KR101924362B1 - Composition for Improving Skin Wrinkles Using an Extract of Germinated Barley - Google Patents

Abstract

The present invention relates to a composition for alleviating skin wrinkles using a germinated barley extract. The composition comprises, as an active ingredient, an extract of a germinated black barley cultivated for 72 hours by using a desalinized lava seawater having the hardness of 400 as cultivation water having an inhibitory activity for collagenase production.

Description

Technical Field [0001] The present invention relates to a composition for improving skin wrinkles using germinated barley extract,

The present invention relates to a composition for improving skin wrinkles using germinated barley extract.

The skin occupies about 16% of the human body and is directly in contact with the external environment and plays an important role in protecting the human body from external harmful factors such as temperature, humidity and ultraviolet rays. The skin consists of epidermis, dermis and subcutaneous tissue under keratin skin. The epidermis is composed of keratinocytes mainly composed of keratinocytes, a thin protective layer composed of melanocytes producing melanin and secreted melanocytes, immune cells such as langerhans cells and Merkel's copuscles To prevent external stimuli and pathogens from infestation, to control body temperature, to maintain moisture and lipid components.

The dermis is a connective tissue beneath the epidermis, mostly composed of a macromolecular network called the extracellular matrix. This extracellular epilepsy is made up of fibroblasts and consists of fibrous proteins such as collagen, elastin and polysaccharides such as hyaluronic acid.

As a fiber component, collagen is the most important protein that maintains skin moisture and maintains skin flexibility and elasticity. It gives strength and tension to the skin, protects skin from external stimuli, occupies 90% of the dermis, (J Am Acad Dermatol, 17 (4): 610-613, 2001), which accounts for 3-4% of the dermal layer and is an important factor in the elasticity of the skin as well as collagen. Most of the collagen is Type I collagen, which is synthesized by fibroblasts and degraded by collagenases such as MMP-1. Examples of the polysaccharide include high molecular substances such as hyaluronic acid, mucopolysaccharide, and proteoclycan, which have strong water retention capability.

 When exposed to aging or ultraviolet rays, the action of fibroblasts and the number of cells decrease, the amount of collagen synthesis decreases, and the action of collagenase such as collagen degradation increases, thereby losing skin moisture and reducing skin flexibility and elasticity.

Currently, the effects of retinoic acid and retinol (J Invest Dermatol 98: 248-260, 1992; Elaine S et al., J Invest Dermatol 96: 975-978, 1991) on retinoic acid and retinol , And dehydroepiandrosterone (Shin MH et al., J Invest Dermatol 124: 315-323, 2005), ginsenoside Rg3 (Kim SW et al., J Soc Cosmet Scientists (Korean Patent No. 101008833), Yeast Extract (Korea Patent No. 100825450), Flavor (Francesco B et al., Int J Pharm 145: 87-94, And wrinkle improvement and skin protection effect of various materials were found. However, some substances have limited use due to low stability or skin irritation resulting in safety problems. Accordingly, the search for excellent wrinkle improving activity and safety has been continued.

The present invention has been completed based on the barley extract germinated in lava sea water based on collagenase inhibitory activity of MMP-1 production and hyaluronic acid production promoting activity having strong water retention ability.

It is an object of the present invention to provide a composition for improving wrinkles using a barley extract germinated in lava sea water.

Other and further objects of the present invention will be described below.

The inventors of the present invention found that germinated barley extract cultivated for 24 to 96 hours by using normal water or desalted lava waters having a water hardness in the range of 100 to 400 as the cultivation waters, Production of collagenase, MMP-1, in human fibroblasts stimulated by UV, according to the hardness of cultured water and cultivation period and in the case of desalted lava sea water, without cytotoxicity against fibroblasts and human keratinocytes And promoted the production of hyaluronic acid in human keratinocytes. Specifically, (a) germinated barley extract cultivated for 72 hours using plain water as cultivation water, (b) germinated barley extract cultivated for 48 hours using desalted lava waters having a hardness of 100 as cultivation water, (c) (D) germinated barley extract cultured for 96 hours using desalted lime waters with a hardness of 200 as a cultivation wastewater, (e) 400 desalted lava waters with a hardness of 400 And (f) cultivation water for 48 hrs., And cultivation for 72 hrs using desalted seawater having a hardness of 400, inhibited the production of collagenase MMP-1. (I) (Ii) germinated barley extracts cultivated for 96 hours using normal water as cultivation water, (iii) desalted seawater having a hardness of 200 and cultivating water was used (Iv) germinated barley extract grown for 48 hours, (iv) germinated barley extract cultivated for 72 hours using desalted lava waters having a hardness of 200 as cultivation water, (v) germinated cultivated for 96 hours with 200 mW of desalted lava water Barley extracts and (vi) Growth of barley extracts grown for 24 to 72 hours using 400 desalted lava waters with a hardness of 400 stimulated hyaluronic acid production.

In view of the above, the present invention provides a method for inhibiting the production of hyaluronic acid, comprising at least one of the germinated barley extracts of (a) to (f) having an activity of inhibiting the production of collagenase, ) To (vi) as a wrinkle-improving composition containing at least one of the extracts of the present invention as an active ingredient.

As used herein, "germinated barley" refers to germinated barley ( Hordeum ) obtained by hydroponic cultivation using general water (water) or desalted lava waters as cultivation water vulgare ), in particular germinated black barley (anthocyanin content of which is markedly higher in normal barley and the seed surface is black). When obtaining such germinated barley, the cultivated water is generally supplied to the plant in a sprinkling manner during the cultivation period and is replaced in order to prevent germination and growth of barley caused by propagation of microorganisms. The supply / replacement cycle of the cultivated water may range from 6 hours to 12 hours.

As used herein, the term "germinated barley extract" refers to an extract of germinated barley, particularly germinated barley, which is a target to be extracted, with water, lower alcohols having 1 to 4 carbon atoms (methanol, ethanol, butanol), methylene chloride, ethylene, acetone, hexane, , Extracts obtained by leaching using ethyl acetate, butyl acetate, N, N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), 1,3-butylene glycol, propylene glycol or a mixed solvent thereof, And extraction with supercritical extraction solvent such as pentane, or a fraction obtained by fractionating the extract. The extraction method is a method of extracting a supernatant from a supercritical fluid, a supercritical fluid, or a supercritical fluid in consideration of the polarity, extraction degree, Any method such as extraction can be applied. In the case of the fractionated extract, a fraction obtained by suspending the extract in a specific solvent and mixing and leaving with a solvent having a different polarity, the crude extract is adsorbed on a column packed with silica gel, and then a hydrophobic solvent, a hydrophilic solvent, Quot; means fractions obtained as a mobile phase. Also, the meaning of the extract includes a concentrated liquid extract or a solid extract in which the extraction solvent is removed by a method such as freeze drying, vacuum drying, hot air drying, spray drying and the like. Preferably an extract obtained by using water, ethanol or a mixed solvent thereof as an extraction solvent.

In this specification, " lava sea water " is defined in Korean Patent No. 0853244 (Application No.: 10-2008-0027861), entitled " Method for producing mineral composition and mineral composition obtained by the method & It is synonymous with the "saltwater underground in the eastern part of Jeju Island" as described. Specifically, "eastern part of Jeju Island" means Jeju-eup, Seongsan-eup and Seokseon-myeon in Jeju-do in administrative district, and "saltwater groundwater" means groundwater containing salinity of a certain amount or more. Specifically, According to the criteria for classification of groundwater in Jeju Island based on the salinity concentration used in the study of the hydrological geology in the eastern part of Cheju Island (Ⅱ), 2003, Kyungwon University, considered as part of this specification), low salt groundwater (electric conductivity of 1,700 ㎲ / cm (17,350 / / cm) and saltwater groundwater (electrical conductivity of 17,350 / / cm or more), but preferably the electrical conductivity is 17,350 / / cm or more or the salt concentration is 30 ‰ ) (Usually, the saline concentration of the seawater is 32 to 35 ‰)

In this specification, " desalted lava sea water " means the result obtained by desalting lava sea water. Desalination of lava seawater can be performed by applying any technique known in the art, such as reverse osmosis, ion exchange resin, electrodialysis, and the like.

In the present specification, the term " active ingredient " alone means an ingredient which exhibits the desired activity or which can exhibit activity together with a carrier which is not itself active.

The composition of the present invention may contain any amount (effective amount) of the active ingredient as long as it can exhibit wrinkle-improving activity and the like which is intended to be treated depending on the purpose of use, formulation, compounding purpose and the like. And will be determined within the range of from 0.001% by weight to 15% by weight, based on the total weight of the composition. Herein, " effective amount " means a dermatologic / pharmacological effect, such as a wrinkle-reducing effect, when the composition of the present invention is administered to a mammal, preferably a human, Refers to the amount of active ingredient contained in the composition of the present invention. Such effective amounts can be determined experimentally within the ordinary skill of those skilled in the art.

The composition of the present invention may contain, in addition to the active ingredient, an addition of similar activities such as enhancement or enhancement of skin wrinkle reducing effect or skin irritation inhibiting activity, skin protecting activity (skin whitening, skin damage suppression by ultraviolet rays, May further comprise any compound or natural extract known to be safe in the art and known to have a corresponding activity in order to improve the dosage, ingestion, and ease of use. These compounds or extracts include the national pharmacopoeia (Korean pharmacopoeia), the national health functional food circulation in Korea (in Korea, the health functional food standard and specification), the functional cosmetics circulation in each country (in Korea, Compounds or extracts listed in the official documents such as "Functional Cosmetics Standards and Test Methods"), compounds or extracts approved by the respective countries in accordance with the laws of the respective countries (in Korea, "Pharmaceutical Affairs Law") regulating the manufacture and sale of pharmaceuticals In accordance with the laws of each country that regulate the manufacture and sale of compounds or extracts and functional cosmetics that are recognized as functioning in accordance with the laws of the respective countries ("Act on Health Functional Foods" in Korea) that regulate the manufacture and sale of functional foods &Quot;).≪ / RTI > Examples of skin whitening ingredients include arbutin, niacinamide, ascorbyl glucoside, alpha-bisabolol and oil soluble licorice (Glycyrrhiza) extract. Examples of the skin wrinkle improving agent include retinol, retinyl palmitate , Adenosine, polyethoxylated amide, and the like. Examples of the ultraviolet protecting component include drometrizol, drometrizol trisiloxane, dipalayyltriolate, Dimethicyldecibenzalmalonate, diethylhexylbutadimido Triazone, and pine bark extract, phosphatidylserine, finger root extract powder, and complex extracts of red ginseng and mountain juice. Examples of the moisturizing component include AP collagen enzyme digesting peptide, collactive collagen peptide, N-acetyl glucosamine, konjac potato extract, dandelion extract, rice bran extract, corn germ extract, low molecular weight collagen peptide, ground extract powder, phosphatidylserine, hyaluronic acid Examples of functional ingredients for improving 'irritable skin condition' include L. sakei Probio 65, gamma linolenic acid-containing oil, L. plantarum CJLP133, and probiotic ATP. Such compounds or natural extracts may be included in the compositions of the present invention in combination with one or more of their active ingredients.

In a specific embodiment, the composition of the present invention can be identified as a food composition.

The food composition of the present invention can be produced in any form and can be used in various forms such as beverages such as tea, juice, carbonated drink, ionic drink, processed oil such as milk and yogurt, gum, rice cake, Korean confectionery, A food, a health food, a food, a tablet, a capsule, a ring, a granule, a liquid, a powder, a slice, a paste, a syrup, a gel, a jelly and a bar.

In addition, the food composition of the present invention may be classified into any product category as long as it meets the laws and regulations on the time of manufacture and distribution in the legal and functional category. For example, it is a health functional food according to the 「Health Functional Food Act」 in Korea, or a food functional food according to the Korean Food Sanitation Law (Food Standards and Specifications) , Special-purpose food, and the like.

The food composition of the present invention may contain food additives in addition to the active ingredients thereof. Food additives are generally understood to be substances that are added to foods and mixed or infiltrated into food in the manufacture, processing or preservation of food, and their safety must be ensured since they are ingested daily with food and for long periods of time. In food additives according to the laws of the respective countries ("Food Sanitation Act" in Korea) regulating the manufacture and distribution of food, food additives with safety are specified in terms of ingredient or function. In the Food Additives Code of Korea (Food Additives Standards and Standards), the food additives are classified into chemical compounds, natural additives and mixed preparations in terms of ingredients. Such food additives are classified into sweeteners, flavors Preservatives, emulsifiers, acidulants, and thickeners.

The sweetener is used for imparting a sweet taste suitable for foods, and both natural and synthetic sweeteners can be used in the composition of the present invention. Preferably, natural sweeteners are used. Examples of natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose and maltose.

Flavors may be used to enhance taste or flavor, both natural and synthetic. Preferably, a natural one is used. When using natural ones, the purpose of nutritional fortification can be performed in addition to the flavor. Examples of natural flavoring agents include those obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or those obtained from green tea leaves, Asiatica, Daegu, Cinnamon, Chrysanthemum leaves and Jasmine. Also, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, banks and the like can be used. The natural flavoring agent may be a liquid concentrate or a solid form of extract. Synthetic flavors may be used depending on the case, and synthetic flavors such as esters, alcohols, aldehydes, terpenes and the like may be used.

As the preservative, calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate and EDTA (ethylenediaminetetraacetic acid) can be used. As the emulsifier, acacia gum, carboxymethyl cellulose, Pectin and the like. As the acidulant, math, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid and the like can be used. The acidulant may be added so that the food composition has a proper acidity for the purpose of inhibiting the growth of microorganisms other than the purpose of enhancing the taste.

Examples of the thickening agent include suspending agents, sedimentation agents, gel-forming agents, bulking agents and the like.

The food composition of the present invention may contain physiologically active substances or minerals which are known in the art and which are stable as a food additive in addition to the above-mentioned food additives in order to supplement and supplement functional and nutritional properties.

Examples of such physiologically active substances include catechins contained in green tea and the like, vitamins such as vitamin B1, vitamin C, vitamin E and vitamin B12, tocopherol, dibenzoyl thiamine, etc. Examples of minerals include calcium preparations such as calcium citrate, magnesium stearate , Iron preparations such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc and the like.

The food composition of the present invention may contain an appropriate amount of the above-mentioned food additives according to the product type so as to achieve the purpose of addition thereof.

With regard to other food additives that may be included in the food composition of the present invention, reference may be made to the Food Code of the respective countries or the Food Additives Code.

In another specific embodiment, the composition of the present invention can be identified as a pharmaceutical composition.

The pharmaceutical composition of the present invention may be prepared into oral formulations or parenteral formulations according to the route of administration by conventional methods known in the art, including pharmaceutically acceptable carriers in addition to the active ingredient. Where the route of administration may be any suitable route including local routes, oral routes, intravenous routes, intramuscular routes, and direct absorption through mucosal tissues, and combinations of two or more routes may be used. An example of a combination of two or more routes is a combination of two or more formulations of the drug according to the route of administration, for example, one drug is administered intravenously and another drug is administered via a local route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration and formulation, and specific reference may be made to the pharmacopoeia of each country, including the " Korean Pharmacopoeia ".

When the pharmaceutical composition of the present invention is prepared into an oral formulation, it may be formulated into powder, granules, tablets, pills, sugar tablets, capsules, solutions, gels, syrups, suspensions, wafers And the like. Examples of suitable carriers include starches such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol and xylitol, corn starch, potato starch and wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Hydroxypropylmethylcellulose and the like; polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol Serol, and the like. In case of formulation, suitable binders, lubricants, disintegrants, coloring agents, diluents and the like may be included as needed. Examples of suitable binders include starch, magnesium aluminum silicate, starch pellets, gelatin, methyl cellulose, sodium carboxymethyl cellulose, polyvinyl pyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax and the like. Examples of the disintegrating agent include starch, methylcellulose, magnesium stearate, magnesium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, Agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt, and the like. Examples of the diluent include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine and the like.

When the pharmaceutical composition of the present invention is prepared into a parenteral dosage form, it may be formulated in the form of an injection, transdermal drug delivery, nasal aspirate and suppository together with a suitable carrier according to methods known in the art. As the carrier suitable for injection preparation, aqueous isotonic solutions or suspensions may be used. Specifically, PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, and isotonic solution such as 5% dextrose may be used . When formulated with a transdermal preparation, it can be formulated in the form of ointments, creams, lotions, gels, external liquids, pastes, liniments, and air-lozenges. Nasal inhalers may be formulated in the form of aerosol sprays using suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc., and when formulated as a suppository, witepsol, tween 61, polyethylene glycols, cacao butter, laurin, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, and sorbitan fatty acid esters.

The formulation of pharmaceutical compositions is well known in the art and can be found, for example, in Remington ' s Pharmaceutical Sciences (19th ed., 1995). This document is considered part of this specification.

The preferred dosage of the pharmaceutical composition of the present invention is 0.001 mg / kg to 10 g / kg per day, preferably 0.001 mg / kg to 1 g / day, depending on the patient's condition, body weight, sex, age, / kg < / RTI > The administration can be carried out once or several times a day. Such dosages should in no way be construed as limiting the scope of the invention.

In another specific embodiment, the composition of the present invention can be identified as a cosmetic composition.

Even when the composition of the present invention is identified as a cosmetic composition, the cosmetic composition may be classified into any product category according to the use of the cosmetic composition. Specifically, the cosmetic composition may contain functional cosmetic products having applications such as improvement of skin troubles and improvement of atopic dermatitis, General cosmetics, and the like. Specific examples of the product form include a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing oil, powder foundation, emulsion foundation, wax Foundation, spray, and the like. In a specific product form, it may be a form of flexible lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

The cosmetic composition of the present invention may contain, in addition to its active ingredient, conventional additives such as stabilizers, solubilizing agents, surfactants, vitamins, colorants and antioxidants, and carriers commonly used in cosmetic compositions.

When the formulation of the present invention is a paste, a cream or a gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may 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 component. In the case of a spray, in particular, / Propane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component. Specifically, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid esters of sorbitan, and the like.

When the formulation of the present invention is a suspension, a carrier, such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component is selected from aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

The cosmetic composition of the present invention can be produced according to a method for producing a cosmetic composition which is usually carried out in the art, except that it contains the effective ingredient showing skin wrinkle improving activity.

As described above, according to the present invention, it is possible to provide a composition for improving skin wrinkles using germinated barley extract.

The composition of the present invention can be commercialized as food or cosmetics.

Fig. 1 shows the results of experiments for inhibiting MMP-1 production.
Fig. 2 shows the results of experiments for promoting hyaluronic acid (HA) production.

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

<Example> Preparation of germinated black barley extract

Example 1: Production of germinated black barley

The seeds of black barley were immersed in purified water for 3 hours at a temperature of 22 to 25 ° C and a humidity of 40 to 60% in a plant growing machine. Then, at 9:00 am and 7:00 pm daily, deionized sea water Mineral water) was cultivated for 24 hours to 96 hours to prevent the growth of microorganisms. The grown black barley was dried in a hot air drier at 40 ° C for 15 hours and then pulverized using a dry mill to be used for extraction.

Example 2: Preparation of germinated black barley extract

The germinated black barley powder obtained in Example 1 was added with 10 times the weight of water, and the mixture was extracted for 24 hours while maintaining the temperature at 80-100 ° C. After filtration, the filtrate was concentrated under reduced pressure and lyophilized to obtain a germinated black barley extract .

<Experimental Example> Experiment on wrinkle improving activity of germinated black barley extract

1. Experimental Method

1.1 Cell culture

(100 IU / mL), streptomycin (100 μg / mL), Supplement (insulin 0.1%, rhFGF-B 0.1%, GA- 1000, 0.1% FBS, 2% FBS), and cultured in an incubator containing 5% carbon dioxide at 37 ° C.

HaCaT, a human keratinocyte, was cultured in an incubator containing DMEM (Dulbecco's Modified Eagle's Medium) containing 10% fetal bovine serum (FBS) at 37 ° C and 5% carbon dioxide.

1.2 Cell toxicity experiments

HaCaT cells were plated at 1.5 × 10 ⁴ cells / well and 0.6 × 10 ⁴ cells / well in 96-well plates at 37 ° C and 5% CO 2 for 24 h. ₂ And cultured in a cell incubator. After 24 hours, the medium was discarded and washed with PBS. Then, HaCaT was transformed into DMEM medium (serum-free medium) containing no FBS, human fibroblasts were transformed into starvation using FBM medium without supplement, The next day, samples were treated at a constant concentration and the cells were cultured for 24 hours. The WST-1 reagent diluted 10-fold in the medium was added to each well and incubated for 2 hours. Absorbance was measured at 450 nm with an ELISA reader. Cell viability was determined as percentage compared to negative control group without sample treatment.

1.3 Quantification of MMP-1

HaCaT cells were plated at 2.5 × 10 ⁵ cells / well in a 6-well plate. Cells were plated at 37 ° C in 5% CO ₂ And cultured in a cell incubator. After 24 hours, the medium was discarded, and the cells were washed with PBS. Then, cells were frozen using DMEM medium (serum-free medium) containing no FBS, and cultured the next day with UV treatment. Human fibroblasts (NHF) were dispensed in 96-well plates at a density of 0.6 × 10 ⁴ cells / well and cultured in a 5% CO ₂ cell incubator at 37 ° C for 24 h. After 24 hours, the cells were turned into a starved state using an FBM medium containing no supplement, and the culture medium of HaCaT, which was UV-stimulated the next day, was treated with human fibroblasts and cultured. EGCG (epigallocatechin gallate) and RA (retinoic acid) were used as positive control. After the experiment using the MMP-1 ELISA kit (R & D System, DY901) according to the manufacturer's protocol, the absorbance at 450 nm was measured with an ELISA reader. The amount of MMP-1 was calculated as a percentage by comparison with the negative control group without sample treatment.

1.4 Quantification of HA (Hyaluronic acid)

HaCaT cells were seeded at 1.5 × 10 ⁴ cells / well in a 96-well plate and cultured in a 5% CO ₂ cell incubator at 37 ° C for 24 h. After 24 hours, the cells were turned into a starvation state using DMEM medium (serum-free medium) not containing FBS, and then the samples were cultured for the next day. RA (retinoic acid) was used as a positive control at this time. After the experiment using the HA ELISA kit (R & D System, DY3614) according to the manufacturer's protocol, the absorbance at 450 nm was measured with an ELISA reader. The amount of HA was calculated as a percentage compared to the negative control group treated with the sample.

2. Experimental results

2.1 Cytotoxicity to HaCaT and NHF cells

The cytotoxicity results for HaCaT and NHF cells are shown in [Table 1] and [Table 2] below, respectively.

The germinated black barley extract did not show specific cytotoxicity on these cells regardless of the germination time and the cultivation water used.

2.2 Quantification of MMP-1

MMP-1 quantification results are shown in [Table 3] and [Figure 1] below. The MMP-1 production was significantly increased 4.4 times in the UV treated group compared to the untreated group, and the inhibitory activity on MMP-1 production of the germinated black barley extract obtained by germination for 72 hours using 400 desalted lava waters with a hardness of 400 , Followed by germinated black barley extracts obtained by germination for 48 hours using 400 desalted lava waters as cultivation water.

2.3 HA quantitation

The HA quantification results are shown in [Table 4] and [Figure 2] below. The growth promoting activity of the germinated black barley extract obtained by germination for 72 hours using 400 desalted lava waters as the cultivation water was the best. Next, the germinated black barley extracts obtained by germination for 96 hours using 400 desalted lava waters with a hardness of 400 were excellent in activity.

Claims (8)

A food composition for improving wrinkles comprising an extract of germinated black barley cultivated for 72 hours by using desalted lava waters having a hardness of 400 and cultivation water having an activity of inhibiting the production of collagenase as an active ingredient.
The method according to claim 1,
Wherein the extract is an extract of water, ethanol or a mixed solvent thereof.
A cosmetic composition comprising germinated black barley extract cultivated for 72 hours using desalted lava waters having a hardness of 400 as cultivation water having an activity of inhibiting the production of collagenase as an active ingredient.
The method of claim 3,
Wherein the extract is an extract of water, ethanol or a mixed solvent thereof.
A food composition for improving wrinkles comprising at least one of the following germinated black barley extracts (i) and (ii) having hyaluronic acid production promoting activity:
(i) germinated black barley extract cultivated for 48 hours using desalted lava waters having a hardness of 200 and cultivation water, and
(ii) Extracts of germinated black barley cultivated for 24 to 72 hours using desalted lava waters having a hardness of 400 as cultivation water.
6. The method of claim 5,
Wherein the extract is an extract of water, ethanol or a mixed solvent thereof.
A cosmetic composition for improving wrinkles comprising at least one of the following germinated black barley extracts (i) and (ii) having hyaluronic acid production promoting activity:
(i) germinated black barley extract cultivated for 48 hours using desalted lava waters having a hardness of 200 and cultivation water, and
(ii) Extracts of germinated black barley cultivated for 24 to 72 hours using desalted lava waters having a hardness of 400 as cultivation water.
8. The method of claim 7,
Wherein the extract is an extract of water, ethanol or a mixed solvent thereof.

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