阅读说明：本技术 包含幼苗人参发酵物的化妆料组合物及其制备方法 (Cosmetic composition comprising young ginseng fermented product and preparation method thereof ) 是由 李银海 李慧子 廉贤淑 金地慧 朴尽五 李知愿 于 2020-09-22 设计创作，主要内容包括：本发明涉及一种制备包含幼苗人参发酵物的化妆料组合物的方法及包含该幼苗人参发酵物的化妆料组合物,所述幼苗人参发酵物是利用作为乳酸菌的鼠李糖乳杆菌(Lactobacillus rhamnosus)使幼苗人参提取物发酵而获得的,所述化妆料组合物具有抗氧化活性效果、皮肤保湿效果或改善皱纹的效果。(The present invention relates to a method for preparing a cosmetic composition comprising a young seedling ginseng ferment obtained by fermenting a young seedling ginseng extract using Lactobacillus rhamnosus (Lactobacillus rhamnosus) as a lactic acid bacterium, the cosmetic composition having an antioxidant activity effect, a skin moisturizing effect, or an effect of improving wrinkles, and a cosmetic composition comprising the young seedling ginseng ferment.)

1. A cosmetic composition comprises fermented young ginseng as an effective ingredient.

2. The cosmetic composition according to claim 1, wherein the young ginseng extract is obtained by fermenting a young ginseng extract using lactic acid bacteria.

3. The cosmetic composition according to claim 2, wherein the lactic acid bacteria are added in an amount of 0.1 to 1.5 parts by weight relative to 100 parts by weight of the extract of young plants of ginseng.

4. The cosmetic composition according to claim 2, wherein the lactic acid bacteria comprise a strain selected from the group consisting of a strain of the genus Lactobacillus (Lactobacillus), a strain of the genus Bifidobacterium (Bifidobacterium), a strain of the genus Streptococcus (Streptococcus), and combinations thereof.

5. The cosmetic composition according to claim 4, wherein the Lactobacillus strain comprises a strain selected from the group consisting of Lactobacillus rhamnosus (L.rhamnosus), Lactobacillus plantarum (L.plantarum), Lactobacillus acidophilus (L.acidophilus), Lactobacillus bulgaricus (L.bulgaricus), Lactobacillus fuscus (L.affusis), Lactobacillus reuteri (L.reuteri), Lactobacillus casei (L.casei) and combinations thereof,

the Bifidobacterium strain comprises a strain selected from Bifidobacterium breve (B.breve), Bifidobacterium longum (B.longum), Bifidobacterium infantis (B.infarnatis), Bifidobacterium bifidum (B.bifidum), Bifidobacterium thermophilum (B.thermophilum), and combinations thereof,

the streptococcus strain comprises a strain selected from the group consisting of streptococcus faecalis (str. faecalis), streptococcus thermophilus (str. thermophilus), and combinations thereof.

6. The cosmetic composition of claim 1, wherein the young ginseng ferment is contained in an amount of 0.001 to 60% by weight in 100% by weight of the cosmetic composition.

7. The cosmetic composition according to any one of claims 1 to 6, wherein the cosmetic composition has one or more effects selected from the group consisting of anti-oxidation, skin moisturizing, and wrinkle improvement.

8. A method of preparing a cosmetic composition comprising the steps of:

extracting the seedling ginseng to obtain a seedling ginseng extract;

fermenting the seedling ginseng extract with lactic acid bacteria to obtain a seedling ginseng ferment.

9. The method of preparing a cosmetic composition according to claim 8, wherein the extraction of the young ginseng comprises the steps of:

adding purified water 8-22 times the volume of the prepared seedling ginseng; and

extracting young ginseng added with the purified water at 50-90 deg.C for 1-3 hr.

10. The method of preparing a cosmetic composition according to claim 8, wherein the lactic acid bacteria comprise a strain selected from the group consisting of a strain of Lactobacillus (Lactobacillus), a strain of Bifidobacterium (Bifidobacterium), a strain of Streptococcus (Streptococcus), and combinations thereof.

11. The method of preparing a cosmetic composition according to claim 10, wherein the Lactobacillus strain comprises a strain selected from the group consisting of Lactobacillus rhamnosus (L.rhamnosus), Lactobacillus plantarum (L.plantarum), Lactobacillus acidophilus (L.acidophilus), Lactobacillus bulgaricus (L.bulgaricus), Lactobacillus fuscus (L.convulsus), Lactobacillus reuteri (L.reuteri), Lactobacillus casei (L.casei), and combinations thereof,

the Bifidobacterium strain comprises a strain selected from Bifidobacterium breve (B.breve), Bifidobacterium longum (B.longum), Bifidobacterium infantis (B.infarnatis), Bifidobacterium bifidum (B.bifidum), Bifidobacterium thermophilum (B.thermophilum), and combinations thereof,

the streptococcus strain comprises a strain selected from the group consisting of streptococcus faecalis (str. faecalis), streptococcus thermophilus (str. thermophilus), and combinations thereof.

12. The method of preparing a cosmetic composition according to claim 8, wherein the fermentation is performed by adding 0.1-1.5 parts by weight of lactic acid bacteria to 100 parts by weight of the young plant ginseng extract, and fermenting at 35-39 ℃ for 3-5 days.

Technical Field

The present invention relates to a cosmetic composition comprising a young seedling ginseng ferment obtained by fermenting a young seedling ginseng extract using lactic acid bacteria, which has an antioxidant activity effect, a skin moisturizing effect, or an effect of improving wrinkles, and a preparation method thereof.

Background

In recent years, the stress on the skin has been increasing with changes in the environment and the life pattern. As a result, various phenomena occur in the skin, such as reduction of moisture in the stratum corneum, increase in roughness, and the like. Therefore, there is an increasing demand for materials having skin moisturizing, skin wrinkle improving, and skin calming effects.

Since the use of conventionally developed chemical substances is limited due to various problems such as toxicity, low activity, limitation of use, and side effects caused by excessive administration, research for extracting active ingredients from natural plants has been actively conducted as part of the development of materials with less possibility of causing side effects. In particular, in recent years, with the spread of the trend of healthy life (well-wearing), the demand for maintaining health and beauty for a long period of time has been increasing, and thus attention has been increasingly paid to natural plant materials and plant materials excellent in physiological activity, which have been passed down by conventional folk remedies, in all fields of foods, cosmetics, residential environments, and the like.

Since natural plant materials are safer and more environmentally friendly than synthetic materials, the use of natural plant materials has been started in the cosmetic industry since a long time ago, and in recent years, cosmetics using plant materials and Chinese herbal medicines having high physiological activities have been dominant. Since plant materials and Chinese herbal medicines are materials that have been traditionally used for hundreds of years and have been verified in terms of safety or effect, they are easily approved and used by consumers, and have an advantage that their effect is not inferior to that of synthetic materials. Further, by subjecting the extract obtained from such plant materials and Chinese herbal medicines to self-fermentation or fermentation by inoculating lactic acid bacteria or yeast, the nutrient components of the plant can be converted into a form which can be easily absorbed, and the components produced by the action of enzymes can express a novel physiological regulation function.

In addition, fermentation (fermentation) refers to a process of decomposing organic substances by enzymes secreted from microorganisms. In addition, the production of useful substances by microorganisms in the course of energy production is also referred to as fermentation. Korean is familiar with this fermentation, and traditional fermented foods such as pickles, fermented soybean paste, and Qingju paste are representative. In this fermentation process, organic matter is completely decomposed by anaerobic respiring microorganisms and other kinds of organic matter are produced, thus generating a small amount of energy. The kind of such fermentation includes lactic acid fermentation, alcohol fermentation, propionic acid fermentation, methane fermentation, etc., depending on the microorganism.

Typical lactic acid bacteria of microorganisms involved in such fermentation decompose sugars such as glucose and lactose to produce organic acids such as lactic acid and acetic acid, and by-products such as amylase, cellulase, lipase, and protease are produced, and the enzymes are converted into low molecules of giant molecules in natural products, so that the enzymes easily permeate the skin.

However, in the case of natural products such as such plant materials and fermentation products of such plant materials, when used at effective concentrations or more in cosmetics or pharmaceuticals, there are many problems in terms of safety, stability, possibility of discoloration, and the like, and satisfactory effects cannot be achieved.

[ Prior art documents ]

[ patent document ]

(patent document 0001) Korean laid-open patent publication No. 10-2019-0037014

Disclosure of Invention

Technical problem to be solved

Accordingly, the present inventors have made an effort to develop a substance having more excellent effects of improving wrinkles, moisturizing skin, improving antioxidant activity, etc. than those of the conventionally known substances, and have confirmed that the antioxidant effect, collagen synthesis promoting effect, and moisturizing effect are improved when ginseng fermentation products of young plants are mixed, thereby completing the present invention.

The present invention is directed to provide a cosmetic composition comprising a young ginseng fermented product obtained by fermenting a young ginseng extract using lactic acid bacteria, which has an antioxidant activity effect, a skin moisturizing effect, or an effect of improving wrinkles, and a preparation method thereof.

The object of the present invention is not limited to the above-described object. The objects of the present invention will become more apparent from the following description and can be achieved by the features and combinations thereof recited in the claims.

Technical scheme

The cosmetic composition according to one embodiment of the present invention comprises a young ginseng fermented product as an effective ingredient.

The seedling ginseng fermented product can be obtained by fermenting a seedling ginseng extract with lactic acid bacteria.

The lactic acid bacteria may be added in an amount of 0.1-1.5 parts by weight relative to 100 parts by weight of the extract of young ginseng.

The young ginseng fermented product may be contained in an amount of 0.001-60 wt% in 100 wt% of the cosmetic composition.

The cosmetic composition may have one or more effects selected from the group consisting of anti-oxidation, skin moisturizing, and wrinkle improvement.

The cosmetic composition may be formulated into any one or more dosage forms selected from the group consisting of a solution, a topical ointment, a cream, a foam, a nourishing lotion, a softening lotion, a pack, a softening lotion, an emulsion, a barrier cream, an essence, a soap, a liquid detergent, a bath agent, a sunscreen cream, a sunscreen oil, a suspension, an emulsion, a paste, a gel, a lotion, a powder, a surfactant-containing cleanser, an oil, a powder foundation, a milky foundation, a wax foundation, a patch, and a spray.

Further, the method of preparing a cosmetic composition according to an embodiment of the present invention may include the steps of: extracting the seedling ginseng to obtain a seedling ginseng extract; and fermenting the seedling ginseng extract with lactic acid bacteria to obtain a seedling ginseng ferment.

The seedling ginseng extract may comprise an extract selected from the group consisting of a hot water extract, a solvent extract, and combinations thereof.

The hot water extraction may comprise the steps of: adding purified water 8-22 times the volume of the prepared seedling ginseng; and extracting the young ginseng added with the purified water at 50-90 ℃ for 1-3 hours.

The solvent may comprise a solvent selected from the group consisting of fermentation alcohols, ethanol, methanol, butanol, propanol, ethyl acetate, isoprene glycol, glycerol, ethylene glycol, propylene glycol, butylene glycol, chloroform, dichloromethane, hexane, acetone, acetonitrile, petroleum ether, diethyl ether, and combinations thereof.

The seedling ginseng extract can be filtered using 0.20-0.50 μm filter paper.

The lactic acid bacteria may comprise a strain selected from the group consisting of a strain of the genus Lactobacillus (Lactobacillus), a strain of the genus Bifidobacterium (Bifidobacterium), a strain of the genus Streptococcus (Streptococcus), and combinations thereof.

The lactobacillus strain may include a strain selected from lactobacillus rhamnosus (l.rhamnous), lactobacillus plantarum (l.plantarum), lactobacillus acidophilus (l.acidophilophilus), lactobacillus bulgaricus (l.bulgaricus), lactobacillus fuscus (l.convulsus), lactobacillus reuteri (l.reuteri), lactobacillus casei (l.casei), and combinations thereof, the bifidobacterium strain may include a strain selected from bifidobacterium breve (b.breve), bifidobacterium longum (b.longum), bifidobacterium infantis (b.infarnentis), bifidobacterium bifidum (b.bifidum), bifidobacterium thermophilum (b.thermophilum), and combinations thereof, and the streptococcus strain may include a strain selected from streptococcus faecalis (str.faecis), streptococcus thermophilus (str.thermophilus), and combinations thereof.

The fermentation may be carried out by adding 0.1-1.5 weight parts of lactobacillus to 100 weight parts of young seedling ginseng extract, and fermenting at 35-39 deg.C for 3-5 days.

Advantageous effects

It was confirmed that the fermented seedling ginseng prepared according to the present invention has low cytotoxicity, and also has moisturizing factor production increasing effect, collagen and collagen fiber production increasing effect, radical scavenging effect, and intracellular active oxygen inhibiting effect. This means that the young ginseng ferment of the present invention can be used as a functional material derived from natural materials for moisturizing, anti-wrinkle and anti-oxidation, and the cosmetic composition comprising the young ginseng ferment of the present invention can be widely used in the cosmetic and food fields for improving skin conditions.

The effects of the present invention are not limited to the effects described above. It is to be understood that the effects of the present invention include all effects that can be inferred from the following description.

Drawings

Fig. 1 is a flow chart illustrating a method of preparing a cosmetic composition according to the present invention.

Fig. 2 is a graph showing the Production amount of hyaluronic acid (HA Production% of control) of the ginseng ferment of seedling of the present invention relative to the control group.

Fig. 3 is a graph showing the amount of collagen in the fermented seedling ginseng of the present invention.

FIG. 4 is a graph showing the degree of fluorescent expression of the NHDF fibroblasts of test example 1-1, which were treated with the fermented product of Panax schinseng of the present invention, by staining the intracellular collagen fibers (collagen fibers) according to the immunofluorescent staining method and observing the cells with a fluorescent microscope.

FIG. 5 is a graph showing radical scavenging activity (%) of the seedling ginseng ferment of the present invention.

FIG. 6 is an image of HaCaT cells of test example 1-1 treated with fermented seedling ginseng according to the present invention, stained for intracellular reactive oxygen species according to the immunofluorescence staining method, and observed for the degree of fluorescence expression using a fluorescence microscope.

Detailed Description

The above objects, other objects, features and advantages of the present invention can be easily understood by the accompanying drawings and the following related preferred embodiments. However, the present invention is not limited to the embodiments described below, but may be implemented as other embodiments. The embodiments described below are provided so that this disclosure will be thorough and complete, and will fully convey the concept of the invention to those skilled in the art.

In the present specification, it is to be understood that the terms "comprises," "comprising," "includes" or "including," and the like, are used for specifying the presence of the stated features, integers, steps, actions, elements, components, or groups thereof, but do not preclude the presence or addition of one or more other features, integers, steps, actions, elements, components, or groups thereof.

Unless otherwise indicated, all numbers, values and/or expressions referring to amounts of ingredients, reaction conditions, polymer compositions and formulations used in the specification are to be understood as being modified in all instances by the term "about" because such numbers are essentially approximations that reflect the various uncertainties in the measurements that occur when such values are obtained from others. Further, where a range of values is disclosed herein, such range is continuous and, unless otherwise stated, includes all values from the minimum value of such range to the maximum value included therein. Further, when such ranges are integers, unless otherwise indicated, all integers from the minimum to the maximum recited, including the maximum.

Hereinafter, the method for preparing the cosmetic composition and the cosmetic composition comprising the fermented young ginseng of the present invention will be described in detail.

Prepared according to one embodiment described aboveCosmetic compositionComprises a young seedling ginseng fermented product as an effective ingredient, preferably, the young seedling ginseng fermented product can be obtained by fermenting a young seedling ginseng extract using lactic acid bacteria. The seedling ginseng of the present invention is a conventional seedling ginseng, generally a ginseng cultivated for 1 to 2 years, and contains saponin components in leaves and stems about 7 to 8 times as much as that of roots, and has advantages of low price compared to conventionally used root ginseng because of its advantages of being producible all year round and shortened cultivation time by hydroponic cultivation, and thus is not particularly limited as long as it is easily used as a raw material for ginseng processed products. In addition, there is no particular limitation as long as the seedling ginseng contains saponins that affect the central nervous system, endocrine system, immune system, metabolismSystemic, etc. exert various effects on body regulation functions, and in addition, the saponin has pharmacological effects such as strong lipolysis ability, promotion of absorption and digestion of nutrients, promotion of metabolism through activation of intracellular enzymes, recovery of physical strength through increase of energy, improvement of fatigue, weakness and anorexia, promotion of serum protein synthesis, etc.

The seedling ginseng extract, the seedling ginseng ferment and the lactic acid bacteria may be the same as or different from those described in the following preparation methods.

The young ginseng fermented product of the present invention may be contained in an amount of 0.001 to 60% by weight, and preferably may be contained in an amount of 0.5 to 10% by weight in 100% by weight of the cosmetic composition. When the content of the ginseng fermented product of young plants is less than 0.001 wt%, there is a disadvantage that efficacy of oxidation resistance, skin moisturizing or wrinkle improvement may be reduced, and when the content of the ginseng fermented product of young plants exceeds 60 wt%, there is a disadvantage that toxicity may be generated.

The cosmetic composition of the present invention may be prepared in any form generally prepared in the art, and may be formulated into, for example, a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleanser, an oil, a powder foundation, a milk foundation, a wax foundation, a spray, etc., but is not limited thereto. More specifically, the cosmetic composition of the present invention may be formulated into the form of a skin softening lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing foam, a cleansing lotion, a pack, a spray or a powder.

As a carrier effective for the cosmetic material contained in the cosmetic composition of the present invention, a carrier generally used in the art may be used depending on the formulation.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide, etc. can be used as a carrier component.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier ingredient, and particularly, when the formulation of the present invention is a spray, a propellant such as chlorofluorocarbon, propane/butane or dimethyl ether may be further contained.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or opacifying agent, such as water, butylene glycol, hexylene glycol, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol (1,3-butyl glycol) oil, glycerin fatty acid ester, polyethylene glycol or sorbitan fatty acid ester, is used as a carrier component.

When the dosage form of the present invention is a suspension, as a carrier ingredient, a diluent in a liquid phase such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, or the like can be used.

When the formulation of the present invention is a surfactant-containing detergent, as a carrier component, fatty alcohol sulfate, fatty alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazoline derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamide betaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, ethoxylated glycerin fatty acid ester, or the like can be used.

The formulation of the cosmetic composition of the present invention may preferably be selected from the group consisting of solutions, emulsions, suspensions and combinations thereof.

The ingredients contained in the cosmetic composition of the present invention include, in addition to the active ingredient and the carrier ingredient, ingredients generally utilized in cosmetic compositions, and may include, for example, conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and perfumes.

Fig. 1 is a flow chart illustrating a method of preparing a cosmetic composition according to the present invention. Referring to fig. 1, the method of preparing a cosmetic composition includes: a step of preparing seedling ginseng S10, a step of extracting the seedling ginseng to obtain a seedling ginseng extract S20, a step of filtering the seedling ginseng extract S30, and a step of fermenting the filtered seedling ginseng extract with lactic acid bacteria to obtain a seedling ginseng ferment S40.

The step S10 of preparing seedling ginseng is a step of pulverizing the seedling ginseng to more easily extract the seedling ginseng. The young ginseng may be the same as or different from that described in the cosmetic composition.

The step of pulverizing the prepared ginseng seedling is not particularly limited as long as it is a conventional pulverizing method for obtaining an extract. However, in order to effectively extract young ginseng, young ginseng may be dried at 10-55 deg.C and then pulverized. When the drying temperature is less than 10 ℃, the ginseng seedling cannot be sufficiently dried, and when the drying temperature exceeds 55 ℃, there is a disadvantage that the effective ingredient is destroyed.

The step S20 of obtaining the seedling ginseng extract is a step of extracting seedling ginseng to obtain a seedling ginseng extract and preparing for fermentation of the seedling ginseng extract. The seedling ginseng extract is not particularly limited as long as it is a conventional seedling ginseng extract for obtaining a seedling ginseng ferment, and may include an extract selected from the group consisting of a hot water extract, a solvent extract, and a combination thereof.

The hot water extraction process for obtaining the hot water extract may comprise the steps of: adding purified water 8-22 times, preferably 18-22 times, the volume of the prepared seedling ginseng; and extracting the young ginseng added with the purified water at 50-90 ℃ for 1-3 hours. Further, when the hot water extraction time is less than 1 hour, the effective components cannot be sufficiently extracted, and thus there is a disadvantage that the efficacy cannot be sufficiently exhibited, and when the hot water extraction time exceeds 3 hours, the extraction efficiency is not high as compared with the time to be put, and thus there is a disadvantage that it is uneconomical. Further, when the hot water extraction temperature is less than 50 ℃, there is a disadvantage that the effective components cannot be completely extracted, and when the hot water extraction temperature exceeds 90 ℃, there is a disadvantage that the effective components are destroyed.

The solvent extraction method for obtaining the solvent extract is not particularly limited as long as it is a conventional extraction method for obtaining a solvent extract. The solvent may comprise a solvent selected from the group consisting of fermentation alcohol, ethanol, methanol, butanol, propanol, ethyl acetate, isoprene glycol, glycerol, ethylene glycol, propylene glycol, butylene glycol, chloroform, dichloromethane, hexane, acetone, acetonitrile, petroleum ether, diethyl ether and combinations thereof, and preferably may be water.

The step S30 of filtering the ginseng seedling extract is a step of filtering the extracted ginseng seedling extract to be in a state of being fermented by lactic acid bacteria. The filtration of the present invention is not particularly limited as long as it is a conventional filtration method for filtering an extract. Preferably, the filter paper may be 0.20 to 0.50 μm, and more preferably, the filter paper may be 0.45 μm, for efficient fermentation using lactic acid bacteria. When the filter paper is less than 0.20 μm, there is a disadvantage that the fermentation metabolites disappear, and when the filter paper exceeds 0.50 μm, bacteria may remain.

The step S40 of obtaining a ginseng seedling fermented product is a step of fermenting the filtered ginseng seedling extract with lactic acid bacteria to obtain a ginseng seedling fermented product. The fermentation method of the present invention is not particularly limited as long as it is a conventional fermentation method of extract, but, preferably, 0.1 to 1.5 parts by weight of lactic acid bacteria may be added and fermented at 35 to 39 ℃ for 3 to 5 days, relative to 100 parts by weight of young ginseng extract, and preferably, 1.0 part by weight of lactic acid bacteria may be added and fermented at 37 ℃ for 3 to 5 days, relative to 100 parts by weight of young ginseng extract, to obtain young ginseng fermented product, in order to enhance antioxidant activity effect, skin moisturizing effect or wrinkle improving effect. When the content of the lactic acid bacteria is less than 0.1 parts by weight, there is a disadvantage in that the fermentation rate is reduced, and when the content of the lactic acid bacteria exceeds 1.5 parts by weight, there is a disadvantage in that excessive fermentation is caused. In addition, when the fermentation temperature is less than 35 ℃ or exceeds 39 ℃, there is a disadvantage that the survival rate of the lactic acid bacteria is reduced. In addition, when the fermentation time is less than 3 days, there is a disadvantage that the content of fermentation metabolites is small, and when the fermentation time exceeds 5 days, there is a disadvantage that it may become fermented vinegar due to excessive fermentation.

The lactic acid bacteria used in the fermentation are conventional lactic acid bacteria that can ferment a ginseng seedling extract to obtain a ginseng seedling fermented product having an excellent antioxidant activity effect, skin moisturizing effect, or wrinkle-improving effect, and for example, the lactic acid bacteria may be one or more strains selected from the genera Lactobacillus (Lactobacillus), Bifidobacterium (Bifidobacterium), and Streptococcus (Streptococcus).

The lactobacillus strains include lactobacillus rhamnosus (l.rhamnous), lactobacillus plantarum (l.plantarum), lactobacillus acidophilus (l.acidophilus), lactobacillus bulgaricus (l.bulgaricus), lactobacillus fuscus (l.convulsus), lactobacillus reuteri (l.reuteri), lactobacillus casei (l.casei), and the like, the bifidobacterium strains include bifidobacterium breve (b.breve), bifidobacterium longum (b.longum), bifidobacterium infantis (b.infarnatis), bifidobacterium bifidum (b.bifidum), and bacillus thermophilus (b.thermophilum), and the like, and the streptococcus strains include streptococcus faecalis (str.faecalis), streptococcus thermophilus (str.thermophilus), and the like.

The lactic acid bacteria are not limited to specific lactic acid bacteria, but are preferably a strain of lactobacillus that can most effectively impart excellent antioxidant activity effect, skin moisturizing effect, or wrinkle-improving effect to young ginseng, more preferably a strain of lactobacillus rhamnosus, and even more preferably a strain of lactobacillus rhamnosus KCTC 5033.

Thereafter, further, the fermented product may be once filtered again to finally obtain a young ginseng fermented product.

Hereinafter, preferred embodiments are proposed to aid understanding of the present invention. However, the following examples are provided only for easier understanding of the present invention, and the contents of the present invention are not limited to the following examples.

Examples

S10: drying seedling Ginseng radix at 50 deg.C, and pulverizing. S20: purified water was added in an amount of 20 times the volume of the prepared ginseng seedling sample, and then hot water extraction was performed at 80 ℃ for 2 hours. S30: after the extraction, filtration was performed using a 0.45 μm filter paper. S40: to the obtained extract, Lactobacillus rhamnosus (Lactobacillus rhamnosus) KCTC5033 was added in an amount of 1%, and then a fermentation process was performed at 37 ℃ for 3-5 days. The lactobacillus rhamnosus KCTC5033 is obtained from a biological resource center. Thereafter, filtration was performed, and the seedling ginseng fermentation product was used as a sample of the present invention.

Test example 1 evaluation of cytotoxicity of fermented product of Panax japonicus (EZ-cytox test (assay))

Test example 1-1 culture of skin cells

In the culture of skin keratinocytes, HaCaT cells, which are skin keratinocytes, obtained from Cell Line Service GmbH, (germany), were used. Dulbecco's Modified Eagle's Medium (DMEM) containing penicillin-streptomycin (penicillin-streptomycin) and Fetal Bovine Serum (FBS) was used at 37 deg.C in 5% CO₂The culture was carried out in a thermostat, and subculture was carried out at intervals of 2 to 3 days.

In addition, in the culture of skin Fibroblasts, Normal Human skin Fibroblasts (NHDF) obtained from Lonza (Lonza Walkersville) as skin Fibroblasts were used, and Fibroblast Basal Medium (FBM) containing hFGF-B, Insulin (Insulin), GA-1000 and Fetal Bovine Serum (FBS) was used at 37 ℃ at 5% CO₂The culture was carried out in a thermostat, and subculture was carried out at intervals of 3 to 5 days.

Test example 1-2 evaluation of cytotoxicity (EZ-cytox test)

The EZ-cytox assay is a representative method for measuring the survival rate of cells by using the principle that water-soluble tetrazolium salt (WST) reacts with Dehydrogenase (Dehydrogenase) of living cells to produce orange water-soluble formazan (formazan).

To confirm toxicity in cells, HaCaT cells and NHDF cells were cultured at 5 × 10 in DMEM medium⁴And 1X 10⁴cells/mL (cells/mL) were distributed in 96-well plates (well plates) and incubated at 37 ℃ in 5% CO₂Under the conditions of (1) for 18 hours. Cultured cells were changed to serum-free (serum-free) medium and treated with various concentrations (0.5%, 1.0%, 2.0%) of the fermented seedling ginseng prepared in the examples. After that, EZ-cytox was added to each well (well) at 37 ℃ in 5% CO₂The reaction was carried out for 30 minutes, and then the absorbance was measured at 450nm using a microplate reader. The control group for cytotoxicity evaluation utilized cells that were not treated with any extract. The average absorbance values of the respective experimental groups were calculated and compared with those of the control group, thereby evaluating the cell survival rate.

[ Table 1]

As shown in the table 1, the fermented ginseng plantlets prepared in the examples were not observed to be cytotoxic in skin keratinocytes and skin fibroblasts at various concentrations.

Test example 2 confirmation of moisturizing Effect of fermented product of Ginseng radix

HaCaT cells were cultured at 1.0X 10⁵cells/mL were distributed in 24-well plates at 37 ℃ in 5% CO₂Under the conditions of (1) for 18 hours. The seedling ginseng fermentations prepared in the examples described were treated with various concentrations (0.5%, 1.0%, 2.0%) in serum-free DMEM instead, and cultured for 24 hours. For the negative control group of the experiment, no treatment with any extract was performed, and for the positive control group, treatment with Retinoic Acid (RA) at a concentration of 10. mu.M was performed. Thereafter, the culture solution of each group was taken, and the transparency was measured using an HA-ELISA kit (Cusabio Biotechnology Co., Ltd.)The amount of produced Hyaluronic Acid (HA) was measured according to the method provided by the manufacturer, and the results are shown in fig. 2 and table 2.

[ Table 2]

Categories	Treatment concentration (%)	Amount of HA formed (%)
			No treatment group	-	100±7.36
Examples	0.5	122±0.6
			Examples	1.0	173±2.6
Examples	2.0	149±12.8
			Retinoic acid	0.001	113±4.4

Referring to fig. 2 and table 2, it was confirmed that seedling ginseng fermentate concentration-dependently increased the production of HA. In addition, it is known that the seedling ginseng ferment more increased the production of hyaluronic acid than retinoic acid as a control group.

Test example 3 confirmation of wrinkle-improving Effect of fermented seedling Ginseng

Experimental example 3-1 confirmation of PIP production Effect

Fibroblast NHDF at 2X 10⁴cells/mL were dispensed in 24-well plates and then cultured under cell culture conditions for 24 hours. After cultivation, the seedling ginseng fermentations prepared in the examples were treated with various concentrations (0.5%, 1.0%, 2.0%) of serum-free FBM and cultured for 24-48 hours. For the negative control group of the experiment, no extract was treated, and for the positive control group, ascorbic acid (ascorbic acid) was treated at a concentration of 10 ppm. After the culture, the supernatant of each group was taken, and with respect to the amount of free procollagen (procollagen) in the medium, the amount of collagen was quantified by measuring absorbance at 450nm using procollagen type I peptide (PIP) EIA kit (Takara Biomedical Co.). The measurement method was performed according to the method provided by the manufacturer. The results are shown in fig. 3 and table 3.

[ Table 3]

Categories	Treatment concentration (%)	Amount of collagen produced (%)
			No treatment group	-	100±1.0
Examples	0.5	139±0.2
			Examples	1.0	168±0.3
Examples	2.0	198±0.3
			Ascorbic acid	0.001	156±1.0

Referring to fig. 3 and table 3, it was confirmed that the seedling ginseng fermentate concentration-dependently increased collagen synthesis.

Test example 3-2 confirmation of collagen fiber (collagen) production Effect

Fibroblast NHDF at 2X 10⁴cells/mL were dispensed in 24-well plates and then cultured under cell culture conditions for 24 hours. After cultivation, the seedling ginseng fermentations prepared in the examples were treated with various concentrations (0.5%, 1.0%, 2.0%) of serum-free FBM and cultured for 24-48 hours. For the negative control group of the experiment, no extract was used for treatment, and for the positive control group, ascorbic acid was used at a concentration of 10 ppm. After the culture, the cells were washed with HEPES-BSS, and then stained for intracellular collagen fibers according to an immunofluorescence staining method. Thereafter, the degree of fluorescence expression was measured by a fluorescence microscope, and the result thereof is shown in fig. 4.

Referring to fig. 4, it was confirmed that the ginseng seedling fermented product concentration-dependently increased the production of collagen fibers.

Therefore, the ginseng seedling fermentate prepared in the examples increases collagen synthesis and collagen fiber production concentration-dependently, which indicates an excellent wrinkle-improving effect.

Test example 4 confirmation of Effect of antioxidant Activity of fermented product of seedling Ginseng radix

Test example 4-1 measurement of antioxidant Effect

The radical scavenging test is a test method utilizing the principle that the absorbance of a stable 2, 2-diphenyl-1-picrylhydrazino (2,2-diphenyl-1-picryl-hydrazyl radial, DPPH) shows the maximum absorbance at 517nm, and the radical DPPH is scavenged by a sample from purple to pale yellow, that is, the absorbance at said wavelength of 517nm decreases as the radical scavenging rate increases.

First, dilution with methanol was performed so that the concentrations of the ginseng seedling fermentations prepared in the examples became 3.125%, 6.25%, 12.5%, 25%, 50%, thereby preparing dilutions of the ginseng seedling fermentations. A sample was prepared by mixing 1mL of a dilution of the seedling ginseng ferment and 1mL of a 0.1mM DPPH (Sigma) solution. The prepared sample was left at room temperature for 15 minutes, and then the absorbance was measured at a wavelength of 517nm using a microplate reader. In this test example, the measurement was performed on a sample of the first control group (1mL of DPPH and 10ppm of ascorbic acid) by the method described above, and the second control group (1mL of DPPH and 1mL of methanol) was used in order to obtain respective color correction values for the sample and the control group. The results of measuring the radical scavenging activity of the ginseng fermentations of young plants are shown in fig. 5 and table 4.

[ Table 4]

Categories	Treatment concentration (%)	DPPH radical scavenging Activity (%)
			No treatment group	-	0±2.0
Examples	3.13	10.3±1.6
			Examples	6.25	17.6±1.8
Examples	12.50	32.4±1.5
			Examples	25.00	52.7±1.4
Examples	50.00	73.3±2.6
			Ascorbic acid	0.001	46.6±7.4

Referring to fig. 5, it was confirmed that the concentration-dependently increased the radical scavenging activity of the ginseng seedling fermentations.

Test example 4-2 measurement of inhibitory Effect of Reactive Oxygen Species (ROS) in cells

To measure changes in intracellular Reactive Oxygen Species (ROS), HaCaT cells were plated at 1.0X 10⁵Cells/well (cells/well) were seeded into 24-well plates and cultured for 24 hours, and then cultured for 24 hours after treatment with various concentrations (0.5%, 1.0%, 2.0%) of the ginseng seedling fermentations prepared in the examples. For cultured cells, washing was performed with PBS, followed by 300. mu.M H₂O₂Treated for 3 hours for further culture. The result of adding 50 μ M of dichlorofluorescein diacetate (DCF-DA), a dye for measuring intracellular Reactive Oxygen Species (ROS), and washing with PBS after 1 hour of incubation, followed by measuring the degree of fluorescence expression using a fluorescence microscope is shown in fig. 6.

Referring to fig. 6, it was confirmed that the seedling ginseng fermentate concentration-dependently inhibited the generation of ROS.

Therefore, the concentration of radical scavenging activity of the young ginseng fermentate increases dependently and the generation of ROS is inhibited, which indicates an excellent antioxidant effect.

As described above, the present inventors prepared ginseng seedling fermentation products and confirmed that the fermentation products have low cytotoxicity and excellent effects of increasing the production of moisturizing factors, increasing the production of collagen and collagen fibers, scavenging free radicals, and inhibiting intracellular reactive oxygen species. This means that the young ginseng ferment of the present invention can be used as a functional material derived from natural materials for moisturizing, anti-wrinkle and anti-oxidation, and the cosmetic composition comprising the young ginseng ferment of the present invention can be widely used in the cosmetic and food fields for improving skin conditions.