KR20180104960A - Composition comprising extracts of banana, cherry, and walnut for anti-skin aging - Google Patents
Composition comprising extracts of banana, cherry, and walnut for anti-skin aging Download PDFInfo
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- KR20180104960A KR20180104960A KR1020170031931A KR20170031931A KR20180104960A KR 20180104960 A KR20180104960 A KR 20180104960A KR 1020170031931 A KR1020170031931 A KR 1020170031931A KR 20170031931 A KR20170031931 A KR 20170031931A KR 20180104960 A KR20180104960 A KR 20180104960A
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/52—Juglandaceae (Walnut family)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
- A61K36/736—Prunus, e.g. plum, cherry, peach, apricot or almond
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
Abstract
The present invention provides a composition for preventing or ameliorating skin damage comprising extracts of bananas, cherries, and walnuts. A cosmetic composition comprising an extract of bananas, cherries, and walnuts according to one aspect of the present invention restores the expression of MT-1, SIRT-1 and catalase which may be degraded by stress hormone, thereby causing skin wrinkles and skin aging Which is useful for preventing or improving skin damage.
Description
To a composition for preventing or ameliorating skin damage, including extracts of bananas, cherries, and walnuts.
The expression of cortisol activating enzyme is increased in aging skin tissue. In particular, it has been reported that the activity of cortisol is increased in aged skin tissues after menopause and in skin tissues where photoaging by ultraviolet light has progressed. Cortisol is a type of stress hormone that is secreted by organs such as the kidneys, heart, lungs and skin in the body when physical damage or mental stress is applied. In particular, cortisol in the skin is involved in the synthesis of collagen. The long-term secretion of cortisol affects collagen metabolism in the body, reducing collagen synthesis, and ultimately causing skin damage such as atrophodermia.
On the other hand, melatonin is a type of neurotransmitter that induces a sleeping surface produced in the pineal gland of the brain. It is produced after a certain period of time after sunlight and has various beneficial effects in the body. In particular, melatonin removes free radicals and inhibits melanin which makes skin darker, thereby contributing to skin health. When melatonin is applied to the skin, it is possible to prevent erythema formation, mitigate atopic dermatitis, Aging effects and the like have been reported
Accordingly, the present inventors developed a mixed extract of bananas, cherries, and walnuts, which is known to be a natural substance that contains melatonin and helps in sleeping, as a skin wrinkle improving material. In addition, it was confirmed that the mixed extract of bananas, cherries, and walnuts can restore the activity of MT-1, SIRT-1 and catalase inhibited by dermal fibroblasts to prevent skin aging, thereby completing the present invention.
One aspect provides a cosmetic composition for preventing or ameliorating skin damage comprising extracts of bananas, cherries, and walnuts.
Another aspect provides a composition for a health functional food for improving skin damage comprising an extract of bananas, cherries, and walnuts.
Another aspect provides a pharmaceutical composition for treating skin damage comprising extracts of bananas, cherries, and walnuts.
One aspect provides a cosmetic composition for improving skin damage comprising an extract of bananas, cherries, and walnuts. Another aspect provides a composition for a health functional food for improving skin damage comprising an extract of bananas, cherries, and walnuts. Another aspect provides a pharmaceutical composition for treating skin damage comprising extracts of bananas, cherries, and walnuts.
The term "skin damage" refers to damage caused by external physical damage, penetration of chemicals, bacteria, fungi, viruses, exposure to ultraviolet rays, skin moisture loss, wrinkles caused by aging, skin pigmentation, Including damage from the clinical and cosmetic point of view of the skin tissue or cells involved.
Banana (Banana) is a generic term for plants belonging to the bananas over par, Musa Safienthum ( Musa) Sapientum ) or Musa paradisia L. ( Musa paradisiaca L.) and the like. It is rich in vitamins A and B, which gives skin radiance and elasticity to prevent aging, and contains tryptophan, which can help in the synthesis of melatonin in the body.
Cherry means a plant belonging to the rose and cherry tree, and Prunus pauciflora Bunge, Prunus avium), pandanus fruit Serra Seuss L. (Prunus cerasus L.) or pandanus fruit Espoo four K.Schneider (Prunus effusa K. Schneider). It contains antioxidant, anti-inflammation and anti-aging effect because it contains a lot of red anthocyanin. It contains melatonin, which can help reduce body fatigue and maintain health.
Walnut (walnut) belongs to the family Phalaenopsis, Juglans nigra L., Juglans regia regia , or plants with the names of Juglans sinensis . It contains Linoleic Acid and Vitamin E to prevent various cardiovascular diseases and to help your skin by antioxidant action. It also contains melatonin and calcium, which is effective in controlling insomnia.
The extracts of bananas, cherries and walnuts may be extracts extracted by solvent from whole, part of, or a material derived from the above ground part of the tree, respectively. The part may be a stem, root, leaf, flower, petal, seed, pulp, fruit skin or fruit of a tree. The whole, part of, or a material derived therefrom of the tree used for the extraction may be ground or chopped or suitably dried.
The extract may be extracted using water, acetone, alcohol, for example, C1-C6 alcohol, C3-C6 polyhydric alcohol, or a mixture thereof as a solvent. The C1-C6 alcohol includes methanol, ethanol, propanol, isopropanol, 1,3-propanediol, butanol, pentanol, or hexanol. The solvent may be, for example, a mixture of water and an alcohol, that is, an aqueous solution of an alcohol. The alcohol concentration of the alcohol aqueous solution may be 1 to 100 (w / w)%, for example 1 to 99.5 (w / w)%, 10 to 100 (w / (W / w)%, 40 to 100 (w / w)%, 50 to 100 (w / w)%, 60 to 100 (w / (W / w)%, 60 to 90 (w / w)%, 60 to 80 (w / w)%, 65 to 75 have. The aqueous alcohol solution may be methanol, ethanol, or an aqueous butanol solution.
The extract may be extracted by a conventional method in the art such as warm extraction, pressure extraction, ultrasonic extraction, hot water extraction, reflux cooling extraction, subcritical extraction, or supercritical extraction. The extraction may be carried out at a temperature of 4 ° C to 70 ° C, for example 4 ° C to 50 ° C, 4 ° C to 40 ° C, 4 ° C to 30 ° C, 10 ° C to 70 ° C, 15 ° C to 70 ° C, 10 C to 50 C, 10 C to 50 C, 4 C to 40 C, 4 C to 30 C, 10 C to 40 C, 10 C to 35 C, or 10 C to 30 C, . The extraction time may vary depending on the selected temperature and extraction method. The extraction time may be 1 to 3 days, 1 to 2 days, 1 hour to 1 day, 5 hours to 3 days, 5 hours to 2 days, 5 hours to 1 day, 10 hours to 3 days, 15 hours to 2 days, 15 hours to 36 hours, 18 hours to 30 hours, 1 day to 3 days, 1 day to 2 days, or 24 hours. For example, the powder of each fruit can be extracted by ultrasonic extraction by immersing in an extraction solvent for 2 to 4 hours, or by immersing in a solvent for 24 to 72 hours at room temperature. The extraction may be repeated one or more times, for example one to five times, and each extraction may be performed in the same way or may be carried out in other ways.
The extraction may further include a step of separating the plant residue and the extract by a known method such as filtration. The extraction may also include removing the solvent from the resulting extract by known methods, such as concentration under reduced pressure. The extraction may also comprise preparing the dried extract by drying, such as nitrogen drying or lyophilization, of the resulting extract. The extract may be in the form of a concentrate or, after drying, if necessary, resolved in an appropriate solvent such as DMSO, or lyophilized and pulverized.
The composition may further comprise a fraction of the extract. The fraction means a substance in which the extract is divided into a part of its components, that is, a fractionated substance, and can be obtained by fractionation according to a method commonly known in the art.
The extract may be an extract obtained by extracting individual extracts of bananas, cherries, and walnuts, respectively, or an extract for a crushed mixture of bananas, cherries and walnuts. When each of bananas, cherries, and walnuts is individually extracted, the extraction method and extraction conditions may be the same or different. The weight ratio of the extracted bananas, cherries and walnuts, or their extracts, can be suitably adjusted according to the purpose. For example, the ratio of the raw material powder to the banana, cherry, and walnut extract may be 1-10: 1 to 10: 1 to 10: 1. Also, for example, the extract of banana, cherry and walnut may have a ratio of the extract of 1 to 20: 1 to 20: 1 to 20, or 1 to 2: 1 to 2: 1 to 2.
The composition may be present in an amount of from 0.001% to 80%, such as from 0.01% to 60%, from 0.01% to 40%, from 0.01% to 30%, from 0.01% to 20% %, 0.01% to 10%, 0.01% to 5%, 0.05% to 60%, 0.05% to 40%, 0.05% to 30%, 0.05% to 20% From 0.05% to 10%, from 0.05% to 5%, from 0.1% to 60%, from 0.1% to 40%, from 0.1% to 30%, from 0.1% to 20% % To 10 wt%, or 0.1 wt% to 5 wt% of banana, cherry, and walnut extract. If it is less than 0.01% by weight in the above range, the effect of improving the skin tends to be insignificant, and it is not preferable that it is more than 60% by weight from the viewpoint of economy.
The composition may contain the extract in an effective amount or as an active ingredient. The effective amount may be appropriately selected depending on the individual. The severity of the disease or condition, the age, weight, health, sex, sensitivity of the individual to the extract, time of administration, route and rate of excretion, duration of administration, factors including other compositions combined or co- May be determined according to factors well known in the art, physiology or medical field.
When the composition is a cosmetic composition, the extract can be used as a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, , A nutritional essence, a pack, a soap, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion, a body cleanser, a suspension, a gel, a powder, a paste, a mask pack or a sheet or an aerosol composition. Compositions of such formulations may be prepared according to methods conventional in the art. The cosmetic composition may further contain preservatives, stabilizers, surfactants, solubilizers, moisturizers, emollients, ultraviolet absorbers, preservatives, bactericides, antioxidants, pH regulators, organic and inorganic pigments, fragrances, have. The amount of the additional component such as the above-mentioned moisturizing agent and the like can be easily selected by those skilled in the art within a range not to impair the purpose and effect of the present invention. The amount of the additional component is 0.001 to 5% by weight, % ≪ / RTI >
The extract may be an external preparation for skin to prevent or improve skin aging. The composition for external application for skin may be prepared in any formulations conventionally produced in the art, and examples thereof include emulsion, cream, lotion, pack, foundation, lotion, essence and hair cosmetic. Specifically, the present invention relates to a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisturizing lotion, a nutrition lotion, a massage cream, a nutritive cream, a moisturizing cream, a hand cream, a foundation, Foams, cleansing lotions, cleansing creams, body lotions and body cleansers.
When the composition is a composition for a health functional food, it can be formulated into a typical health functional food formulation known in the art. The composition for health functional foods may be prepared by conventional formulations such as powders, granules, tablets, pills, capsules, suspensions, emulsions, syrups, And may be manufactured in the form of any health food such as candy, snacks, confectionery, pizza, ramen, other noodles, dairy products including gums, jellies, ice cream, various soups, drinks, tea, drinks, alcoholic beverages and vitamin complexes . A pharmaceutically acceptable carrier or additive may be used for the formulation of the health food, and any carrier or additive known to be usable in the art for the preparation of the formulation to be prepared may be used. As the above-mentioned additives, there can be used various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, Carbonating agents, and the like. In addition, it may contain natural fruit juice, fruit juice beverage and flesh for the production of vegetable beverages. These additive components may be used independently or in combination, and the proportion of the additive may be 0.001 to 5 wt%, specifically 0.01 to 3 wt%, based on the total weight of the composition.
The content of the extract in the health functional food composition may be suitably determined according to the intended use (prevention or improvement). Generally, it may contain 0.01 to 15% by weight of the total food, and when it is prepared as a beverage, it may contain 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 mL.
The beverage may further contain ingredients other than the above extract, and may further contain various flavors or natural carbohydrates commonly used in beverages. Examples of the natural carbohydrate include conventional sugars such as monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugars such as xylitol, sorbitol, erythritol, Of sugar alcohols may be contained. In addition, as a flavoring agent, a natural flavoring agent (e.g., tau Martin, stevia extract, etc.) and a synthetic flavoring agent (e.g., saccharin, aspartame, etc.) may be contained. The ratio of the natural carbohydrate may be generally about 1 to 20 g, preferably about 5 to 12 g per 100 mL of beverage.
The composition may be a pharmaceutical composition. The composition may comprise a pharmaceutically acceptable diluent or carrier. The diluent may be lactose, corn starch, soybean oil, microcrystalline cellulose, or mannitol, the lubricant may be magnesium stearate, talc, or combinations thereof. The carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low substituted hydroxy cellulose, or combinations thereof. The disintegrant may be carboxymethylcellulose calcium, starch glycolate sodium, calcium monohydrogen phosphate anhydrate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or combinations thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.
The inventors of the present invention have found that when extracting natural products known to have a sleeping effect for preventing or improving aging, the mixed extract of bananas, cherries, and walnuts is used as an antioxidative factor related to skin aging, catalase and aging-related factor SIRT-1 , And MT-1, which is a melatonin receptor, and thus completed the present invention.
In addition, in the case of the mixed extract of banana, cherry and walnut, the recovery effect of MT-1, SIRT-1 and catalase reduced by the stress hormone was remarkably increased compared with the single extract. In view of this, it can be seen that the mixed extract of bananas, cherries and walnuts shows a synergistic effect more than a simple combination in comparison with the extracts of bananas, cherries and walnuts alone, and that synergy effect can prevent or improve skin aging or wrinkles.
Compositions comprising extracts of bananas, cherries, and walnuts are useful for restoring the expression of MT-1, SIRT-1 and catalase to prevent or ameliorate skin damage.
1 shows the effect of restoring the expression of MT-1 in banana extract, cherry extract, walnut extract and mixed extract thereof.
FIG. 2 shows the effect of restoring the expression of SIRT-1 in banana extract, cherry extract, walnut extract and mixed extract thereof.
FIG. 3 shows the effect of restoring expression of catalase in banana extract, cherry extract, walnut extract and mixed extract thereof.
Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.
Comparative Example 1: Preparation of banana extract
Banana (Musa 100 g of raw material powder for fruit of Sapientum were put into separate containers and immersed in 1 L of a 70% by weight aqueous ethanol solution as an extraction solvent. After that, the mixture was allowed to stand at room temperature for 24 hours to obtain an extract. The obtained extract was filtered using a filter paper, and the filtrate was concentrated under reduced pressure using a rotary evaporator to obtain a final banana extract. The final extract was dissolved in DMSO (dimethyl sulfoxide) at a concentration of 1% by weight and used in the following Experimental Examples.
Comparative Example 2: Preparation of cherry extract
Cherry ( Prunus pauciflora Bunge ) were put into separate containers and immersed in 1 L of a 70 wt% aqueous ethanol solution as an extraction solvent. After that, the mixture was allowed to stand at room temperature for 24 hours to obtain an extract. The obtained extract was filtered using a filter paper, and the filtrate was concentrated under reduced pressure using a rotary evaporator to obtain a final cherry extract. The final extract was dissolved in DMSO at a concentration of 1% by weight and used in the following experimental examples.
Comparative Example 3: Preparation of walnut extract
Walnuts ( Juglans) nigra L.) seeds were placed in separate containers and immersed in 1 L of a 70 wt% aqueous ethanol solution as an extraction solvent. After that, the mixture was allowed to stand at room temperature for 24 hours to obtain an extract. The obtained extract was filtered using a filter paper, and the filtrate was concentrated under reduced pressure using a rotary evaporator to obtain a final walnut extract. The final extract was dissolved in DMSO at a concentration of 1% by weight and used in the following experimental examples.
Example 1: Preparation of a Banana, Cherry and Walnut Mixture Extract
The extracts of each of the bananas, cherries, and walnuts obtained in Comparative Example 1-3 were mixed in a ratio of 1: 1: 1 in the ratio of banana: cherry: walnut (weight ratio). Mixed extracts were dissolved in DMSO (0.33% by weight of bananas, cherry and walnut extracts, 1% by weight of mixed extracts, 10 μg / mL).
Experimental Example One: MT -One, SIRT -One, Catalase ( catalase) Recovery efficacy experiment
MT-1, SIRT-1, and catalase recovery effects of the extracts of Comparative Examples 1-3 and Example 1 were confirmed. To this end, human fibroblasts were inoculated into dexamethasone-treated neuroblast cultures at a density of 4 × 10 5 / well and cultured in a 5% CO 2 incubator at 37 ° C. for 24 hours.
Specifically, neuroblasts were treated with 1 μM dexamethasone and cultured for 24 hours to obtain a culture solution. Human dermal fibroblasts were inoculated into the culture at a density of 4 x 10 < 5 > / well. Then, 1 mL (10 μg / mL) of each of Comparative Example 1-3 and Example 1 was added and cultured in a 5% CO 2 incubator at 37 ° C. for 24 hours. 100 μL of lysis buffer was added to extract proteins. The Bradford assay was performed on the extracted proteins and the protein was quantified by measuring the absorbance at 595 nm wavelength. In addition, SDS-PAGE was performed and the protein band was transferred to PVDF (Polyvinylidene fluoride, Bio-Rad, USA), and the protein expression level was confirmed by ECL (Enhanced chemiluminescence, ThermoScientific, USA) solution.
As a result, as shown in Figs. 1 to 3, when the extract of Example 1 was treated, it showed the highest MT-1, SIRT-1, and catalase recovery effect. In addition, each protein was measured as shown in Table 1 below.
Correction value
As shown in the above results, it was confirmed that each of the banana, cherry or walnut extracts exhibited a synergistic effect when added as a mixed extract, rather than when added to a cell culture.
Experimental Example 2: Depending on the mixing ratio of banana, cherry and walnut extracts MT -One, SIRT -1, catalase recovery efficacy experiment
The extracts of each of the bananas, cherries and walnuts obtained in the above Comparative Example 1-3 were mixed in a ratio of 1: 1: 1, 2: 1: 1, 1: 2: 1 or 1: 1: 2 And used in the following experimental examples. Mixed extracts were dissolved in DMSO (1% total mixed extract, 10 μg / mL). Then, the amount of protein was quantified by the same method as Experimental Example 1.
The results are shown in FIG. 4 and Table 2 below.
Correction value
1: 1: 1
2: 1: 1
1: 2: 1
1: 1: 2
1 to 2: 1 to 2: 1 to 2 as compared with the ratio of 1: 1: 1 as shown in the above results. In particular, when the bananas, cherry and walnut extracts were mixed in a ratio of 1: 2: 1 , It was confirmed that the anti-aging effect due to MT-1 and catalase recovery was greatest.
Formulation Example 1: Manufacture of Toner
The toner preparation containing the extract of Example 1 was prepared in a conventional manner according to the compositional components and composition ratios shown in Table 3 below.
Formulation Example 2: Preparation of lotion agent
Lotion preparations containing the extracts obtained in Example 1 were prepared according to the conventional methods according to the composition components and composition ratios shown in Table 4 below.
Formulation Example 3: Manufacture of cream agent
The creams containing the extracts obtained in Example 1 were prepared according to the conventional methods according to the compositional components and composition ratios shown in Table 5 below.
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