KR102110902B1 - Composition for anti-oxidation, anti-inflammation, anti-diabetes, whitening, anti-wrinkle, or anti-gout comprising extract of Ruby-S apple peel - Google Patents

Abstract

The present invention relates to a composition for antioxidant, anti-inflammatory, anti-diabetes, whitening, wrinkle improvement, and gout suppression, which includes a ruby apple extract as an active ingredient, Ruby-S, a new breeding variety of the present invention. Malus pumila Mill.) Apple peel extract has an antioxidant effect by inhibiting the DPPH, ABTS, and TBARs radicals and increasing PF, and inhibiting tyrosinase to suppress melanin production, thereby exhibiting a skin whitening effect. In addition, by inhibiting elastase activity, the elastin molecule form is maintained, collagenase activity is inhibited, wrinkle improvement effect is exhibited, and hyaluronidase activity is inhibited, thereby suppressing inflammatory reactions. In addition, the ruby apple peel extract exhibits an inhibitory effect on gout (arthritis) by inhibiting the activity of the xanthin oxidase enzyme, and a pharmaceutical composition showing an inhibitory effect on type 2 diabetes by inhibiting the enzyme activities of α-amylase and α-glucosidase. It can be usefully used as a health functional food composition, and a cosmetic composition.

Description

Antioxidant, anti-inflammatory, anti-diabetic, whitening, anti-wrinkle, or gout inhibiting composition containing ruby apple peel extract as an active ingredient {Composition for anti-oxidation, anti-inflammation, anti-diabetes, whitening, anti-wrinkle, or anti-gout comprising extract of Ruby-S apple peel}

The present invention relates to a composition for antioxidant, anti-inflammatory, anti-diabetes, whitening, wrinkle improvement, or gout suppression, which contains the extract of the apple extract of Ruies as an active ingredient.

Apple, one of Korea's leading fruits ( Malus pumila Mill.) As the income level improves due to economic growth, consumption patterns are changing to qualitative rather than quantitative. 'Fuji', commonly called 'busa'(Busa; Malus domestica Borkh.) is the most representative apple variety, accounting for 29.5% of the total domestic fruit production in 2012, and is mainly exported to Taiwan. About 50% of domestic apples are consumed in the year, and the rest are stored and consumed in the following year. Fuji apples exported after cold storage have been deteriorated in quality, and exports have been very limited since February of the following year. In addition, the recognition and price are much lower than that of Japanese apples, so that Korean apples are still recognized as medium in quality. However, after the Japanese nuclear accident in 2011, as consumer concerns about food safety grew, imports of Japanese agricultural products from Taiwan, Korea, China, and the United States decreased, and import restrictions on Japanese agricultural products were implemented in 37 countries as of April 2014. Is becoming. In China, strict measures are being taken to ban the importation of agri-food from regions that are considered safe in Japan. It was necessary to develop high-quality apples that can be exported to various countries, mainly in China, by taking advantage of the food safety and price competitiveness of Koreans as an opportunity to change foreign trade in agricultural and marine products after the nuclear accident in Japan.

Apples are taxonomically perennial woody plants that are grown in the Northern Hemisphere of Europe, Asia, and North America. In the private sector, apples are used as medicines for beauty, vomiting, hookworm, sperm, and constipation. One of the types of apples, eungeum, governs cows, gwakran, and abdominal pain, removes phlegm, and treats dysentery. The undercooked persimmon has a strong taste but is used as a medicine. However, if you eat a lot, you get a lot of sleep, a phlegm occurs, and a boil occurs. Cider wine made by fermenting apples has a unique flavor and is a drink that helps digestion after eating. According to local medicine, if you make apple cider vinegar by oxidizing fermentation a little more and apply it to burns, it will stop the pain immediately, and if you apply apple vinegar to varicose veins on the affected area day and night, the topic will be much smaller after 1 month. Apples prove their effectiveness not only through folk remedies, but also through research. Apple fruit has good organoleptic properties and nutritionally contains physiologically active substances such as vitamins, minerals, potassium, sodium, and polyphenolic compounds, in addition to palatable components such as organic acids and sugars. Among them, various polyphenol substances such as procyanidin, chlorogenic acid, caffeic acid, epicatechin, catechin, p-coumaroyl qunic acid, rutin, phloridzin, and quercetin are skin aging and obesity caused by oxidative damage of biological components by free radicals. , Inflammation, arteriosclerosis, diabetes, hypertension, and other cancers, it is known to be effective in preventing degenerative diseases (Yun HJ, Lim SY, Hur JM, Jeong JW, Yang SH, Kim DH. 2007. Korean J Food Preserv 14: 239-246, Hwang IW, Kim CS, Chung SK. 2011. Korean J Food Preserv 18: 700-705 .).

Physiologically active substances are widely distributed in animals and plants, and plants contain polyphenol-based antioxidants in cells to protect themselves from UV-induced oxidation, auto-oxidation, and damage from insects. Flavonoids and acids Phenolic compounds are known to have various physiological activities such as antioxidant, anti-allergy, anti-diabetes, anti-cancer, anti-inflammatory, whitening, wrinkle improvement. Recently, many researches and developments have been made in the field of functional foods and functional cosmetics for physiological activity and excellent functional substances derived from proven medicinal plant resources, and search for medicinal parts such as medicinal plants, wild vegetables, herbal medicines, fruits, etc. Therefore, it can be said that developing as a health functional food or food supplementary material is also very meaningful in terms of efficient use of natural resources and contribution to the development of new food resources and new materials.

On the other hand, Rubies (Ruby-S; Malus pumila Mill.) Apples are new breeds that were nurtured by breeding 'Sansa' varieties in Japanese varieties 'Alps Autome' varieties, ripening in late August and weighing 90g (usually Apple size is 270g ~ 300g) and is only the size of a table tennis ball. Alps Autome, a small apple that has been distributed in the domestic market, has a lot of fruits and ripened fruits at the time of ripening, and has a short taste, but the ruby developed by supplementing this has a fruit size of 50 g compared to Alps Otome. Because it is much larger, it has more portions to eat, and the sugar content is high and the acidity is low (13.9 ° Brix sugar, 0.49% acidity), so it tastes better. It is particularly resistant to anthracnose and has few fruits before harvest.

Accordingly, the present inventors attempted to develop a high value-added functional material through analysis of the effects of antioxidant, anti-inflammatory, anti-diabetic, whitening, wrinkle improvement, and pain suppression for apples of the new breed of ruby varieties bred in Korea.

The present inventors confirmed that apples of the new breed of ruby varieties bred in Korea have superior antioxidant, anti-inflammatory, anti-diabetic, whitening, wrinkle improvement, and pain-suppressing effects compared to adjuvant varieties, especially in the peel extract of ruby apples. The present invention was completed by discovering the most remarkable effect.

In order to achieve the above object, the present invention is an antioxidant for containing a ruby apple peel extract as an active ingredient; Anti-inflammatory; For skin whitening; Wrinkle improvement; Or it provides a pharmaceutical composition for the prevention or treatment of gout or diabetes.

In addition, the present invention is an antioxidant for containing the extract of the apples of the ruby as an active ingredient; Anti-inflammatory; For skin whitening; Wrinkle improvement; Or to provide a health functional food composition for preventing or improving gout or diabetes.

In addition, the present invention provides a cosmetic composition for anti-oxidation, anti-inflammatory, skin whitening, or wrinkle improvement, which includes the extract of the apple extract of Ruies as an active ingredient.

In one embodiment of the present invention, the composition is 1,1-diphenyl-2-picrylhydrazyl (1,1-diphenyl-2-picrylhydrazyl; DPPH) radical inhibition, 2,2'-azinobis- (3 -Ethylbenzothiazoline-6-sulphonic acid) (2,2'-Azinobis- (3-ethylbenzothiazoline-6-sulfonic acid); ABTS) radical inhibition, Thiobarbituric acid reactive substances (TBARs) inhibition Alternatively, an antioxidant protection factor (PF) may be increased.

In another embodiment of the present invention, the composition may inhibit hyaluronidase.

In another embodiment of the present invention, the composition may inhibit tyrosinase.

In another embodiment of the invention, the composition may inhibit elastase (elastase) or collagenase (collagenase).

In another embodiment of the present invention, the composition may inhibit xanthine oxidase.

In another embodiment of the present invention, the composition may inhibit α-amylase (α-Amylase) or α-glucosidase (α-Glucosidase).

In another embodiment of the present invention, the extract of the apple of the ruby is water, C ₁ to C ₄ lower alcohol, n-hexane, ethyl acetate, acetone, butyl acetate, 1,3-butylene glycol, methylene chloride, And it may be extracted with a solvent selected from the group consisting of these mixed solvents.

In addition, the present invention is an antioxidant comprising the step of administering the composition to the subject; Anti-inflammatory; Skin whitening; wrinkle improvement; Or provide methods for preventing or treating gout or diabetes.

In addition, the present invention is the antioxidant of the composition; Anti-inflammatory; Skin whitening; wrinkle improvement; Or provides prevention or treatment for gout or diabetes.

The new growth breed of the present invention, Ruby-S ( Malus pumila Mill.) Apple peel extract has antioxidant effects by inhibiting DPPH, ABTS, and TBARs radicals and increasing PF, and inhibiting melanin production by inhibiting tyrosinase. Skin whitening effect. In addition, by inhibiting elastase activity, the elastin molecule form is maintained, collagenase activity is inhibited, wrinkle improvement effect is exhibited, and hyaluronidase activity is inhibited, thereby suppressing inflammatory reactions. In addition, the extract of the apple peel of Rubyis exhibits an inhibitory effect on gout (arthritis) by inhibiting the activity of the xanthin oxidase enzyme, and an inhibitory effect on type 2 diabetes by inhibiting the enzyme activities of α-amylase and α-glucosidase.

Therefore, it is expected that the peel extract of the ruby apple according to the present invention can be usefully used as a pharmaceutical composition, a health functional food composition, and a cosmetic composition.

1 is a view showing the phenolic content in the water and ethanol extract of the peel (peel), fruit (fruits), and whole apples of the ruby according to an embodiment of the present invention.
Figure 2 is a view showing the ABTS radical scavenging ability in the peel, pulp, whole apple extract of Ruies according to an embodiment of the present invention.
Figure 3a is a view showing the results of measuring the respective DPPH activity for the comparison of the physiological activity of the phenolic component and the solid content of the ruby apples according to an embodiment of the present invention.
Figure 3b is a view showing the ABTS radical scavenging ability for comparison of the physiological activity of the phenolic component and the solid content of the ruby apples according to an embodiment of the present invention.
Figure 4a is a view showing the phenolic content in the extract of each solvent type of the ruby apple peel according to an embodiment of the present invention.
Figure 4b is a view showing the phenolic content in the extract according to the ethanol concentration of the ruby apple peel according to an embodiment of the present invention.
Figure 5a is a view showing the results of measuring the PF of each phenolic concentration of water and ethanol extract of the ruby apple peel and adjuvant apple peel according to an embodiment of the present invention.
Figure 5b is a view showing the results of measuring the TBARs for each phenolic concentration of water and ethanol extract of the ruby apple peel and adjuvant apple peel according to an embodiment of the present invention.
6 is a view showing the hyaluronidase inhibitory effect of phenolic concentrations of water and ethanol extracts of the ruby apple peel and the adjuvant apple peel according to an embodiment of the present invention.
7 is a view showing the tyrosinase inhibitory effect of phenolic concentrations of water and ethanol extracts of a ruby apple peel and an adjuvant apple peel according to an embodiment of the present invention.
Figure 8a is a diagram showing the elastase inhibitory effect of each phenolic concentration of water and ethanol extract of the ruby apple peel and adjuvant apple peel according to an embodiment of the present invention.
Figure 8b is a view showing the collagenase inhibitory effect of each phenolic concentration of water and ethanol extract of the ruby apple peel and adjuvant apple peel according to an embodiment of the present invention.
9 is a view showing the inhibitory effect of xanthine oxidase for each phenolic concentration of water and ethanol extracts of ruby apple peel and adjuvant apple peel according to an embodiment of the present invention.
Figure 10a is a view showing the inhibitory effect of α-amylase by phenolic concentration of water and ethanol extract of the ruby apple peel and the adjuvant apple peel according to an embodiment of the present invention.
10B is a view showing the inhibitory effect of α-glucosidase by phenolic concentration of water and ethanol extracts of ruby apple peel and adjuvant apple peel according to an embodiment of the present invention.

The present invention can be applied to various transformations and may have various embodiments, and specific embodiments will be illustrated in the drawings and described in detail in the detailed description. However, this is not intended to limit the present invention to specific embodiments, and should be understood to include all conversions, equivalents, and substitutes included in the spirit and scope of the present invention. In the description of the present invention, when it is determined that detailed descriptions of related well-known technologies may obscure the subject matter of the present invention, detailed descriptions thereof will be omitted.

Hereinafter, the present invention will be described in detail.

The present invention is an antioxidant for containing the extract of the extract of the apples of the apple as an active ingredient; Anti-inflammatory; For skin whitening; Wrinkle improvement; Or it provides a pharmaceutical composition for the prevention or treatment of gout or diabetes.

The term "prevention" as used in the present invention means any action that suppresses the disease or delays the onset by administration of the pharmaceutical composition according to the present invention.

The term "treatment" as used in the present invention means all actions in which symptoms of disease are improved or advantageously changed by administration of a pharmaceutical composition according to the present invention.

In the present invention, the "extract" is an extract obtained by the extraction treatment of the russ apple peel, a diluent or concentrate of the extract, a dried product obtained by drying the extract, a crude or purified product of the extract, or a mixture thereof, etc. , And extracts of all formulations that can be formed using the extracts themselves and the extracts.

In the extract of the russ apple peel of the present invention, the method for extracting the russ apple peel is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method may include a hot water extraction method, an ultrasonic extraction method, a filtration method, a reflux extraction method, etc. These may be performed alone or in combination of two or more methods.

In the present invention, the type of the extraction solvent used to extract the russ apple peel is not particularly limited, and according to a conventional method known in the art for extracting an extract from a natural product, that is, at a normal temperature and pressure. Under conditions, it can be extracted using a conventional solvent. For example, in the present invention, the extract of ruby apple peel is water, lower alcohol having 1 to 4 carbon atoms, n-hexane, ethyl acetate, acetone, butyl acetate, 1,3-butylene glycol, methylene chloride, and mixed solvents thereof It may be extracted using one or more solvents selected from the group consisting of.

The prepared extract can then be filtered or concentrated or dried to remove the solvent, and all filtered, concentrated and dried. For example, filtration may use a filter paper or a vacuum filter, concentration may be performed under a reduced pressure concentrator, drying may be a freeze drying method, but is not limited thereto.

In addition, the present invention is an antioxidant comprising the step of administering the composition to the subject; Anti-inflammatory; Skin whitening; wrinkle improvement; Or provide methods for preventing or treating gout or diabetes.

As used herein, the term "administration" means providing the subject with the composition of the invention in any suitable way.

As used herein, the term "individual" refers to a subject in need of treatment of a disease, and more specifically, human or non-human primate, mouse, dog, cat, horse, and cow, etc. It means mammal.

In addition, the present invention is the antioxidant of the composition; Anti-inflammatory; Skin whitening; wrinkle improvement; Or provides prevention or treatment for gout or diabetes.

In the present invention, "pharmaceutical composition" means that it is prepared for the purpose of preventing or treating a disease, and can be used by being formulated in various forms according to a conventional method. For example, powders, granules, tablets, capsules, suspensions, emulsions, syrups, etc. can be formulated into oral dosage forms, creams, gels, patches, sprays, ointments, warnings, lotions, linen agents, pasta agents, or It can be formulated and used in the form of external preparations such as cataplasma, suppositories, and sterile injectable solutions.

The pharmaceutical composition according to the present invention may further include suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions. At this time, carriers, excipients, and diluents that may be included in the composition include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium Silicates, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include at least one excipient in the extract, for example, starch, calcium carbonate, sucrose ) Or by mixing lactose, gelatin, and the like. In addition, lubricants such as magnesium styrene talc are used in addition to simple excipients. Liquid preparations for oral use include suspensions, intravenous solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients, such as wetting agents, sweeteners, flavoring agents, fragrances, and preservatives, can be included. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used.

The pharmaceutical composition of the present invention may be administered orally or parenterally (eg, intravenously, subcutaneously, intraperitoneally, or topically) according to a desired method, and the dosage may be adjusted according to the patient's condition, weight, and disease. It depends on the degree, drug type, route of administration, and time, but can be appropriately selected by those skilled in the art.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "a pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is the type of patient's disease, severity, activity of the drug, Sensitivity to the drug, time of administration, route of administration and rate of discharge, duration of treatment, factors including co-drugs, and other factors well known in the medical field can be determined. The pharmaceutical composition according to the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, which can be easily determined by those skilled in the art. The administration may be administered once a day, or may be divided into several times.

In addition, the present invention is an antioxidant for containing the extract of the apples of the ruby as an active ingredient; Anti-inflammatory; For skin whitening; Wrinkle improvement; Or to provide a health functional food composition for preventing or improving gout or diabetes.

As used herein, the term “improvement” refers to any action that at least reduces the severity of parameters associated with the condition being treated, eg, symptoms.

In the present invention, "health functional food composition" is characterized by being formulated as one selected from the group consisting of tablets, pills, powders, granules, powders, capsules, and liquid formulations, including one or more of carriers, diluents, excipients, and additives. Is done.

Foods that can be added to the ruby apple peel extract of the present invention include various foods, powders, granules, tablets, capsules, syrups, drinks, gums, teas, vitamin complexes, and health functional foods. Additives that may be further included in the present invention include natural carbohydrates, flavoring agents, nutrients, vitamins, minerals (electrolytes), flavoring agents (synthetic flavoring agents, natural flavoring agents, etc.), colorants, fillers, pactic acid and salts thereof, Alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, antioxidants, glycerin, alcohol, carbonic acid, and one or more components selected from the group consisting of flesh can be used. Examples of the natural carbohydrates described above include monosaccharides, such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, etc .; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, natural flavoring agents (taumatine, stevia extract (for example, rebaudioside A, glycyrrhizine, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used.

In addition to the above, the composition according to the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring and neutralizing agents, pactic acid and salts thereof, alginic acid and salts thereof, organic acids, and protection It may contain sex colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohol, carbonic acid used in carbonated beverages, and the like.

In addition, the composition according to the present invention may contain flesh for the preparation of natural fruit juices and vegetable drinks. These ingredients can be used independently or in combination.

Specific examples of the carrier, excipient, diluent and additive include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium phosphate, calcium silicate, Group consisting of microcrystalline cellulose, polyvinylkyrrolidone, cellulose, polyvinylpyrrolidone, methylcellulose, water, sugar syrup, methyl hydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate, and mineral oil It is preferred that at least one selected from is used.

In addition, the present invention provides a cosmetic composition for anti-oxidation, anti-inflammatory, skin whitening, or wrinkle improvement, which includes the extract of the apple extract of Ruies as an active ingredient.

The cosmetic composition containing the extract of ruby apple peel of the present invention can be used in various ways, such as anti-oxidation of skin, anti-inflammatory, whitening, wrinkle improvement and cosmetics. The products to which the composition can be added include, for example, cosmetics such as various creams, lotions, and skins, and cleansing agents, face washes, soaps, treatments, and cosmetics.

The cosmetic composition of the present invention includes a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingo lipids and seaweed extract.

The water-soluble vitamin may be any compound that can be blended into cosmetics, but preferably vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, vitamin H, etc. In addition, their salts (thiamine hydrochloride, sodium ascorbate salt, etc.) or derivatives (ascorbic acid-2-sodium phosphate salt, ascorbic acid-2-magnesium phosphate salt, etc.) can also be used in the water-soluble vitamins that can be used in the present invention. Is included. The water-soluble vitamin can be obtained by a conventional method such as a microorganism conversion method, a purification method from a culture of microorganisms, an enzyme method, or a chemical synthesis method.

The oil-soluble vitamin may be any compound that can be blended into cosmetics, but preferably vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol), and the like. , Their derivatives (ascorbyl palmitate, ascorbyl stearate, ascorbine dipalmitate, dl-alpha tocopherol acetate, dl-alpha tocopherol acetate nicotine E, DL-pantothenyl alcohol, D-pantothenyl alcohol, pantothenylethyl Ether, etc.) are also included in the oil-soluble vitamin used in the present invention. Oil-soluble vitamins can be obtained by conventional methods such as microbial transformation, microbial culture purification, enzyme or chemical synthesis.

The polymer peptide may be any compound as long as it can be blended into cosmetics, and preferably collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, keratin, and the like. The polymer peptide can be purified by a conventional method such as a purification method from a culture medium of microorganisms, an enzymatic method or a chemical synthesis method, or can be purified and used from natural substances such as dermis such as pigs and cows and silkworm silkworms.

The polymer polysaccharide may be any compound as long as it can be blended into cosmetics. Preferably, hydroxyethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate, or a salt thereof (sodium salt, etc.) may be mentioned. For example, chondroitin sulfate or a salt thereof can be used after being purified from mammals or fish.

The sphingo lipid may be any compound as long as it can be blended into cosmetics, but preferably, ceramide, phytosphingosine, sphingoglycolipid, and the like. Sphingo lipids are usually purified from mammals, fish, shellfish, yeast or plants by conventional methods or can be obtained by chemical synthesis.

The seaweed extract may be anything as long as it can be blended into cosmetics, but preferably, brown seaweed extract, red seaweed extract, green algae extract, etc., and further, callaginan, arginic acid, sodium arginate purified from these seaweed extracts, Potassium arginate and the like are also included in the seaweed extract used in the present invention. Seaweed extract can be obtained by purification from seaweed by a conventional method.

The cosmetic composition of the present invention may be blended with the above essential ingredients as well as other ingredients normally blended into the cosmetic composition as necessary.

Other ingredients that may be added include fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavors, And blood circulation accelerators, cooling agents, limiting agents, and purified water.

Examples of the fats and oils component include ester fats and fats, hydrocarbon fats and fats, silicone fats and fats, fluorine fats, animal fats, and vegetable fats and oils.

Examples of ester oils and fats include tri2-ethylhexanoate glyceryl, 2-ethylhexanoate cetyl, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, and stearic acid. Butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacinate, adipine Diisopropyl acid, isoalkyl neopentanoate, tri (caprylic, capric acid) glyceryl, tri2-ethylhexanoate trimethylolpropane, triisostearate trimethylolpropane, tetra2-ethylhexanoate pentaerythritol , Cetyl caprylate, decyl laurate, hexyl laurate, decyl myristate, myristyl myristate, cetyl myristate, stearyl stearate, decyl oleate, ricinooleic acid , Isostearyl laurate, isotridecyl myristate, isocetyl palmitate, octyl stearate, isocetyl stearate, isodecyl oleate, octyldodecyl oleate, octyldodecyl oleate, isopropyl isoleate, 2 -Sethostearyl ethylhexanoate, stearyl 2-ethylhexanoate, hexyl isostearate, ethylene glycol dioctanoate, ethylene glycol dioleate, propylene glycol dicaprate, propylene glycol di (caprylic, capric acid), dica Propylene glycol triacrylate, neopentyl glycol dicapric acid, neopentyl glycol dioctanoate, glyceryl tricaprylic acid, glyceryl triundecylate, glyceryl triisopalmitate, glyceryl triisostearate, octyl dodecyl neopentane , Isostearyl octanoate, octyl isononanoate, hexyldecyl neodecanoate, octyldodecyl neodecanoate, isocetyl isostearate, isostearyl isostearate, Octyldecyl stearate, polyglycerin oleic acid ester, polyglycerin isostearic acid ester, triisocetyl citrate, triisoalkyl citrate, triisooctyl citrate, lauryl lactate, myristyl lactate, cetyl lactate, octyldecyl lactate, trityl citrate Ethyl, acetyl triethyl citrate, acetyl tributyl citrate, trioctyl citrate, diisostearyl malate, 2-ethylhexyl hydroxystearic acid, di2-ethylhexyl succinate, diisobutyl succinate, diisopropyl sebacate, Dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, phytsteryl isostearate, phytyl oleate, 12-Stealoylhydroxystearate isocetyl, 12-stealoylhydroxystearate stearyl, 12-stealo And ester systems such as isostearyl monohydroxystearate.

Examples of hydrocarbon-based fats and oils include hydrocarbon-based fats and oils such as squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybutene, microcrystalline wax, and petrolatum.

Examples of silicone oils and fats include polymethylsilicone, methylphenylsilicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane / methylcetyloxysiloxane copolymer, dimethylsiloxane / methylstearoxylsiloxane copolymer, and alkyl And modified silicone oils and amino-modified silicone oils.

Perfluoropolyether etc. are mentioned as a fluorine-type oil and fat.

As animal or vegetable oil, avocado oil, almond oil, olive oil, sesame oil, rice bran oil, new flower oil, soybean oil, corn oil, rapeseed oil, almond oil, palm kernel oil, palm oil, castor oil, sunflower oil, grape seed oil , Cottonseed Oil, Palm Oil, Cucumin Nut Oil, Wheat Germ Oil, Rice Germ Oil, Shea Butter, Moon Seed Colostrum, Marker Demi Nut Oil, Meadow Home Oil, Yolk Yolk Oil, Uji, Horse Oil, Mink Oil, Orange Rape Oil, Jojoba Oil , Candeler wax, carnauba wax, liquid lanolin, hardened castor oil and other animal or plant fats.

Examples of the moisturizing agent include water-soluble low molecular moisturizing agents, oil-soluble molecular moisturizing agents, water-soluble polymers, and fat-soluble polymers.

As a water-soluble low-molecular moisturizer, serine, glutamine, sorbitol, mannitol, sodium pyrrolidone-sodium carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (polymerization degree n = 2 or more), polypropylene Glycol (polymerization degree n = 2 or more), polyglycerin B (polymerization degree n = 2 or more), lactic acid, lactate, and the like.

Examples of fat-soluble low-molecular moisturizers include cholesterol and cholesterol esters.

Water-soluble polymers include carboxyvinyl polymer, polyasparaginate, tragacanth, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitin, chitosan, dextrin, etc. Can be.

Examples of the fat-soluble polymer include polyvinylpyrrolidone-eicosen copolymer, polyvinylpyrrolidone-hexadecene copolymer, nitrocellulose, dextrin fatty acid ester, and polymer silicone.

Examples of the emollient agent include long-chain acyl glutamate cholesteryl ester, hydroxystearate cholesteryl, 12-hydroxystearic acid, stearic acid, rosin acid, lanolin fatty acid cholesteryl ester, and the like.

Examples of the surfactant include nonionic surfactants, anionic surfactants, cationic surfactants and amphoteric surfactants.

Examples of the nonionic surfactant include self-emulsifying glycerin monostearate, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene) sorbitan fatty acid ester, POE sorbit fatty acid ester, and POE. Glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hardened castor oil, POE castor oil, POE ㆍ POP (polyoxyethylene ㆍ polyoxypropylene) copolymer, POE ㆍ POP alkyl ether, polyether-modified silicone, lauric acid And alkanolamides, alkylamine oxides, and hydrogenated soybean phospholipids.

Examples of the anionic surfactant include fatty acid soap, alpha-acyl sulfonate, alkyl sulfonate, alkyl allyl sulfonate, alkyl naphthalene sulfonate, alkyl sulfate, POE alkyl ether sulfate, alkylamide sulfate, alkyl phosphate, POE alkyl phosphate, and alkylamide phosphate. , Alkyloyl alkyl taurine salts, N-acyl amino acid salts, POE alkyl ether carboxylate salts, alkyl sulfosuccinate salts, sodium alkyl sulfoacetate salts, acylated hydrolyzed collagen peptide salts, perfluoroalkyl phosphate esters, and the like. .

Examples of the cationic surfactant include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, and chloride. And benzalkonium, diethylaminoethyl stearate, dimethylaminopropylamide stearate, and quaternary ammonium salts of lanolin derivatives.

Examples of the amphoteric surfactant include carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amidesulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, and amideamine type. And amphoteric surfactants.

Examples of organic and inorganic pigments include silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, bengala, clay, bentonite, titanium film mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, and aluminum oxide , Inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine, chromium oxide, chromium hydroxide, calamine, and complexes thereof; Polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluorine resin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene ㆍ styrene copolymer, And organic pigments such as silk powder, cellulose, CI pigment yellow, and CI pigment orange, and composite pigments of inorganic pigments and organic pigments.

Examples of the organic powder include metal soaps such as calcium stearate; Alkyl phosphate metal salts such as sodium cetyl acid zinc, lauryl acid zinc, and calcium lauryl acid; Acylamino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc, and N-lauroylglycine calcium; Amide sulfonic acid polyvalent metal salts such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; N such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoylazine, N-alpha-paritoylolnitin, N-alpha-lauroylarginine, N-alpha-cured Uji fatty acid acyl arginine -Acyl basic amino acids; N-acylpolypeptides, such as N-lauroylglycylglycine; Alpha-amino fatty acids such as alpha-amino caprylic acid and alpha-amino lauric acid; And polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene-styrene copolymer, and ethylene tetrafluoride.

Examples of ultraviolet absorbers include paraaminobenzoic acid, ethyl paraaminobenzoate, amyl paraaminobenzoate, octyl paraaminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomentyl salicylate, benzyl cinnamate , Paramethoxy cinnamic acid-2-ethoxyethyl, paramethoxy cinnamic acid octyl, diparamethoxy cinnamic acid mono-2-ethylhexaneglyceryl, paramethoxy cinnamic acid isopropyl, diisopropyl-diisopropyl cinnamic acid ester mixture, uro Canonic acid, ethyl urocanate, hydroxymethoxybenzophenone, hydroxymethoxybenzophenone sulfonic acid and salts thereof, dihydroxymethoxybenzophenone, dihydroxymethoxybenzophenone disulfonate sodium, dihydroxybenzophenone , Tetrahydroxybenzophenone, 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino-p- (carbo-2'-ethylhexyl-1'-oxy) -1 , 3,5-triazine, 2- (2-hydroxy Ci-5-methylphenyl) benzotriazole etc. are mentioned.

As a fungicide, hinokitiol, triclosan, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, zincfilionone, benzalkonium chloride, photosensitive No. 301, mononitrowire sodium, and undeciric acid.

Examples of antioxidants include butyl hydroxyanisole, propyl gallic acid, and elisoric acid.

Examples of the pH adjusting agent include citric acid, sodium citrate, malic acid, sodium malate, phthalic acid, sodium phthalate, succinic acid, sodium succinate, sodium hydroxide, and sodium monohydrogen phosphate.

Examples of the alcohol include higher alcohols such as cetyl alcohol.

In addition, the compounding components that may be added in addition are not limited thereto, and any of the above components can be blended within a range not impairing the objects and effects of the present invention, but preferably 0.01-5% by weight based on the total weight, More preferably, it is compounded in a 0.01-3% weight percentage.

The cosmetic composition of the present invention may take the form of solutions, emulsions, viscous mixtures, and the like.

The components included in the cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the extract as an active ingredient, and for example, conventional adjuvants such as stabilizers, solubilizers, vitamins, pigments, and fragrances And carriers.

The cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, milk, cream, lotion, pack, foundation, lotion, cosmetic liquid, hair cosmetic, and the like.

Specifically, the cosmetic composition of the present invention is a skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, foundation, essence, nutrition essence, Contains formulations of pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion and body cleanser.

When the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. Can be.

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, and in the case of a spray, additionally chlorofluorohydrocarbon, propane / Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, suspensions such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or trakant, etc. can be used.

When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide as a carrier component Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linoline derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

In one embodiment of the present invention, as a result of comparing the physiological activity of the extract of each portion (shell, pulp, whole apple) of the Ruies apple, it was confirmed that the peel extract of the Ruies apple has the highest physiological activity (Example 2) Reference).

In an embodiment of the present invention, as a result of DDPH radical and ABTS radical inhibitory effects for comparison of the physiological activity of solid and phenolic of the extract of Ruies apple, it was confirmed that the phenolic compounds of the Ruies apple peel are directly involved in physiological activity. (See Example 3).

In one embodiment of the present invention, the extraction yield of phenolic compounds by solvent type and concentration of Ruies apples was compared.

Extraction is a method of separating physiologically active substances and functional substances contained in a sample according to the characteristics of each solvent, and the setting of the solvent used during extraction has an important effect on the stages before the physiological activity experiment. Hot water extraction using pure distilled water has no problem of harmfulness to the solvent, high yield, and is widely used for extraction of water-soluble substances. In the case of ethanol extraction, which is a polar solvent, purification is easy after extraction, and tannin, saponin, and organic acid of plants It is mainly used for extracting various useful ingredients such as.

As a result of measuring the phenolic content by extracting the peel of the Ruies apple by type of solvent, it showed a high extraction yield in the order of water, methanol, and ethanol. In the extract by ethanol concentration, the highest extraction yield of the phenolic content from 40% ethanol extract It was shown (see Example 4).

In one experimental example of the present invention, the antioxidant effect of the extract of the Ruies apple peel was confirmed.

Antioxidants neutralize them before free radicals that cause harmful reactions in cells in the body attack DNA or oxidize lipids. Therefore, antioxidants are known as one of the functional or bioactive substances that humans are currently focusing on, which can prevent food deterioration and prevent aging in the human body and prevent adult diseases. Antioxidants have high reactivity and react with harmful substances in the body, preventing the main substances in the cell from chain reaction by free radicals to protect the cells. Antioxidant enzymes such as superoxide dismutase (SOD), catalase, glutathione peroxidase (GSH-Px), and glutathione S-transferase (GST) are present in the body and vitamin C, an antioxidant that acts as an antioxidant or free radical scavenger with low molecular weight. Several minerals, including vitamin E, β-carotin, carotenoids, flavonoids, and selenium, are known to protect the body.

As a result of measuring PF and TBARs in order to evaluate the antioxidant effect of the extract of the Ruies apple peel, it was confirmed that the Ruies apple peel extract showed higher antioxidant activity compared to the adjuvant apple peel extract (see Experimental Example 1).

In one experimental example of the present invention, the anti-inflammatory effect of the extract of the Ruies apple peel was confirmed.

The hyaluronic acid (HA) of the polymer inhibits the phagocytic ability of macrophage, which is an important factor in the formation of inflammation, whereas the degradation product of HA or low-molecular HA increases inflammation, fibrosis, and collagen deposition during wound healing. In addition, hyaluronidase is generally present in lysosomes in an inactive form to maintain homeostasis, but is activated when an inflammatory disease such as physical injury or rheumatism develops and is involved in vascular permeability and inflammatory reactions. It has also been reported to be involved in cancer cell metastasis.

As a result of measuring the hyaluronidase inhibitory effect in order to evaluate the anti-inflammatory effect of the extract of the Ruies apple peel, it was confirmed that the hyaluronidase inhibitory effect of the Ruis apple peel extract was superior to that of the adjuvant peel extract, indicating an excellent anti-inflammatory effect (Experimental Example) 2).

In one experimental example of the present invention, the whitening effect of the extract of the Ruies apple peel was confirmed.

Melanin (melanin) is a generic term for black or brown pigments present in tissues such as the skin or eyes of various animals. In particular, melanin is an important factor that determines the skin color of a person, and the melanin expression gene varies according to race, and accordingly, the amount of melanin cells is regulated to determine the skin color. The function of melanin in the skin is a UV protection function, which maintains the body temperature of the skin and protects the skin from UV rays, but excessive melanin production causes spots, freckles, and skin spots, and further contributes to skin cancer. Currently, in the field of cosmetics, substances targeting tyrosinase inhibition are being developed to prevent pigmentation due to imbalance of the melanin formation process, and representative examples include kojic acid.

As a result of measuring tyrosinase inhibitory effect that causes pigmentation due to imbalance of melanin formation process in order to evaluate the skin whitening effect of ruby apple peel extract, the ruby apple peel extract exhibits a higher tyrosinase inhibitory effect than adjuvant apple peel extract. It was confirmed that the skin whitening effect was excellent (see Experimental Example 3).

In one experimental example of the present invention, it was confirmed the wrinkle improvement effect of the extract of the Ruies apple peel.

Structural protein elastin decreases as skin aging progresses, and the biosynthesis of type-1 collagenase increases according to the degree of skin aging, leading to decomposition of matrix proteins such as collagen fibers and elastic fibers in the dermis, thereby reducing skin elasticity. Knitting and causing the formation of skin wrinkles. Elastase present in the neutrophil granulocytes of the human body is an enzyme that breaks down the skin elastic fibers (elastin), and inhibition of elastase means improvement of skin wrinkles.

In addition, Collagen has the function of inducing mechanical firmness of skin and resistance of connective tissue, tissue power, cell division and differentiation, and internal factors such as reduction of cell activity due to natural aging, increased stress due to various harmful environments, and ultraviolet irradiation It is decomposed by external factors such as photoaging. It has been reported that this decrease in collagen decreases the elasticity of the skin and causes wrinkles and sagging.

As a result of measuring the elastase and collagenase inhibitory effect to evaluate the wrinkle improvement effect of the extract of ruby apple peel, it was confirmed that the ruby apple peel extract exhibited a higher elastase and collagenase inhibitory effect compared to the adjuvant apple peel extract, thus showing excellent wrinkle improvement effect. It was done (see Experimental Example 4).

In one experimental example of the present invention, it was confirmed that the ventilation suppression effect of the extract of the Ruies apple peel.

Gout is a disease that causes recurrent paroxysmal inflammation as the uric acid concentration in the body increases due to various causes, and as a result, excessively accumulated uric acid crystallizes, forming a pointed shape and accumulating in the joints and surrounding tissues. Gout is a disease that is well known for its direct relevance to uric acid, and gout is associated with hyperuricemia, and refers to when the urate in the blood is greater than 6.8 mg / dL (404 μmol / L). Uric acid is the end product of purine metabolism in the body. During the metabolic process in the body, hypoxanthine and xanthine produce uric acid that can induce gout by the action of XOase, and xanthine oxidase plays a decisive role in generating gout. It is known as an enzyme. In other mammals, an enzyme called urate oxidase is present, which is converted into more soluble allantoin and excreted, but in humans it is mainly formed in the liver by uric acid, filtered by the kidneys, and excreted mainly in urine. In the recent study, uric acid has been highlighted in relation to cardiovascular diseases including hypertension, kidney disease, abdominal obesity, and metabolic diseases such as diabetes.

As a result of measuring the inhibitory effect of xanthine oxidase in order to evaluate the gout suppressing effect of the extract of ruby apple peel, it was confirmed that the ruby apple peel extract exhibited the inhibitory effect of xanthine oxidase that did not appear in the adjuvant apple peel extract, indicating that it has a gout inhibitory effect ( See Experimental Example 5).

In one experimental example of the present invention, the anti-diabetic effect of the extract of the Ruies apple peel was confirmed.

Starch, the main species of carbohydrate, is hydrolyzed to glucose by α-amylase in the intestine and maltase in the small intestine, and eventually absorbed into the bloodstream, leading to an increase in blood glucose level and insulin level. Therefore, when the action of enzymes such as α-amylase can be restricted and regulated, the digestion rate of starch is delayed, which affects the distribution of absorption kinetics and blood sugar levels, and thus can be applied to patients with diabetes or obesity symptoms caused by abnormal control of carbohydrates. Therefore, it can increase the therapeutic effect of diet.

In addition, α-Glucosidase is present in the small intestine chorionic membrane and decomposes disaccharides or polysaccharides into smaller monosaccharides to promote the absorption of sugar and cause blood sugar synergy. Particularly, after 30 minutes after eating, absorption of sugar is promoted to increase blood sugar. In diabetic patients, hyperactivity of hyperglycemia and hyperglycemia continues to occur. Diseases such as retinopathy occur.

As a result of measuring α-amylase and α-glucosidase inhibitory effects in order to evaluate the anti-diabetic effect of the extract of Rubies apple peel, the extract of Rubies apple peel has a higher α-amylase and α-glucosidase inhibitory effect than the adjuvant apple peel extract. It was confirmed that the anti-diabetic effect was excellent (see Experimental Example 6).

Hereinafter, preferred embodiments and experimental examples of the present invention will be described in detail with reference to the accompanying drawings. However, these Examples and Experimental Examples are only for illustrating the present invention, and the scope of the present invention will not be interpreted as being limited by these Examples and Experimental Examples.

Example  1. Material preparation

1-1. Ruby's  Apple sample preparation

The test material used was a new breeding cultivar, cultivated by breeding using Alpine Otome varieties (parents) and Sansa varieties (headquarters) as a mating combination at the Apple Research Institute in Gunwi-gun, Gyeongbuk. Samples were harvested from the apple tree of Rubies, and then the foreign matter was removed and tested on the skins, pulp and whole apples constituting the apple. Peel, pulp, and whole apple samples were lyophilized (freeze dryer, FD8518, Ilshinbiobase, Yangju, Korea) to remove moisture, crushed with 40 mesh, stored at a low temperature of 4 ° C and used as samples.

1-2. Ruby's  From an apology phenolic  Extract preparation

Preparation of extracts for functional food and functional cosmetic activity analysis was performed by extracting 1 g of sample powder of ruby apples in 200 ml of distilled water in the case of hot water extract, heating the mixture to 100 ml, cooling it, and extracting with stirring for 24 hours. In the case of the ethanol extract, 100 ml of ethanol at a concentration of 10 to 100% was added to 1 g of the Rubies apple powder, and the mixture was stirred and extracted in a shaking incubator at 4 ° C. for 24 hours. Each extract contains Whatman No. 1 After filtration with filter paper (Whatman inc., Piscataway, New jersey, USA), concentrate on a rotary vacuum evaporator (Eyela NE, Tokyo, Japan) as needed and store at a low temperature in a refrigerator at 4 ° C and adjust the concentration of phenolic compounds in the sample. 25, 50, 75, 100 μg / ml or 50, 100, 150, 200 μg / ml phenolics concentrations, respectively, were used in the experiment.

Example  2. Ruby's  Comparison of Physiological Activities of Extracts by Parts of Apple (Peel, Pulp, Whole Apple)

As a result of measuring the phenolic content by extracting the skin, pulp, and whole apples of the new-growing varieties Ruies apples with water and 60% ethanol by the method of Example 1, it shows the highest phenolic content in the peel as shown in FIG. And whole apples and pulp. Therefore, it was speculated that the physiological activity of the bark extract would be the highest.

As a result of comparing the ABTS radical scavenging ability, which indicates water-soluble antioxidant capacity, to evaluate the antioxidant effect of the peel, pulp, and whole apple extracts of Ruies apples, compared with BHT, a synthetic antioxidant used as a positive control, as shown in FIG. 2. The highest efficacy was found in the peel of ruby apples, and the antioxidant activity was low in pulp and whole apples.

According to the above results, when comparing the peel, pulp, and whole apples of the ruby apples, the phenolic content was high in the peels and whole apples, and it was confirmed that the peeling activity was higher in the antioxidant activity than in the flesh. Therefore, in the subsequent experiments, experiments were conducted on the skin with high efficacy.

Example  3. Ruby's  Comparison of physiological activity between solid and phenolic in apple extract

In order to compare the bioactive efficacy of the solid content and the phenolic component of Ruies apple, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical inhibitory effect was confirmed. The DPPH radical inhibitory effect was in accordance with the method of Blois, and 3 mL of 60 μM DPPH was added to 0.5 mL of a sample of the russ apple peel extract prepared in Example 1, followed by vortexing, standing for 15 minutes, and absorbance at 517 nm was measured. .

As a result, as shown in Fig. 3A, the solid content of 100 μg / mL of the water and ethanol extracts of the Ruies apple peel showed electron donating ability of 16.25 and 19.19%, respectively, but 88.34 and 90.38% at 100 μg / mL phenolic concentration, respectively. It was confirmed that it exhibits the activity of phenolic compounds by showing the electron donating ability of.

In addition, the 2,2'-Azinobis- (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical inhibitory effect was measured to compare the physiological activity of solids and phenolic components. The ABTS radical inhibitory effect is based on the method of Fellegrini et al., After mixing 7 mM ABTS and 140 mM K2S2O8 in 5 mL: 88 μL and leaving it in a dark place for 12 to 16 hours, mixing it with ethanol in a ratio of 1:88 to 734 nm Using the ABTS solution adjusted so that the absorbance value of the control was 0.7 ± 0.02, 50 μL of the sample solution and 1 mL of the ABTS solution were mixed for 30 seconds, followed by incubation for 2.5 minutes to measure the absorbance at 734 nm.

As a result, as shown in Fig. 3b, water and ethanol extracts of the Ruis apple peel showed an electron donating ability of 3.47 and 3.42%, respectively, at a solid concentration of 100 µg / mL, and 98.46 and 95.66%, respectively, at a concentration of 100 µg / mL phenolic. It was confirmed that the activity of the apple was dominated by the phenolic compound contained in the solid content.

Therefore, as a result of the comparative experiments on the physiological activity of solids and phenolic components, it was confirmed that the phenolic compounds of the ruby apple peel were directly involved in the physiological activity, and the experiment was performed by extracting the phenolic compound of the rust of the ruby apple.

Example  4. Ruby's  Comparison of phenolic compound extraction yield by solvent type and concentration of apple

4-1. Quantification of total phenolic compounds content

For total phenolic quantification, 1 mL of 95% ethanol and 5 mL of distilled water were added to 1 mL of extract according to the method of Folin and Denis, 1 mL of 1 N Folin-ciocalteu reagent 0.5 mL was mixed well, allowed to stand for 5 minutes, and 1 mL of Na2CO3 was added to absorbance 725 Measured within 1 hour at nm, the amount was converted from a standard curve using gallic acid.

4-2. Ruby's  Comparison of phenolic compound extraction yield by solvent type and concentration of apple

As a result of measuring the phenolic content by extracting the peel of the ruby apple of Example 1 according to the type of the solvent, as shown in Figure 4a, water, methanol, ethanol, acetone, butanol, ethyl acetate in the order of 5.53, 5.29, 4.99, 2.82 , 2.75, and 2.35 mg / g. Among water, methanol and ethanol, which showed relatively high extraction yields, water and ethanol extracts, which are harmless to the human body and showed relatively high phenolic contents, were tested.

In addition, the extract according to the concentration of ethanol in the shell showed the highest extraction yield of phenolic content at 8.76 mg / g in 40% ethanol extract as shown in FIG. 4B.

Therefore, the experiment was conducted by extracting the apple peel with water and 40% ethanol as a solvent, and the phenolic content was adjusted to 25-200 µg / mL for reproducibility of the experiment.

Experimental Example  One. Ruby's  Confirm the antioxidant effect of apple peel extract

1-1. Antioxidant protection factor ( PF ) Measure

In order to evaluate the antioxidant effect of the extract of Ruies apple peel, 11-linked β-carotine, composed of 11 double bonds, reacts very easily with peroxyl radicals. PF, which exhibits fat-soluble antioxidant capacity, was measured using the properties acting as. PF measurement is based on the method of Andarwulan and Shetty, 1 mL of a solution of 10 mg β-carotene dissolved in 50 mL of chloroform is placed in a water container for evaporator, distilled chloroform in a water bath at 40 ℃, and then 20 μL linoleic acid, 184 μL To make an emulsion by adding tween 40 and 50 mL H ₂ O ₂ , mix 100 μL of the ruby apple peel extract sample prepared in Example 1 with 5 mL emulsion, mix well with vortex and react for 30 minutes at 50 ℃. After cooling, the absorbance at 470 nm was measured.

As a result, as shown in Fig. 5a, water and ethanol of 1.56 ~ 1.87 and 1.39 ~ 1.77 PF were shown at 25-100 μg / mL phenolic concentrations of the extract of ruby shell, respectively, and water and ethanol of the skin of the adverb variety used as a control The extract showed 1.22 ~ 1.37 PF and 1.18 ~ 1.41 PF at 25 ~ 100 ㎍ / mL phenolic concentration, respectively, and it was confirmed that the extract of Ruies showed better fat-soluble antioxidant ability than the apple.

1-2. Thiobarbituric  acid reactive substances ( TBARs ) Inhibition effect measurement

The inhibitory effect of TBARs showing another fat-soluble antioxidant ability was measured using the extract of Ruies bark. The effect of inhibiting TBARs was based on the method of Buege and Aust, making an emulsion with 1% linoleic acid and 1% Tween 40, mixing 0.8 mL of the emulsion with 0.2 mL of the Ruies apple peel extract sample prepared in Example 1, and then 50 ° C. water The reaction was performed for 10 hours in a bath. 2 mL of TBA regent was added to 1 mL of the reaction solution, followed by boiling for 15 minutes, cooling for 10 minutes, centrifugation at 1,000 rpm for 15 minutes, standing at room temperature for 10 minutes, and absorbance of the supernatant at 532 nm was measured.

As a result, as shown in Fig. 5b, the water and ethanol extracts of the ruby shells exhibited activity of 37.69 to 80.17% and 19.59 to 49.29%, respectively, and the water and ethanol extracts of the adjuvant varieties used as controls were 32.60 to 55.40, respectively. % And 21.02 ~ 75.41%.

Therefore, from the above results, it was confirmed that the extract of ruby apple peel has higher antioxidant activity than the control apple apple.

Experimental Example  2. Ruby's  Confirmation of anti-inflammatory effect of apple peel extract

In order to confirm the anti-inflammatory effect of the extract of Ruies apple peel, the inhibitory effect of Hyaluronidase (HAase), an enzyme related to inflammation, was confirmed.

Hyaluronidase inhibitory effect was measured at 600 nm by absorbing N-acetyl-glucosamine formed from sodium-hyaluronic acid (HA) into a glucoxazoline derivative and developing with ρ-dimethylaminobenzaldehyde (DMAB) according to the methods of Dorfman and OTT.

As a result, as shown in FIG. 6, the water extract of the Ruiesus bark showed a HAase inhibitory effect of 1.16 to 6.37% at a phenolic concentration of 50 to 200 μg / mL, and the water extract of the bark variety bark used as a control was 50 to It showed the inhibitory effect of HAase of 0.03 ~ 1.79% at phenolic concentration of 200 μg / mL.

In addition, the ethanol extract of the Ruies bark showed an inhibitory effect of 2.36 to 31.87% at a phenolic concentration of 50 to 200 μg / mL, and an ethanol extract of the adverb variety was 1.29 to 12.72% at a phenolic concentration of 50 to 200 μg / mL. By showing the inhibitory effect of, it was confirmed that the hyaluronidase inhibitory effect of the extract of Ruies was superior to the adjuvant variety.

Experimental Example  3. Ruby's  Confirm the whitening effect of apple peel extract

In order to confirm the whitening effect of the extract of the russ apple peel, the inhibitory effect of tyrosinase, which causes pigmentation due to the imbalance of the melanin formation process, was measured.

The whitening effect is 250 U / mL mushroom tyrosinase (Sigma-Aldrich Co.) 0.1 in a mixture of 2.3 mL of 0.1 M sodium phosphate buffer (pH 6.8) and 0.4 mL of the substrate solution 1.5 mM L-tyrosine solution according to Vincent and Hearing's method. Add 0.2 mL of each sample of 50 ~ 200 μg / mL phenolic compounds concentration of mL and the ruby apple peel extract prepared in Example 1, and add 0.2 mL of distilled water instead of the sample to the control for 20 minutes at 37 ° C. Then, absorbance was measured at 475 nm.

 As a result, as shown in FIG. 7, the tyrosinase inhibitory effect was not observed in the phenolic concentration of 50-200 μg / mL in the water extract of the Ruiesus bark, and the tyrosinase inhibitory effect was not observed in the bark water extract of the adjuvant variety used as a control. .

However, in the ethanol extract of the Ruies bark, the tyrosinase inhibitory effect was 5.82 to 17.50% at a phenolic concentration of 50 to 200 μg / mL, and the adverb ethanol extract was very high at 2.69 to 5.23% at a phenolic concentration of 50 to 200 μg / mL. It exhibited a low inhibitory effect, and it was confirmed that the tyrosinase inhibitory effect of the extract of Ruies bark was better than that of the adjuvant variety.

Experimental Example  4. Ruby's  Confirmation of wrinkle improvement effect of apple peel extract

4-1. elastase  Check the inhibitory effect

In order to confirm the anti-wrinkle effect of the extract of Rubies apple peel, the inhibitory effect of elastase, an enzyme that produces skin wrinkles, was measured by decomposing elastin, which maintains the skin elasticity of the human body.

The elastase inhibitory effect was 1.0 U / mL in a mixture of 0.1 mL of the substrate solution 0.8 mM N-succinyl- (Ala) 3-ρ-nitroanilide solution in 1 mL of 0.2 M Tris-HCl buffer (pH 8.0) according to the method of Kraunsoe et al. Porcine pancreatic elastase (PPE) (Sigma-Aldrich Co.) 0.1 mL of the enzyme solution and 0.1 mL of each sample of 50-200 μg / mL phenolic compounds concentration of the Ruies apple peel extract prepared in Example 1 above were added and a sample was added to the control group. Instead, 0.1 mL of distilled water was added to react at 25 ° C for 20 minutes, and the amount of ρ-nitroaniline was measured at absorbance at 410 nm.

As a result, as shown in FIG. 8A, the water extract of the Ruiesus bark showed an elastase inhibitory effect of 4.07 to 6.70% at a phenolic concentration of 50 to 200 μg / mL, and the water extract of the bark varieties used as a control was 50 to Elastase inhibitory effect of 0.00 ~ 5.35% was exhibited at the phenolic concentration of 200 μg / mL.

In addition, the ethanol extract of the Ruies bark showed an elastase inhibitory effect of 15.83 to 41.95% at a phenolic concentration of 50 to 200 µg / mL, and the ethanol extract of the bark peel was 8.02 to 32.10% at a phenolic concentration of 50 to 200 µg / mL. It exhibited the elastase inhibitory effect of, it was confirmed that the elastase inhibitory effect of the extract of the ruby shell compared to the adjuvant varieties.

4-2. Collagenase  Check the inhibitory effect

In order to confirm the anti-wrinkle effect of the extract of Ruies apple peel, the inhibitory effect of collagenase, an enzyme that breaks down collagen, was measured.

The collagenase inhibitory effect was 4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-D-Arg (0.3 mg / mL) by adding 4 mM CaCl ₂ to 0.1 M Tris-HCl buffer (pH 7.5) according to the method of Wunsch and Heidrich. 0.2 mg / mL collagenase (Sigma-Aldrich Co.) in a mixture of 0.25 mL of dissolved substrate solution and 0.1 mL of each sample solution at a concentration of 50-200 μg / mL phenolic compounds of the Rubies apple peel extract prepared in Example 1 above. ) After adding 0.15 mL and allowing to stand at room temperature for 20 minutes, 0.5 mL of 6% citric acid was added to stop the reaction, 2 mL of ethyl acetate was added, and absorbance was measured at 320 nm.

As a result, as shown in Figure 8b, the water extract of the Ruies bark showed a collagenase inhibitory effect of 12.60 to 69.85% at a phenolic concentration of 50 to 200 μg / mL, and the water extract of the bark varieties used as a control was 50 to 200 At phenolic concentration of ㎍ / mL, collagenase inhibitory effect was 25.89 ~ 59.73%.

In addition, the ethanol extract of the Ruies bark showed a very high collagenase inhibitory effect of 35.01 to 80.73% at a phenolic concentration of 50 to 200 µg / mL, and the ethanolic extract of an adjuvant shell ethanol extract at 47.79 to phenolic concentration of 50 to 200 µg / mL. It showed 79.05% inhibitory effect, so that the extract of Ruby's bark was similar or somewhat superior to each concentration section compared to the adjuvant variety.

Therefore, from the above results, it was confirmed that the extract of the Ruis apple peel had a higher skin wrinkle improvement effect than the control group adjuvant peel extract.

Experimental Example  5. Ruby's  Confirming the effect of suppressing gout of apple peel extract

In order to confirm the gout suppressing effect of the extract of the Rubies apple peel, the inhibitory effect of xanthin oxidase producing uric acid, a pain-inducing substance, was measured.

The inhibitory effect of Xanthine oxidase (XOase) was based on the method of Stirpe and Corte, 0.1 mL of enzyme solution (0.05 U / 0.1 mL) in 3 mL of 2 mM xanthine substrate solution dissolved in 0.1 M potassium phosphate buffer (pH 7.5) and Example 1 above. 0.3 mL of the sample of the extract of the apple extract of ruby prepared in, added 0.3 mL of distilled water instead of the sample to the control, reacted at 37 ° C. for 5 minutes, added 1 mL of 20% TCA as a termination reagent, and centrifuged the reaction solution to remove protein And the resulting uric acid was measured at 292 nm absorbance.

As a result, as shown in FIG. 9, the xanthine oxidase inhibitory effect was not observed in the extract of the Ruiesus husk, and the xanthine oxidase inhibitory effect of 27.91 to 28.66% at the phenolic concentration of 50-100 μg / mL was observed in the Ruiesus husk ethanol extract. Shown. The inhibitory effect on xanthine oxidase was not observed in the water and ethanol extracts of bark varieties used as a control.

Therefore, it was judged that the ventilation inhibitory substance that was not present in the apple of the existing adverb variety was present in the new varieties of apples, and the effect of breed improvement was obtained.

Experimental Example  6. Ruby's  Apple peel extract Anti-diabetes  Check effect

6-1. Confirmation of α-amylase inhibitory effect

In order to confirm the anti-diabetic effect of the extract of Ruies apple peel, α-amylase inhibitory effect was measured.

The α-amylase inhibitory effect was in accordance with the methods of Davidson and Parish, 5 g of agar and 5 g of soluble starch were dissolved in distilled water, boiled, sterilized at 121 ° C for 15 minutes, poured into 15 ml of petridish and hardened, and then 10 mm in diameter. Raise the disc paper and mix 0.8 ml (1,000 U / mL) of the sample solution of the Ruies apple peel extract prepared in Example 1 and 0.2 ml (1,000 U / mL) of the enzyme solution, respectively, and dispense them on the disc paper. After incubation for 3 days in C, 3 mL of I ₂ / KI (5 mM I ₂ in 3% KI) was added, followed by color development for 15 minutes to calculate the area of the clear zone to measure the inhibition rate.

As a result, as shown in FIG. 10A, in the water extract of Ruies bark, the α-amylase inhibitory effect was 1.92 ~ 3.64% at a phenolic concentration of 50-200 μg / mL, and the ethanol extract of Ruies bark was 50-200 μg / It showed an inhibitory effect of α-amylase of 2.33 ~ 2.71% at phenolic concentration of mL.

On the other hand, the bark water extract of the adjuvant variety used as a control showed an inhibitory effect of α-amylase of 1.19 to 2.02% at a phenolic concentration of 50 to 200 μg / mL, and 0.84 to 0.91% α-amylase of the adjuvant skin ethanol extract. By showing the inhibitory effect, it was confirmed that the extract of Ruies bark exhibited a somewhat superior α-amylase inhibitory effect compared to the adjuvant bark extract.

6-2. α- glucosidase  Check the inhibitory effect

In order to confirm the anti-diabetic effect of the extract of Ruies apple peel, α-glucosidase inhibitory effect was measured.

The α-glucosidase inhibitory effect is based on Tibbot and Skadsen's method, dissolving ρ-nitrophenol-α-Dglucopyranoside (PNPG) in 50 mM sodium succinate buffer (pH 6.8) to make a substrate at a concentration of 1 mg / mL, and substrate 1 After mixing mL and 0.1 mL of enzyme solution, 0.1 mL of distilled water was added to the control group, 0.1 mL of a sample of 200 μg / mL phenolic concentration of the ruby apple peel extract prepared in Example 1 was added to the reaction group, and reacted at 37 ° C for 30 minutes. Color development was performed by adding 0.1 mL of 1 N NaOH to measure absorbance at 400 nm.

As a result, as shown in Fig. 10b, the water extract of the Ruiesus bark showed a high α-glucosidase inhibitory effect of 65.56 to 94.93% at a phenolic concentration of 50 to 200 μg / mL, and the ethanolic extract of the Ruisus shell 50 to 200 It showed very high α-glucosidase inhibitory effect of 96.62 ~ 100.00% at phenolic concentration of ㎍ / mL.

On the other hand, the bark water extract of the adjuvant variety used as a control showed an inhibitory effect of α-glucosidase of 0.00-63.33% at a phenolic concentration of 50-200 ㎍ / mL, and phenolic of 50-200 ㎍ / mL in the ethanol extract of the adjuvant skin It showed that the α-glucosidase inhibitory effect was found at 62.41 to 95.56% at the concentration, and it was confirmed that the extract of Ruies bark exhibited a better α-glucosidase inhibitory effect than the adjuvant variety even at low concentrations.

Therefore, from the above results, it can be confirmed that the ruby apple peel extract has a strong anti-diabetic effect by inhibiting the activity of the carbohydrate-degrading enzyme very strongly compared to the adjuvant skin extract.

Since the specific parts of the present invention have been described in detail above, it is obvious to those skilled in the art that such specific techniques are only preferred embodiments, and the scope of the present invention is not limited thereby. something to do. Therefore, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (17)

delete delete delete delete delete delete delete delete A health functional food composition for wrinkle improvement, comprising a ruby apple peel extract as an active ingredient.
The method of claim 9,
The composition is 1,1-diphenyl-2-picrylhydrazyl (1,1-diphenyl-2-picrylhydrazyl; DPPH) radical inhibition, 2,2'-azinobis- (3-ethylbenzothiazoline-6- Sulfonic acid) (2,2'-Azinobis- (3-ethylbenzothiazoline-6-sulfonic acid); ABTS) radical inhibition, Thiobarbituric acid reactive substances (TBARs) inhibition or antioxidant protection factor (Antioxidant protection) Health functional food composition, characterized by increasing the factor (PF).
The method of claim 9,
The composition is characterized in that it inhibits hyaluronidase (hyaluronidase), health functional food composition.
The method of claim 9,
The composition is characterized in that it inhibits tyrosinase (tyrosinase), health functional food composition.
The method of claim 9,
The composition is characterized in that it inhibits elastase (elastase) or collagenase (collagenase), health functional food composition.
The method of claim 9,
The composition is characterized in that it inhibits xanthine oxidase (xanthine oxidase), health functional food composition.
The method of claim 9,
The composition is characterized in that it inhibits α-amylase (α-Amylase) or α-glucosidase (α-Glucosidase), health functional food composition.
The method of claim 9,
The ruby apple peel extract is from the group consisting of water, C ₁ to C ₄ lower alcohol, n-hexane, ethyl acetate, acetone, butyl acetate, 1,3-butylene glycol, methylene chloride, and mixed solvents thereof. Characterized in that extracted with a selected solvent, health functional food composition.
A cosmetic composition for wrinkle improvement, comprising a ruby apple peel extract as an active ingredient.

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