KR102154140B1 - Composition comprising tulip leaf extract, amorphophallus paeoniifolius hydrolysis extract and parrot ferment extract - Google Patents

Abstract

The present invention relates to a cosmetic composition and/or a food composition comprising a tulip leaf extract, a hydrolyzed extract of elephant food and a fermented Schotia brachypetala extract. The composition may be used as a composition for moisturizing skin and delaying aging.

Description

A composition comprising tulip leaf extract, hydrolyzed extract of Old Yakic potato, and fermented parrot extract {COMPOSITION COMPRISING TULIP LEAF EXTRACT, AMORPHOPHALLUS PAEONIIFOLIUS HYDROLYSIS EXTRACT AND PARROT FERMENT EXTRACT}

The present invention relates to a cosmetic composition and/or a food composition comprising a tulip leaf extract, an Old Yak potato hydrolyzed extract, and a parrot tree fermented extract. The composition may be used as a composition for improving skin moisturization and aging.

Skin aging is an intrinsic (chronological aging) phenomenon that occurs naturally due to the decrease in physiological functions of the living body as age increases, and photoaging in which the appearance or function of the skin changes as the skin is repeatedly exposed to light. have. However, recently, research has been reported that as the earth's temperature rises, the increase in skin temperature due to the heat of light is also an important cause of skin aging. It is proved that as the skin is exposed to heat and the skin temperature rises, collagen and elastin, which are major components of the skin, decrease, and the decomposition of the component protein is accelerated to promote skin aging, such as wrinkles. However, up to now, sunscreens cannot solve the rise in skin temperature, which is an important cause of skin aging.

On the other hand, as various studies reveal the process of controlling skin temperature, attempts to lower the skin temperature are underway. According to a recent study, it has been found that transient receptor potential channels (TRP channels) play an important role in the process of temperature signaling, and phosphatidylinositol diphosphate and diacylglycerol (phosphatidylinositol diphosphate) and diacylglycerol (via G-protein) play an important role. diacylglycerol), protein kinase C.

The TRP channel, a temporary sensory receptor, is a non-specific cation channel that is widely expressed in the skin. It is mainly responsible for the influx of calcium ions into cells, and functions for various senses such as light stimulation, mechanical stimulation, temperature stimulation, chemical stimulation, and osmotic pressure change. This is known. Among them, TRPV1 (Transient Receptor Potential Vanilloid1) is a thermal receptor and a pungent receptor that functions as a sensor for harmful stimuli such as ultraviolet rays. That is, since TRPV1 can be a target for preventing skin photoaging caused by exposure to ultraviolet rays, etc., many studies are being conducted as a skin temperature control mechanism to prevent and prevent skin aging.

Therefore, as described above, in order to solve the increase in skin temperature, which is an important cause of skin aging, it is necessary to discover natural materials that do not have toxicity and side effects, and that can control skin temperature.

Korean Patent Publication No. 10-2015-0090014

An object of the present invention is to provide a cosmetic composition comprising a tulip leaf extract, an Old Yak potato hydrolyzed extract and a parrot tree fermented extract.

Another object of the present invention is to provide a food composition comprising a tulip leaf extract, an Old Yak potato hydrolyzed extract and a parrot tree fermented extract.

One aspect of the present invention for achieving the above object relates to a cosmetic composition comprising a tulip leaf extract, an Old Yak potato hydrolyzed extract, and a parrot tree fermented extract.

In the present invention, "tulip ( Liriodendron tulipifera )" is a plant belonging to the Magnoliaceae group and is known as yellow poplar. Tulip leaves and stems contain l-tuliposide A, B, and C, and are known to have antibacterial effects. In addition, it contains a large amount of flavonoid components, and is known to have antioxidant and anticancer effects. In the present invention, among tulips, a leaf extract known to be excellent in antioxidant and antibacterial activity is used.

The "extract" of the present invention is an extract obtained by extracting tulip leaves, a dilution or concentrate of the extracts, a dried product obtained by drying the extract, a preparation or purified product of the extract, or a mixture thereof, the extract itself, and It includes extracts of all formulations that can be formed using the extract.

The extract of the present invention may be extracted from natural, hybrid or variant plants of the tulip, and may also be extracted from plant tissue culture.

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

The type of the extraction solvent used in the present invention is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the extraction solvent include water, C ₁ to C ₄ anhydrous or lower alcohol, a mixed solvent of water and lower alcohol, acetone, 1,3-butylene glycol, ethyl acetate, selected from the group consisting of chloroform It can be used alone or in combination of two or more.

More specifically, ethanol may be used as the extraction solvent in the present invention. A solvent extract may be prepared by extracting the tulip at least once using the solvent, and a dried extract obtained by distilling the solvent extract under reduced pressure and then freeze drying or spray drying may be prepared. In a specific example of the present invention, a liquid extract was obtained using ethanol as an extraction solvent (Example 1-1)

In the present invention, the extract obtained by heat extraction or cold needle extraction is filtered to remove suspended solid particles, for example, nylon, etc. to filter the particles, or after filtration using a freeze filtration method, etc., and then used as such, or freeze-dried, hot-air drying, It can be used after drying by spray drying or the like.

In the present invention, “Old Yak Potato ( Amophophallus rivieri Durieu )” is also called Konjac Potato, and is a perennial grass of the Young Man family , native to Indonesia. Old Yak potatoes are rich in dietary fiber and are known to help intestinal action, constipation, and metabolic syndrome, and are known to be effective in preventing diabetes and controlling cholesterol levels by inhibiting the absorption of glucose. In particular, it is known to have an excellent effect of inhibiting melanin secretion and preventing aging because it is rich in ceramide ingredients.

The hydrolyzed extract of Old Yak Potato refers to an extract obtained by using a hydrolysis reaction including a hydrolyzed enzyme in Old Yak Potato. Specifically, the Old Yak potato hydrolyzed extract is obtained by inoculating a hydrolytic enzyme, and the hydrolytic enzyme is any one selected from the group consisting of cellulase, xylanase, and mannase. I can.

In one embodiment of the present invention, after hot water extraction of Old Yak potatoes, cellulase hydrolase was treated to obtain a hydrolyzed extract of Old Yak potatoes (Example 1-2).

In the present invention, "parrot tree ( Butea monosperma )" is a legume plant that includes orange flowers of about 6 to 12 m in height. Parrot tree exhibits various effects depending on the site. The roots show night blindness, hemorrhoids, ulcers, tumors, and pain relief effects, and flowers are effective in purifying body wastes, tonics, leprosy, skin diseases, gout, and burns. It is effective in dysfunction and edema.

The parrot tree fermented extract of the present invention refers to a composition obtained by inoculating the parrot tree extract and fermenting the strain.

In the present invention, "fermentation" refers to a process of decomposing organic matter using enzymes possessed by the fermentation strain itself. It is known that raw materials that have undergone a fermentation process through a fermentation strain have an effect of at least two to tens of times greater than before fermentation. Metabolites that have undergone fermentation contain various amino acids, organic acids, and antioxidants that are good for the skin to promote skin metabolism and smooth the skin texture. In addition, through the fermentation process, the particles become smaller and toxicities such as heavy metals are decomposed, so that the absorption rate is good, and skin troubles and allergic side effects are alleviated. Furthermore, by fermentation using beneficial bacteria such as lactic acid bacteria and yeast, it is possible to improve the efficacy of promoting and/or promoting beneficial bacteria in the ingredients.

Specifically, the parrot tree fermentation extract may be obtained by inoculating mushroom mycelium. More specifically, the mushroom mycelium may be a mycelium made of shiitake and shiitake mushrooms as strains, but is not limited thereto, and various mushroom mycelium may be used.

In an embodiment of the present invention, a parrot tree fermented extract was prepared by treating the mushroom mycelium with the extract obtained by hot water extraction of the parrot tree (Example 1-3).

In the present invention, the inventors prepared a mixed composition comprising the tulip leaf extract, the Old Yakic potato hydrolyzed extract, and the parrot tree fermented extract in a certain ratio, and found out through specific experiments that the composition has excellent efficacy in moisturizing skin and improving aging. I did. Accordingly, a composition comprising the tulip leaf extract, hydrolyzed extract of Old Yakic potato, and fermented parrot tree extract is intended to be used in a cosmetic composition.

In an embodiment of the present invention, as a result of evaluating cytotoxicity to skin keratinocytes, in order to confirm the skin stability of the cosmetic composition comprising the tulip leaf extract, the Old Yakic potato hydrolysis extract, and the parrot tree ferment extract, skin keratinocytes It was confirmed that the stability was high (Table 1).

In the present invention, the term “skin moisturizing” includes both supplementing by supplying moisture to the skin or protecting the skin from escaping moisture. In general, the amount of moisture in the skin is about 70% in the dermis layer, but decreases toward the epidermis layer to become about 10 to 30% in the stratum corneum. When the moisture content of such a cell layer is less than 10%, the skin becomes rough and the body protective function is weakened, resulting in aging, and moisture supply and moisturization are important for skin protection.

In an embodiment of the present invention, the effect of increasing the production amount of hyaluronic acid and aquaporin 3, which are skin moisturizing factors, was confirmed, and in particular, all of the tulip leaf extract, the hydrolyzed extract of Old Yak and the parrot tree fermented extract were included. It was confirmed that the composition exhibited excellent skin moisturizing effect compared to when using the composition of one or two extract combinations (Tables 2 and 3).

In the present invention, "aging" is a process in which the organs of the human body gradually deteriorate structurally and functionally with age. In the case of aging, there are aging due to highly reactive free radicals generated during normal metabolism and aging due to genetic causes. As for the aging phenomenon, various phenomena such as dry skin symptoms appear due to decrease in sebum amount, intercellular lipid, collagen, etc., wrinkle formation and elasticity decrease due to decrease in dermal cells, and skin protection ability decrease.

The skin aging improvement is to prevent or improve skin aging that is promoted due to such external or internal causes, and may improve skin aging through antioxidant action and/or skin wrinkle improvement.

In the present invention, “antioxidation” refers to an action of inhibiting oxidation, and in the human body, an oxidation promoting substance and an oxidation inhibitor are in balance, but due to various factors, the balance is lost and the direction of promoting oxidation When inclined toward, oxidative stress is induced in a living body, leading to cell damage and pathological diseases. Reactive oxygen species (ROS), which are the direct cause of oxidative stress, are chemically unstable and highly reactive, so they can easily react with various biomaterials such as DNA, proteins, lipids, and carbohydrates. Attacks cause irreversible damage to cells and tissues, or cause mutations, cytotoxicity and cancer, and are also a direct cause of aging. By removing or reducing these reactive oxygen species, antioxidant effects are obtained, preventing aging and maintaining health.

In one embodiment of the present invention, the antioxidant effect was confirmed by measuring free radical scavenging activity, and in particular, it was confirmed that the antioxidant effect became remarkable by hydrolysis and fermentation of Old Yak potato and parrot tree, respectively (Table 4).

In the present invention, "skin wrinkles" refers to fine lines caused by deterioration of the skin, which may be caused by genes, reduction of collagen and elastin present in the skin dermis, and external environment.

In the present invention, "skin wrinkle improvement" refers to suppressing or inhibiting the generation of wrinkles on the skin, or alleviating wrinkles that have already been generated.

In one embodiment of the present invention, when a cosmetic composition comprising a tulip leaf extract, an Old Yakic potato hydrolyzed extract, and a parrot tree fermented extract is applied, it exhibits excellent collagenase inhibitory activity, thereby showing a substantially remarkably excellent skin wrinkle improvement effect. It was confirmed that it can be (Table 5).

The cosmetic composition of the present invention is a solution, external ointment, cream, foam, nutrient lotion, flexible lotion, pack, softening water, emulsion, makeup base, essence, soap, liquid detergent, bathing agent, sunscreen cream, sun oil, suspension, emulsion It can be prepared in a formulation selected from the group consisting of liquid, paste, gel, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, patch, and spray, but is limited thereto. no.

The cosmetic composition of the present invention may additionally include one or more cosmetically acceptable carriers blended in general skin cosmetics, and as common ingredients, for example, oil, water, surfactants, moisturizers, lower alcohols, thickeners , A chelating agent, a colorant, a preservative, a fragrance, and the like may be appropriately blended, but are not limited thereto.

The cosmetically acceptable carrier included in the cosmetic composition of the present invention varies depending on the formulation of the cosmetic composition.

When the formulation of the present invention is an ointment, paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. May be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc. may be used as a carrier component. In particular, in the case of a spray, additional chlorofluorohard It may include a propellant such as locarbon, propane/butane or dimethyl ether, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent may be used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, Propylene glycol, 1,3-butyl glycol oil, and the like may be used, and in particular, cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan May be used, but is not limited thereto. These may be used alone or in combination of two or more.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, micro Crystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used, but are not limited thereto. These may be used alone or in combination of two or more.

In addition, the composition of the present invention may be used as a transdermal administration method such as direct application or spraying on the skin, and the administration route of the composition of the present invention may be administered through any general route as long as it can reach the target tissue.

The amount of use of the composition of the present invention may be appropriately adjusted according to individual differences or formulations such as age and degree of lesions, and may be used from one week to several months by applying one or several times a day to the skin.

Another aspect of the present invention relates to a food composition comprising a tulip leaf extract, an Old Yak potato hydrolyzed extract, and a parrot tree fermented extract.

Description of the "tulip leaf extract", "old yak potato hydrolyzed extract" and "parrot tree fermented extract" are the same as described above.

Specifically, the Old Yakic potato hydrolyzed extract is obtained by inoculating a hydrolytic enzyme, and the hydrolytic enzyme is any one selected from the group consisting of cellulase, xylanase, and mannase. I can.

In addition, specifically, the parrot tree fermentation extract may be obtained by inoculating cultured mushroom mycelium.

In addition, specifically, the food composition may be a food for skin moisturizing or skin aging improvement.

The “skin moisturizing” and “skin aging improvement” are already the same as described above.

Foods to which the tulip leaf extract of the present invention, the hydrolyzed extract of Old Yakic potato, and the fermented parrot tree extract can be added include meat, bread, sausage, chocolate, snacks, candy, confectionery, ramen, pizza, other noodles, gum, ice cream Dairy products, including various soups, beverages, tea, vitamin complexes, and health supplements.

The type of food may specifically be a health functional food. The health functional foods include various nutrients, vitamins, minerals (electrolytes), synthetic flavors and flavoring agents such as natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, organic acids, protective colloidal growth It may contain an agent, a pH adjuster, a stabilizer, a preservative, glycerin, alcohol, a carbonation agent used in carbonated beverages, and the like. These components may be used alone or in combination, and the proportion of these additives is generally selected from 0.001 to 50 parts by weight per total weight of the composition.

The health functional food is a food that emphasizes the biological regulation function of food, and is a food that has added value to act and express it for a specific purpose by using physical, biochemical, and bioengineering methods. Ingredients of these health functional foods are designed and processed to fully exert the body's control functions related to body defense, regulation of body rhythm, prevention and recovery of diseases to the living body, and food additives, sweeteners, or functional foods acceptable as foods. It may contain raw materials.

The health functional food may be any one formulation selected from the group consisting of tablets, granules, powders, capsules, liquid solutions, and pills, but is not limited thereto. Specifically, the health functional food in the form of a tablet is a composition comprising a tulip leaf extract, an Old Yakic potato hydrolyzed extract, and a parrot tree fermented extract, an excipient, a binder, a disintegrant, and a mixture of other additives, granulated by a conventional method, and then activated. Compression molding, or the like can be prepared by directly compression molding the mixture. In addition, the health functional food in the form of a tablet may contain a bitter agent or the like as necessary, and may be coated with a suitable coating agent if necessary.

Among the capsule-type health functional foods, the hard capsule is a mixture with additives such as a composition and excipients containing tulip leaf extract, Old Yak potato hydrolyzed extract, and parrot tree ferment extract in a conventional hard capsule, or granular or coated granules thereof It can be prepared by filling water. Soft capsules may be prepared by filling a mixture with additives such as a composition including a tulip leaf extract, an Old Yakic potato hydrolyzed extract, and a parrot tree fermented extract, and an excipient in a capsule base such as gelatin. The soft capsules may contain plasticizers such as glycerin or sorbitol, colorants, preservatives, and the like, if necessary.

The ring-shaped health functional food may be prepared by molding a mixture of a tulip leaf extract, a hydrolyzed extract of Old Testament potato, and a parrot tree fermented extract, an excipient, a binder, a disintegrant, etc. It is also possible to coat the coat with a white sugar or other suitable coat, or with starch, talc, or a suitable material.

The granular health functional food may be prepared in granular form by a suitable method, including a composition including tulip leaf extract, hydrolyzed extract of Old Yakic potato, and fermented parrot tree extract, excipients, binders, and disintegrants. It may contain flavoring agents, bittering agents, and the like.

The definition of terms for the excipients, binders, disintegrants, lubricants, bittering agents, flavoring agents, and the like are known in the art, and may include the same or similar functions such as functions.

In addition, the type of food may be a food additive, and the food additive refers to a substance used by a method such as adding, mixing, infiltrating or adding to food for manufacturing, processing, or preserving food. The food additives include natural products and synthetic products, and can be classified according to functions and uses. Currently, 370 kinds of chemical synthetic products and 50 kinds of natural additives are approved as food additives in Korea. Depending on the purpose, preservatives, fungicides, antioxidants, coloring agents, coloring agents, bleaching agents, seasonings, sweeteners, flavoring agents, expanding agents, strengthening agents, and improving agents , Emulsifiers, thickeners and stabilizers, coating agents, gum base agents, defoaming agents, solvents, releasing agents, insect repellents, quality improvers and other additives for food manufacturing.

The form of the food additive may include a powder, granule, tablet, capsule or liquid form, and specifically may be a capsule form, but is not limited to the form.

When used as a food composition comprising the tulip leaf extract of the present invention, the Old Yakic potato hydrolyzed extract and the parrot tree fermented extract, the tulip leaf extract, the Old Yakic potato hydrolyzed extract and the parrot tree fermented extract were added as it is, or other foods or foods It can be used together with an ingredient, and can be used suitably according to a conventional method. The mixing amount of the tulip leaf extract, the hydrolyzed extract of Old Yakic potato, and the parrot tree fermented extract may be appropriately determined according to the purpose of use (prevention, health or improvement, therapeutic treatment).

The cosmetic composition comprising the tulip leaf extract, the hydrolyzed extract of Old Yakic potato, and the parrot tree fermented extract of the present invention has excellent skin moisturizing and aging improvement effects, and it is useful as a cosmetic composition and food composition for skin moisturizing or aging improvement. high.

The effects of the present invention are not limited to the above effects, and should be understood to include all effects that can be deduced from the configuration of the invention described in the detailed description or claims of the present invention.

Hereinafter, the present invention will be described in detail by examples. However, the following examples are only illustrative of the present invention, and the present invention is not limited by the following examples.

Example 1. Composition containing tulip leaf extract, hydrolyzed extract of Old Yakic potato, and fermented parrot tree extract

1-1. Manufacture of tulip leaf extract

Each tulip leaf was washed with purified water, dried, and coarsely pulverized. 100 g of the pulverized product was mixed with 1 L of 70% ethanol at a ratio of 10% by weight. The mixed raw materials were immersed and extracted at room temperature for 7 days, and then filtered through 250 mesh and 0.5 μm filter paper to obtain a liquid extract. The obtained liquid extract was used in a rotary vacuum concentrator and a freeze dryer to obtain a tulip leaf extract.

1-2. Manufacture of hydrolyzed extract of Old Yak potato

The Old Yak potatoes were washed with purified water, dried, and 20 times the weight of purified water was added as an extraction solvent, and an extractor equipped with a cooling condenser (Cosmos-660, heated to 100℃ in Kyungseo Machinery) was extracted three times for 2 hours each. For cellulase activity as a degrading enzyme, an acetate buffer (pH 4.0) and a sodium hydroxide buffer (pH 10.0) were added to adjust the pH to 3-4. 1% by weight of cellulase was added to the cooling condenser. In an extractor (Cosmos-660, Kyungseo Machinery), the reaction was carried out for 2-6 hours at 200 rpm at 50° C. After that, the reaction solution was allowed to stand at room temperature for 3 days, the precipitate was filtered through a 300 mesh filter paper, and the precipitate was adventek No. 5 Filtered twice with filter paper and Whatman GFC 150 mm filter paper, and concentrated at 50℃ using a vacuum concentrator, and then used a spray dryer (B-290, BUCHI) to inlet temperature of 180℃ and inhaler ( Aspirator) efficiency of 100% (35 cm ² / hour), Pump efficiency of 25% (7.5 ml / min), Nozzle Cleaner 4 and Flow meter (Rotameter): 30 mm (357 liters / hour) Dry under conditions to obtain the title extract powder.

1-3. Parrot tree ferment extract manufacturing

First, using shiitake mushrooms and shiitake mushrooms as strains, they were used by subculturing them on a plate medium of Potato Dextrose Agar (PDA, DIFCO, USA). Shiitake and shiitake mushroom mycelium were cultured in PDA medium at a temperature of about 30° C. for 10 days. The obtained mushroom mycelium was cut with an inoculation knife and inoculated in Potato Dextrose Broth medium and cultured with shaking. After 4 days of culture, each cell was treated 5 times for 1 second using a Waring blender, and 10 mL was again inoculated into Potato Dextrose Broth (100 mL) to prepare mushroom mycelia.

Each parrot tree was washed with purified water, dried, and coarsely pulverized, followed by high-pressure hot water extraction at 121°C for 15 minutes with 20 times the extraction solvent (purified water). The extracted parrot tree extract was centrifuged to recover the extracted supernatant, and the extract was filtered through a filter having a size of 0.2 μm to prepare parrot tree extracts, respectively. The extracted and filtered parrot tree extract was used after lyophilization.

Then, a parrot tree fermented extract was prepared using the cultured mushroom mycelium. After mixing the parrot tree extract with the mushroom mycelium, it was fermented by incubating for 5 days at a temperature of 27 ~ 30 ℃. The fermented product was recovered, mixed with 10 times the weight of 1,3-butylene glycol and purified water 3: 7 (w/w) in a mixed solvent, immersed for 3 days, and boiled for 8 hours at 100℃ in an extractor equipped with a cooling condenser. Extracted. After filtering using filter paper (Qualitive Filter Papers NO.2), it was obtained as a filtrate.

1-4. Preparation of complex extract containing tulip leaf extract, hydrolyzed extract of Old Yakic potato, and fermented parrot tree extract

A composite extract was prepared by mixing the tulip leaf extract, hydrolyzed extract and parrot tree fermented extract prepared in 1-1, 1-2 and 1-3 above in a ratio of 1:1.

Comparative Example 1. Composition containing tulip leaf extract

Each tulip leaf was washed with purified water, dried, and coarsely pulverized. 100 g of the pulverized product was mixed with 1 L of 70% ethanol at a ratio of 10% by weight. The mixed raw materials were immersed and extracted at room temperature for 7 days, and then filtered through 250 mesh and 0.5 μm filter paper to obtain a liquid extract. The obtained liquid extract was used in a rotary vacuum concentrator and a freeze dryer to obtain a tulip leaf extract.

Comparative Example 2. Composition containing hydrolyzed extract of Old Yakic potato and fermented parrot tree extract

2-1. Manufacture of hydrolyzed extract of Old Yak potato

The Old Yak potatoes were washed with purified water, dried, and 20 times the weight of purified water was added as an extraction solvent, and an extractor equipped with a cooling condenser (Cosmos-660, heated to 100℃ in Kyungseo Machinery) was extracted three times for 2 hours each. For cellulase activity as a degrading enzyme, an acetate buffer (pH 4.0) and a sodium hydroxide buffer (pH 10.0) were added to calibrate the pH to 3 to 4. 1% by weight of cellulase was added and an extractor with a cooling condenser ( Cosmos-660, Kyungseo Machinery) reacted for 2 to 6 hours at 200 rpm at 50° C. After that, the reaction solution was allowed to stand at room temperature for 3 days, the precipitate was filtered through a 300 mesh filter paper, and the precipitate was filtered with an Edbentech No. 5 filter paper and watts. Only GFC was filtered twice with 150 mm filter paper, and concentrated at 50℃ using a vacuum concentrator, and then inlet temperature 180℃ and aspirator efficiency using a spray dryer (B-290, BUCHI). 100% (35 cm ² /hour), Pump efficiency 25% (7.5 ml/min), Nozzle Cleaner 4 and Rotameter: Dry under the conditions of 30 mm (357 liters/hour) To obtain the title extract powder.

2-2. Parrot tree ferment extract manufacturing

First, using shiitake mushrooms and shiitake mushrooms as strains, they were used by subculturing them on a plate medium of Potato Dextrose Agar (PDA, DIFCO, USA). Shiitake and shiitake mushroom mycelium were cultured in PDA medium at a temperature of about 30° C. for 10 days. The obtained mushroom mycelium was cut with an inoculation knife and inoculated in Potato Dextrose Broth medium and cultured with shaking. After 4 days of culture, each cell was treated 5 times for 1 second using a Waring blender, and 10 mL was again inoculated into Potato Dextrose Broth (100 mL) to prepare mushroom mycelia.

Each parrot tree was washed with purified water, dried, and coarsely pulverized, followed by high-pressure hot water extraction at 121°C for 15 minutes with 20 times the extraction solvent (purified water). The extracted parrot tree extract was centrifuged to recover the extracted supernatant, and the extract was filtered through a filter having a size of 0.2 μm to prepare parrot tree extracts, respectively. The extracted and filtered parrot tree extract was used after lyophilization.

Then, a parrot tree fermented extract was prepared using the cultured mushroom mycelium. After mixing the parrot tree extract with the mushroom mycelium, it was fermented by incubating for 5 days at a temperature of 27 ~ 30 ℃. The fermented product was recovered, mixed with 10 times the weight of 1,3-butylene glycol and purified water 3: 7 (w/w) in a mixed solvent, immersed for 3 days, and boiled for 8 hours at 100℃ in an extractor equipped with a cooling condenser. Extracted. After filtering using filter paper (Qualitive Filter Papers NO.2), it was obtained as a filtrate.

2-3. Preparation of complex extract containing hydrolyzed extract of Old Yakic potato and fermented parrot tree extract

A composite extract was prepared by mixing the hydrolyzed extract of Old Yak potato and the fermented parrot tree extract prepared in 2-1 and 2-2 at a ratio of 1:1.

Comparative Example 3. A composition comprising a tulip leaf extract, an old yak potato extract and a parrot tree extract.

Tulip leaf extract was prepared in the same manner as in Comparative Example 1.

In the case of Old Yak potato extract, wash the Old Yak potato with purified water, dry it, add 20 times the weight of purified water as an extraction solvent, and heat it to 100℃ in an extractor with a cooling condenser (Cosmos-660, heated to 100℃ in Kyungseo Machinery, 3 times a total of 2 hours). Extracted.

In the case of the parrot tree extract, each parrot tree was washed with purified water, dried, and coarsely pulverized, followed by high-pressure hot water extraction at 121°C for 15 minutes with 20 times the extraction solvent (purified water). The extracted parrot tree extract was centrifuged to recover the extracted supernatant, and the extract was filtered through a filter having a size of 0.2 μm to prepare parrot tree extracts, respectively. The extracted and filtered parrot tree extract was used after lyophilization.

Thereafter, a composite extract was prepared by mixing the tulip leaf extract, the Old Yak potato extract, and the parrot tree extract in a ratio of 1:1.

Experimental Example 1. Skin keratinocyte culture

HaCaT cells, which are skin keratinocytes, are Dr. Pre-sale from CG Hyun (Jeju National University, Korea) and using Dulbecco's Modified Eagle's Medium (DMEM) medium containing 100 units/ml penicillin-streptomycin and 10% fetal bovine serum (FBS) at 37°C, It was cultured in a 5% CO ₂ incubator, and subculture was performed every 3 to 4 days.

Experimental Example 2. Cytotoxicity evaluation (EZ-cytox assay)

To confirm the skin stability of Example 1, cytotoxicity evaluation was performed using the EZ-cytox assay method. EZ-cytox assay is a representative method of measuring cell viability using the principle of producing orange water-soluble formazan by reacting with dehydrogenase of living cells.

Specifically, in order to confirm the toxicity in skin keratinocytes, HaCaT cells were dispensed into a 96-well plate at 5 X 10 ⁴ cells/mL using DMEM medium supplemented with 10% FBS, under conditions of 37°C and 5% CO _2. After reacting for 18 hours at, the absorbance was measured at 450 nm using a microplate reader. The average absorbance value for each sample group was obtained, and the cell viability was evaluated by comparing it with the absorbance value of the control group. The results are shown in Table 1.

Cell viability (% control) Concentration (ug/ml) 0 50 100 125 Example 1 100 110 122 133

As shown in Table 1, a composition comprising a tulip leaf extract, an Old Yakic potato hydrolysis extract, and a parrot tree ferment extract (Example 1) showed excellent cell survival regardless of concentration, and toxicity to skin keratinocytes. It was confirmed that the stability was high because there was little.

Experimental Example 3. Confirmation of skin moisturizing effect

3-1. Confirmation of the effect of increasing the amount of moisturizing factor hyaluronic acid produced

In order to confirm the moisturizing effect of Example 1 and Comparative Examples 1 to 3, an experiment was performed to confirm the effect of increasing the amount of hyaluronic acid produced.

Specifically, HaCaT cells were dispensed into a 24-well plate at 1.0 × 10 ⁵ cells/mL, and cultured for 18 hours at 37°C and 5% CO ₂ . It was exchanged for serum-free DMEM, treated with Example 1 and Comparative Examples 1 to 3, and cultured for 24 hours. Thereafter, the culture medium was removed, centrifuged at 15,000 rpm for 5 minutes, and the supernatant was removed and stored frozen (-20°C) until quantification. The enzyme immunoassay (ELISA: Enzyme-Linked Immunosorbent Assay) was performed using a hyaluronic acid ELISA kit (Elabscience Biotechnology Co., Ltd), and was carried out by the method provided by the manufacturer. The results are shown in Table 2.

Hyaluronic acid production increase (%) - density % Control - 100 Example 1 50 156.0 Comparative Example 1 50 100.4 Comparative Example 2 50 76.0 Comparative Example 3 50 120.5

As shown in Table 2, the composition (Example 1) comprising a tulip leaf extract, an Old Yakic potato hydrolyzed extract, and a parrot tree fermented extract (Example 1) was a tulip leaf alone extract (Comparative Example 1) or an Old Yakic potato hydrolyzed extract and parrot tree It was confirmed that the amount of hyaluronic acid produced was significantly increased compared to the composition containing the fermented extract (Comparative Example 2).

This suggests that the composition including the tulip leaf extract, the hydrolyzed extract of Old Yakic potato, and the parrot tree fermented extract has excellent skin moisturizing effect.

3-2. Confirmation of the effect of increasing the amount of moisturizing factor Aquaporin 3 (Aquaporin3, AQP3) produced

In order to confirm the moisturizing effect of Example 1 and Comparative Examples 1 to 3, an experiment was performed to confirm the effect of increasing the amount of aquaporin 3 produced.

HaCaT cells were dispensed into a 24-well plate at 1.0 × 10 ⁵ cells/mL and cultured for 18 hours at 37°C and 5% CO ₂ . It was exchanged with serum-free DMEM, and the sample was treated and incubated for 24 hours. After removing the culture medium for each group and washing with PBS, PBS containing no drugs that may affect protein quantification was treated. The cells were lysed by repeating incubation at low and room temperature, and then proteins were collected. Quantification was performed using the AQP3-ELISA kit (Elabscience Biotechnology Co., Ltd.) and was performed by the method provided by the manufacturer. The results are shown in Table 3.

Aquaporin 3 (AQP3) production increase (%) - density % Control - 100 Example 1 50 180 Comparative Example 1 50 114.6 Comparative Example 2 50 91.5 Comparative Example 3 50 125.6

As shown in Table 3, it was confirmed that the composition (Example 1) comprising a tulip leaf extract, an Old Yak potato hydrolyzed extract, and a parrot tree ferment extract significantly increased the amount of aquaporin 3 factor produced. In particular, it was confirmed that the composition (Comparative Example 2) containing the hydrolyzed extract of Old Yakic potato and the fermented parrot tree had a slight skin moisturizing effect, while the skin moisturizing effect was remarkably increased by mixing the tulip leaf extract.

Through this, it was confirmed that the composition comprising the tulip leaf extract, the hydrolyzed extract of Old Yakic potato and the fermented parrot tree extract of the present invention is effective in moisturizing the skin.

Experimental Example 4. Antioxidant effect confirmation

4-1. DPPH scavenging activity confirmation

The antioxidant activity of Example 1 and Comparative Examples 1 to 3 was confirmed. For the evaluation of antioxidant activity, DPPH (1,1-diphenyl1-2-picry1hydrazy1) was used to measure the free radical scavenging activity. Specifically, after adding 20 μL of each of the extracts of Example 1 and Comparative Examples 1 to 3 to 180 μL of the DPPH solution, blocking the light from the outside and reacting at room temperature for 30 minutes, and then measuring the absorbance at 517 nm to measure the absorbance of the control group. Antioxidant activity was shown as a reduction rate. At this time, as a control for determining the rate of decrease in absorbance, ascorbic acid, an antioxidant, was used, and the same amount as the sample was added and measured in the same manner. Then, DPPH scavenging activity was calculated according to Equation 1 below.

[Equation 1]

DPPH scavenging activity (%) = [1-(absorbance of the sample treatment group in the Example or Comparative Example / absorbance of the sample untreated group)] X 100

DPPH elimination (%) Concentration (ug/ml) 125 250 500 Example 1 7.9 27.0 49.5 Comparative Example 1 5.5 20.5 39.8 Comparative Example 2 7.0 24.4 42.5 Comparative Example 3 7.4 25.6 43.0

As shown in Table 4, it can be seen that both Example 1 and Comparative Examples 1 to 3 exhibit DPPH scavenging activity. In particular, it was confirmed that the anti-wrinkle effect was remarkably increased by including the hydrolyzed extract of Old Yakic potato and the fermented parrot extract.

Experimental Example 5. Wrinkle improvement effect confirmation

The collagenase inhibition activity of Example 1 and Comparative Examples 1 to 3 was evaluated.

Specifically, by adding 4 mM CaCl ₂ to 0.1 M tris-HCl buffer (pH 7.5), 0.3 mg/mL of 4-phenylazobenzyl oxycarbonyl-Pro-Leu-Gly-Pro- 10 μL of a sample was mixed with 125 μL of a substrate solution in which D-Arg (sigma, USA) was dissolved.

To the mixture was added 0.075 mL of 1 mg/mL Clostridium histolyticum collagenase collagenase (Sigma-Aldrich Co, Louis, MO, USA). After standing at room temperature for 20 minutes, 0.5 mL of 6% citric acid was added to stop the reaction, and 2 mL of ethyl acetate was added, and the absorbance was measured at 320 nm with a spectrophotometer (UV-1650PC, Shimadzu, Kyoto, Japan).

The collagenase inhibitory activity was calculated by the ratio of the sample untreated according to the following equation, and the results are shown in Table 5.

[Equation 2]

Collagenase inhibitory activity (%) = 100-[(absorbance of strain culture solution treated group/absorbance of strain culture solution untreated group) X100]

Collagenase inhibitory activity (%) Concentration (μg/ml) 125 250 500 Example 1 48.6 55.7 67.6 Comparative Example 1 44.9 48.5 51.5 Comparative Example 2 44.9 47.0 50.2 Comparative Example 3 45.5 49.8 52.2

As shown in Table 5, the composition (Example 1) comprising a tulip leaf extract, an Old Yakic potato hydrolyzed extract, and a parrot tree fermented extract (Example 1) was a tulip leaf alone extract (Comparative Example 1), an Old Yakic potato hydrolyzed extract and a parrot tree It was confirmed that the fermentation extract (Comparative Example 2) and tulip leaf extract, Old Yak potato extract, and parrot tree extract (Comparative Example 3) showed remarkably superior collagenase inhibitory activity.

Preparation Example 1. Preparation of an essence containing a cosmetic composition comprising a tulip leaf extract of the present invention, a hydrolyzed extract of Old Yakic potato, and a parrot tree fermented extract

A cosmetic composition of an essence formulation was prepared as shown in Table 6 below (unit: wt%).

Raw material name Content (% by weight) Cetearyl alcohol 2 Cetearyl glucoside One Jojoba seed oil 5 Silica 0.5 glycerin 10 Example 1 3 to 4 Purified water To 100

Preparation Example 2. Preparation of a cream containing a cosmetic composition comprising the tulip leaf extract of the present invention, the hydrolyzed extract of Old Yakic potato, and the fermented parrot tree extract

As shown in Table 7 below, a cream-formed cosmetic composition was prepared (unit: wt%).

Composition Content (% by weight) Example 1 3 to 4 Stearic acid 15 ethanol One Potassium hydroxide 0.7 glycerin 5 Propylene glycol 3 antiseptic Appropriate amount incense Appropriate amount Purified water to 100

Preparation Example 3. Preparation of a mask pack containing a cosmetic composition comprising a tulip leaf extract, an Old Yak potato hydrolyzed extract and a parrot tree fermented extract of the present invention

As shown in Table 8 below, a mask pack was prepared using the cosmetic composition (unit: wt%).

Composition Content (% by weight) Example 1 3 to 4 Polyvinyl alcohol 15 Cellulose gum 0.15 glycerin 3.0 PEG 1500 2.0 Cyclomextrin 0.15 DL-Panlenol 0.4 Allantoin 0.1 Monoammonium glycyrrhizinate 0.3 Nicotinamide 0.5 ethanol 6.0 PEG 40 hydrogenated castor oil 0.3 Preservatives, fragrances, and pigments Appropriate amount Distilled water to 100

The above description of the present invention is for illustrative purposes only, and those of ordinary skill in the art to which the present invention pertains can understand that it is possible to easily transform it into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not limiting. For example, each component described as a single type may be implemented in a distributed manner, and similarly, components described as being distributed may also be implemented in a combined form.

The scope of the present invention is indicated by the claims to be described later, and all changes or modified forms derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention.

Claims (8)

As a cosmetic composition for skin moisturizing and skin aging improvement comprising a tulip leaf extract, an Old Yakic potato hydrolyzed extract, and a parrot tree fermented extract,
The hydrolyzed extract of Old Yak potatoes is obtained by inoculating cellulase, a cosmetic composition. delete The method of claim 1,
The parrot tree fermentation extract is obtained by inoculating cultured mushroom mycelium, cosmetic composition. delete As a food composition for skin moisturizing and skin aging improvement comprising a tulip leaf extract, an Old Yakic potato hydrolyzed extract and a parrot tree fermented extract,
The hydrolyzed extract of Old Yak potatoes is obtained by inoculating cellulase, food composition. delete The method of claim 5,
The parrot tree fermentation extract is obtained by inoculating cultured mushroom mycelium, food composition. delete