KR20220080587A - Method for extracting okra using process of osmotic pressure dehydration by salt and molecular compression dehydration, and cosmetic composition for anti-aging prepared thereby - Google Patents

Abstract

It relates to a method for extracting okra using osmotic dehydration and molecular compression dehydration, and according to a method according to an aspect, it is possible to exhibit a remarkably excellent extraction yield by solving the problems of the existing plant dehydration extraction method, and excellent quality okra extract can be obtained. In addition, in the case of a cosmetic composition comprising an okra extract prepared by the above method, it may exhibit a remarkably excellent skin wrinkle improvement effect, skin elasticity improvement effect, skin itch improvement effect, or skin moisturizing effect.

Description

The method for extracting okra using osmotic pressure and molecular compression dehydration process by salt, and the cosmetic composition for anti-aging prepared thereby thereby}

It relates to a method for extracting okra using osmotic dehydration and molecular compression dehydration, and to a cosmetic composition comprising the okra extract prepared thereby.

A method of extracting a plant generally used includes a hot water extraction method using high-temperature purified water, a solvent extraction method using a solvent such as ethanol or butylene glycol, and the like.

The hot water extraction method has the advantage that the extraction method is convenient and the yield is high, but there is a disadvantage that the intrinsic nutrients of the raw material may be destroyed by heat. In addition, the solvent extraction method has a disadvantage that raw materials and solvents discarded after extraction may adversely affect the environment.

In general, osmotic dehydration refers to a method in which raw materials are dehydrated in such a way that water moves from a high concentration to a low concentration or from a low concentration to a high concentration due to the difference in osmotic pressure at the cell membrane boundary when the raw material is immersed in a high-concentration liquid phase. In the case of osmotic dehydration, the dehydration phenomenon is stopped when the concentration inside and outside the cell membrane of the raw material is balanced. Although this has the advantage that the initial dehydration rate is fast, salts or saccharides derived from hypertonic solution are introduced into the intracellular space and remain in the cells, and there is a disadvantage that the final amount of dehydration is small. In addition, osmotic dehydration is suitable for stored foods such as pickles, but when restoring the dehydrated tissue, the cell membrane of the dehydrated tissue is destroyed, and the useful components of the dehydrated tissue are reduced, thereby reducing the quality and the texture of the dehydrated tissue is similar to that of living things. There is a downside to being different.

Under this background, the present inventors have developed a method of extracting okra in an eco-friendly manner without destroying the nutrients inherent in the raw material while compensating for the disadvantages of the existing process and increasing the extraction yield, and By confirming the remarkably excellent skin wrinkle improvement effect, skin elasticity improvement effect, skin itch improvement effect, or skin moisturizing effect of the cosmetic composition, the present application was completed.

Republic of Korea Patent No. 2155897

One aspect comprises the steps of extracting okra using an osmotic dehydration extraction method by mixing salts and okra; And to provide a method for extracting okra comprising the step of mixing okra and a polymer material and extracting the okra using a molecular compression dehydration extraction method.

Another aspect is to provide an okra extract prepared by the above method.

Another aspect is to provide a cosmetic composition comprising the okra extract.

Other objects and advantages of the present application will become more apparent from the following detailed description in conjunction with the appended claims and drawings. Content not described in this specification will be omitted because it can be sufficiently recognized and inferred by those skilled in the technical field or similar technical field of the present application.

Each description and embodiment disclosed in this application may also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in the present application fall within the scope of the present application. In addition, it cannot be seen that the scope of the present application is limited by the detailed description described below.

One aspect comprises the steps of extracting okra using an osmotic dehydration extraction method by mixing salts and okra; And it provides a method of extracting okra comprising the step of mixing the okra and a polymer material and extracting the okra using a molecular compression dehydration extraction method.

The term, "okra", refers to a perennial plant of the dicotyledonous plant Dicotyledonous family Malvaceae mallow family. The scientific name of the okra may be Hibiscus esculentus. The okra, the leaves are alternate phyllotaxis, long stalks, heart-shaped, divided into about 3 to about 10, may represent the appearance of sawtooth in forked pieces.

The okra may mean an unprocessed raw material, and may be frozen or refrigerated okra. In addition, the okra may be a fruit, flower, leaf, root, or combination thereof of okra, and may mean the whole plant. In one embodiment, the okra may be minced. As the okra is minced, it is possible to increase the surface area that can be in contact with the dehydration solution containing the salt or polymer material, so that extraction can be carried out effectively.

The term "salt" refers to a compound that is an ionic substance in which an anion of an acid and a cation of a base are coupled by electrostatic attraction. The salt is not particularly limited in its kind. Specifically, the salt may be a sodium salt, a potassium salt, a calcium salt, or a combination thereof. Preferably, the salt may be sodium chloride, sodium lactate, sodium acetate, potassium chloride, calcium chloride, or a combination thereof. The sodium chloride may be rock salt, purified salt, sea salt, magnetic salt, prepared salt, or a combination thereof.

The term, “dehydration extraction” refers to discharging water and oil-soluble components of a plant tissue, which is a raw material, to the outside of the original material. The dehydration extraction may be osmotic dehydration extraction or molecular compression dehydration extraction.

The osmotic dehydration extraction may refer to discharging water or oil-soluble components within the cell to the outside of the cell by using osmotic pressure, which is a force caused by a concentration gradient inside and outside the cell. In the osmotic dehydration extraction, phase equilibrium is achieved from a high concentration to a low concentration or from a low concentration to a high concentration due to the difference in osmotic pressure at the cell membrane boundary, and the dehydration phenomenon may be stopped when the concentration inside and outside the cell membrane is balanced.

In the step of mixing the okra and the salt to extract the okra using an osmotic dehydration extraction method, the salt and the okra may be mixed in a weight ratio of 5 to 100: 100. Specifically, in the extraction using the osmotic dehydration extraction method, the salt and the okra 5 to 90, 5 to 80, 5 to 70, 5 to 60, 5 to 50, 5 to 40, 5 to 30, 5 to 20, 5 to 10, 10 to 100, 10 to 90, 10 to 80, 10 to 70, 10 to 60, 10 to 50, 10 to 40, 10 to 30, 10 to 20, 20 to 100, 20 to 90, 20 to 80, 20 to 70, 20 to 60, 20 to 50, 20 to 40, 20 to 30, 30 to 100, 30 to 90, 30 to 80, 30 to 70, 30 to 60, 30 to 50, It may be mixed in a weight ratio of 30 to 40, 40 to 100, 50 to 90, or 60 to 80: 100, and preferably mixed in a weight ratio of 20 to 40: 100.

According to one embodiment, in the step of extracting using the osmotic dehydration extraction method, when the salt and the okra are mixed in a weight ratio of less than 10: 100, the amount of dehydration decreases and the extraction yield is reduced or included in the extract The amount of oil-soluble ingredients used may be reduced. In addition, in the extraction using the osmotic dehydration extraction method, when the salt and the okra are mixed in a weight ratio of more than 100: 100, the solubility of the salt decreases, and the extraction yield is reduced by the undissolved salt or , The cell tissue of the okra is damaged, and polymer components such as enzymes in the cell may leak out of the cell. Therefore, the finally obtained extract may show deterioration in quality, such as containing impurities or browning by enzymes. In addition, it may be difficult to obtain and use the final extract due to the undissolved salt.

In the step of extracting the okra by using the osmotic dehydration extraction method by mixing the okra and the salt, the mixing time of the okra and the salt may be about 15 hours to about 32 hours. Specifically, the mixing time is about 15 hours to about 31 hours, about 15 hours to about 30 hours, about 16 hours to about 32 hours, about 16 hours to about 31 hours, about 16 hours to about 30 hours, about 17 hours. to about 32 hours, from about 17 hours to about 31 hours, from about 17 hours to about 30 hours, from about 18 hours to about 32 hours, from about 18 hours to about 31 hours, or from about 18 hours to about 30 hours. Preferably, the mixing time may be about 18 hours to about 30 hours.

According to one embodiment, in the step of extracting using the osmotic dehydration extraction method, when the mixing time of the okra and the salt is less than about 15 hours, the amount of dehydration decreases and the extraction yield decreases, or the oil-soluble component contained in the extract amount may decrease. In addition, in the step of extracting using the osmotic dehydration extraction method, if the mixing time of the okra and the salt exceeds about 32 hours, dehydration is stopped due to the concentration equilibrium that occurs as dehydration proceeds, so that the extraction yield is no longer This may not increase. In addition, when the okra is exposed to the salt for a long time, the cell tissue of the okra may be damaged, and polymer components such as enzymes in the cell may leak out of the cell. Therefore, the finally obtained extract may show deterioration in quality, such as containing impurities or browning by enzymes.

In addition, the step of extracting the okra by using the osmotic dehydration extraction method by mixing the okra and the salt may further include removing the salt from the obtained dehydrated extract.

The method of extracting the okra may include extracting the okra using a molecular compression dehydration extraction method by mixing the okra and a polymer material.

The molecular compression dehydration extraction may refer to discharging water or oil-soluble components in cells to the outside of the cells by using the cytolytic action of a polymer.

The term "Cytorrhysis" means permanent damage to the plant cell wall due to the loss of positive pressure inside the cell. In the cytosis, a polymer material having a size larger than the pores of the plant cell wall cannot penetrate into the cell wall and diffusion pressure is applied from the outside of the cell, so that the cell is contracted and crushed, and dehydration may occur.

In the case of the molecular compression dehydration extraction using the cytosis, it is similar to the conventional osmotic dehydration in that it dehydrates using a solute outside the cell, but in the conventional osmotic dehydration, the solute can move through the pores of the cell wall, The polymer material used in torsis is larger than the pores of the cell wall, so there is a difference that it stays only in the extracellular tissue. Therefore, in the conventional osmotic dehydration, dehydration is stopped when the concentration gradient disappears as the concentration of solute inside and outside the cell becomes the same, whereas in the case of the molecular compression dehydration extraction using cytosis, the concentration gradient inside and outside the cell is continuously maintained. A large amount of water can be dehydrated than osmotic dehydration, and thus the extraction yield or content of oil-soluble components of the obtained extract can be significantly increased.

It is preferable that the polymer material has a size larger than the pores or pores of the cell wall of plant cells, and has a high solubility in water, so that a high concentration aqueous solution can be prepared. do.

The polymer material may be at least one selected from the group consisting of polyethylene glycol (PEG), arabic gum, dextrin, and arabinogalactan, but is not limited thereto. Preferably, the polymer material may be maltodextrin.

The molecular weight of the polymer material may be about 500 or more to about 10000 or less. Specifically, the molecular weight of the polymer material is about 500 or more to about 8000 or less, about 500 or more to about 6000 or less, about 500 or more to about 4000 or less, about 500 or more to about 2000 or less, about 500 or more to about 1000 or less, about 1000 or more to about 10000 or less, about 1000 or more to about 8000 or less, about 1000 or more to about 6000 or less, about 1000 or more to about 4000 or less, about 1000 or more to about 2000 or less, about 2000 or more to about 10000 or less, about 2000 or more to about 8000 or less, about 2000 or more to about 6000 or less, about 2000 or more to about 4000 or less, about 4000 or more to about 10000 or less, about 4000 or more to about 8000 or less, about 4000 or more to about 6000 or less, about 6000 or more to about 10000 or less, about 6000 or more to about 8000 or less, or about 8000 or more to about 10000 or less. Preferably, the molecular weight of the polymer material may be about 2000 or more to about 6000 or less.

According to one embodiment, when the molecular weight of the polymer material is less than about 500, the polymer material is smaller in size than the pores or pores of the cell wall of the okra cell and can penetrate into the cell, in this case, the concentration inside and outside the cell within a short time As the gradient is lost and dehydration is stopped, the amount of dehydration may decrease. Due to this, the extraction yield of the obtained extract and the content of oil-soluble components may decrease. In addition, when the molecular weight of the polymer material exceeds about 10000, the size of the polymer material is excessively larger than the pores or pores of the cell wall of the okra cell, so that cytosis is not easily induced, and the amount of dehydration may be reduced. Due to this, the extraction yield of the obtained extract and the content of oil-soluble components may decrease.

In the step of extracting the okra by using a molecular compression dehydration extraction method by mixing the okra and the polymer material, the polymer material may be adhered to or applied to the tissue surface of the okra by a cytolytic action. Therefore, even if the water discharged from the okra increases and the dehydrated liquid outside the okra tissue is diluted, the concentration gradient between the okra tissue surface and the inside of the tissue can be maintained by the polymer material attached or applied to the okra tissue surface. . Due to this, dehydration may occur continuously, the amount of dehydration or extraction yield may increase, and the content of oil-soluble components contained in the obtained extract may increase. In addition, since the polymer material is attached to or applied to the tissue surface of the okra by cytosis, it is possible to prevent the okra tissue from being directly exposed to air, thereby reducing the browning phenomenon caused by oxidation. Due to this, the problem of the existing dehydration extraction method, that is, as dehydration from plant tissue continues, concentration equilibrium is induced inside and outside the plant tissue and dehydration is stopped. It is possible to solve the problem that a large amount of polymer material was required for this purpose.

In the step of extracting the okra by using a molecular compression dehydration extraction method by mixing the okra and the polymer material, the polymer material and the okra may be mixed in a weight ratio of 10 to 300: 100. Specifically, in the step of extracting using the molecular compression dehydration extraction method, the polymer material and the okra 10 to 250, 10 to 200, 10 to 150, 10 to 100, 10 to 50, 50 to 300, 50 to 250, 50 to 200, 50 to 150, 50 to 100, 100 to 300, 100 to 250, 100 to 200, 100 to 150, 150 to 300, 150 to 250, 150 to 200, 200 to 300, 200 to 250, Alternatively, it may be mixed in a weight ratio of 250 to 300: 100, and preferably mixed in a weight ratio of 50 to 150: 100.

According to one embodiment, in the step of extracting using the molecular compression dehydration extraction method, when the polymer material and the okra are mixed in a weight ratio of less than 10: 100, the amount of dehydration decreases and the extraction yield decreases, or the extract The amount of oil-soluble ingredients contained in it may be reduced. In addition, in the extraction using the molecular compression dehydration extraction method, when the polymer material and the okra are mixed in a weight ratio of more than 300: 100, the solubility of the polymer material decreases, The extraction yield may be reduced or the cell tissue of the okra may be damaged, and polymer components such as enzymes in the cell may be leaked out of the cell. Therefore, the finally obtained extract may show deterioration in quality, such as containing impurities or browning by enzymes. In addition, it may be difficult to obtain and use the final extract due to the undissolved polymer material.

In the step of extracting the okra by using a molecular compression dehydration extraction method by mixing the okra and the polymer material, the mixing time of the okra and the polymer material may be about 15 hours to about 32 hours. Specifically, the mixing time is about 15 hours to about 31 hours, about 15 hours to about 30 hours, about 16 hours to about 32 hours, about 16 hours to about 31 hours, about 16 hours to about 30 hours, about 17 hours. to about 32 hours, from about 17 hours to about 31 hours, from about 17 hours to about 30 hours, from about 18 hours to about 32 hours, from about 18 hours to about 31 hours, or from about 18 hours to about 30 hours. Preferably, the mixing time may be about 18 hours to about 30 hours.

According to one embodiment, in the step of extracting using the molecular compression dehydration extraction method, when the mixing time of the okra and the polymer material is less than about 15 hours, the amount of dehydration decreases and the extraction yield is reduced or included in the extract The amount of oil-soluble ingredients used may be reduced. In addition, in the step of extracting using the molecular compression dehydration extraction method, if the mixing time of the okra and the polymer material exceeds about 32 hours, the dehydration is stopped by the concentration equilibrium that occurs as the dehydration proceeds, so that more Ideally, the extraction yield may not increase. In addition, when the okra is exposed to the polymer material for a long time, the cell tissue of the okra may be damaged, and polymer components such as enzymes in the cell may leak out of the cell. Therefore, the finally obtained extract may show deterioration in quality, such as containing impurities or browning by enzymes.

In addition, the step of extracting the okra by using the molecular compression dehydration extraction method by mixing the okra and the polymer material may further include the step of removing the polymer material from the obtained dehydrated extract.

The method of extracting the okra may include a first dehydration step and a second dehydration step. Specifically, when the first dehydration step is a step of extracting using an osmotic dehydration extraction method, the second dehydration step may be a step of extracting using a molecular compression dehydration extraction method. In addition, when the first dehydration step is a step of extracting using a molecular compression dehydration extraction method, the second dehydration step may be a step of extracting using an osmotic dehydration extraction method.

In one embodiment, the method of extracting the okra is a primary dehydration step of mixing okra and salt to proceed with osmotic dehydration extraction; and a secondary dehydration step of performing molecular compression dehydration extraction by mixing the primary dehydrated okra and a polymer material. In addition, the method includes a primary dehydration step of mixing okra and a polymer material to extract molecular compression dehydration; and a secondary dehydration step of osmotic dehydration extraction by mixing the primary dehydrated okra and salt.

According to one embodiment, compared to the okra extract obtained through a single extraction method in which only the osmotic dehydration extraction or the molecular compression dehydration extraction is performed alone, a complex extraction method in which the second dehydration step is performed after the first dehydration step That is, in the case of an okra extract obtained through a complex extraction method that proceeds by combining the extraction using the osmotic dehydration extraction method and the extraction using the molecular compression dehydration extraction method, the extraction yield or content of oil-soluble components This can increase significantly. In addition, compared with the okra extract obtained through the single extraction method, in the case of the okra extract obtained through the complex extraction method, quality, preservation power, skin wrinkle improvement effect, skin elasticity improvement effect, skin itch improvement effect, or skin moisturizing effect may increase significantly.

Compared to the single extraction method, in the case of performing the complex extraction method, in the step of extracting the okra using a molecular compression dehydration extraction method by mixing the okra and the polymer material, the polymer material is applied to the tissue surface of the okra. It may be attached or applied with higher efficiency. This can maintain the concentration gradient between the surface of the okra tissue and the inside of the tissue for a longer time, and can cause the cytosis phenomenon to occur more efficiently. Due to this, the dehydration time may increase, the dehydration amount or extraction yield may further increase, and the content of the oil-soluble component contained in the obtained extract may further increase. Therefore, the problem of the existing dehydration extraction method through the complex extraction method, that is, as dehydration continues from the plant tissue, concentration equilibrium is induced inside and outside the plant tissue and dehydration is stopped, the amount of dehydration and the extraction yield are inevitably reduced, It is possible to solve the problem that a large amount of polymer material was required to increase the extraction yield.

In addition, compared to the single extraction method, when the complex extraction method is performed, the polymer material is attached or applied to the tissue surface of the okra with higher efficiency, thereby more strongly inhibiting damage to the okra tissue. By more strongly preventing the outflow of polymer components such as enzymes in cells from the dehydrated tissue, or more strongly preventing the air exposure of the okra tissue, browning caused by polymer components or oxidation can be further reduced. For this reason, the okra extract obtained through the complex extraction method may exhibit better quality.

The method of extracting the okra includes the steps of extracting using an osmotic dehydration extraction method, followed by extraction using a molecular compression dehydration extraction method; Alternatively, after the extraction using the molecular compression dehydration extraction method, the extraction may be performed in the order of the extraction using the osmotic dehydration extraction method.

In one embodiment, when the method of extracting okra proceeds in the order of the extraction using the molecular compression dehydration extraction method after the step of extracting using the osmotic dehydration extraction method, compared to the case of proceeding in the reverse order The extraction yield may be further increased. When the osmotic dehydration extraction is pre-treated, the polymer material can be more efficiently attached or applied to the cell wall softened through the osmotic dehydration extraction, and thereby, the cytosis phenomenon in the post-treated molecular compression dehydration extraction It may occur more efficiently and the extraction yield may be further increased.

In addition, in one embodiment, when the method of extracting okra proceeds in the order of the extraction using the molecular compression dehydration extraction method after the extraction using the osmotic dehydration extraction method, the initial dehydration rate is fast There may be advantages.

In the step of extracting using the osmotic dehydration extraction method of the method of extracting the okra or the extraction using the molecular compression dehydration extraction method, the salt or the polymer material may be mixed with the okra in a solid or solution state have.

The solid state may mean a suspension or a crystalline state or a powder state in which a polymer material is mixed with water in a saturated state or more, and some are dispersed in a solid state of crystals or powders.

In the step of extracting using the osmotic dehydration extraction method of the method of extracting the okra or the extraction using the molecular compression dehydration extraction method, when the salt or the polymer material is mixed with the okra in a solid state, first the The salt or the polymer material may be dissolved by moisture on the surface of the okra tissue to cause dehydration. As the dehydration proceeds, the salt or polymer material is continuously dissolved by the water leaking out of the okra tissue, so the okra tissue is placed in a saturated solution throughout the dehydration process, so that the dehydration can be continued effectively. For this reason, it is possible to efficiently dehydrate okra using a minimum amount of salt or high molecular material, thereby increasing the amount of dehydration or extraction yield, and increasing the content of oil-soluble components contained in the obtained extract.

In addition, in each of the above steps, when the salt or the polymer material is mixed with the okra in a solid state, the oil-soluble component extracted from the okra tissue is not diluted and hardly undergoes changes due to enzymes or oxidation. It can be extracted while retaining the intrinsic properties of oil-soluble components. Due to this, the obtained extract may contain oil-soluble components of high quality, and may exhibit more excellent efficacy.

In addition, in each of the above steps, when the salt or the polymer material is mixed with the okra in a solid state, the concentration of the salt or the polymer material is maintained almost in a saturated state and the water activity is lowered, so that the Corruption of the dehydrated extract is prevented, and an extract of excellent quality that does not deteriorate without a separate process such as addition of a preservative or sterilization treatment can be obtained, and the obtained extract can be stored in a fresh state for a long time.

In the step of extracting using the osmotic dehydration extraction method of the method of extracting the okra or the extraction using the molecular compression dehydration extraction method, the salt or the polymer material and the okra may be mixed at room temperature. The room temperature may mean an indoor or outdoor temperature without additional temperature treatment. Specifically, the room temperature may mean about 10 ℃ to about 40 ℃. The room temperature is about 10 °C to about 35 °C, about 10 °C to about 30 °C, about 10 °C to about 25 °C, about 10 °C to about 20 °C, about 10 °C to about 15 °C, about 15 °C to about 40 °C , from about 15 °C to about 35 °C, from about 15 °C to about 30 °C, from about 15 °C to about 25 °C, from about 15 °C to about 20 °C, from about 20 °C to about 40 °C, from about 20 °C to about 35 °C, or It may mean about 20 ℃ to about 30 ℃.

The method of extracting the okra may further include filtering or centrifuging the extract obtained in the extracting using the osmotic dehydration extraction method or the extracting using the molecular compression dehydration extraction method. In the step of filtration or centrifugation, the dehydrated tissue may be separated from the obtained extract. For example, the dehydrated tissue contained in the extract obtained in the first dehydration step may be separately separated in the filtration or centrifugation step and used in the second dehydration step. In addition, in the filtration or centrifugation step, the dehydrated tissue, undissolved salt or polymer material, or impurities may be separated and removed from the extract obtained in the extraction step.

The step of extracting using the osmotic dehydration extraction method of the method of extracting okra or the step of extracting using the molecular compression dehydration extraction method is a dehydration aid, additive, or pH adjusting agent in the salt or a mixture of the polymer material and the okra. By adding additionally and leaving it to stand, it may be to increase the dehydration efficiency. The dehydration aid may be a low molecular weight dehydration aid, and the additive may be salt, sugar, polysaccharide, sugar alcohol, or dietary fiber. In addition, the pH adjusting agent may be an inorganic acid such as hydrochloric acid, sulfuric acid, and caustic soda, an organic acid such as alkali and acetic acid, citric acid, malic acid tartaric acid, or salts thereof. In one embodiment, in each step, a pH adjuster may be additionally mixed to reduce quality change due to a change in pH that may occur during dehydration, such as a change in pH due to addition of a salt or a polymer material.

Another aspect provides an okra extract prepared by the method of extracting the okra.

In one embodiment, the effective concentration of the okra extract may be about 1 to 300 μg / mL. Specifically, the effective concentration of the okra extract is about 1-250, about 1-200, about 1-150, about 1-100, about 10-300, about 10-250, about 10-200, about 10-150, or about 10 to 100 μg/mL.

In one embodiment, the okra extract may inhibit collagenase (MMP-1), which is a collagen hardening enzyme or a collagen degrading enzyme. The okra extract may exhibit an anti-aging effect, improving skin wrinkles, improving skin elasticity, or improving skin elasticity by inhibiting the enzyme.

In one embodiment, the okra extract may increase the expression of the COL1A1 gene, which is a type 1 collagen gene. The okra extract may exhibit an anti-aging effect by increasing the expression of the gene, thereby improving skin wrinkles, improving skin elasticity, or improving skin elasticity.

In one embodiment, the okra extract may increase the expression of hyaluronic acid synthase (Hyaluronan synthase 3, HAS3). The okra extract may exhibit a skin moisturizing effect by increasing the expression of the enzyme.

In one embodiment, the okra extract may inhibit the expression of Thymic stromal lymphopoietin (TSLP) gene, which is an itch-inducing gene. The okra extract may exhibit an effect of improving skin itch by suppressing the expression of the gene.

In one embodiment, the pH of the okra extract is about 3 or more to about 7 or less, preferably, the pH of the okra extract may be about 5. If the pH of the okra extract is less than about 3, there may be restrictions on the use of the cosmetic composition, and if the pH is greater than about 7, there may be a problem of inhibiting enzyme activity.

Among the terms or elements mentioned in the extract, those mentioned in the description of the method are understood as being mentioned in the description of the method above.

Another aspect provides a cosmetic composition comprising an okra extract prepared by the method of extracting the okra.

The cosmetic composition may exhibit an effect of improving skin wrinkles, improving skin elasticity, improving skin itchiness, or improving skin moisture.

Accordingly, the cosmetic composition may be an anti-aging cosmetic composition.

The term "wrinkle" refers to fine lines caused by deterioration of the skin. The wrinkles are caused by genes, a decrease in collagen present in the dermis of the skin, external environment, aging, and the like.

The term "wrinkle improvement" means inhibiting or inhibiting the formation of wrinkles on the skin or alleviating wrinkles already formed. It includes alleviating skin wrinkles by improving or increasing skin elasticity, and may include improving skin wrinkles by reducing the pore area.

The term "skin moisturizing" refers to any action that maintains skin moisture or prevents moisture loss.

The cosmetic composition may be a composition for cosmetic raw materials used in the manufacture of cosmetics. In addition, the cosmetic composition may be a cosmetic composition having a final cosmetic formulation.

In one embodiment, the cosmetic composition may further include an antioxidant, a preservative, a thickener, or a humectant.

The antioxidant is a substance added to prevent automatic oxidation by the action of oxygen, which is easy to occur in cosmetic compositions, and for example, butylhydroxyanisole, propyl gallic acid, elisorbic acid, and the like may be used.

The preservative refers to any raw material that dissolves in the oil phase and exhibits antiseptic activity, and the natural raw material may include an essential oil extracted by steam distillation having antibacterial and anti-inflammatory effects. For example, pentylene glycol, hexanediol, ethylhexyl glycerin, caprylyl glycol, glycerin, butylene glycol, propylene glycol, etc. belonging to phenoxyethanol, ethanol and polyhydric alcohols, etc. Organic acids [acetic acid, butyric acid, citric acid, palmitic acid, oxalic acid, tartaric acid, glycolic acid, lactic acid ( lactic acid), maleic acid, etc.], or dipotassium glycyrrhizate.

The moisturizing agent is a material that can prevent the drying of the skin and maintain the flexibility and elasticity of the skin, and includes polyols, amino acids, or sugars. The polyols may include alcohols such as glycerin, propylene, butylene glycol, and alcohol derivatives such as panthenol. Monosaccharides such as glucose, trehalose, xylitol, and erythritol, polysaccharides such as chitosan and hyaluronic acid, and ammonium salts such as polyquaternium-51 may be used as the amino acids and sugars.

The thickener is a substance that improves the viscosity, texture, etc. of the cosmetic composition, for example, cellulose gum, xanthan gum, genlan gum, agar, tamarind gum, guar gum, gum arabic, cetyl alcohol, stearyl alcohol, cetyl alcohol Tearyl alcohol, sodium chloride, polyquaternium-7, etc. can be used.

The cosmetic composition may be prepared in any formulation conventionally prepared in the art. Specifically, the formulation of the cosmetic composition is a solution, emulsion, suspension, softening lotion, nutrient lotion, massage cream, nutrient cream, pack, gel, skin adhesion type cosmetics, lipstick, powder, makeup base, foundation, shampoo , conditioner, scalp cleaner, scalp lotion, scalp cream, scalp pack, scalp gel, scalp ointment, scalp gel, body cleanser, soap, toothpaste, mouthwash, paste, lotion, ointment, gel, cream , a patch, a spray, or a spray, but is not limited thereto.

The carrier included in the cosmetic composition may be selectively used according to the formulation of the cosmetic. For example, when manufacturing cosmetics in the form of ointments, pastes, creams, or gels, wax, paraffin, starch, tracanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc. may be used alone or in combination. When manufacturing powder or spray-type cosmetics, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, chlorofluorohydrocarbon, propane/butane, dimethyl ether, etc. are used alone as carrier components. Or they can be used in combination. When preparing cosmetics in the form of solutions or emulsions, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, cottonseed oil, peanuts as carrier ingredients Oil, corn germ oil, olive oil, castor oil, sesame oil, glycerol fatty ester, polyethylene glycol, or fatty acid ester of sorbitan may be used alone or in combination. In the case of manufacturing a cosmetic in the form of a suspension, as a carrier component, water, ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide , bentonite, agar, tracanth, and the like may be used alone or in combination. In the case of manufacturing soap-type cosmetics, alkali metal salts of fatty acids, fatty acid hemiester salts, fatty acid protein hydrolyzates, isethionate, lanolin derivatives, aliphatic alcohols, vegetable oils, glycerol, sugar, etc. are used alone as carrier components. may be used alone or in combination.

Among the terms or elements mentioned in the composition, those mentioned in the description of the methods and extracts are understood to be as mentioned in the description of the methods and extracts above.

According to the method of extracting okra according to an aspect, it is possible to exhibit a remarkably excellent extraction yield by solving the problems of the existing plant dehydration extraction method. Specifically, according to the method of extracting okra according to an aspect, it is possible to obtain the maximum dehydration effect while using a minimum amount of salt or polymer material, so that the extraction yield can be significantly increased, as well as oil-soluble components included in the extract The amount of can be significantly increased, and it is possible to prepare an extract of excellent quality in which the intrinsic properties of oil-soluble components are not destroyed.

In addition, in the case of an okra extract prepared by the method of extracting okra of one aspect or a cosmetic composition comprising the same, it may exhibit a remarkably excellent skin wrinkle improvement effect, skin elasticity improvement effect, skin itch improvement effect, or skin moisturizing effect. .

1 is a schematic diagram showing steps of a method of extracting okra of an aspect.
2 is a graph showing the activity of collagen hardening enzyme when fibroblasts are treated with an okra extract of one aspect.
3 is a graph showing the expression level of collagenase (MMP-1) when fibroblasts are treated with an okra extract of one aspect.
4 is a graph showing the expression level of type 1 collagen gene when human fibroblasts are treated with an okra extract of one aspect.
5 is a graph showing the HAS3 gene expression level when human epidermal cells are treated with an okra extract of one aspect.
6 is a graph showing the level of itchiness in the human epidermal cells when one aspect of the okra extract is treated.

Hereinafter, the present invention will be described in more detail through experimental examples and examples. However, these Experimental Examples and Examples are for illustrative purposes of the present invention, and the scope of the present invention is not limited to these Experimental Examples and Examples.

In addition, unless specifically defined herein, all scientific and technical terms used herein may have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

Example 1. Obtaining Okra Extract Using Osmotic Dehydration Extraction and Molecular Compression Dehydration Extraction

1.1 Preparation of Okra

Okra was prepared in order to obtain an okra extract using the osmotic dehydration extraction method, the molecular compression dehydration extraction method, or a combination thereof.

Specifically, the size of the raw okra was about 1.0 to 200.0 mm in size, and preferably okra having a size of about 1.0 to 5.0 mm was used. Taking care not to damage the cell membrane of the okra, the polymer material was cut to a size within about 2 mm in an easy direction so that the polymer material could move to the maximum on the surface area of the okra.

1 is a schematic diagram showing steps of a method of extracting okra of an aspect.

Using the steps shown in FIG. 1, the osmotic dehydration extraction method using salt, the molecular compression dehydration extraction method using a polymer material, and the extraction method combining them were compared.

1.2 Osmotic dehydration extraction method using salt

In the osmotic dehydration extraction method using a salt, sodium chloride was used as the salt.

Osmotic dehydration extraction method (salt: raw material W/W, %) treatment process Preparation Example 1 Preparation 2 Preparation 3 Preparation 4 Preparation 5 Preparation 6 weight ratio 10: 100 30: 100 50 : 100 100 : 100 150 : 100 200 : 100

Table 1 shows the weight ratio of salts and okra fragments used in each preparation by the osmotic dehydration extraction method.

Preparation Examples 1 to 6 were obtained by mixing the salt and the okra fragment in the same weight ratio as shown in Table 1 and leaving it at room temperature for about 48 hours.

1.3 Molecular compression dehydration extraction method using polymer materials

In molecular compression dehydration extraction, a water-soluble solid polymer having a molecular weight of about 3,000 to 4,500 was used, and specifically, arabic gum, PEG, or maltodextrin was primarily selected. Among them, maltodextrin most suitable for combination with the osmotic dehydration extraction method was used.

Molecular compression dehydration extraction method (maltodextrin: raw material W/W, %) treatment process Preparation 7 Preparation 8 Preparation 9 Preparation 10 Preparation 11 Preparation 12 weight ratio 50 : 100 100 : 100 200 : 100 300 : 100 400 : 100 500 : 100

Table 2 shows the weight ratio of maltodextrin and okra fragments used in each preparation example by the molecular compression dehydration extraction method.

By mixing maltodextrin and okra fragments in the same weight ratio as shown in Table 2 and leaving them at room temperature for about 48 hours, Preparation Examples 7 to 12 were obtained.

1.4 Extraction method using osmotic dehydration extraction and molecular compression dehydration extraction

The method of extracting by combining the osmotic dehydration extraction method and the molecular compression dehydration extraction method precedes the osmotic dehydration extraction step, then performs the molecular compression dehydration extraction step, precedes the molecular compression dehydration extraction step, and then osmotic dehydration extraction The steps were divided by the way they were carried out. Each experimental condition was comparatively analyzed at intervals of about 6 hours up to about 48 hours, and extracted at room temperature conditions.

Processing time (hr) extraction process Preparation 13 Preparation 14 Preparation 15 Preparation 16 Preparation 17 Preparation 18 Preparation 19 osmotic pressure
preprocessing extraction
(30: 100
W/W, %) osmotic pressure 6 12 18 24 30 36 42 molecular compression 42 36 30 24 18 12 6

Processing time (hr) extraction process production example
20 Preparation 21 Preparation 22 Preparation 23 Preparation 24 Preparation 25 Preparation 26 molecular compression
preprocessing extraction
(100 : 100 W/W, %) molecular compression 6 12 18 24 30 36 42 osmotic pressure 42 36 30 24 18 12 6

Table 3 is This is a table showing the conditions for each treatment time in a complex process that precedes the osmotic dehydration extraction step and then performs the molecular compression dehydration extraction step. The weight ratios in Table 3 represent the weight ratio of salt to raw material.

Table 4 is a table showing the conditions for each treatment time in a complex process that precedes the molecular compression dehydration extraction step and then performs the osmotic dehydration extraction step. The weight ratios in Table 4 represent the weight ratios of maltodextrin to raw material.

As shown in Tables 3 and 4, Preparation Examples 13 to 19 preceding the osmotic dehydration extraction step and Preparation Examples 20 to 26 preceding the molecular compression dehydration extraction step were prepared by varying the treatment time in the complex process.

Example 2. Evaluation of extraction yield of okra according to each extraction method

2.1 Extraction yield and solubility of osmotic dehydration extraction method

Table 5 is a table showing the yield of osmotic dehydration extraction according to the weight ratio. In Table 5 below, the extraction yield is the yield including the weight of the dissolved salt, and the extraction yield excluding the raw material is the yield excluding the weight of the dissolved salt, or when it is insoluble and cannot be obtained, the yield is compared to the difference in weight of the raw material. , in solubility, - means impossible to obtain, O means complete dissolution, △ indicates incomplete dissolution, and X means insoluble.

Yield (%) according to weight ratio (salt: raw material W/W, %) treatment process Preparation Example 1 Preparation 2 Preparation 3 Preparation 4 Preparation 5 Preparation 6 osmotic pressure 10: 100 30: 100 50 : 100 100 : 100 150 : 100 200 : 100 extraction yield 36.34 51.38 54.81 22.91 - - Excluding raw materials
extraction yield 29.97 38.89 42.21 31.73 32.52 30.8 Solubility O O △ X X X

As a result, as shown in Table 5, when the mixing weight ratio of the salt and the raw material was 50: 100 (W/W, %), the optimal extraction yield was obtained, but there was a problem in that the salt was incompletely dissolved. In addition, when the weight ratio exceeds 100: 100, the salt was not completely dissolved. The extraction yield, excluding the weight of the dissolved salt in the extract, also showed the highest yield when the mixed weight ratio of the salt and the raw material was 50: 100, but as described above, incomplete dissolution occurred when the weight ratio was 50: 100, and the weight ratio In the case of 100: 100, most of the salts were not dissolved and remained. If the salt remains undissolved, there is a problem in that the extract cannot be collected due to the remaining salt. Therefore, in the mixing weight ratio of the salt and the raw material, the 30: 100 (W/W, %) ratio of Preparation Example 2 was determined as the optimal ratio.

2.2 Extraction yield and solubility of molecular compression dehydration extraction method

Table 6 is a table showing the yield of molecular compression dehydration extraction according to weight ratio. In Table 6 below, the extraction yield is the yield including the weight of the dissolved dextrin, the extraction yield excluding the raw material is the yield excluding the weight of the dissolved dextrin, or when it is insoluble and cannot be obtained, the yield comparing the weight difference of the raw material In the solubility, - means impossible to obtain, O means complete dissolution, △ indicates incomplete dissolution, and X means insoluble.

Yield (%) according to weight ratio (maltodextrin: raw material W/W, %) treatment process Preparation 7 Preparation 8 Preparation 9 Preparation 10 Preparation 11 Preparation 12 molecular compression 50 : 100 100 : 100 200 : 100 300 : 100 400 : 100 500 : 100 extraction yield 61.86 75.66 56.81 - - - Excluding raw materials
extraction yield 42.79 51.32 60.44 53.48 48.25 48.99 Solubility O O △ X X X

As a result, as shown in Table 6, in the case of molecular compression dehydration extraction using dextrin, until the mixing weight ratio of dextrin and raw material is 100: 100 (W/W, %), the dextrin is completely dissolved to effectively obtain an extract. However, when the weight ratio exceeds 200: 100, the dextrin is not dissolved and agglomerate occurs, and when the weight ratio is 300: 100, there is a problem in that it is difficult to collect the extract.

When the extraction yield was compared, when the mixed weight ratio of dextrin and raw material was 100: 100 (W/W, %), it was confirmed that up to 75.66% was extracted, but when the weight ratio was 200: 100, the aggregation of dextrin caused Undissolved dextrin remained and the extraction yield decreased, and when the weight ratio was 300: 100, there was a problem in that it was impossible to obtain an extract.

The extraction yield excluding raw materials was found to be 51.32% when the weight ratio was 100: 100, and 60.44% when the weight ratio was 200: 100. Therefore, when the weight ratio is 200: 100, the extraction yield may be the highest, but it is difficult to remove the undissolved dextrin due to the aggregation of dextrin, and it is in the form of a highly viscous extract, so it may be difficult to recover the extract And it was found. Therefore, in the molecular compression dehydration extraction process of okra using dextrin, it was found that the mixing weight ratio of dextrin and the raw material was 100: 100 (W/W, %) of Preparation Example 8 as the optimal ratio.

2.3 Extraction Yield of Osmotic Dehydration Extraction and Molecular Compression Dehydration Extraction Combination Extraction Method

In order to analyze the yield and solubility in the case of extraction by combining osmotic dehydration extraction and molecular compression dehydration extraction method, the extraction yield for each treatment time when osmotic dehydration extraction was pre-treated or molecular compression dehydration extraction was pre-treated was analyzed.

Table 7 shows the extraction yield for each treatment time of the osmotic dehydration extraction pretreatment complex process. The extraction yield is the yield including the weight of the dissolved salt and polymer material, the extraction yield excluding the raw material is the yield excluding the weight of the dissolved salt and the polymer material, or the yield compared to the difference in the weight of the raw material when it cannot be obtained because it is insoluble is shown.

okra Treatment time (hr) / Yield (%) extraction process Preparation 13 Preparation 14 Preparation 15 Preparation 16 Preparation 17 Preparation 18 Preparation 19 Osmotic pretreatment Osmotic pressure (30 : 100 W/W, %) 6 12 18 24 30 36 42 Molecular compression (100 : 100 W/W, %) 42 36 30 24 18 12 6 extraction yield 69.1 70.9 78.2 82.2 86.7 58.6 43.9 Extraction yield without raw materials 53.92 54.17 60.93 65.1 69.6 44.76 41.11

Table 8 shows the extraction yield for each treatment time of the molecular compression dehydration extraction pretreatment complex process.

okra Treatment time (hr) / Yield (%) extraction process Preparation 20 Preparation 21 Preparation 22 Preparation 23 Preparation 24 Preparation 25 Preparation 26 Molecular compression pretreatment Molecular compression (100 : 100 W/W, %) 6 12 18 24 30 36 42 Osmotic pressure (30 : 100 W/W, %) 42 36 30 24 18 12 6 extraction yield 36.5 55.8 83.7 81.3 77.8 71.3 69.4 Extraction yield without raw materials 40.93 46.32 62.58 60.03 55.83 49.01 50.66

As a result, as shown in Table 7, in the case of pretreatment of the osmotic dehydration extraction process, the treatment of about 18 hours of molecular compression dehydration extraction after about 30 hours of osmotic dehydration extraction showed the highest extraction yield. In addition, as shown in Table 8, in the case of the molecular compression dehydration extraction pretreatment, it showed the highest extraction yield when the molecular compression dehydration extraction was performed for about 18 hours followed by the osmotic dehydration extraction for about 30 hours. In addition, it was confirmed that the efficiency was lowered when the dextrin of the molecular compression dehydration extraction process was not sufficiently dissolved than when the salt of the osmotic dehydration extraction process was not sufficiently dissolved during extraction. In addition, the method of pre-treating osmotic dehydration extraction in the two complex processes in which the same raw material and raw materials were treated showed a relatively high extraction yield compared to the method of pre-treating molecular compression dehydration extraction, and both complex processes were extracted in a single process It was confirmed that the yield was higher than the yield.

This result is thought to be because dextrin, a polymer material, efficiently adhered to the cell wall softened during osmotic dehydration extraction, and the cytosis phenomenon occurred more efficiently. By suppressing the concentration equilibrium that occurs, it means that the disadvantage of the existing process, which requires a large amount of raw material to be diluted by the dehydration solution, has been improved.

Next, the okra extract according to Example 1, specifically, the okra extract under the maximum yield condition among the osmotic dehydrated extracts by salt (Preparation Example 2), and the okra extract under the maximum yield condition among the molecularly compressed dehydrated extracts (Preparation Example 8) ), and to compare the efficacy in skin cells of the okra extract (Preparation Example 17) in the maximum yield condition among the extracts by the complex process of osmotic dehydration extraction and molecular compression dehydration extraction, wrinkle improvement, elasticity improvement, moisturizing, and itching Improvement efficacy was evaluated.

Example 3. Efficacy evaluation of okra extract

3.1 Evaluation of the inhibitory efficacy of Lysyl oxidase

In order to confirm the wrinkle suppression, elasticity improvement, or anti-aging effect of the okra extract of one aspect, the collagen curing enzyme (Lysyl oxidase) inhibitory effect of the okra extract of one aspect was evaluated.

An experimental group was prepared to compare the inhibitory effect on the collagen sclerosing enzyme of Preparation Examples 2 and 8 extracted by a single method and Preparation Example 17 extracted by combining osmotic dehydration extraction and molecular compression dehydration extraction method. Specifically, i) about 25 mM medium with glucose without okra extract, ii) about 50 mM medium with glucose without okra extract, iii) about 10 μg/mL of okra extract of Preparation Example 2 and about 50 mM of glucose a medium comprising, iv) a medium comprising about 10 μg/mL of the okra extract of Preparation Example 8 and about 50 mM glucose, v) a medium comprising about 10 μg/mL of the okra extract of Preparation Example 17 and about 50 mM glucose, vi) a medium containing about 100 μg/mL of the okra extract of Preparation Example 2 and about 50 mM glucose, vii) a medium containing about 100 μg/mL of the okra extract of Preparation Example 8 and about 50 mM glucose, viii) Preparation Example A medium containing about 100 μg/mL of the okra extract of 17 and about 50 mM of glucose, and ix) a medium containing about 50 mM of aminopropionitrile (Beta-Aminopropionitrile, BAPN) and glucose was prepared.

Hs68 fibroblasts were aliquoted in a 6-well plate by the number of about 3.5x10 ⁵ per well, and then cultured for about 24 hours in an incubator at about 37°C and about 5% CO ₂ conditions. Next, the cultured medium of the fibroblasts was replaced with the medium of the six experimental groups, and further cultured for about 24 hours. After about 24 hours, about 1 ml of the medium was collected from each well, and then, only the supernatant was recovered by centrifugation at a speed of about 13000 rpm at about 4° C. for about 5 minutes. Next, about 50 ul of the supernatant and about 50 ul of the preparation buffer of the Lysyl oxidase activity assay kit (ab112139, abcam) were put into a 96 well black well and reacted at about 37°C for about 30 minutes. Then, the absorbance at Ex/Em = 540/590 nm was measured using a Victor 3 plate reader. By calculating the percentage using only about 25 mM glucose medium as a negative control, the collagen sclerosis activity of cells in each experimental group was shown.

2 is a graph showing the activity of collagen hardening enzyme when fibroblasts are treated with an okra extract of one aspect.

As a result, as shown in FIG. 2, when about 100 ug/ml of the okra extract of Preparation Example 17 was treated, the inhibitory effect of the collagen hardening enzyme was the highest. These results show that the okra extract extracted by combining the osmotic dehydration extraction and the molecular compression dehydration extraction method has a better collagen sclerosing enzyme inhibitory effect than the okra extract by a single method, thereby further increasing collagen production, It means that it can exhibit excellent wrinkle suppression, elasticity improvement, or anti-aging effect.

3.2 MMP-1 inhibitory ability evaluation

In order to confirm the wrinkle suppression, elasticity improvement, or anti-aging effect of the okra extract of one aspect, the inhibitory ability of the okra extract of one aspect on collagenase (MMP-1), which is an enzyme decomposing collagen, was evaluated.

An experimental group was prepared to compare the MMP-1 gene expression inhibitory ability of Preparation Examples 2 and 8 extracted by a single method and Preparation Example 17 extracted by combining osmotic dehydration extraction and molecular compression dehydration extraction methods. Specifically, i) a UV-irradiated group that did not irradiate UV rays, ii) a UV-irradiated group that did not include okra extract, iii) a UV-irradiated group that was cultured with epigallocatechin gallate (EGCG), iv ) UV irradiation group cultured including about 10 μg/mL of okra extract of Preparation Example 2, v) UV-irradiated group cultured with about 10 μg/mL of okra extract of Preparation Example 8, vi) Okra of Preparation Example 17 UV-irradiated group cultured with about 10 µg/mL of the extract, vii) UV-irradiated group cultured with about 100 µg/mL of the okra extract of Preparation Example 2, viii) About 100 µg/mL of the okra extract of Preparation Example 8 A UV-irradiated group incubated including, and ix) a UV-irradiated group incubated with about 100 μg/mL of the okra extract of Preparation Example 17 was prepared.

Specifically, in the experimental groups, the fibroblast cell line Hs68 was dispensed in a 6-well plate by the number of about 3x10 ⁵ per well, and then cultured for about 24 hours in an incubator at about 37°C and about 5% CO ₂ conditions. Next, after removing the medium and adding DPBS, the remaining cell groups except for the UVB non-irradiated group were irradiated with UVB of about 20 mJ/cm ² . Next, the EGCG or okra extract was added and further cultured for about 24 hours.

After the additional culture, fibroblasts were obtained from each experimental group. From the cells, RNA was isolated using Trizol (RNA iso, DAKARA, Japan), and then RNA was quantified at 260 nm using nanodrop, and cDNA was synthesized in an amplifier using about 2 μg of each RNA. (C1000 Thermal Cycler, Bio-Rad, USA). Finally, a real-time polymerase chain reaction is performed in a real-time PCR machine using a mixture in which the target protein MMP-1 primer and the cyanine dye cybergreen (SYBR Green supermix, Applied Biosystems, USA) are added to the synthesized cDNA. The expression level of the MMP-1 gene was evaluated. The real-time polymerase chain reaction is, after activation of the polymerase at about 94°C for about 5 minutes, the reaction at about 95°C for about 30 seconds, the reaction at about 60°C for about 30 seconds, and the reaction at about 72°C for about 30 seconds. About 40 cycles were carried out under the reaction conditions. In addition, the expression level of the gene was finally analyzed through correction for the β-actin gene.

Table 9 shows the primer sequences used in the real-time polymerase chain reaction to evaluate the expression level of the MMP-1 gene.

Primer order MMP-1 F 5'-CGAATTGCCGACAGAGATGA-3' (SEQ ID NO: 1) R 5'-GTCCCTGAACAGCCCAGTACTT-3' (SEQ ID NO: 2) COL1A1 F 5'-GAGGGCCAAGACGAAGACATC-3' (SEQ ID NO: 3) R 5'-CAGATCACGTCATCGCACAAC-3' (SEQ ID NO: 4) HAS3 F 5'-CTTAAGGGTTGCTTGCTTGC-3' (SEQ ID NO: 5) R 5'-GTTCGTGGGAGATGAAGGAA-3' (SEQ ID NO: 6) TSLP F 5'-GCTATCTGGTGCCCAGGCTAT-3' (SEQ ID NO: 7) R 5'-CGACGCCACAATCCTTGTAAT-3' (SEQ ID NO: 8) β-actin F 5'-GGCCATCTCTTGCTCGAAGT-3' (SEQ ID NO: 9) R 5'-GAGACCTTCAACACCCCAGC-3' (SEQ ID NO: 10)

3 is a graph showing the expression level of collagenase (MMP-1) when fibroblasts are treated with an okra extract of one aspect.

As a result, as shown in FIG. 3 , it was confirmed that the expression level of MMP-1 significantly decreased in the treatment group cultured including the okra extract of Preparation Example 17, whereas the expression level of MMP-1 increased rapidly in the UV irradiation group. These results show that the okra extract extracted by combining the osmotic dehydration extraction and molecular compression dehydration extraction method has a better collagenase (MMP-1) inhibitory effect than the okra extract by a single method, thereby further increasing collagen production, , which means that, due to this, more excellent wrinkle suppression, elasticity improvement, or anti-aging effect can be exhibited.

3.3 Wrinkle Inhibition Effect - Confirmation of COL1A1 Expression

In order to confirm the wrinkle suppression, elasticity improvement, or anti-aging effect of the okra extract of one aspect, the expression level of COL1A1 was evaluated whether the okra extract of one aspect increases the production of collagen closely related to wrinkle formation and aging. It was confirmed by measuring.

An experimental group was prepared to compare the collagen production effect of Preparation Examples 2 and 8 extracted by a single method and Preparation Example 17 extracted by combining osmotic dehydration extraction and molecular compression dehydration extraction method. Specifically, i) fibroblasts without okra extract, ii) fibroblast test group containing epigallocatechin gallate (EGCG), iii) okra extract of Preparation Example 2 containing about 10 μg/mL a fibroblast test group, iv) a fibroblast test group comprising about 10 μg/mL of the okra extract of Preparation Example 8, v) a fibroblast test group comprising about 10 μg/mL of the okra extract of Preparation Example 17, vi) Preparation Example 2 A fibroblast test group containing about 100 µg/mL of the okra extract of vii) a fibroblast test group containing about 100 µg/mL of the okra extract of Preparation Example 8, and ix) about 100 µg/mL of the okra extract of Preparation Example 17 An experimental group containing fibroblasts was prepared.

Specifically, human fibroblasts were inoculated in a 6-well cell culture dish at a number of about 4X10 ⁵ per well, and then cultured for about 24 hours in an incubator at about 37 °C and about 5% CO ₂ conditions. After washing with phosphate buffered saline solution (PBS), the okra extract of Preparation Examples 2, 8, or 17 was treated in DMEM medium without FBS in each well and further cultured for about 24 hours. Next, RNA was isolated from the cells of each experimental group using trizol (RNA iso, DAKARA, Japan), and then RNA was quantified at 260 nm using nanodrop. cDNA was synthesized in an amplifier using about 2 μg of RNA extracted from each experimental group (C1000 Thermal Cycler, Bio-Rad, USA). The expression level of the COL1A1 gene was evaluated by conducting real-time polymerase chain reaction in a real-time PCR machine using a mixture of the synthesized cDNA containing the target protein type 1 collagen, COL1A1, primer, and the cyanine dye, cybergreen. .

The sequence and reaction conditions of the primers were the same as in Example 3.2, and the expression level of the gene was finally analyzed through correction for the β-actin gene.

Figure 4 is a graph showing the expression level of type 1 collagen gene when human fibroblasts are treated with an okra extract of one aspect.

As a result, as shown in FIG. 4 , it was confirmed that the expression of type 1 collagen gene was most increased when human fibroblasts were treated with the okra extract of Preparation Example 17. These results indicate that the okra extract extracted by combining the osmotic dehydration extraction and the molecular compression dehydration extraction method may have a better collagen-generating effect than the okra extract by a single method, and thereby, more excellent anti-wrinkle, improved elasticity, or anti-wrinkle effect. This means that it may exhibit an aging effect.

3.4 Moisturizing effect

In order to confirm the moisturizing effect of the okra extract of one aspect, the expression level of hyaluronic acid synthase (Hyaluronan synthase 3, HAS3) generated in human epidermal cells was measured.

An experimental group was prepared to compare the effect of increasing the expression of hyaluronic acid synthase of Preparation Example 17 extracted by combining Preparation Examples 2 and 8 extracted by a single method and osmotic dehydration extraction and molecular compression dehydration extraction method. Specifically, i) human epidermal cell experimental group without okra extract, ii) human epidermal cell experimental group containing retinoic acid (RA), iii) human epidermal cell containing about 10 μg/mL of okra extract of Preparation Example 2 Experimental group, iv) Human epidermal cell experimental group containing about 10 μg/mL of okra extract of Preparation Example 8, v) Human epidermal cell experimental group containing about 10 μg/mL of Okra extract of Preparation 17, vi) of Preparation 2 Human epidermal cell test group containing about 100 µg/mL of okra extract, vii) human epidermal cell test group containing about 100 µg/mL of okra extract of Preparation Example 8, and ix) about 100 µg/mL of okra extract of Preparation Example 17 A human epidermal cell test group containing

The cells of the test group were aliquoted in a 6 well plate and cultured for about 24 hours. Next, after washing with phosphate buffered saline solution (PBS), each experimental group was added to DMEM medium without FBS and further cultured for about 24 hours. After that, RNA was isolated from the cells of each experimental group using trizol (RNA iso, DAKARA, Japan), and RNA was quantified at 260 nm using nanodrop. Next, cDNA was synthesized in an amplifier using about 2 μg of quantified RNA (C1000 Thermal Cycler, Bio-Rad, USA). Finally, the expression level of HAS3 was evaluated by conducting real-time polymerase chain reaction in a real-time PCR machine using a mixture of HAS3 primer and cyanine dye, cybergreen, added to the synthesized cDNA.

The sequence and reaction conditions of the primers were the same as in Example 3.2, and the expression level of the gene was finally analyzed through correction for the β-actin gene.

5 is a graph showing the HAS3 gene expression level when human epidermal cells are treated with an okra extract of one aspect.

As a result, as shown in FIG. 5 , it was confirmed that the HAS3 expression increase effect was significant in the okra extract-treated experimental group of Preparation Example 17 compared to the untreated group or the okra extract-treated experimental group of Preparation Examples 2 or 8. These results mean that the okra extract extracted by combining the osmotic dehydration extraction and molecular compression dehydration extraction method has a better skin moisturizing effect than the okra extract by a single method.

3.5 Anti-itch effect

In order to confirm the itch inhibitory effect of the okra extract of one aspect, the expression level of Thymic stromal lymphopoietin (TSLP) gene, which is an itch-inducing gene that occurs in human epidermal cells, was measured.

An experimental group was prepared to compare the itch inhibitory effect of Preparation Examples 2 and 8 extracted by a single method and Preparation Example 17 extracted by combining osmotic dehydration extraction and molecular compression dehydration extraction method. Specifically, i) the human epidermal cell test group that did not induce itch and did not include the okra extract, ii) the human epidermal cell test group that caused itching and did not include the okra extract, iii) induced itching, and Preparation Example 2 Human epidermal cell experimental group comprising about 10 μg/mL of okra extract iv) induce itch, human epidermal cell experimental group containing about 10 μg/mL of okra extract of Preparation Example 8, v) induce itch, Preparation Example The human epidermal cell test group containing about 10 μg/mL of the okra extract of 17, vi) induces itch, and the human epidermal cell test group containing about 100 μg/mL of the okra extract of Preparation Example 2, vii) induces itching, Human epidermal cell test group containing about 100 μg/mL of the okra extract of Preparation Example 8, viii) itch, and the human epidermal cell experimental group containing about 100 μg/mL of the okra extract of Preparation Example 17, and ix) itching induced, and an experimental group of human epidermal cells containing dexamethasone (DEX) was prepared.

The experimental group was incubated for about 24 hours. Next, after washing with phosphate buffered saline solution (PBS), and treating each well in DMEM medium without FBS, about 10 ug/ml of Poly I:C and about 10 ng/ml of Recombinant human IL-4 was treated, and it was further cultured for about 4 hours while inducing itching. After that, RNA was isolated from the cells of each experimental group using trizol (RNA iso, DAKARA, Japan), and RNA was quantified at 260 nm using nanodrop. cDNA was synthesized in an amplifier using about 2 μg of quantified RNA from each experimental group (C1000 Thermal Cycler, Bio-Rad, USA). Finally, the expression level of TSLP was evaluated by conducting a real-time polymerase chain reaction in a real-time PCR machine using a mixture of TSLP primer and cyanine dye, cybergreen, added to the synthesized cDNA.

Primer sequences and reaction conditions were the same as in Example 3.2, and the expression level of the gene was finally analyzed through correction for the β-actin gene.

6 is a graph showing the level of itchiness in the human epidermal cells when one aspect of the okra extract is treated.

As a result, as shown in FIG. 6 , it was confirmed that the skin itch inhibitory effect was significantly increased in the okra extract-treated experimental group of Preparation Example 17 compared to the untreated group or the okra extract-treated experimental group of Preparation Examples 2 or 8. These results mean that the okra extract extracted by combining the osmotic dehydration extraction and molecular compression dehydration extraction method is more effective in inhibiting skin itch than the okra extract by a single method.

<110> Intercare <120> Method for extracting okra using process of osmotic pressure dehydration by salt and molecular compression dehydration, and cosmetic composition for anti-aging prepared thereby <130> PN136081KR <160> 10 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> MMP-1 Primer_F <400> 1 cgaattgccg acagagatga 20 <210> 2 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> MMP-1 Primer_R <400> 2 gtccctgaac agcccagtac tt 22 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> COL1A1 Primer_F <400> 3 gagggccaag acgaagacat c 21 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> COL1A1 Primer_R <400> 4 cagatcacgt catcgcacaa c 21 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> HAS3 Primer_F <400> 5 cttaagggtt gcttgcttgc 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> HAS3 Primer_R <400> 6 gttcgtggga gatgaaggaa 20 <210> 7 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TSLP Primer_F <400> 7 gctatctggt gcccaggcta t 21 <210> 8 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> TSLP Primer_R <400> 8 cgacgccaca atccttgtaa t 21 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> B-actin Primer_F <400> 9 ggccatctct tgctcgaagt 20 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> B-actin Primer_R <400> 10 gagaccttca acaccccagc 20

Claims (10)

Extracting the okra using an osmotic dehydration extraction method by mixing okra and salt; and
A method of extracting okra, comprising the step of mixing okra and a polymer material and extracting the okra using a molecular compression dehydration extraction method. The method according to claim 1, wherein the method comprises: extracting using the molecular compression dehydration extraction method after the extraction using the osmotic dehydration extraction method; Or, the method of extracting okra, which proceeds in the order of the extraction using the osmotic dehydration extraction method after the extraction using the molecular compression dehydration extraction method. The method according to claim 1, wherein the polymer material is at least one selected from the group consisting of PEG (polyethylene glycol), gum arabic (arabic gum), dextrin, and arabinogalactan. The method according to claim 1, wherein in the extraction using the osmotic dehydration extraction method, the salt and the okra are mixed in a weight ratio of 5 to 100: 100. The method of extracting okra. The method according to claim 1, wherein in the extraction using the molecular compression dehydration extraction method, the polymer material and the okra are mixed in a weight ratio of 10 to 300: 100. The method of claim 1, wherein the mixing time of the okra and the salt is 18 to 30 hours. The method according to claim 1, wherein the mixing time of the okra and the polymer material is 18 to 30 hours. The method according to claim 1, wherein the method further comprises the step of filtering or centrifuging the extract obtained in the step of extracting using the osmotic dehydration extraction method or extracting using the molecular compression dehydration extraction method, How to extract okra. A cosmetic composition comprising an okra extract prepared by the method of any one of claims 1 to 8. The cosmetic composition according to claim 9, wherein the cosmetic composition exhibits a skin wrinkle improvement effect, a skin elasticity improvement effect, a skin itch improvement effect, or a skin moisturizing improvement effect.

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