KR102530638B1 - Skin useful composition for antiaging - Google Patents

Abstract

The present invention discloses a skin-useful composition for anti-aging, which has collagenase inhibitory activity and elastase inhibitory activity, and exhibits ability to improve skin wrinkles and elasticity. The composition is obtained by preparing a dry solid product using terminalia bellerica , amla ( Amla, Phyllanthus emblica ), triphala ( Triphala ) and papaya ( Carica papaya ) and supercritical extraction using ethanol and a supercritical fluid. will be.

Description

Skin oil composition for anti-aging {SKIN USEFUL COMPOSITION FOR ANTIAGING}

The present invention relates to a skin-useful composition for anti-aging, which has collagenase inhibitory activity and elastase inhibitory activity and can express skin wrinkles and elasticity improvement ability.

With the development of medical technology and the improvement of living standards, the preference for skin wrinkle improvement, elasticity, and skin whitening is increasing, and the cosmetics market is also expanding.

The skin is composed of epidermis, dermis, and subcutaneous tissue under keratin to protect the human body from external harmful factors such as temperature, humidity, and ultraviolet rays in direct contact with the external environment.

With aging or exposure to ultraviolet rays, the amount of collagen synthesis decreases due to the action of fibroblasts and the decrease in the number of cells, and the action of collagenase and elastase, which decompose collagen, increases, resulting in loss of skin moisture and skin Flexibility and elasticity are reduced.

The most important environmental factor causing such skin aging is ultraviolet rays. When the skin is exposed to ultraviolet rays, metabolism harmful to the skin is activated, and chain scission and abnormal cross-linking of collagen and elastin are induced, resulting in skin tissue damage and skin wrinkles.

Therefore, a substance having an activity capable of inhibiting collagenase or elastase may have a skin wrinkle improvement effect (Photochem, acd Phptobiol. 2001. 74:283-287).

Terminalia bellerica is a deciduous tree belonging to the Terminalia family and has an antiviral effect against various pathogenic bacteria. There are only a number of reports on its antibacterial activity against Escherichia coli and Staphylococcus aureus, but it improves skin wrinkles or elasticity. There is nothing disclosed or disclosed about Barhera ( Terminalia bellerica ) with respect to performance for.

Accordingly, while the present inventors were researching a composition for improving skin wrinkles and elasticity, while improving skin wrinkles from a mixture using Bahera, Amla ( Amla, Phyllanthus emblica ), Triphala and Papaya (Carica papaya) The present invention was completed by confirming that the anti-aging performance that also improves elasticity can be expressed.

An object of the present invention is to provide a skin-useful composition that simultaneously expresses skin wrinkles and elasticity improvement using a supercritical extract of Terminalia bellerica .

The above and other objects of the present invention can all be achieved by the present invention described below.

In order to achieve the above object, the present invention

Bahera ( Terminalia bellerica ), Amla ( Amla, Phyllanthus emblica ), Triphala ( Triphala ) and Papaya ( Carica papaya ) The supercritical extract obtained by extracting using ethanol and supercritical fluid to the dried solids was used as an active ingredient. It provides a skin useful composition for improving skin wrinkles and elasticity comprising.

The supercritical fluid may be carbon dioxide, nitrogen, or a mixture thereof.

The supercritical extract may be a 70% ethanol supercritical extract of dried solids using Terminalia bellerica , Amla, Phyllanthus emblica , Triphala and Carica papaya .

The barhera ( Terminalia bellerica ), amla ( Amla, Phyllanthus emblica ), triphala ( Triphala ) and papaya ( Carica papaya ) are barhera (a): amla (b): triphala (c): papaya (c) It may satisfy a weight ratio of 1:0.5-1.5:0.5-1.5:0.5-1.5.

The composition may be a cosmetic composition.

According to the present invention, by maximizing the total polyphenol content and total flavonoid content, radical scavenging activity, SOD (superoxide dismutase)-like activity, antioxidant effect and whitening effect by xanthine oxidase inhibitory activity, and inhibitory activity against collagenase It is possible to provide a composition for preventing and treating skin wrinkles and elasticity improvement, which contains, as an active ingredient, a supercritical extract derived from Bahera, which simultaneously expresses skin wrinkle and elasticity improvement effects by inhibitory activity on elastase.

1 is a schematic diagram of a supercritical extraction device used in one embodiment of the present invention.
Figure 2 shows the total polyphenol content and total flavonoid content measurement results for three types of samples (TATP-3) obtained in Example 1 and samples (TATP-1, TATP-2) obtained in Comparative Examples 1 and 2 It is a graph for comparison.
Figure 3 is a graph comparing DPPH and ABTS radical scavenging activities for three types of samples (TATP-3) obtained in Example 1 and samples (TATP-1 and TATP-2) obtained in Comparative Examples 1 and 2. .
4 is a graph comparing the whitening activity of three types of samples (TATP-3) obtained in Example 1 and samples (TATP-1 and TATP-2) obtained in Comparative Examples 1 and 2.
5 is a graph comparing cytotoxicity by concentration for the sample (TATP-3) obtained in Example 1.

Hereinafter, the present invention will be described in more detail to aid understanding of the present invention.

Terms or words used in this specification and claims should not be construed as being limited to their usual or dictionary meanings, and given that the inventors may appropriately define the concept of terms in order to best describe the invention. Therefore, it should be interpreted as meaning and concept consistent with the technical spirit of the present invention.

As confirmed in the following Examples and Experimental Examples, the present invention is a solid phase using Terminalia bellerica , Amla, Phyllanthus emblic a, Triphala and Papaya ( Carica papaya ) When the dried product is prepared and the supercritical extract derived from Bahera is prepared using the fluid, the total polyphenol content and total flavonoid content are maximized to show radical scavenging activity, SOD (superoxide dismutase)-like activity, and xanthine oxidase inhibitory activity A composition for preventing and treating skin wrinkles and elasticity improvement by simultaneously expressing antioxidant effect, whitening effect, and skin wrinkle and elasticity improvement effect due to collagenase inhibitory activity and elastase inhibitory activity can provide

Ethanol-free supercritical extract or 100% ethanol supercritical extract maximizes the total polyphenol content and total flavonoid content, resulting in antioxidant and whitening effects by radical scavenging activity, SOD (superoxide dismutase)-like activity, and xanthine oxidase inhibitory activity , And did not show the effect of improving skin wrinkles and elasticity due to the inhibitory activity on collagenase and the inhibitory activity on elastase at the same time, and also not shown in the experimental example below, but barhera ( Terminalia bellerica ) content , Amla ( Phyllanthus emblica ) content, and triphala ( Triphala ) and papaya ( Carica papaya ) Total polyphenol content and total flavonoid content even when the weight ratio of 1:0.5-1.5:0.5-1.5 is not satisfied Radical scavenging activity by maximizing the content, SOD (superoxide dismutase)-like activity, antioxidant effect and whitening effect due to xanthine oxidase inhibitory activity, and skin wrinkles due to collagenase inhibitory activity and elastase inhibitory activity and the effect of improving elasticity was not shown at the same time or the effect was reduced.

By maximizing the total polyphenol content and total flavonoid content of the present invention, radical scavenging activity, SOD (superoxide dismutase)-like activity, antioxidant effect and whitening effect by xanthine oxidase inhibitory activity, and inhibitory activity against collagenase and ella A composition that simultaneously implements the effect of improving skin wrinkles and elasticity by the inhibitory activity on stease is provided based on these experimental results, and the skin-useful composition for improving skin wrinkles and elasticity of the present invention is Barhera ( Terminalia bellerica ) Amla ( Phyllanthus emblica ), Triphala ( Triphala ), and Papaya ( Carica papaya ) were mixed in a weight ratio of 1:0.5 to 1.5:0.5 to 1.5:0.5 to 1.5, and then to the obtained solid phase product. It is characterized in that it contains a supercritical extract obtained using a fluid as an active ingredient.

The skin-useful composition for improving skin wrinkles and elasticity of the present invention preferably contains terminalia bellerica , amla ( Amla, Phyllanthus emblica ), triphala ( Triphala ) and papaya ( Carica papaya ) in a ratio of 1:0.5 to 1.5: 0.5 ~ 1.5: It is characterized by including a supercritical extract obtained by using ethanol and a fluid as an active ingredient in a solid phase product obtained after mixing at 0.5 ~ 1.5.

In the present description, unless otherwise specified, an active ingredient means a component that exhibits the desired activity alone or that can exhibit activity in combination with a carrier that is not active by itself.

Improvement in the present description includes alleviation of symptoms of skin wrinkles, alleviation of sagging symptoms of skin elasticity, suppression of expression, delay of expression, and elimination of expression of such symptoms.

In the present description, unless otherwise specified, an extract refers to an extract obtained by using a supercritical fluid such as carbon dioxide or pentane and ethanol, or a fraction obtained by fractionating the extract, and the extraction method refers to the extraction method of the active substance. Arbitrary methods such as cooling, reflux, heating, ultrasonic radiation, and supersonic extraction may be applied in consideration of polarity, degree of extraction, and degree of preservation. In the case of a fractionated extract, the fraction or crude extract obtained by suspending the extract in a specific solvent, mixing with a solvent of different polarity, and standing still is adsorbed on a column filled with silica gel, etc. It means including the fraction obtained by.

In the present disclosure, the shape of the solid material is not particularly limited, and may be in the form of pills, granules, or powder.

The solid product may be obtained in the form of a solid product after thoroughly washing Terminalia bellerica , Amla, Phyllanthus emblica , Triphala and Carica papaya . After cleaning, some of the moisture is removed by drying in an arbitrary method such as natural drying (shade drying) or hot air drying (in the form of pulverized material so that the moisture content of the solid material is less than 10% (w/w)), and then supercritical can be used for extraction.

Furthermore, the pulverized product can be used as it is or in a dried state after steaming step by step, and it is preferable to use it in a dried state after steaming step by step because it can provide usefulness of storage and increase effect of active ingredients.

In the present disclosure, the supercritical extraction may be performed by various known methods as long as the extraction using a supercritical fluid is not particularly limited, and may be performed according to the schematic diagram shown in FIG. 1 as an example.

In the present description, a supercritical fluid is a fluid in a state above the critical temperature and critical pressure, and is also referred to as a concentrated gas in the sense of emphasizing the solvency of the fluid. In general, matter has three phases, namely solid, liquid, and gas, according to changes in temperature and pressure. It reaches a fourth state, the so-called supercritical state, in which it no longer changes to solid, liquid, or gas.

Supercritical Fluid Extraction (hereinafter also referred to as 'SFE') using a supercritical fluid has several advantages because it has the characteristics of a complex technology in which the principles of distillation and extraction are applied together.

Specifically, supercritical fluids can be set under any conditions from high density to low density by manipulating pressure and temperature, so selectivity such as fractionation and separation is excellent, so that high-purity products can be obtained, and extraction solvents can be almost completely removed without loss. It can be recovered, and a purified product without residual solvent can be obtained.

In addition, the supercritical fluid has low viscosity and good permeability to the sample, so the extraction efficiency is high and the diffusion coefficient is high, so the extraction speed is fast. It has many advantages, such as the large difference in density from the supercritical fluid and the low viscosity of the supercritical fluid, such that it is easy to separate the solvent from the extraction residue.

However, since a high-pressure device must be used, there is a disadvantage in that facility and maintenance costs are high, so extraction using a supercritical fluid is known to be economical only when it is performed with high efficiency.

The supercritical fluid of the present description is not particularly limited, but carbon dioxide may be used as a non-limiting example. Carbon dioxide has a critical pressure of 73.8 bar and a critical temperature of 31 °C, which is relatively low compared to other fluids, making it easy to create supercritical conditions, non-toxic, and economical.

As a non-polar solvent, supercritical carbon dioxide is widely used for extraction of low-polar substances such as oil and fat. For example, it is possible to easily induce a change in the polarity of the supercritical fluid by partially adding a polar substance such as ethanol, so that the solvent can be appropriately adjusted to be used for extraction of various fats and oils.

In addition, even under supercritical conditions, the properties of the supercritical fluid can be changed depending on the conditions of temperature and pressure. For example, when the supercritical carbon dioxide is extracted under low-pressure conditions, the solubility of the active ingredient decreases and the concentration of sugar in the extract increases, and even when extracted under high-pressure conditions suitable for the active ingredient extraction conditions, If separated by fractionation, the concentration of the active ingredient is higher than that of the extract in the second half. In this extraction method, it is possible to adjust the content of free fatty acids and the content of active ingredients by adjusting the fractionation time. There is a way to lower the degree of concentration by mixing the sections.

The free fatty acid content of raw materials varies depending on the country of origin or cropping, and since raw materials with low acid values are usually formed at high prices, they are less competitive as raw materials. In addition, since there is no raw material without free fatty acids, free fatty acids are concentrated in the extract concentration section, so even if a product is made by setting a standard for using raw materials with a low acid value, it is easy to meet the acid value standard while satisfying the active ingredient content at a certain concentration or higher. It's not work. Accordingly, in order to have industrial stability in the process of producing a high-concentration fraction using supercritical carbon dioxide, a technique for removing free fatty acids from the concentrated fraction is required.

In the supercritical extraction method, the degree of content of the constituents varies depending on the extraction conditions and the time of fractionation. This is because the solubility of each substance is different depending on the supercritical condition. However, since substances with similar solubility are extracted with similar tendencies depending on the extraction condition change or fractionation time point, it is difficult to separate them completely, although there are differences depending on the conditions or degree of fractionation.

According to the present invention, it is possible to provide an extract in which the content of active ingredients is maximized and the content of free fatty acids is lowered.

Both the active ingredient and free fatty acid are components that dissolve in supercritical fluid, and the solubility of the active ingredient and free fatty acid component differs depending on the supercritical condition, and even under the same condition, the content ratio of the early and second half of extraction is different. When supercritical conditions are set to low-density conditions, relatively active ingredients and free fatty acids are extracted in a high ratio in the extract, which is not desirable in terms of extraction efficiency because it lowers the overall extraction efficiency. When obtained by separating the first half and the second half of extraction under oil and fat extraction conditions, the active ingredient and free fatty acid are concentrated in the early part of extraction. By adjusting the fractionation time point, an extract enriched with active ingredients can be prepared.

In order to reduce the free fatty acid content of the fraction ?? Since mixing of fractions with low content is unavoidable, when raw materials with high free fatty acids are used or ?? In the case of high concentration, it is difficult to provide the adjustment of the fractionation time to the setting of the fractionation section.

Therefore, a post-treatment process that removes free fatty acids but does not affect the efficacy without loss of active ingredients is required. It is more preferable to use a fine filtration membrane of 1.0 μm or less, preferably 0.45 μm or less, to provide more excellent efficacy.

Hereinafter, with reference to FIG. 1, a method for preparing an extract using supercritical fluid extraction according to the present description will be described in detail. The drawing shown is a schematic diagram showing an example of an extraction process, and the invention of the present disclosure is not construed as being limited by FIG. 1 .

The solid dry matter is filled in the extractor 1, and the supercritical fluid is supplied to the lower end of the extractor 1 through the heat exchanger 9 using the fluid pump 7. At this time, the extractor 1 may be installed in two or more, and the pump 7 may supply the supercritical fluid supplied to the extractor 1 to adjust the pressure in the extractor, and the heat exchanger 9 may be used. By controlling the temperature of the fluid, the state of the fluid in the extractor 1 can be changed and/or maintained in a supercritical state. In addition, the supercritical fluid is easily induced to change the polarity of the supercritical fluid by flowing 70% ethanol through the regulator 8, so that the extractor 1 can extract various oils and fats.

As a specific example, it is preferable to maintain the temperature in the extractor 1 at 60° C. or less, or 45 to 55° C., since it can provide stability in extraction of related substances.

The supercritical fluid supplied to the extractor (1) is supplied at a flow rate of, for example, 250 ml/min or less, contacts the filled solid dry matter to extract the extract from the solid dry matter, and rises while containing the extract to the outside of the extractor (1). The mixture of ethanol, supercritical fluid, and extract is adiabatically expanded while being reduced to 70 bar or less, or 45 to 55 bar via a back pressure regulator (2), and can be transferred to the separator (3).

At this time, the time required for extraction is not limited thereto, but may be 3 hours or less, or 2 hours or less.

In the separator (3), the extracted extract and the fluid are separated, and the separated fluid is liquefied through the cooler (4) and stored in the storage tank (5) for reuse, and is circulated in the storage tank (5) for fluids such as carbon dioxide. In addition to the supplied fluid, a fluid of the same type or a different type may be supplemented from the outside so as to compensate for the loss of fluid occurring in the entire process.

The fluid stored in the reservoir 5 is pressurized through the pump 7 and changed to a supercritical state in the regulator 8, and then supplied to the extractor 1 again through the heat exchanger 9 so that it can be circulated.

In addition, the cooler 4 can be adjusted to less than 0 degrees, specifically -1 ° C or less, more preferably -1 to -3 ° C.

The extract separated in the separator (3) can be fractionated and commercialized (10) as needed. At this time, two or more fractionation reservoirs (not shown) may be installed, and a valve may be installed at each inlet of the extract into the fractionation reservoir to control the opening and closing of the valve according to various conditions to obtain extracts with various physical properties at the same time. can

If necessary, it may be sent from the fractionation storage tank to a maturation tank (not shown) for the maturation process, and transferred to a purification tank (not shown) in a matured and refined state for storage.

The process may be continuously performed from the solid dried material to extraction exhibiting desired physical properties, and the number of devices and valves may be appropriately adjusted as needed.

Specifically, the supercritical extract manufacturing method according to the present disclosure uses ethanol and a supercritical fluid to clean Terminalia bellerica , Amla ( Amla, Phyllanthus emblica ), and Triphala ( Triphala ) and papaya ( Carica papaya ). extracting an extract from the solid or dried product obtained after washing; and separating the supercritical fluid from the mixture of the ethanol, the supercritical fluid, and the extract, and fractionating the extract.

Hereinafter, a supercritical extraction method according to the present invention will be described in more detail.

preprocessing step

In the present description, a pretreatment step may be included prior to the supercritical extraction step. There is no particular limitation on the solid material used in this description, and the pretreatment step is a treatment for removing harmful substances such as microorganisms, residual pesticides and preservatives on the surface of the solid material or its dried material, and increasing the contact area of the supercritical fluid. processing may be included.

The pretreatment method of this embodiment is not particularly limited, but one or more of ultraviolet irradiation, heat treatment, peeling, grinding, and roasting may be used.

When grinding, the particle size is preferably within the range of 2 mm or less, or 0.1 mm to 2 mm, since it can increase the surface area and increase the extraction efficiency.

If necessary, it may be pulverized after drying at 95° C. or less, or 50 to 90° C. for 36 hr or more, or 48 hr or more.

Extraction stage of solids

Subsequently, the supercritical extract may be extracted from the solid by filling the extractor with the solid (or the pretreated solid) and supplying ethanol and a supercritical fluid to the extractor filled with the solid.

At this time, the extraction step of the supercritical extract of the present description may be carried out under conditions of 100 bar or more and 40 to 90 ° C., specifically under pressure conditions of 100 to 500 bar, more specifically 100 to 200 bar, It can be carried out at a temperature condition of 60 ° C, specifically 45 to 55 ° C, and the S / F ratio of the supercritical fluid (S / F ratio: means kg of supercritical fluid used per kg of filling material used for extraction) ) may be greater than 0 to 30, specifically may be greater than 0 to 20.

Extraction efficiency of the supercritical extract can be improved within the above range.

The supercritical fluid supplied to the extractor 1 is supplied at a flow rate of 250 ml/min or less, specifically 150 ml/min or less, more preferably 20 to 60 ml/min through the fluid pump 7, and additionally a controller Through (8), 70% ethanol is supplied at a flow rate of 0.1 to 3.0 mL/min, or 0.5 to 1.5 mL/min to contact the solid dry matter to extract the extract from the solid dry matter, and rise while containing the extract to extract the extractor ( 1) Discharged to the outside, the mixture of ethanol, supercritical fluid and extract may be transferred to a separator (3) to be described later via a back pressure regulator (2).

At this time, the time required for extraction is not limited thereto, but may be 3 hours or less, or 2 hours or less.

Separation and fractionation step of supercritical extract

Subsequently, the mixture of the supercritical fluid and the extract can be separated in the separator 3, which can be performed at 40 to 70 bar and 20 to 50 ° C., specifically at 50 to 60 bar and 30 to 40 ° C. It can be.

In addition to the active ingredient, the composition of the present invention may be prepared by including a skin whitening component, a skin wrinkle improvement component, a skin elasticity improvement component, a UV protection component, a skin moisturizing component, etc. known in the art in order to reinforce or add skin-related activities. can Specifically, skin whitening ingredients include arbutin, niacinamide, ascorbyl glucoside, alpha-bisabolol, oil soluble licorice (Glycyrrhiza) Extract, etc., and skin wrinkle improvement ingredients include retinol and retinyl. palmitate, adenosine, polyethoxylated amide, and the like, and examples of ultraviolet protection components include drometrizole, drometrizole trisiloxane, digaloyltrioleate, dimethicodyethylbenzalmalonate, diethylhexyl part and complex extracts such as tamidotriazone and pine bark extract, phosphatidylserine, finger root extract powder, and complex extracts such as red ginseng and cornus officinalis. In addition, as moisturizing ingredients, AP collagen enzyme decomposition peptide, Collactive collagen peptide, N-acetylglucosamine, konjac potato extract, complex extracts such as dandelion, rice bran extract, corn germ extract, low molecular weight collagen peptide, weed extract powder, phosphatidylserine, hyaluronic acid, etc. can be heard For other skin whitening ingredients, wrinkle improvement ingredients, UV protection ingredients, and skin moisturizing ingredients, the Functional Cosmetics Code according to the Cosmetics Act (“Functional Cosmetics Standards and Test Methods” in the Ministry of Food and Drug Safety Notice), and Health Functional Foods under the Health Functional Food Act It is possible to refer to the official (MFDS notice "Health functional food standards and specifications). These ingredients may be included in the composition of the present invention one or more together with the active ingredient.

The composition of the present invention may contain the active ingredient in an arbitrary amount (effective amount) as long as it can exhibit a skin wrinkle improvement effect, antioxidant effect, etc. depending on the product type, product formulation, etc., but a typical effective amount is the total weight of the composition It will be determined within the range of 0.001% by weight to 15% by weight as a reference. Here, "effective amount" means that when the composition of the present invention is administered to a mammal, preferably a human to which it is applied, during the administration period according to the advice of medical experts, etc., the effect of improving skin wrinkles, improving skin elasticity, antioxidant effect, etc. It refers to the amount of the active ingredient included in the composition of the present invention that can exhibit medical/cosmetic effects. Such an effective amount can be determined empirically within the ordinary skill of the skilled artisan.

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

Even when the composition of the present invention is identified as a cosmetic composition, the cosmetic composition may be classified as an arbitrary product in terms of its use and law, and specifically, it may be a functional cosmetic having a purpose such as skin whitening, a non-functional general cosmetic, etc. .

In product form, it may take any product form, specifically, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax It can be formulated as a foundation, spray, and the like. In a specific product form, it may be a softening lotion, nutrient lotion, nutrient cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder formulation.

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

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane / May contain a propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing 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 ,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol, fatty acid esters of sorbitan, and the like can be used.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, or polyoxyethylene sorbitan ester, microcrystals Star cellulose, aluminum metahydroxide, bentonite, agar, and the like can be used.

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

The cosmetic composition of the present invention may be prepared according to a method for preparing a cosmetic composition conventionally performed in the art, except for containing an active ingredient exhibiting skin wrinkle improvement activity, skin elasticity improvement activity, and the like.

In another specific aspect, the composition of the present invention can be regarded as a food composition.

The food composition of the present invention can be prepared in any form, for example, beverages such as tea, juice, carbonated beverages, and ionic beverages, processed oils such as milk and yogurt, chewing gum, rice cakes, traditional Korean snacks, bread, snacks, noodles, etc. It can be manufactured into health functional food preparations such as food, tablet, capsule, pill, granule, liquid, powder, flake, paste, syrup, gel, jelly, bar, etc. In addition, the food composition of the present invention may take any product classification as long as it conforms to the enforcement regulations at the time of manufacture / distribution in functional classification. For example, it may be a health functional food according to the Health Functional Food Act, or soybean milk, fermented beverages, special purpose food, etc. according to each food type in the Food Code (MFDS notification, food standards and specifications) of the Food Sanitation Act.

The food composition of the present invention may include food additives in addition to the active ingredient. Food additives are generally understood as substances that are added to, mixed with, or infiltrated into food in manufacturing, processing, or preserving food, and since they are taken daily and for a long period of time with food, their safety must be guaranteed. According to the Food Additive Code (Ministry of Food and Drug Safety Notice, Food Additive Standards and Specifications) under the Food Sanitation Act, safety-guaranteed food additives are classified into chemical synthetic products, natural additives, and mixed formulations and are limitedly regulated.

These food additives can be classified into sweeteners, flavors, preservatives, emulsifiers, acidulants, thickeners, etc. in terms of functionality.

Sweeteners can be used in amounts that give the food a moderately sweet taste, and can be natural or synthetic. Preferably, a natural sweetener is used, and examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavors may be used to improve taste or aroma, and both natural and synthetic flavors may be used. Preferably, it is the case of using a natural one. In case of using natural ones, in addition to flavor, the purpose of enhancing nutrition can also be combined. As a natural flavoring agent, it may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, etc., or obtained from green tea leaves, roundworms, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo, etc. can also be used. Natural flavors can be liquid concentrates or solid extracts. In some cases, synthetic flavors may be used, and synthetic flavors may include esters, alcohols, aldehydes, terpenes, and the like.

As preservatives, sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. may be used, and as emulsifiers, acacia gum, carboxymethylcellulose, xanthan gum, pectin and the like, and examples of the acidulant include acid salt, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, and the like. Acidulants may be added to the food composition to have an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to improving taste.

As the thickening agent, a suspending agent, a sedimentation agent, a gel forming agent, a swelling agent, and the like may be used.

In addition to the food additives described above, the food composition of the present invention may include physiologically active substances or minerals known in the art and whose stability is guaranteed as food additives for the purpose of supplementing or reinforcing functionality and nutrition.

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

The food composition of the present invention may include the above-mentioned food additives in an appropriate amount to achieve the purpose of addition according to the type of product.

Regarding other food additives that may be included in the food composition of the present invention, reference may be made to the Food Codex or Food Additives Codex.

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

The pharmaceutical composition of the present invention may be formulated into an oral formulation or a parenteral formulation according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. Here, "pharmaceutically acceptable" means that it does not inhibit the activity of the active ingredient and does not have toxicity more than is adaptable to the subject of application (prescription).

When the pharmaceutical composition of the present invention is prepared as an oral dosage form, powder, granule, tablet, pill, dragee, capsule, liquid, gel, syrup, suspension, wafer according to a method known in the art together with a suitable carrier. It can be prepared in formulations such as Examples of suitable pharmaceutically acceptable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, and wheat starch, cellulose, methylcellulose, ethyl cellulose, Celluloses such as sodium carboxymethylcellulose and hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil etc. are mentioned. In the case of formulation, if necessary, diluents and/or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants may be included.

When the pharmaceutical composition of the present invention is prepared as a parenteral formulation, it may be formulated in the form of an injection, transdermal administration, nasal inhalation, and suppository along with a suitable carrier according to a method known in the art. In the case of formulation as an injection, suitable carriers include sterile water, ethanol, polyols such as glycerol or propylene glycol, or mixtures thereof, preferably Ringer's solution, triethanolamine-containing PBS (phosphate buffered saline) or sterile water for injection , isotonic solutions such as 5% dextrose, etc. may be used. When formulated as a transdermal formulation, it may be formulated in the form of an ointment, cream, lotion, gel, external agent, pasta, liniment, or air roll. In the case of nasal inhalation, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc., and suppository bases include witepsol, twin (tween) 61, polyethylene glycols, cacao butter, laurin paper, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearates, sorbitan fatty acid esters, and the like may be used.

Formulation of pharmaceutical compositions is known in the art, and may be specifically referred to in Remington's Pharmaceutical Sciences (19th ed., 1995). These documents are considered as part of this specification.

The preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, body weight, sex, age, severity of the patient, and route of administration. /kg range. Administration can be done once a day or divided into several times. These dosages should not be construed as limiting the scope of the present invention in any respect.

The cosmetic composition, food composition, and pharmaceutical composition of the present invention may be prepared according to the preparation method of the composition commonly performed in the art, except for including active ingredients known in the art.

In describing the skin-useful composition for improving skin wrinkles and elasticity of the present description, other conditions or equipment not explicitly described may be appropriately selected within the range commonly practiced in the art and are not particularly limited. specify

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily practice the present invention. However, the present invention may be embodied in many different forms and is not limited to the embodiments described herein.

[Example]

<Example 1> Preparation of supercritical Bahera ethanol extract

(1) Barhera ( Terminalia bellerica ), Amla (Amla, Phyllanthus emblica), Triphala (Triphala), and Papaya (Carica papaya) were thoroughly washed, dried at 70° C. for 48 hr, and pulverized into a size of 2 mm or less. The pulverized raw materials were mixed at a constant weight (100 g: 100 g: 100 g: 100 g).

(2) The mixed solid dry material was filled in the extractor 1 of FIG. 1 below, and the supercritical fluid was supplied to the lower end of the extractor 1 through the heat exchanger 9 using the pump 7.

Specifically, solid dry matter filled by supplying the supercritical fluid supplied to the extractor (1) at a flow rate of about 40 ml/min for 2 hours while maintaining the extractor (1) at 45 to 55 ° C. and 100 to 200 bar. The process of extracting the extract from the solid dry matter in contact with was performed 4 times. 70% ethanol was flowed into the extractor (1) through the regulator (8) at a flow rate of 1.0 mL/min.

(3) Then, it rises with the extract and is discharged out of the extractor (1), and the mixture of ethanol, supercritical fluid and extract is reduced to about 50 bar via a back pressure regulator (2) and adiabatic expansion and transferred to the separator (3).

In the separator (3), the extracted extract, ethanol, and fluid are separated, and the separated fluid is liquefied through the cooler (4) adjusted to -1 ° C., stored in the storage tank (5), and can be reused. Fluids such as carbon dioxide In addition to the fluid circulated and supplied to the reservoir 5, the same or different fluids are replenished from the outside to compensate for the loss of fluid occurring in the entire process, and the fluid stored in the reservoir 5 is pumped through the pump 7. It was pressurized and changed to a supercritical state, and was supplied to the extractor 1 again through the heat exchanger 9 and circulated.

The extract separated in the separator (3) was filtered through a 0.45 μm membrane filter and then concentrated in vacuum and at room temperature for 3 hours to prepare a supercritical Bahera ethanol extract (sample TATP-3).

<Comparative example>

<Comparative Example 1> Preparation of Supercritical Bahera Extract

(1) Barhera ( Terminalia bellerica ), Amla (Amla, Phyllanthus emblica), Triphala (Triphala), and Papaya (Carica papaya) were thoroughly washed, dried at 70° C. for 48 hr, and pulverized into a size of 2 mm or less. The pulverized raw materials were mixed at a constant weight (100 g: 100 g: 100 g: 100 g).

(2) The mixed solid dry material was filled in the extractor 1 of FIG. 1 below, and the supercritical fluid was supplied to the lower end of the extractor 1 through the heat exchanger 9 using the pump 7.

Specifically, solid dry matter filled by supplying the supercritical fluid supplied to the extractor (1) at a flow rate of about 40 ml/min for 2 hours while maintaining the extractor (1) at 45 to 55 ° C. and 100 to 200 bar. The process of extracting the extract from the solid dry matter in contact with was performed 4 times.

(3) Then, it rises with the extract and is discharged out of the extractor (1), and the mixture of the supercritical fluid and the extract is adiabatically expanded while being reduced to about 50 bar via a back pressure regulator (2), Transferred to (3).

In the separator (3), the extracted extract and the fluid are separated, and the separated fluid is liquefied through the cooler (4) adjusted to -1 ° C and stored in the storage tank (5) for reuse, and the storage tank for fluids such as carbon dioxide In (5), the same or different types of fluids are replenished from the outside to make up for the loss of fluids occurring in the entire process, in addition to the fluids supplied by circulation, and the fluid stored in the reservoir 5 is pressurized through the pump 7 It was changed to a supercritical state and supplied to the extractor 1 again through the heat exchanger 9 and circulated.

The extract separated in the separator (3) was filtered through a 0.45 μm membrane filter and then concentrated in vacuum and at room temperature for 3 hours to prepare a supercritical Bahera extract (sample TATP-1).

<Comparative Example 2> Preparation of Supercritical Bahera Pure Ethanol Extract

(1) Barhera ( Terminalia bellerica ), Amla (Amla, Phyllanthus emblica), Triphala (Triphala), and Papaya (Carica papaya) were thoroughly washed, dried at 70° C. for 48 hr, and pulverized into a size of 2 mm or less. The pulverized raw materials were mixed at a constant weight (100 g: 100 g: 100 g: 100 g).

(2) The mixed solid dry material was filled in the extractor 1 of FIG. 1 below, and the supercritical fluid was supplied to the lower end of the extractor 1 through the heat exchanger 9 using the pump 7.

Specifically, solid dry matter filled by supplying the supercritical fluid supplied to the extractor (1) at a flow rate of about 40 ml/min for 2 hours while maintaining the extractor (1) at 45 to 55 ° C. and 100 to 200 bar. The process of extracting the extract from the solid dry matter in contact with was performed 4 times. 100% ethanol was flowed into the extractor (1) through the regulator (8) at a flow rate of 1.0 mL/min.

(3) Then, it rises with the extract and is discharged out of the extractor (1), and the mixture of ethanol, supercritical fluid and extract is reduced to about 50 bar via a back pressure regulator (2) and adiabatic expansion and transferred to the separator (3).

In the separator (3), the extracted extract, ethanol, and the fluid are separated, and the separated fluid is liquefied through the cooler (4) adjusted to -1 ° C., stored in the storage tank (5), and can be reused. Fluids such as carbon dioxide In addition to the fluid circulated and supplied to the reservoir 5, the same or different fluids are replenished from the outside to compensate for the loss of fluid occurring in the entire process, and the fluid stored in the reservoir 5 is pumped through the pump 7. It was pressurized and changed to a supercritical state, and was supplied to the extractor 1 again through the heat exchanger 9 and circulated.

The extract separated in the separator (3) was filtered through a 0.45 μm membrane filter and then concentrated in vacuum and at room temperature for 3 hours to prepare a supercritical Bahera pure ethanol extract (sample TATP-2).

<Experimental example>

<Experimental Example 1> Total polyphenol and total flavonoid content test

Total polyphenol and total flavonoid content tests were performed on the samples obtained in Example 1 and the samples obtained in Comparative Examples 1 and 2.

(1-1) Total polyphenol content

Take 100 mg each of the samples (TATP-1 to TATP-3) and dilute to 100 mL with 80% ethanol to use as the test solution. Separately take 100 mg of gallic acid and make 100 mL with 80% ethanol. Take 0.1, 0.2, 0.5, 1.0 mL of this solution, dilute it to 5 mL, and use this solution as the standard solution. After adding 100 μL and 100 μL of 10% sodium carbonate to the e-tube for the test and standard solutions, add 100 μL of Folin-Ciocalteu reagent, mix them using a vortex for 30 seconds, leave them in the dark for 30 minutes, and measure the absorbance of the reaction solution. Measured at 750 nm. A calibration curve of the standard solution was drawn for the measured absorbance, and the total polyphenol content was expressed as mg GAE/g in FIG. 2 below.

(1-2) Total flavonoid content

Take 100 mg each of the samples (TATP-1 to TATP-3) and dilute to 100 mL with 80% ethanol to use as the test solution. Separately take 100 mg of quercetin and make 100 mL with 80% ethanol. Take 0.1, 0.2, 0.5, 1.0 mL of this solution, dilute it to 5 mL, and use this solution as the standard solution. Add 500 μL of the sample and standard solutions, 100 μL of 10% aluminum nitrate, and 100 μL of 1 M potassium acetate into an e-tube and mix. After 40 minutes of mixing, the absorbance is measured by measuring the absorbance at 415 nm using a UV-vis spectrophotometer. A calibration curve of the standard solution was drawn for the measured absorbance, and the total flavonoid content was expressed in mg QE/g as shown in FIG. 2 below.

As shown in FIG. 2 below, in the case of sample TATP-3 according to Example 1, the highest polyphenol content was found to be 195.7 mgGAE/g.

On the other hand, in the case of sample TATP-1 according to Comparative Example 1 without using a co-solvent for the supercritical fluid, the polyphenol content was confirmed to be 95.2 mgGAE/g, and in Comparative Example 2 using 100% concentration ethanol as a co-solvent In the case of the sample TATP-2 according to the analysis, the polyphenol content was analyzed to be 143.8 mgGAE/g, indicating that the polyphenol content increased when a co-solvent of an appropriate concentration for the supercritical fluid was used.

In addition, in the case of sample TATP-3 according to Example 1, the highest flavonoid content was found to be 97.7 mgQE/g, whereas in the case of sample TATP-1 according to Comparative Example 1 in which no co-solvent was used for the supercritical fluid, the flavonoid content was found to be 97.7 mgQE/g. The content was confirmed to be 42.4 mgQE/g, and in the case of sample TATP-2 according to Comparative Example 2 using 100% concentration of ethanol as a co-solvent, the flavonoid content was analyzed to be 54.1 mgQE/g, indicating the same tendency as the polyphenol content. analyzed.

<Experimental Example 2> Antioxidant activity test 1

Antioxidant tests on the samples obtained in Example 1 and the samples obtained in Comparative Examples 1 and 2 were carried out to evaluate the radical scavenging ability of the sample by the ABTS method and the DPPH method.

(2-1) ABTS radical scavenging activity

Take 100 mg each of the samples (TATP-1 to TATP-3), dilute to 100 mL with water, and use it as the test solution. 7 mM ABTS and 2.45 mM potassium persulfate are mixed and reacted at room temperature for 12 hours in the dark to form ABTS cations. Thereafter, ethanol was added to adjust the absorbance value at 734 nm to be 0.70 ± 0.02.

Add 100 μL of the test solution and 100 μL of the prepared ABTS solution to a 96-well plate, react at room temperature for 7 minutes, and measure at 734 nm. Compared with the blank test solution, the ABTS clearance rate was obtained as a percentage (%) and is shown in FIG. 3 below.

(2-2) DPPH radical scavenging activity

Take 100 mg each of the samples (TATP-1 to TATP-3), dilute to 100 mL with water, and use it as the test solution. Add 100 μL of the test solution and 100 μL of 0.2 mM DPPH to a 96-well plate, and after 30 minutes, measure the absorbance at 517 nm using a microplate reader. Compared to the blank test solution, the DPPH radical scavenging rate was obtained in percentage (%) and is shown in FIG. 3 below.

As shown in FIG. 3 below, in the case of sample TATP-3 according to Example 1, as a result of DPPH radical analysis, it can be seen that it exhibits the highest antioxidant activity of 68.3% at a concentration of 2.0 mg/mL.

On the other hand, in the case of sample TATP-2 according to Comparative Example 1 in which no co-solvent was used in the supercritical fluid, the antioxidant capacity was 53.7% at a concentration of 2.0 mg/mL, and the co-solvent was used but 100% ethanol was used. In the case of sample TATP-2 according to Comparative Example 2, it was analyzed to exhibit poor antioxidant activity compared to Example 1 at 61.3% at a concentration of 2.0 mg/mL. All test materials were analyzed for scavenging activity in a concentration-dependent manner, and all were found to be lower than the control ascorbic acid.

In addition, in the case of sample TATP-3 according to Example 1, the ABTS radical analysis result was found to be the highest at 2.0 mg/mL concentration of 84.9%, whereas sample TATP according to Comparative Example 1 without using a co-solvent for supercritical fluid In the case of -1, it was confirmed as 57.9% at a concentration of 2.0 mg / mL, and in the case of sample TATP-2 according to Comparative Example 2 using 100% concentration of ethanol as a co-solvent, it was analyzed at 64.7% at a concentration of 2.0 mg / mL and the DPPH of It was analyzed to show the same tendency as the experimental results.

<Experimental Example 3> Antioxidant activity test 2

As an antioxidant test on the sample obtained in Example 1 and the sample obtained in Comparative Examples 1 and 2, SOS-like activity and xanthine oxidase inhibitory activity were tested.

(3-1) SOS-like activity

Samples (TATP-1 to TATP-3) are diluted in water to a certain concentration and used as test solutions. To 0.2 mL of the sample solution, add 2.6 mL of tris-HCl buffer corrected to pH 8.5 and 0.2 mL of 7.2 mM pyrogallol, and react at 25 °C for 10 minutes. Add 0.1 mL of 1 N HCl to the reacted solution to stop it. The amount of oxidized pyrogallol is determined by measuring the absorbance at 420 nm. Compared to the blank test solution, the SOD-like activity was obtained in percentage (%) and is shown in Table 1 below.

(3-2) Xanthine oxidase inhibitory activity

Samples (TATP-1 to TATP-3) are used as test solutions by diluting test materials (TATP-1 to 3) in water at a certain concentration. Add 0.6 mL of 0.1 M potassium phosphate buffer (pH 7.5) and 0.2 mL of 1 mM xanthine to 1.0 mL of the test solution. Then, 0.1 mL of 0.2 U/mL xanthine oxidase was added to stop the reaction. The resulting uric acid is obtained by measuring the absorbance at 292 nm. Compared with the blank test solution, the Xanthine oxidase inhibitory activity was obtained in percentage (%) and is shown in Table 1 below.

*Control: ascorbic acid

As shown in Table 1, in the case of sample TATP-3 according to Example 1, it can be seen that the SOS-like activity analysis shows the highest activity at 2.0 mg/mL concentration of 38.8%.

On the other hand, in the case of sample TATP-2 according to Comparative Example 1 in which no co-solvent was used in the supercritical fluid, an activity of 27.5% was exhibited at a concentration of 2.0 mg/mL, and a co-solvent was used but 100% ethanol was used. In the case of sample TATP-2 according to Comparative Example 2, it was analyzed to exhibit poor activity compared to Example 1 at 35.6% at a concentration of 2.0 mg/mL. All test materials were analyzed for scavenging activity in a concentration-dependent manner, and all were found to be lower than the control ascorbic acid.

In addition, in the case of sample TATP-3 according to Example 1, the xanthine oxidase inhibitory activity was confirmed to be the highest at 2.0 mg/mL concentration of 41.3% as a result of analysis, whereas Comparative Example 1 without using a co-solvent for supercritical fluid In the case of sample TATP-1 according to, it was confirmed as 33.6% at a concentration of 2.0 mg / mL, and in the case of sample TATP-2 according to Comparative Example 2 using 100% concentration of ethanol as a co-solvent, it was found to be 35.1% at a concentration of 2.0 mg / mL. It was analyzed and analyzed to show the same tendency as the SOS-like activity test result.

<Experimental Example 4> Whitening activity test

A trypsinase inhibitory activity test was performed as a whitening test on the samples obtained in Example 1 and the samples obtained in Comparative Examples 1 and 2.

Specifically, the samples (TATP-1 to TATP-3) are diluted in water at a certain concentration and used as a test solution. Add 0.5 mL of 175 mM sodium phosphate buffer (pH 6.8) and 0.2 mL of 10 mM L-DOPA (3,4-dihydroxy-L-phenyl-alanine) to 0.1 mL of the test solution. Thereafter, 0.2 mL of a 110 U/mL solution was added and reacted at 25 °C for 2 minutes. The resulting DOPA chrome was obtained by measuring the absorbance at 475 nm, and the tyrosinase inhibition rate was obtained as a percentage (%) compared to the blank test solution, and is shown in FIG. 4 below.

As shown in FIG. 4 below, in the case of sample TATP-3 according to Example 1, as a result of the trypsinase inhibitory activity assay, it can be seen that it exhibits the highest activity of 33.7% at a concentration of 2.0 mg/mL.

On the other hand, in the case of sample TATP-2 according to Comparative Example 1 in which no co-solvent was used in the supercritical fluid, an activity of 23.2% was exhibited at a concentration of 2.0 mg/mL, and a co-solvent was used but 100% ethanol was used. In the case of sample TATP-2 according to Comparative Example 2, it was analyzed to exhibit poor activity compared to Example 1 at 26.1% at a concentration of 2.0 mg/mL. All test materials were analyzed for scavenging activity in a concentration-dependent manner, and all were found to be lower than the control ascorbic acid.

<Experimental Example 5> Anti-wrinkle evaluation test

Collagenase inhibitory activity and elastase inhibitory activity were tested for anti-wrinkle evaluation on the samples obtained in Example 1 and Comparative Examples 1 and 2.

(5-1) collagenase inhibitory activity

Samples (TATP-1 to TATP-3) are diluted in water to a certain concentration and used as test solutions. Add 4 mM calcium chloride to 0.1 M tris-HCl buffer (pH 7.5) and add 0.2 mL of the test solution to 0.5 mL of a solution of 4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-DArg (0.3 mg/mL) dissolved in it. Thereafter, 0.3 mL of 200 U/mL collagenase type I was added and reacted at room temperature for 20 minutes. 0.5 mL of 5% citric acid was added as a reaction stop solution, 1 mL of ethyl acetate was added, and the absorbance was measured at 320 nm, and the collagenase inhibition rate was obtained in percentage (%) compared to the blank test solution, and is shown in Table 2 below.

(5-2) Elastase inhibitory activity

Samples (TATP-1 to TATP-3) are diluted in water to a certain concentration and used as test solutions. After adding 50 μg/mL of pancreatic solution, 100 μg/mL of N-succinyl-(LAla)3-p-nitroanilide (1 mg/mL) dissolved in 50 mM tris-HCl buffer (pH 8.6) was added and reacted for 30 minutes After measuring the absorbance at 410 nm, the elastase inhibition rate was obtained as a percentage (%) by comparing with the blank test solution, and it is shown in Table 2 below.

*Control: ascorbic acid

As shown in Table 2, it can be seen that sample TATP-3 according to Example 1 exhibits the highest inhibitory activity of 58.1% at a concentration of 2.0 mg/mL as a result of analysis of collagenase inhibitory activity.

On the other hand, in the case of sample TATP-2 according to Comparative Example 1 in which no co-solvent was used in the supercritical fluid, the inhibitory activity was 41.3% at a concentration of 2.0 mg/mL. In the case of sample TATP-2 according to Comparative Example 2, it was analyzed to exhibit poor inhibitory activity compared to Example 1 at 53.3% at a concentration of 2.0 mg/mL. All test materials were analyzed for scavenging activity in a concentration-dependent manner, and all were found to be lower than the control ascorbic acid.

In addition, in the case of sample TATP-3 according to Example 1, the elastase inhibitory activity was confirmed to be the highest at 2.0 mg/mL concentration of 48.6% as a result of analysis, whereas in Comparative Example 1 without using a co-solvent for supercritical fluid Sample TATP-1 according to Comparative Example 2 using 100% ethanol as a co-solvent was found to be 41.4% at 2.0 mg/mL concentration, and 42.9% at 2.0 mg/mL concentration for sample TATP-2 according to Comparative Example 2 using 100% ethanol as a co-solvent. It was analyzed to show the same tendency as the collagenase inhibitory activity test result.

<Experimental Example 6> safety test

In order to evaluate the safety of the samples obtained in Example 1, MTT assay, a representative cytotoxicity test, was used. Quantification was determined by modifying the Mosmann method.

Specifically, HaCaT cells were divided into 1Х10 ⁴ cell/ml and cultured for 24 hours, then replaced with a new medium supplemented with diluted samples at concentrations of 0.5, 1.0, 1.5, and 2.0 mg/ml, and cultured for another 24 hours. did 20 μl of EZ-Cytox was added to each well, and after 1 hour of incubation in a 37 °C, 5% CO ₂ incubator, the absorbance was measured with an ELISA reader (Epoch ^TM 2, BioTek, USA) at 450 nm.

Cell viability was calculated using Equation 1 below, and the results are shown in FIG. 5 below.

[Equation 1]

Cell viability (%) = [(Exp. - Blank)/Control] Х 100

(In Equation 1, Exp: absorbance of extract containing cells, Blank: absorbance of extract without cells, Control: absorbance of distilled water containing cells.)

As shown in FIG. 5, the cytotoxicity of the extract in cells was 0.5 mg/g, 1.0 mg/g, 1.5 mg/g, and 2.0 mg/g based on cell viability (cell viability: 100%) of the untreated group. As a result of the experiment, no cytotoxicity was observed for all samples (TATP-1 to TATP-3) at all concentrations. Therefore, the safety of the extract according to Example 1 was confirmed in HaCaT cells.

In conclusion, when the supercritical extract obtained by using the fluid in the solid phase dry matter using Bahera (Terminalia bellerica), Amla (Amla, Phyllanthus emblica), Triphala and Papaya (Carica papaya) is included as an active ingredient , The obtained composition maximizes the total polyphenol content and total flavonoid content, resulting in radical scavenging activity, SOD (superoxide dismutase)-like activity, antioxidant effect and whitening effect by xanthine oxidase inhibitory activity, and inhibitory activity against collagenase It was confirmed that it is suitable for cosmetic compositions, food compositions, and other pharmaceutical compositions by providing an anti-aging effect by providing an effect of improving skin wrinkles and elasticity by inhibiting activity against and elastase.

1 ... extractor 2 ... pressure regulator 3 ... separator 4 ... cooler 5 ... reservoir
6...Fluid supply tank 7...Fluid pump 8...Regulator 9...Heat exchanger
10...product 11...fluid discharge 12...drain

Claims (6)

Bahera ( Terminalia bellerica ), Amla ( Amla, Phyllanthus emblica ), Triphala ( Triphala ) and Papaya ( Carica papaya ) The supercritical extract obtained by extracting using ethanol and supercritical fluid to the dried solids was used as an active ingredient. wherein the supercritical extract is a 70% ethanol supercritical extract of solid phase dry matter using Terminalia bellerica , Amla, Phyllanthus emblica , Triphala and Carica papaya skin A skin-useful composition for improving wrinkles and elasticity. According to claim 1,
The supercritical fluid is carbon dioxide, nitrogen or a mixture thereof skin oil composition for improving skin wrinkles and elasticity. delete According to claim 1,
The bahera ( Terminalia bellerica ), Amla ( Amla, Phyllanthus emblica ), Triphala ( Triphala ), and papaya ( Carica papaya ) have a bahera (a): amla (b): the sum of triphala and papaya (c) is 1 : 0.5 ~ 1.5: 0.5 ~ 1.5 to satisfy the weight ratio skin useful composition for improving skin wrinkles and elasticity. According to claim 1,
The barhera ( Terminalia bellerica ), Amla ( Amla, Phyllanthus emblica ), Triphala ( Triphala ), and papaya ( Carica papaya ) are 1:0.5 to 1.5: skin wrinkles that satisfy the weight ratio of 0.5 to 1.5:0.5 to 1.5 A skin-useful composition for improving elasticity. According to claim 1,
The composition is a skin oil composition for improving skin wrinkles and elasticity that is a cosmetic composition.

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