KR102436798B1 - Methods for manufacturing of coix lacryma-jobi and scoria complex ferment with improved skin sedation and anti-aging effects and cosmetic composition using same - Google Patents

Abstract

Disclosed are a fermented radish extract and a cosmetic composition containing the same, which reduces toxic substances and the like of natural products, and has anti-wrinkle and antioxidant effects. The present invention provides a fermented adil radish extract comprising a barley radish extract and volcanic clusters.

Description

BACKGROUND OF THE INVENTION Field of the Invention }

The present invention relates to a fermented barley radish extract and a cosmetic composition containing the extract, and more particularly, to a fermented product obtained by fermenting the barley radish extract using a microorganism and a cosmetic composition containing the same.

Fermentation refers to the process in which enzymes possessed by microorganisms change raw materials to produce beneficial ingredients. During the fermentation process, active ingredients are extracted and the content of functional substances increases.

And fermentation technology has overlap in meaning with terms such as bioconversion, biosynthesis, and biocatalyst, and refers to a technology for producing a desired product from a precursor using the enzymatic function of microorganisms. Fermentation technology used in the cell or enzyme stage is the main process for the production of useful substances and constitutes the current biological industry. If fermented food uses a one-dimensional fermentation technology that utilizes naturally occurring microorganisms, fermented cosmetics require more advanced technology. Bioconversion fermentation technology, which was developed and laid the foundation for cosmetic functional materials industry through advanced fermentation production technology that can mass-produce only useful physiologically active substances, is positioning itself as the third generation fermentation technology.

Since Koreans use fermented foods such as kimchi and soybean paste a lot, the concept of fermentation is very familiar, and cosmetics using fermented raw materials are also used without objection. Also, when a new material is developed using fermentation technology for the general public, it is known that it shows completely new effects from existing materials or that many harmful ingredients are changed to safe ingredients, so the preference for fermented cosmetics is high. Although not inferior to the level, it is known that many fermented materials currently applied to cosmetics use natural fermentation or a fermentation method close to natural aging.

The recent research trend in domestic fermentation technology is known to use natural fermentation or a fermentation method close to natural aging. Although it takes a long time to naturally produce ingredients useful for Fermentation has disadvantages in that it is difficult to obtain fermentation products of constant quality for each production batch, and quality control is also difficult.

The Korean fermented cosmetics market has been growing at a high rate of 40% over the past three to four years, and is expected to reach 450 billion won in 2012, 650 billion won in 2013, and 1 trillion won (estimated) in 2014. are doing

Fermented products can be broadly classified into fermentation filtrate and fermentation lysate. is a method in which microorganisms are killed by using heat, pH, and enzyme (Lysozyme) high pressure after fermentation is completed.

Despite the increasing preference for fermented cosmetics and the launch of many products, one of the biggest reasons for not choosing fermented cosmetics or fermented raw materials for cosmetics is the specific odor of fermented products, and the specific odor of fermented products is The odor of the medium being added is the cause.

The technology currently used to remove the peculiar odor of these fermented products utilizes the adsorption of fragrance using activated carbon after fermentation, the method of adding another strong fragrance to mask the fragrance (fragrance masking), and the method of using a medium from which the characteristic odor has been removed. However, it is difficult to see it as a perfect solution due to reasons such as price and process difficulties.

Scoria refers to volcanic soil with high porosity, which is made of sintered bodies of porous volcanic rock, volcanic sand, and other volcanic ash mixed at high temperature among various substances ejected when a volcano erupts. Oreum) and is distributed throughout Jeju in Korea. In terms of color, it is largely divided into reddish brown, yellowish brown, black and dark gray colors, and reddish-brown clusters are most commonly distributed. In Jeju Island, a high-temperature and humid environment since ancient times, volcanic stone clusters were used on the roof and inside and outside walls of houses to keep them warm, insulated, soundproof, and desiccant. It has been widely used, and has recently been developed as a soil improver, fertilizer additive, feed additive, natural dye, cosmetic raw material, etc. In particular, although it has effects such as antibacterial, anti-inflammatory, and moisturizing effects, systematic research on it is lacking.

Laid-open Patent No. 10-2017-0038679 and Patent Publication No. 10-2015-0015089 disclose a cosmetic composition using volcanic clusters, but do not disclose the content of removing fermented odors using volcanic clusters for fermentation. not.

Registered Patent No. 10-1695403 discloses a method for producing soju using volcanic clusters and the soju, but it is not about cosmetics, and Patent Publication No. 10-2011-0046600 relates to a method for producing black garlic using scoria. It uses volcanic clusters, but it is not about cosmetics.

Therefore, the present invention has been devised to solve the above problems, and there is no objection due to odor, toxic substances of natural products, etc. are reduced, skin wrinkle improvement and antioxidant effect of fermented barley radish extract and a cosmetic composition containing the same would like to provide

In order to solve the above problems, the present invention provides a fermented adil radish extract comprising a barley radish extract and volcanic clusters.

In addition, the fermented barley radish extract provides a fermented barley radish extract, characterized in that it is fermented in a medium containing the volcanic cluster by inoculating the adul radish extract into microorganisms.

In addition, the microorganism Lactobacillus ( Lactobacillus ) or Bacillus ( Bacillus ) It provides a fermented barley radish extract, characterized in that it belongs to the genus.

In addition, the medium containing the volcanic clusters provides a fermented adil radish extract, characterized in that the volcanic clusters are 0.1 to 10% by weight relative to the total medium composition.

In addition, the medium containing the volcanic clusters provides a fermented adil radish extract, characterized in that the volcanic clusters are 0.1 to 10% by weight relative to the total medium composition.

In addition, it provides a cosmetic composition, characterized in that it contains 0.1 to 90% by weight of the fermented barley radish extract relative to the total cosmetic composition.

In addition, the cosmetic composition provides a cosmetic composition, characterized in that for wrinkle improvement and antioxidant.

In addition, the cosmetic composition is a softening lotion, astringent lotion, nutrient lotion, eye cream, serum, nutrition cream, massage cream, cleansing cream, cleansing lotion, cleansing foam, cleansing water, powder, essence, pack, hair tonic, hair treatment , It provides a cosmetic composition for antioxidant and inflammation relief, characterized in that it has a formulation of shampoo or rinse.

According to the present invention, by including the volcanic clusters used during fermentation of the barley radish extract as a component of the medium, the fermentation odor generated during fermentation is removed, and the fermented material is stably maintained so that there is no objection when applied to the skin, skin improvement effect and It is possible to provide a fermented barley radish extract and a cosmetic composition containing the same, which has an antioxidant effect, maintains the uniformity and stability of quality by preventing the propagation of harmful bacteria, and the fermented material is stably maintained.

1 is a table schematically illustrating a method for producing a fermented yulmu extract containing volcanic clusters of the present invention;
Figure 2a is a graph relating to the cell viability of the extract fermented barley radish in Experimental Example 2 of the present invention,
Figure 2b is a graph relating to the cell viability of the non-fermented extract of barley radish in Experimental Example 2 of the present invention,
Figure 2c is a graph relating to the cell viability of the fermented sulfur extract in Experimental Example 2 of the present invention,
Figure 2d is a graph relating to the cell viability of the non-fermented sulfur extract in Experimental Example 2 of the present invention,
Figure 3 is a view showing the skin aging improvement efficacy of the extract fermented barley radish in Experimental Example 3 of the present invention,
Figure 4a is a graph relating to the inhibition of nitrogen monoxide production of the extract fermented barley radish in Experimental Example 4 of the present invention;
Figure 4b is a graph relating to the inhibition of nitrogen monoxide production of non-fermented extract of barley radish in Experimental Example 4 of the present invention;
Figure 4c is a graph relating to the inhibition of nitric oxide production of the fermented sulfur extract in Experimental Example 4 of the present invention;
Figure 4d is a graph relating to the inhibition of nitrogen monoxide production of non-fermented sulfur extract in Experimental Example 4 of the present invention.

Hereinafter, the present invention will be described in detail through preferred embodiments. Prior to this, the terms or words used in the present specification and claims should not be construed as being limited to conventional or dictionary meanings, and the inventor should properly understand the concept of the term in order to best describe his invention. Based on the principle that it can be defined, it should be interpreted as meaning and concept consistent with the technical idea of the present invention. Accordingly, the configuration of the embodiments described in the present specification is merely the most preferred embodiment of the present invention and does not represent all of the technical spirit of the present invention, so various equivalents and modifications that can be substituted for them at the time of the present application It should be understood that there may be In addition, throughout the specification, when a part "includes" a certain component, it means that other components may be further included, rather than excluding other components, unless otherwise stated.

The present inventors have repeatedly studied diligently to provide a fermented barley radish extract and a cosmetic composition containing the same, which removes the fermented odor generated during fermentation of the barley radish extract, and maintains the fermented material stably so that there is no objection when applied to the skin. It was discovered that the efficacy of natural products is improved by the mineral and nutritional components of volcanic clusters at the same time suppressing the strong smell of natural products generated during fermentation of natural products, thereby enhancing the efficacy of natural products, leading to the present invention.

Accordingly, the present invention discloses a fermented adil radish extract comprising a barley radish extract and volcanic clusters.

In the present invention, the fermented barley radish extract can be fermented in a medium containing the volcanic cluster by inoculating the microorganisms with the barley radish extract.

Coix lacryma-jobi is an annual plant of the Grape family, with a height of 1-1.5 m. When used as a herbal medicine, it is called Ui-in (薏苡仁). In addition, flowers bloom in the leaf axils in July, and the fruits are oval-shaped fruits, and the seeds are used for food and medicine. In addition, the raw leaves are used as a tea substitute, and the roots are used to treat jaundice and neuralgia.

Recently, it has been found to have an anticancer effect, and adil radish has an excellent diuretic effect and has an excellent effect in discharging inflammation and wastes from the body. Potassium component is also known to help improve edema symptoms by helping to discharge accumulated waste products in the blood vessels and body.

In particular, adil radish contains a large amount of coixeloride, which is known to have an excellent effect on alleviating skin troubles, so it is known to have excellent efficacy in removing blemishes such as spots and freckles as well as skin troubles. It also has an excellent moisturizing effect, which helps to keep skin elastic and healthy.

In the present invention, the adul radish has a moisturizing effect that supplies moisture to the skin or prevents excessive evaporation of moisture from the skin, and in particular, bio-components are produced by highly reactive active oxygen or free radical peroxides generated inside the body from harmful environments. It can inhibit oxidation or denaturation, which is the main cause of skin aging, and also has a whitening effect that inhibits the auto-oxidation of DOPA by scavenging free radicals and suppresses the pigmentation of the skin due to the formation of excessive melanin pigment in the skin. can indicate

In the present invention, the barley radish extract may be prepared using a conventional extraction method known in the art, for example, a solvent extraction method. The extraction solvent used in the preparation of the mixed extract using the solvent extraction method is water, a lower alcohol having 1 to 4 carbon atoms (eg, methanol, ethanol, propanol and butanol) or a mixture thereof, which is a hydrous lower alcohol, propylene glycol, 1, 3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane and mixtures thereof may be selected from the group consisting of, and among them, water, alcohol, or mixtures thereof. it is preferable When water is used as the extraction solvent, the water is preferably hot water. In addition, when alcohol is used as the extraction solvent, the alcohol is preferably a lower alcohol having 1 to 4 carbon atoms, and the lower alcohol is more preferably selected from methanol or ethanol.

In addition, when an aqueous alcohol is used as the extraction solvent, the alcohol content is preferably 10 to 90%, more preferably 50 to 80%. On the other hand, in the present invention, it is apparent to those skilled in the art that an extract exhibiting substantially the same effect can be obtained using other extraction solvents as well as the above-described extraction solvent. In addition, the mixed extract in the present invention includes not only the extract using the above-described extraction solvent, but also extracts obtained through other conventional extraction methods or extracts subjected to purification and fermentation processes. For example, reduced pressure with carbon dioxide, extraction by supercritical extraction by high temperature, extraction by extraction using ultrasound, separation using an ultrafiltration membrane having a constant molecular weight cut-off value, various chromatography (size, charge, hydrophobicity or affinity The active fraction obtained through various purification and extraction methods additionally performed, such as separation by a method prepared for separation according to sex) or an extract using a fermentation product using a natural state or various microorganisms, is also included in the extract of the present invention.

The extraction method by the supercritical extraction method by the reduced pressure by carbon dioxide and the high temperature refers to the supercritical fluid extraction method, and in general, the supercritical fluid is a liquid having a gas when the gas reaches a critical point under high temperature and high pressure conditions, and It has gaseous properties and chemically has a polarity similar to that of a non-polar solvent. Due to these properties, supercritical fluids are used for the extraction of fat-soluble substances (J Chromatogr A 1998; 479:200-205). Carbon dioxide is a supercritical fluid device. It becomes a supercritical fluid with both liquid and gas properties as the pressure and temperature reach the critical point due to the operation of When the supercritical carbon dioxide passes through an extraction vessel containing a certain amount of the sample, the fat-soluble substance contained in the sample is extracted into the supercritical carbon dioxide. After extracting the fat-soluble substance, if supercritical carbon dioxide containing a small amount of cosolvent is passed through the sample remaining in the extraction container again, components that were not extracted with pure supercritical carbon dioxide alone can be extracted. The supercritical fluid used in the supercritical extraction method of the present invention can effectively extract the active ingredient by using supercritical carbon dioxide or a mixed fluid in which a cosolvent is additionally mixed with carbon dioxide. As the cosolvent, one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether may be used.

In addition, the ultrasonic extraction method is an extraction method using energy generated by ultrasonic vibration, and ultrasonic waves can destroy an insoluble solvent contained in a sample in an aqueous solvent, and due to the high local temperature generated at this time, Since the kinetic energy of the reactant particles is increased, sufficient energy required for the reaction is obtained, and the high pressure is induced by the impact effect of ultrasonic energy to increase the mixing effect of the material and the solvent contained in the sample, thereby increasing the extraction efficiency. The extraction solvent that can be used for the ultrasonic extraction method may be one or a mixture of two or more selected from the group consisting of chloroform, ethanol, methanol, water, ethyl acetate, hexane and diethyl ether. The extracted sample is vacuum filtered to recover the filtrate, then removed by a vacuum evaporator, and an extract can be obtained through a conventional extract preparation method of freeze-drying.

In addition, the Yulmu extract according to the present invention may include an extract that has undergone a fermentation process.

In the present invention, the microorganism may be a microorganism belonging to the genus Lactobacillus or Bacillus, and preferably may be Bacillus subtilis (Apep Microorganism Jeju 170201, A&Pep Co., Ltd., self-isolated).

The genus Lactobacillus is a bacterium that produces a large amount of lactic acid by fermenting sugars to obtain energy. Morphologically, it is a Gram-positive spore-free bacillus and has polymorphism, and the size is 0.5 to 0.9 × 1 to 11 μm. It shows various forms, and there are strains showing the Coryne type. The arrangement of the fungus is single-stranded, double-stranded, single-stranded and single-chained. Most are immobile and usually catalase negative. This genus enjoys growth in an environment with little oxygen and produces lactic acid from various sugars. It is known that many of them are resistant to acidity (pH 3 to 4), their nutritional requirements are very complex, and they require many amino acids or vitamins other than sugars, so that they cannot grow unless micronutrients are added depending on the species or strain.

The genus Bacillus is an aerobic or facultative anaerobic bacillus widely distributed in nature and is involved in denitrification, iron reduction, manganese oxidation, and the like. Depending on the environmental conditions, structural changes occur in the cells to form spores (spores). Spores have strong resistance to heat, radiation, and chemicals, and contain a large amount of calcium salts of dipicoic acid, which are not present in vegetative cells. It is classified into several species according to the characteristics of the cell size, spore shape and location in the cell, the use and product of sugar, the presence or absence of nitrate reduction, and the presence or absence of growth under high salt concentration. Representative species are Bacillus subtilis , anthrax, and cereus (the causative agent of food poisoning). In general terms, it can also refer to bacilli, or the whole of bacteria.

The fermentation refers to a process in which enzymes possessed by microorganisms change raw materials to create beneficial ingredients, and during the fermentation process, active ingredients are extracted and the content of functional substances increases.

As mentioned in the background art, the volcanic cluster refers to volcanic soil having a high porosity consisting of a calcined body calcined at high temperature by mixing porous volcanic rock, volcanic sand, and other volcanic ash among various materials ejected when a volcano erupts. The volcanic cluster is a kind of mineral, and the components include SiO ₂ , Al ₂ O ₃ , Fe ₂ O ₃ , etc. These components are the main raw material for deodorizing components and have a deodorizing function of over 90%, and these minerals are odor and volatile in the volcanic cluster. It is used in industries such as biofilters because it removes odors by adsorbing organic substances.

In addition, since the volcanic cluster has an antibacterial function, it supplies a lot of minerals and nutrients to the growth of beneficial bacteria during fermentation, while antibacterial action against harmful bacteria prevents the reproduction of harmful bacteria during long-term fermentation.

In addition, since the volcanic cluster contains a large amount of useful components such as minerals, it can be mixed with the adulphorus extract during fermentation to improve the efficacy of the adil radish extract.

The medium may contain 0.1 to 10% by weight of the image cluster relative to the entire batch composition, preferably 0.1 to 5% by weight, and more preferably 0.1 to 3% by weight.

If the content of volcanic clusters in the medium is less than 0.1% by weight relative to the total composition, fermentation odor generated during fermentation of natural products cannot be effectively removed, and when the content of volcanic clusters exceeds 10% by weight relative to the total composition, the effect is low This may not be economical.

In addition, in the present invention, when the adsorbate extract is inoculated into microorganisms and added to the medium containing the volcanic clusters, the adil radish extract may be 5 to 20 times, preferably 7 to 13 times, compared to the content of the volcanic clusters.

If the content of the adil radish extract is less than 5 times compared to the content of volcanic clusters contained in the medium, the anti-wrinkle and antioxidant efficacy of the fermented adil radish extract may be reduced, If it exceeds 20 times the content, all fermentation odors may not be removed.

As another aspect of the present invention, there is provided a cosmetic composition comprising 0.1 to 90% by weight of the fermented barley radish extract relative to the total cosmetic composition.

The cosmetic composition has anti-wrinkle and anti-oxidation effects, and the content of the fermented barley radish extract is preferably 0.5 to 80% by weight, more preferably 1 to 70% by weight relative to the total cosmetic composition.

If the fermented barley radish extract is less than 0.1% by weight compared to the total cosmetic composition, wrinkle improvement and antioxidant efficacy may be low, and if the fermented admiral radish extract exceeds 90% by weight compared to the total cosmetic composition, the effect is low and economical can't

The antioxidant cosmetic composition according to the present invention may be applied in a gel type, skin type, cream type, ointment type, etc., but is not limited thereto. The above composition may be suitably prepared by a known method by adding a suitable conventional softening agent, emulsifying agent, thickening agent or other substances known in the art depending on its type.

The gel-type composition may be prepared by adding an emollient such as trimethylolpropane, polyethylene glycol or glycerin, a solvent such as propylene glycol, ethanol, and isocetic alcohol, and purified alcohol.

The skin type composition includes fatty alcohols such as stearyl alcohol, myristyl alcohol, behenyl alcohol, arachidyl alcohol, isostearyl alcohol, isocetyl alcohol, butylene glycol, glycerin, allantoin, methyl paraben, EDTA- It can be prepared by adding 2-sodium, xanthan gum, dimethicone, polyethylene glycol-60 hydrogenated castol oil, polysorbate 60 and purified water.

The cream-type composition includes fatty alcohols such as stearyl alcohol, myristyl alcohol, behenyl alcohol, arachidyl alcohol, isostearyl alcohol, isocetyl alcohol, lecithin, phosphatidylcholine, phosphatidylethanolamine, sofphatzylserine, and sofphatidyl. Lipids such as inositol; It can be prepared by adding a solvent such as propylene glycol and purified water.

The ointment-type composition may be prepared by adding a softening agent, an emulsifying agent, and waxes such as microcrystalline lead, paraffin, ceresin, beeswax, beeswax, and vaseline.

The cosmetic composition of the present invention may further contain one or more skin wrinkle-improving ingredients exhibiting the same or similar function in addition to the active ingredient. The skin wrinkle improvement component may be any one or more selected from the group consisting of vitamin C, retinoic acid, TGF, animal placental-derived protein, betulinic acid, and chlorella extract, but is not limited thereto. In addition, the cosmetic composition of the present invention may further contain excipients including fluorescent substances, fungicides, hydrotropes, humectants, fragrances, fragrance carriers, proteins, solubilizers, sugar derivatives, sunscreens, vitamins, plant extracts, etc. can

In the present invention, the cosmetic composition includes softening lotion, astringent lotion, nourishing lotion, eye cream, serum, nourishing cream, massage cream, cleansing cream, cleansing lotion, cleansing foam, cleansing water, powder, essence, pack, hair tonic, hair treatment It can be formulated as a conditioner, shampoo or rinse.

The cosmetic composition may be formulated by a conventional method. In the formulation of external skin preparations, reference may be made to the contents disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA, and in the formulation of cosmetic compositions, the International cosmetic ingredient dictionary, 6th ed (The cosmetic, Toiletry and Fragrance Association) , Inc, Washington, 1995) may be referred to.

Specifically, the cosmetic composition can be prepared in general emulsified formulations and solubilized formulations. For example, lotion, such as a softening lotion or a nourishing lotion; emulsions such as facial lotions and body lotions; creams such as nourishing creams, moisturizing creams, and eye creams; essence; makeup ointment; spray; gel; pack; sunscreen; makeup base; foundations such as liquid type, solid type or spray type; powder; makeup removers such as cleansing creams, cleansing lotions, and cleansing oils; Or it may be formulated as a cleaning agent such as a cleansing foam, soap, body wash, but is not limited thereto. In addition, the external preparation for skin may be formulated as an ointment, patch, gel, cream or spray, but is not limited thereto.

In the cosmetic composition, in addition to the essential components in each formulation, other components may be appropriately blended within the range that does not impair the purpose according to the present invention, depending on the type of formulation or purpose of use.

The cosmetic composition may include a conventionally acceptable carrier, for example, oil, water, surfactant, humectant, lower alcohol, thickener, chelating agent, colorant, preservative, fragrance, etc. may be appropriately mixed, but is limited thereto not.

The acceptable carrier may vary depending on the formulation. For example, when formulated as an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide or their Mixtures may be used.

When the cosmetic composition is formulated as a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, or mixtures thereof may be used as a carrier component, and in the case of a spray, chlorofluoro It may further comprise a propellant such as hydrocarbon, propane, butane or dimethyl ether.

When the cosmetic composition is formulated as a solution or emulsion, a solvent, solubilizer, or emulsifier may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl benzoate, propylene glycol, 1,3-Butylglycol oil may be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol fatty esters, fatty acid esters of polyethylene glycol or sorbitan may be used.

When the cosmetic composition is formulated as a suspension, a liquid diluent such as water, ethanol or propylene glycol as a carrier component, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester; Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.

The cosmetic composition is a fatty substance, organic solvent, solubilizer, thickening agent, gelling agent, softening agent, antioxidant, suspending agent, stabilizing agent, foaming agent, and fragrance commonly used in the industry depending on the quality or function of the final product. , surfactants, water, ionic or nonionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, commonly used in cosmetics It may additionally contain adjuvants commonly used in the field of cosmetology or dermatology, such as any other ingredients. However, the adjuvant and its mixing ratio may be appropriately selected so as not to affect the desirable properties of the cosmetic composition according to the present invention.

The fermented barley radish extract prepared according to the present invention is fermented using a medium to which volcanic clusters are added during fermentation, thereby removing the fermented odor that occurs during fermentation, and has anti-wrinkle and antioxidant effects, and a cosmetic composition containing the same In addition, it does not smell of fermentation, and contains anti-wrinkle and antioxidant effects.

1 is a table schematically illustrating a method for manufacturing a fermented product containing volcanic clusters.

Referring to FIG. 1 , in the fermented barley radish extract according to the present invention, microorganisms suitable for fermentation are first separated. Thereafter, the adul radish is extracted, and a medium containing volcanic clusters is prepared, and then the barley extract is added to the medium containing volcanic clusters to inoculate the microorganisms.

Hereinafter, specific examples according to the present invention will be described.

Example

The raw radish, the raw material of the fermented product, was coarsely pulverized to 20 mesh or less, 100 g of 70 (w/v)% ethanol was added to 100 g of the sample, extracted for 72 hours, and then filtered (Whatman No 2, Maidstone, England). The filtered ethanol extract was concentrated under reduced pressure (N1000S-WD, Eyela Co., Tokyo, Japan) and then freeze-dried (FDU-1100, Eyela Co., Tokyo, Japan) to prepare an ethanol extract.

The microorganism used for this fermentation was Bacillus subtilis ( Apep Microorganism Jeju 170201) . Microorganisms isolated from the applicant, ANPEP Co., Ltd. were used. Pre-culture was incubated for 24 hours in a shaking incubator at 37°C in nutrient broth (Difco, Detroit, MI, USA). Fermentation was carried out in a medium containing 5% by weight of barley radish extract, 0.5% by weight of volcanic cluster, 0.6% by weight of glucose, 0.3% by weight of yeast extract, and 0.1% by weight of soytone. The medium was inoculated with Bacillus subtilis, and the fermentation was carried out by culturing with shaking at 37° C. for 3 days. After fermentation, the fermented product was filtered using a 0.2 μm filter to prepare a fermented barley radish extract.

comparative example One

The raw radish, the raw material of the fermented product, was coarsely pulverized to 20 mesh or less, 100 g of 70 (w/v)% ethanol was added to 100 g of the sample, extracted for 72 hours, and then filtered (Whatman No 2, Maidstone, England). The filtered ethanol extract was concentrated under reduced pressure (N1000S-WD, Eyela Co., Tokyo, Japan) and then freeze-dried (FDU-1100, Eyela Co., Tokyo, Japan) to prepare a barley radish extract.

comparative example 2

In the above example, except that the microorganism was not inoculated and thus not fermented, a non-fermented extract of barley radish was prepared in the same manner.

comparative example 3

A hwangjeong extract was prepared in the same manner as in Comparative Example 1, except that Hwangjeong was used instead of Adulmu in Comparative Example 1.

comparative example 4

A fermented sulfur extract was prepared in the same manner as in Example, except that yellow rice was used instead of barley radish in the above example.

comparative example 5

In the above example, a non-fermented product of extracting sulfur extract was prepared in the same manner, except that yellow rice was used instead of barley radish and was not fermented because microorganisms were not inoculated.

comparative example 6

A turmeric herb extract was prepared in the same manner as in Comparative Example 1, except that turmeric vinegar was used instead of the adul radish in Comparative Example 1.

comparative example 7

An extract of Byunhaengcho was prepared in the same manner as in Comparative Example 1, except that Byunhaengcho was used instead of Adulmu in Comparative Example 1.

Experimental example One

In order to measure the antioxidant capacity of the extract prepared in Examples 1 to 7 and the extract prepared in Examples 1 to 7, the experiment was conducted according to the following experimental method, and the results are shown in Table 1 below.

Experimental method

The free radical scavenging activity showed the reducing power against the stable radical DPPH (Sigma Co., St. Louis, MO, USA) and the antioxidant power using the ABTS (2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radical. The reducing power for DPPH was measured by adding 200 μL of a 0.15 mM DPPH solution in which 800 μL of a 10-fold dilution of each sample in 99% methanol was dissolved in methanol to prepare each sample, and then left at room temperature for 30 minutes and then at 517 nm. Absorbance was measured The DPPH radical scavenging activity was calculated according to the following equation, and vitamin C was used for comparison.

The radical scavenging activity of each sample was expressed as the RC50 value, which is the concentration of the sample required to reduce the absorbance of the control to which the sample was not added to 1/2.

DPPH radical scavenging activity (%) = [100-(S/C×100)]

S: Absorbance after sample group reaction - Absorbance before sample group reaction

C: Absorbance after control reaction - Absorbance before control reaction

Antioxidative activity measurement using ABTS (2,2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) radical was measured by 100-fold, 50-fold, 10-fold, and 5-fold each sample in 99 (w/v)% methanol. 800 μL of the diluted dilution was performed by the ABTS+·cation decolorization assay method.

7mM ABTS (Sigma Co., St. Louis, MO, USA) and 2.45mM potassium persulfate (Sigma Co., St. Louis, MO, USA) were mixed to a final concentration and kept at room temperature in a dark place for 24 hours. After leaving for a while to form ABTS+·, it was diluted with phosphate buffered saline (PBS, pH 7.4) so that the absorbance value at 732 nm was 0.70 (±0.02). After adding 10 μL of each of the samples prepared above to 990 μL of the diluted solution and leaving it for exactly 1 minute, absorbance was measured, and vitamin C was used for comparison.

The radical scavenging activity of each sample was expressed as the RC50 value, which is the concentration of the sample required to reduce the absorbance of the control to which the sample was not added to 1/2.

Referring to Table 1 above, it can be seen that the antioxidant activity of the adil radish extract fermented product (Example) is superior to that of the adil radish extract (Comparative Example 1), and is significantly improved when compared to the non-fermented adil radish extract (Comparative Example 2). . In addition, it can be seen that the antioxidative ability of the adorum extract is better when comparing the non-fermented, non-fermented adil radish extract (Comparative Example 2) with the admiral radish extract (Comparative Example 1). From this, it can be seen that the antioxidant ability is improved through fermentation. .

In addition, it can be seen that, in the case of the fermented extract of barley radish (Example), the antioxidant activity is very excellent compared to the case of using other herbal medicines.

Experimental Example 2

In order to confirm the cell viability of the extracts prepared in the extracts prepared in Examples 2 (NFYM), 4 (NHJ) and 5 (NFHJ) and fermented Yulmu extract prepared in Example, microglia cell line (microglia cell line) ( BV-2 cells), the experiment was carried out according to the following experimental method, and the results are shown in FIGS. 2A to 2D .

Experimental method

BV-2 cells were prepared with 10 wt% fetal bovine serum (FBS; Gibco, BRL, USA), 100 μg/mL penicillin (Gibco, BRL, USA), and 100 μg/mL streptomycin ( Gibco, BRL, USA) was added using RPMI1640 medium (Gibco, BRL, USA) 5% by weight CO ₂ Subculture in the presence of 37 ℃ incubator once every 2-3 days.

3-[4,5-Dimethylthiazol-2-yl]-2,5 to determine the effect of LPS and ramie extract on cell survival in LPS (lipopolysaccharide)-stimulated BV-2 cells -diphenyl-tetrazolium bromide (MTT) assay was used. Cells (1×10 ⁵ cell/mL) were aliquoted into a 96-well plate by 180 μL and cultured in a CO ₂ incubator for more than 12 hours, and then the samples were treated by concentration and cultured for 24 hours. After incubation, the culture medium was removed, and 200 μL of a medium containing 0.5 mg/mL MTT was added, and then cultured for 4 hours to reduce MTT. After that, the culture medium was removed and formazone crystals produced by adding 100 μL of dimethyl sulfoxide (DMSO) were dissolved, and the absorbance was measured at 540 nm using an ELISA reader (Bio-Rad Co., USA). measured. Cell viability was expressed as a percentage (%) compared to the control.

Figure 2a is a graph relating to the cell viability of the fermented barley radish extract in Experimental Example 2 of the present invention, Figure 2b is a graph relating to the cell viability of the non-fermented barley radish extract in Experimental Example 2 of the present invention, Figure 2c is an experiment of the present invention In Example 2, it is a graph relating to the cell viability of the fermented sulfur extract, and FIG. 2d is a graph relating to the cell viability of the non-fermented sulfur extract in Experimental Example 2 of the present invention.

Referring to Figures 2a to 2d, LPS, the extract prepared in Comparative Examples 2 (NFYM), 4 (NHJ) and 5 (NFHJ) prepared in the above example, LPS, the extract prepared in the above Example As a result of treatment by concentration, when 100 ng/mL of LPS was treated alone, there was no significant difference compared to the control group. In addition, even when treated together, it was confirmed that the cell viability was more than 95% in the 20-200-fold dilution, and there was no effect on the cell viability.

Experimental Example 3

In order to confirm the skin aging improving efficacy of the fermented radish extract (FYM) prepared in the above Example, an MMP-1 activity inhibition test was conducted according to the following experimental method, and the results are shown in FIG. 3 .

Experimental method

Human dermal fibroblasts (HDF) were cultured in a medium supplemented with 10% fetal bovine serum (Sigma) in DMEM (Dulbecco's modified Eagle's media, Sigma) at 37°C and 5% CO2 conditions. . After culturing the cells at a concentration of 2X10 ⁵ cells/well in a 24-well plate and confirming cell adhesion, the control group was treated with nothing and only a solvent was added, and the sample prepared in Example 10, 25, 50 was added to the experimental group. , treated to a concentration of 100 μg/mL, treated with retinol to a concentration of 10 μg/ml in the control group, and cultured for 2 days. After 2 days, the medium was collected and the inhibitory ability of MMP-1 was measured using Matrix Metalloproteinase-1 (MMP-1), Human, biotrak, and ELISA kit (GE healthcare RPN2610).

Figure 3 is a view showing the skin aging improvement efficacy of the extract fermented barley radish in Experimental Example 3 of the present invention.

Referring to FIG. 3, it was confirmed that the expression level of MMP-1 was decreased in most of the experimental groups compared to the control group, retinol, and it was confirmed that there was a difference depending on the combination and composition of functional raw materials in the composition even within the experimental group. When the fermented barley radish extract (Example) was pretreated at a concentration of 25, 50, and 100 μg/mL for 24 hours, the protein expression level of MMP-1 by UV was measured by western blot. It was confirmed that the MMP-1 protein expression was inhibited from the concentration. Therefore, it can be confirmed that, in the case of the fermented barley radish extract (Example), wrinkle generation due to the reduction of collagen by UV can be prevented by inhibiting the degradation of collagen even at a low concentration.

Experimental Example 4

In order to confirm the anti-inflammatory efficacy of the extracts prepared in the fermented yarrow radish extract (FYM), Comparative Examples 2 (NFYM), 4 (FHJ) and 5 (NFHJ) prepared in the above Example, the experiment was carried out according to the following experimental method. , the results are shown in Figs. 4a to 4d.

Experimental method

The change after administration of ethanol extract and ferment extract for inhibition of nitric oxide (NO) production, known as an important factor in the recent inflammatory response, was confirmed. To measure the inhibition of NO production in LPS-activated RAW 2647 cells, samples were treated at different concentrations and treated with LPS (100 ng/mL) for 24 h. After that, NO was measured using the griess reagent system, and the cell culture supernatant and griess reagent were mixed 1:1 in a 96-well plate and reacted for 10 minutes, followed by an ELISA microplate reader (Infinite 200 pro, TECAN). , Mnnedrf, Switzerland) was used to measure the absorbance at 540 nm.

Figure 4a is a graph relating to the inhibition of nitrogen monoxide production of the fermented barley radish extract in Experimental Example 4 of the present invention, Figure 4b is a graph relating to the inhibition of nitrogen monoxide production of the non-fermented barley radish extract in Experimental Example 4 of the present invention, Figure 4c is the present invention A graph on the inhibition of nitrogen monoxide production in the fermented sulfur extract in Experimental Example 4, FIG. 4D is a graph on the inhibition of nitrogen monoxide production in the non-fermented sulfur extract in Experimental Example 4 of the present invention.

4A to 4D, it was confirmed that NO production was inhibited in a concentration-dependent manner in all the experimental groups, and it can be confirmed that the fermented adil radish extract (Example) exhibits the best efficacy.

As such, the fermented barley radish extract prepared according to the present invention and the cosmetic composition containing the same are fermented using volcanic clusters, so that the fermentation odor generated during fermentation is not removed, so there is no objection to use on the skin, and contained in the volcanic clusters. Nutrients are contained in the fermented adil radish extract, so that the efficacy contained in the adil radish extract can be improved, and it is possible to provide a fermented adil radish extract having anti-wrinkle and antioxidant effects and a cosmetic composition containing the same.

The preferred embodiment of the present invention has been described in detail above. The description of the present invention is for illustration, and those skilled in the art to which the present invention pertains will understand that other specific forms can be easily modified without changing the technical spirit or essential features of the present invention.

Accordingly, the scope of the present invention is indicated by the claims described below rather than the above detailed description, and all changes or modifications derived from the meaning, scope, and equivalent concept of the claims are included in the scope of the present invention. should be interpreted

Claims (8)

delete As a fermented barley radish extract comprising a barley radish extract and volcanic clusters,
The fermented barley radish extract is fermented in a medium containing the volcanic cluster by inoculating the barley radish extract into microorganisms,
The microorganism is Bacillus subtilis Apep Microorganism Jeju 170201 strain, characterized in that the extract fermented barley radish. delete 3. The method of claim 2,
The medium containing the volcanic clusters is a fermented barley radish extract, characterized in that 0.1 to 10% by weight of the volcanic clusters compared to the total medium composition. delete A cosmetic composition, characterized in that it contains 0.1 to 90% by weight of the fermented barley radish extract of any one of claims 2 and 4 based on the total cosmetic composition. 7. The method of claim 6,
The cosmetic composition is a cosmetic composition, characterized in that for wrinkle improvement and antioxidant. 7. The method of claim 6,
The cosmetic composition is a softening lotion, astringent lotion, nourishing lotion, eye cream, serum, nourishing cream, massage cream, cleansing cream, cleansing lotion, cleansing foam, cleansing water, powder, essence, pack, hair tonic, hair treatment, shampoo Or a cosmetic composition for antioxidant and inflammation relief, characterized in that it has a rinse formulation.

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