable for long-term use.

Test example 14

Effect of inhibiting IL-8 production

One day before the experiment, skin keratinized epithelial cells (NHEK, Normal human skin keratinocyte; Purchase-Lonza) were fractionated into 5X 10 cells on a 96-well plate⁴Cell number/well, then 5% CO at 37 ℃₂Was cultured in an incubator for 24 hours. After 24 hours, the cells were washed 2 times with PBS and replaced with serum free KBM (serum free Keratinocyte base media). Ginsenoside Y was treated in each well at the concentration of table 19 below and reacted for 30 minutes, followed by treatment with PGSA (10 μ g/ml), PGSA (50 μ g/ml) or PGSA (50 μ g/ml) + LPS (1 μ g/ml), respectively. Among them, PGSA (peptidoglycan from s. aureus) is a peptidoglycan (peptidoglycan) extracted from staphylococcus, and it is known that PGSA is a main structure of cell wall of gram-positive (+) bacteria, and cell membrane components of bacteria cause inflammation, and particularly 90% of allergic patients cause secondary infection by staphylococcus, which is disclosed. It is also known that Lipopolysaccharide (LPS) is leatherThe major constituent of cell membranes of the negative (-) Langerhans is responsible for the initiation of inflammation.

At 37 deg.C, 5% CO₂After 24 hours of culture in the incubator, the culture broth was subjected to ELISA for Interleukin-8 (Interleukin-8, IL-8), and the results are shown in Table 19 below. ELISA was carried out by the manufacturer (BD science).

Watch 19

Components	1L-8 secretion (pg/ml)
		Untreated Control group (Control)	935.12
PGSA(10μg/ml)	4812.60
		PGSA(50μg/ml)	5895.08
PGSA(50μg/ml)+LPS(1μg/ml)	6814.91
		Ginsenoside Y (5ppm)	1436.43
Ginsenoside Y (25ppm)	1204.19
		Ginsenoside Y (50ppm)	1012.73

As can be confirmed from table 19 above, ginsenoside Y can significantly reduce and inhibit IL-8 secretion increased by PGSA and LPS. Therefore, the composition for external preparation for skin of the present invention can significantly reduce the secretion of IL-8 increased by PGSA and LPS, and thus can provide an excellent anti-inflammatory effect.

Test example 15

Evaluation of relief from itching

One day before the experiment, keratinocytes (cell line name: HaCaT, Purchase: ATCC) were aliquoted to 4X 10 cells on 96-well plates⁴Cell number/well, then 5% CO at 37 ℃₂Was cultured in an incubator for 24 hours. After 24 hours, the 96-well plate was washed twice with Hank's Balanced Salt Solution (HBSS), after which reaction buffer (2. mu.M Fluo-4-AM, 20% polyetheracid (pluranic acid), 2.5mM 4- (dipropylsulfamoyl) benzoic acid (probenecid)) was added to the cells. At 37 deg.C, 5% CO₂The cells were reacted in the incubator of (1) for 30 minutes, and after reaction for 30 minutes at normal temperature, washed twice with HBSS buffer, and treated with ginsenoside Y at the concentration (%) in Table 20 below.

After 10 minutes of reaction, 2U/ml Trypsin (Trypsin) or 5. mu.M PAR-2 active peptide (SLIGKV) was treated and intracellular Ca was measured within 80 seconds²⁺The change in concentration. Intracellular Ca²⁺The change in concentration was measured using Flexstation3(Flexstation 3: multifunctional microplate reader, USA). After treating 2U/ml trypsin or 5. mu.M PAR-2 active peptide, the difference between the maximum and minimum values obtained by measuring the contraction (flex) within 80 seconds was calculated and compared with the difference between the minimum and maximum values obtained by treating 2U/ml trypsin or 5. mu.M PAR-2 active peptide, and the inhibition (%) of calcium ion entry into the cells is shown in Table 20 below.

Watch 20

From the above table 20, it can be confirmed that the amount of calcium ions of trypsin or PAR-2 active peptide entering into the cells is decreased according to the treatment of ginsenoside Y, and the calcium ions entering into the cells are significantly decreased with increasing concentration of ginsenoside Y.

Therefore, the composition for external use on skin comprising ginsenoside Y of the present invention can effectively inhibit PAR-2 activity inducing itch, thereby providing excellent anti-itch effects.

Dosage form example 4 and comparative dosage form example 5

The following composition of table 21 produced a shampoo. Specifically, a surfactant and ethylene glycol distearate were added to purified water, and the mixture was heated to 80 ℃ to be uniformly dissolved, and then slowly cooled to 40 ℃ by stirring, and the active ingredient according to the present invention, a preservative, a viscosity modifier, a perfume, and a hair conditioner were added to the above mixture and mixed, and then cooled to room temperature by stirring, thereby completing the production.

TABLE 21

Test example 16

Test for dandruff-reducing Effect

24 men of 19 to 35 years old with more dandruff were selected and divided into two groups of 12 men, and the shampoos of formulation example 4 and comparative formulation example 5 were used for one month by the following method, and then the dandruff reduction rate was measured.

Before the start of the test, hair was washed with a general shampoo, two days of accumulated dandruff after the hair washing was collected, and the weight of the collected dandruff was compared with the weight of the dandruff accumulated for 2 days after the end of the test after the hair washing with the shampoos of dosage form example 4 and comparative dosage form example 5 every two days. At this time, the accumulated dandruff was directly collected from the scalp by using a vacuum suction apparatus, and the dandruff reduction rate was determined according to the following equation 5, and the results are shown in table 22 below.

Mathematical formula 5

TABLE 22

As can be seen from table 22 above, dosage form example 4 containing ginsenoside Y showed an excellent effect of preventing dandruff.

Test example 17

Efficacy test for preventing scalp pruritus

Male and female 24 persons 25 to 45 years old with severe scalp itching were selected and divided into two groups of 12 persons, and they were allowed to use the shampoos of formulation example 4 and comparative formulation example 5 every 3 days for 2 weeks, after which the effect of preventing scalp itching was evaluated by the following evaluation criteria, and the results are shown in table 23 below.

[ evaluation standards ]

TABLE 23

Classification	Dosage form example 4	Comparative formulation example 5
			Scalp pruritus eliminating effect	4	2.3

As is clear from table 23 above, dosage form example 4 containing ginsenoside Y showed a superior effect of preventing scalp itching.

Test example 18

Hair follicle papilla cell proliferation effect

Keratins constituting hair are produced from hair root keratinocytes (keratinocytes) differentiated from hair papilla cells. To evaluate the dermal papilla Cell activity of the present composition, DP6(rat immobilized dermal papilla Cell) Cell line (Wendy Filsell, Journal of Cell Science 107,1761-1772(1994)) was used in the present invention. The hair papilla cell line is a cell line which is isolated from the hair root of male PVG rat beard and cultured by microdissection (microdissection), and is added into DMEM (Dulbecco's modified Eagle's medium, Gibco BRL, Gaithersburg, Md., USA) containing FBS (Fetal bovine serum, Fetal bone serum) and maintained at 5% CO₂And cultured in an incubator at 37 ℃ for 24 hours. DP6 was added to a 96-well plate and cultured in an incubator at 37 ℃ for 24 hours, after which ginsenoside Y was treated to concentrations of 5ppm, 10ppm and 20ppm, respectively. After 24 hours of drug treatment, the cell proliferation potency was determined using the WST-1 kit (Roche). The results are given in Table 24 below.

Watch 24

Classification	Cell proliferation potency (%)
		Untreated Control group (Control)	100
Ginsenoside Y (5ppm)	123
		Ginsenoside Y (10ppm)	131
Ginsenoside Y (20ppm)	146

From the above table 24, it can be confirmed that when ginsenoside Y is treated, proliferation of dermal papilla cells is increased, and there is a difference and a significant increase according to the concentration.

Test example 19

Evaluation of Effect of increasing Potassium channel Activity

Minoxidil as a therapeutic agent for alopecia is a well-known potential mitochondrial potassium ion channel opener (K)_ATPchannel openers) are representative drugs for the treatment of androgenic alopecia. To evaluate the mechanism of this minoxidil, the following test methods were used: treatment of blockages K from fibroblasts of the dermis constituting the scalp_ATPTolbutamide (SIGMA ALDRICH, T0891) in the channel, thereby inhibiting cell proliferation, and then the potassium channel is reopened to restore cell proliferation.

In order to evaluate the function of the potassium channel opener of the present composition, NIH3T3(Mouse embryo cell line) cell strain as a fibroblast was used in the present invention. The cell line was a cell line obtained by naturally immortalizing a fibroblast 3T3 protocol (protocol) isolated from NIH Swiss mouse embryo (Swiss mouse embryo). The cell line was added to DMEM (Dulbecco's modified Eagle's medium, Gibco BRL, Gaithersburg, Md., USA) containing FBS, and maintained at 5% CO₂And cultured in an incubator at 37 ℃ for 24 hours. NIH3T3 was added to a 96-well plate and incubated in an incubator at 37 ℃ for 24 hours, after which tolbutamide was treated, and after 10 minutes minoxidil as a positive control group was treated to 10 μ M and ginsenoside Y was treated to concentrations of 2.5ppm, 5ppm and 10 ppm. After 48 hours of drug treatment, the cell proliferation potency was determined using the WST-1 kit (Roche). The results are given in Table 25 below.

TABLE 25

Classification	Cell proliferation potency (%)
		Untreated Control group (Control)	100
Minoxidil	132
		Ginsenoside Y (2.5ppm)	126
Ginsenoside Y (5ppm)	128
		Ginsenoside Y (10ppm)	133

As can be seen from table 25 above, when ginsenoside Y was treated, proliferation of fibroblasts was recovered and cell proliferation ability increased with the concentration of the treated ginsenoside Y, and it could be confirmed that when ginsenoside Y was treated to 10ppm, proliferation of fibroblasts was recovered to the level of minoxidil.

Test example 20

Effect test of ginsenoside Y on promoting melanogenesis

Melanocytes (melan-a) were divided into 50,000 cells/well in 24-well micro plates (24-well micro plate) in a medium prepared by adding 5% fetal bovine serum, 100IU penicillin G and 0.2. mu.M TPA to RPMI medium. The next day, ginsenoside Y was treated in the cells of the divided plant as a test substance at a concentration of 10ppm or 50ppm, 0.1% DMSO was treated as a negative control group, and 100 μ M IBMX was treated as a positive control group, followed by culture at 37 ℃ for 3 days. After incubation, the wells were washed with PBS and 100. mu.l of 1N NaOH was added to dissolve melanin in the cells. The absorbance of the solubilized melanin was measured in 405nm using a microplate reader. The results of comparing the effect of ginsenoside Y on the promotion of melanin production with that of the control group are shown in table 26 below.

Watch 26

Sample (I)	Amount of melanin synthesized (%)
		DMSO(0.1％)	100
IBMX(100μM)	120
		Ginsenoside Y (10ppm)	108
Ginsenoside Y (50ppm)	118

As shown in table 26 above, ginsenoside Y promotes melanin synthesis in melanocytes, thereby increasing melanin production, and thus exhibits an excellent melanin production promoting effect.

Test example 21

Effect of ginsenoside Y on promoting expression of MITF and tyrosinase in melanocytes

Using 501mel cell line, the protein was obtained after dividing into 500,000 cells/well in 6 microwell plates (6-well microtiterplate), treating 0.1% DMSO as a negative control, treating 100. mu.M IBMX as a positive control, treating ginsenoside Y as a test group to 10ppm, and culturing at 37 ℃ for 24 hours, 48 hours, and 72 hours. The protein thus obtained was subjected to western blotting using antibodies against MITF and tyrosinase. The value of the negative control group was set to 100, and the results after western blotting were compared with 100 and shown in the following table 27.

Watch 27

It can be confirmed from table 27 above that ginsenoside Y can increase the expression of MITF and tyrosinase proteins in melanocytes.

Test example 22

Evaluation of antibacterial ability of ginsenoside Y

In order to evaluate the antibacterial ability of ginsenoside Y, an antibacterial experiment was performed. The specific experimental method is as follows.

The strains of Staphylococcus aureus (Staphylococcus aureus), Escherichia coli (Escherichia coli), Pseudomonas aeruginosa (Pseudomonas aeruginosa) used for the experiments were cultured in Tryptic Soy Broth, while the strains of Candida albicans (Candida albicans), Aspergillus niger (Aspergillus niger) were cultured in Sabouraud's Dextrose Broth. The culture broth was diluted to 1/100 (Candida albicans was 1/10) in each medium, and the diluted solution was used as a test bacterial solution. Aspergillus niger is prepared by preparing into 2 × 10⁸A spore suspension of cfu/ml was used as the test bacterial liquid.

To 15ml of each medium, 0.15ml of test bacterial suspension was added, and the uniformly mixed solution was used as a dilution solution.

In the first row of a 96well plate (96well plate) 16. mu.l of sample and 184. mu.l of dilution solution were added. To the remaining wells 100. mu.l of the diluted solution was added. After the mixed solution in the first row was mixed well, 100. mu.l of the mixed solution was added to the second row and mixed well, and then 100. mu.l of the mixed solution was added to the third row, and diluted twice by the above method.

Staphylococcus aureus (Staphylococcus aureus), Escherichia coli (Escherichia coli), Pseudomonas aeruginosa (Pseudomonas aeruginosa) were cultured in a thermostat at 32 ℃ while Candida albicans and Aspergillus niger (Aspergillus niger) were cultured in a thermostat at 25 ℃.

After 48 hours, the proliferation of the bacteria was confirmed by the suspension degree and microscope, and the Minimum Inhibitory Concentration (MIC) was determined, and the results are shown in Table 28 below.

Watch 28

As shown in table 28 above, it was confirmed that ginsenoside Y showed antibacterial ability against various strains, from which it can be predicted that ginsenoside Y can be used as a natural preservative or antibacterial agent in a composition.

Claims (3)

1. Use of a skin external composition containing ginsenoside Y as a main active ingredient in the preparation of a cosmetic composition for improving acne.

2. Use according to claim 1, for providing an antibacterial effect against acne pathogenic bacteria.

3. The use according to claim 1, for inhibiting the production of lipids.