KR20130094043A - A cosmetic composition comprising subcritical water extracts of laminaria japonica - Google Patents

Abstract

PURPOSE: Subcritical water extracts of laminaria japonica is provided to improve an esterase inhibition activity, a collagen synthesis capability and an anti-wrinkle activity and prevent side effects by using marine algae and an aqueous solvent, thereby having a marked effect in an industrial aspect. CONSTITUTION: A cosmetic composition for wrinkle protection or wrinkle improvement includes the subcritical water extracts of laminaria japonica which are extracted with the subcritical water of 3 MPa and 180-240°C. The cosmetic composition for wrinkle protection or wrinkle improvement adds water in crushed laminaria japonica and includes the subcritical water extracts of laminaria japonica which are characterized by being extracted under 3 MPa and 180-240°C as an active ingredient. A manufacturing method of the subcritical water extracts of laminaria japonica having a wrinkle protection activity and a wrinkle improvement activity comprises: a step of manufacturing crushed laminaria japonica by pulverizing laminaria japonica; a step of adding water in the crushed laminaria japonica; and a step of manufacturing the subcritical water extract of laminaria japonica by reacting the crushed laminaria japonica with water at a condition of 3 MPa and 180-240°C.

Description

TECHNICAL FIELD [0001] The present invention relates to a cosmetic composition containing an extract of kelp starch as an active ingredient,

The present invention relates to a cosmetic composition comprising a natural substance having a skin care function as an active ingredient.

Human skin is the largest organ that makes up the human body that protects the various organs inside the skin from the outside. It is a barrier function, temperature control function, and excretion function that prevents the outflow of moisture and useful components from outside the skin. And various physiological roles such as respiratory function.

However, with age, the skin's physiological function decreases and the skin's thickness becomes thin, which not only reduces its function as a skin barrier, but also internal factors such as external factors such as ultraviolet rays, air pollution, heating and cooling facilities, and hormonal changes according to excessive fatigue, stress, and age. Drying of the skin is caused by a factor, and such drying of the skin also causes the skin to lose its physiological function, causing skin irritation and damage to occur easily even by a small external stimulus.

In general, the growth stage can easily respond to various aging factors from the inside and outside by the skin's various defense systems, but as the age increases, the antioxidant system is destroyed by active oxygen species generated by ultraviolet rays, etc. The defense is significantly reduced. As described above, when oxidative stress is applied to the skin when defense ability is lowered, lipid peroxidation, protein oxidation, chain cutting and abnormal crosslinking of collagen and elastin, elastic fibers, cleavage of hyaluronic acid chain, DNA oxidation, etc. It results in damage to biological components. Due to the damage of the biological component, the initial symptoms of the skin wrinkles, such as sagging, roughening, dryness of the skin is caused, resulting in the formation of skin wrinkles.

In the past, cosmetics, which were previously thought to be used by women, are used not only as a beauty expression tool but also as household goods for men and women of all ages, and as the industry develops, the demand for high-quality and diversification of cosmetics increases. As a result, there is a growing interest in functional products that expect special effects or efficacy.

The functional cosmetics currently recognized in Korea include whitening products that help whitening the skin, wrinkle improvement products that help improve the wrinkles of the skin, and sunscreen products that help to burn the skin finely or protect the skin from ultraviolet rays. .

In general, such functional cosmetics are interpreted as cosmetics that affect the structure or function of the body as well as cosmetic effects, and prevent, improve and treat. In particular, recently, as the sales proportion of functional cosmetics in the total sales of cosmetics significantly increased, researches to find new efficacy ingredients on the human body and use them effectively on the skin have been actively conducted.

In particular, with the recent increase in the elderly population due to the aging of the population and the improvement of the quality of life due to the economic development, there is an increasing demand for products that can suppress or control the wrinkles of the skin, which were naturally associated with aging. . In particular, due to the increase in social activity opportunities of the elderly population, the interest in beauty of the elderly population is increasing, while the factors for stimulating skin and promoting aging, such as ultraviolet rays, are increasing. The demand for and use of functional cosmetics is increasing significantly.

In relation to the wrinkle improvement of the skin, the cause of the loss of the elasticity of the skin is loose, that is, the cause of wrinkles formed in the skin appearance can be divided into the generation of wrinkles by natural aging and the generation of wrinkles by UV exposure (photoaging). As can be seen, the mechanism of wrinkle formation due to two causes is not yet accurately identified. Skin aging, which is closely related to the wrinkles, can also be classified into two types, including natural aging, ie, physiological aging over time and exposure to ultraviolet light, including wind, temperature, humidity, tobacco smoke, and pollution. There is extrinsic aging due to financial factors.

Common causes of these two wrinkles, namely, skin changes due to natural aging and photoaging, are the reduction and alteration of collagen and elastin, the major constituent proteins in the dermis, and the reduction of hyaluronic acid, the substrate of the dermis. It is a decrease in elasticity. In particular, according to the research results so far, it is known that the reduction and modification of the main proteins collagen and elastin in the dermis are directly involved in the generation of wrinkles and the deterioration of skin elasticity.

The collagen is a major component of the extracellular matrix and is present in extracellular epilepsy as a major substrate protein produced in skin fibroblasts. It is also an important protein, accounting for about 30% of the total weight of biological proteins, and has a solid triple helix structure. Collagen forms most of the organic material of the skin, tendons, bones and teeth, especially in the dermis of bone and skin. In most other structures, it exists as a fibrous inclusion.

The main function of the collagen is to impart strength and tension to the skin to secure the mechanical strength of the skin, to induce connective tissue resistance, to bind tissue, to support cell adhesion, to divide and differentiate cells (when organic growth or wound healing). It is known as induction. In addition, collagen plays an important role in wound healing, and it is known that the wound can be quickly recovered without scar by promoting the synthesis of collagen in the damaged epithelium.

In particular, the reduction of collagen, which accounts for 90% of the dermis, is reported to be closely related to aging of the skin. More specifically, the synthesis and degradation of extracellular matrix such as collagen in vivo is properly regulated, but the progress of aging or UV irradiation promotes the expression of matrix metalloproteinase (MMP), an enzyme that degrades collagen. In addition, due to the collagen breakdown by the collagenase, the elasticity of the skin is lowered and wrinkles are formed.

In addition, there are various kinds of skin aging inhibitors known to date depending on the mechanism of skin aging inhibition. Representative examples include collagen production promoters, elastin activators, hyaluronic acid production increase agents, free radical scavengers, cell proliferators, photoaging inhibitors, keratinocyte proliferation inhibitors, humectants, harmful active oxygen production inhibitors, cell regeneration materials and the like.

In addition, a method of using an external composition for skin such as cosmetic or ointment containing collagen as a method for improving skin wrinkles has been proposed. This is suggested to take advantage of the wrinkle improvement and skin recovery effect of collagen, but it is difficult to absorb the percutaneous absorption of collagen, which is a high molecular material, can not expect a moisturizing effect or wound healing effect, so it is not possible to exhibit the essential skin function and wound healing function Pointed out.

It is proposed to solve the conventional problems, a method of preventing the formation of wrinkles by antioxidants such as vitamin C, vitamin E or carotenoids that inhibit the formation or reaction of free radicals, increasing collagen biosynthesis in the dermis such as retinol The method, the method of injecting collagen into the skin, or the method of injecting botulinum toxin into the skin has been proposed.

However, the vitamin C, vitamin E, carotenoids and retinol all have the disadvantage of being easily broken at room temperature, the retinol is known to have a disadvantage that the side effects on the skin when the treatment concentration is increased. In addition, collagen or botulinum toxin has been pointed out that there is a limit to the amount of use or the effect is insignificant due to safety problems such as irritation and redness when applying skin, so that the effect of improving skin function or wound healing cannot be expected.

In relation to the raw materials of functional cosmetics, recent researches related to functional materials are focused on natural materials derived from natural products mainly in conjunction with environmentally friendly hot air, and mainly on herbs or herbal medicines used as medicines. Research is ongoing.

In addition, with respect to such natural materials, studies on materials used mainly as herbs and herbal medicines have been conducted, and studies on seaweeds have been conducted only in limited areas in some areas.

Therefore, in consideration of the geographical environment of Korea, which is rich in marine resources, the necessity of research on the materials of functional cosmetics, especially wrinkle prevention and improvement effect from marine resources including seaweeds, is increasing.

In this regard, the necessity and urgency of research and development using marine resources of Korea including the marine resources of Jeollanam-do region is further increased to utilize abundant seaweed resources in Jeollanamdo region of Korea to develop new materials, .

KR 0750870 B KR 0879283 B KR 20010010368 A KR 20030076829 A KR 20120000251 A

The present invention is to improve the problems of the prior art as described above, an object of the present invention to provide a cosmetic composition comprising a natural substance as an active ingredient.

In order to achieve the above object, the present invention provides a cosmetic composition comprising an extract of Kelpia tincture as an active ingredient. Specifically, the kelp kidney extract extract of the present invention may be a kelp kid extract extract obtained by extracting 3 MPa and 180 DEG C to 240 DEG C, and more specifically, the kelp extract extract at 3 MPa and 200 DEG C To 220 < 0 > C. More specifically, the kelp kidnote extract may be extracted with an aside factor of 3 MPa and 210 < 0 > C.

In addition, the present invention relates to a method for producing kelp ( Laminaria japonica ), and extracting the mixture under the conditions of 3 MPa and 180 ° C to 240 ° C. The cosmetic composition may be a cosmetic composition for preventing or improving wrinkles containing the tincture extract factor as an active ingredient.

In addition, the present invention relates to a method for producing an extract of kelp starch having the highest anti-wrinkle activity from kelp and the highest extraction yield.

Specifically, the method for preparing the kelp starch extract comprises the steps of: grinding kelp ( Laminaria japonica ) to prepare a tuna crush; Adding water to the kelp pulverized product; And a step of reacting the kelp pulverized product and water at 3 MPa and a temperature of 180 ° C to 240 ° C to prepare a kelp kidney extract extract, wherein the kelp extract has a wrinkle preventing or improving ability.

More specifically, the amount of water added to the kelp crushed product is 500 parts by weight to 1,500 parts by weight based on 100 parts by weight of the crushed margarine product. The reaction conditions for preparing the kelp craving factor extract are 3 MPa and 200 to 220 Lt; 0 > C.

More specifically, the step of preparing the tuna crushing product is performed by drying the crushed tuna ( Laminaria japonica ), followed by pulverization. The amount of water to be added to the crushed tangle may be 900 weight% To 1, 100 parts by weight, and the step of preparing the kelp starch extract may be carried out at 3 MPa and 210 캜 for 1 minute.

The present inventors have been studying marine resources in Jeollanamdo region where researches have been limited, unlike the land plants which have been variously studied in the past. Among them, kelp, one of the abundant marine resources in Korea, The inventors of the present invention have confirmed that the functionality can be improved compared to the conventional solvent extraction method and further studies have been conducted to find that the efficacy associated with wrinkle improvement is the most excellent wrinkle improving activity under the specific conditions, that is, 3 MPa and 210 캜, Starch inhibitory activity and collagen aggregation ability, and the activity is lowered at the temperature lower or higher than the above-described temperature, and the extraction yield is also the highest. Thus, the present invention has been completed.

In the present invention, the cosmetic composition is a composition used in the human body to clean and beautify the human body to add charm and brighten the appearance, or to maintain or promote the health of skin and hair. Detergents and cosmetics (cosmetics) The concept includes.

In the present invention, the active ingredient means a component of the cosmetic composition applied to the skin condition to provide a prevention, improvement or supplementary effect of skin wrinkles.

In the present invention, the supercritical fluid is when the material is at a critical temperature where the two states of liquid and gas are indistinguishable from each other and the vapor pressure at this time, i.e., at a temperature and pressure higher than its critical point. A fluid that is in a state above the supercritical point. In addition, in the present invention, the sub-supercritical fluid refers to a fluid having almost the same characteristics as the supercritical fluid. In addition, subcritical extraction means the extraction method using a subcritical fluid.

The subcritical fluid may be water, carbon dioxide (CO ₂ ), ethylene, ethane, propane, acetylene or ammonia, preferably economical, free of harmful substances, safe to dispose of and environmentally friendly, Water which confirms the condition capable of maximizing the extraction efficiency, more preferably, the submergence coefficient. The subcount means water of subcritical condition.

Hereinafter, the present invention will be described in more detail.

The present invention relates to a cosmetic composition comprising an extract of tallow starch as an active ingredient

The kelp ( Laminaria japonica) is a kelp (Laminaria japonica) is a marine algae belonging to galjo plants statement (Phaeophyta) take crude (Phaeophyceae) kelp (Laminariales) dasimagwa (Laminariaceae), are classified in appearance to the leaf top, stem, root, into phyllodes blowing strip The central part of the foliage forms a thick middle part, and the middle part has a vertical flaw. The stem is usually circumferential and upright, and the root represents the dendritic shape branched from the bottom of the stem.

The distribution area of the kelp is the Pacific coast of Korea, Japan or the Kamchatka peninsula, and has been widely used for edible purposes in Korea, Japan, and China since ancient times.

The kelp crab index extract was prepared from the sea tangle ( Laminaria japonica ) < / RTI >

In addition, the kelp kidnote extract may be used at sub-critical conditions of 1 MPa to 5 MPa and 180 ° C to 240 ° C or 2 MPa to 4 MPa and 200 ° C to 220 ° C or 3 MPa and 200 ° C to 210 ° C, To 210 < 0 > C for 1 minute.

In addition, the kelp starch extract may be obtained by adding water to the sea tangle powder, and extracting it under the conditions of 3 MPa and 180 ° C to 240 ° C.

The kelp limulus extract may be extracted with an asymptotic modulus of 3 MPa and 200 ° C to 220 ° C, or with an asymptotic modulus of 3 MPa and 210 ° C.

The condition of the subcritical water extraction method for preparing the subcritical water extract may be 1 MPa to 5 MPa or 2 MPa to 4 MPa or 2.5 MPa to 3.5 MPa or 3 MPa, the temperature of the subcritical water May be 180 ° C to 240 ° C or 190 ° C to 230 ° C or 200 ° C to 220 ° C or 210 ° C.

The subcritical extraction method for preparing the subcritical water extract may be performed for 5 seconds to 10 minutes, or 10 seconds to 5 minutes, or 30 seconds to 3 minutes or 45 seconds to 2 minutes or 1 minute.

In particular, the extract of the invention has an excellent elastase inhibitory activity and collagen synthesis ability as compared with the solvent extract, and particularly has excellent elastase inhibitory activity at a pressure of 3 MPa and at 150 DEG C or 270 DEG C at 210 DEG C, Collagen synthesis ability was confirmed.

The subcritical water extract may optionally add a filtration process, a concentration process or a drying process after the subcritical extraction, the filtration process may be performed using filter paper, the concentration process may be carried out through a reduced pressure concentration, etc. The drying process may be performed by a method such as hot air drying or freeze drying.

As described above, when inhibiting elastase and inducing collagen synthesis, it was confirmed that the effect of preventing or improving wrinkles was excellent, and as a result, it was confirmed through experiments that the cytotoxicity was lower than other extraction conditions, The cosmetic composition of the present invention can be applied to skin requiring a wrinkle-improving activity because seaweed, a natural substance, that is, a substance made of sea tangle and water is an effective ingredient and has no side effects and is safe

Therefore, the cosmetic composition of the present invention can be a cosmetic composition for improving wrinkles, a cosmetic composition for preventing or improving wrinkles, or a cosmetic composition for anti-wrinkles.

The cosmetic composition may contain 0.0001% by weight to 50% by weight, preferably 0.001% by weight to 30% by weight, of the kelp kidney extract extract on the skin. That is, the content of the tallow starch extract should be at least a minimum content level in order to achieve skin-related functionality, specifically, a wrinkle-preventing effect. However, considering the possibility of application to various formulations, Can be freely adjusted within a range. At this time, the content of the active ingredient and the content of the additional component may be appropriately adjusted within the above range according to the content of the components contained in the formulation or cosmetic composition.

The cosmetic composition of the present invention may be provided for use in skin external ointments, basic cosmetics, makeup cosmetics, body cosmetics or shaving cosmetics, and the like, and examples of the basic cosmetics include essences, lotions, creams, gels, lotions, packs, massages, and the like. There may be creams, latexes, and the like, and examples of the makeup cosmetics may include foundation, lipstick, eyeshadow, eyeliner, mascara, eyebrow pencil, makeup base, and the like, and body cosmetics include soap, liquid cleaner, and bathing agent. , Sun screen cream, sun oil, and the like, and shaving cosmetics include aftershave and shaving cream.

The cosmetic composition of the present invention may contain, in addition to the kelp extract as an active ingredient, a functional substance having a wrinkle-improving effect or a functional substance having another function specifically, a whitening effect, a UV blocking or moisturizing effect, A diluent, and the like may be further included.

For example, the cosmetic composition may be additionally added to the perfume, pigments, fungicides, antioxidants, preservatives, moisturizers, thickeners, inorganic salts or synthetic polymer materials to improve physical properties. In addition to the blending components that may be added, it may be a fat or oil component, an emollient agent, a surfactant, an organic pigment, an inorganic pigment, an organic powder, an ultraviolet absorber, a pH adjuster, an alcohol, a blood circulation accelerator, a cooling agent, a limiting agent, or purified water.

In addition, the cosmetic composition may further comprise one or more selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids and seaweed extract.

The compounding component which may be added other than the said active ingredient is not limited to this, Any said component can be mix | blended within the range which does not impair the objective and effect of this invention.

In addition, the cosmetic composition may take the form of a solution, an emulsion, a viscous mixture, and the like, and any formulation commonly prepared in the art, for example, emulsion, cream, lotion, pack, foundation, lotion, beauty liquid, hair cosmetic, etc. It may be a formulation.

More specifically, the cosmetic composition of the present invention is a skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisturizing lotion, nutrition lotion, massage cream, nutrition cream, moisturizing cream, hand cream, foundation, essence, nutrition essence And formulations of packs, soaps, cleansing foams, cleansing lotions, cleansing creams, body lotions and body cleansers.

For example, when the formulation of the present invention is a paste, cream or gel, the carrier component is animal fiber, vegetable fiber, wax, paraffin, starch, trakant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide. This can be used.

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 the carrier component, and especially when the formulation of the present invention is a spray, additionally chlorofluoro Propellants such as rohydrocarbon, propane / butane or dimethyl ether.

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

When the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolenic derivatives or ethoxylated glycerol fatty acid esters.

3 MPa and 180 ° C to 240 ° C, or 3 MPa and 200 ° C to 220 ° C, or 3 MPa, as compared to kelp extract extracts obtained by other subcritical extraction conditions or kelp solvent extracts prepared by other extraction methods, The sea tangle extracted with an index of 3 MPa and 210 DEG C ( Laminaria japonica ) extract factor exhibits an excellent effect of inhibiting elastase inhibition activity and collagen synthesis and is safe from side effects since it is a substance derived from a natural substance and thus can be used as an effective ingredient of cosmetics.

In this respect, the present invention may relate to a method for producing wrinkle-improving activity, that is, wrinkle-preventing or improving ability of kelp extract.

The extract prepared by the process of the present invention has the best anti-wrinkle activity, and the production method of the present invention has the advantage that the extraction yield is the highest as compared with other extraction methods or other extraction conditions.

Specifically, the method for preparing the kelp starch extract comprises the steps of: grinding kelp ( Laminaria japonica ) to prepare a tuna crush; Adding water to the kelp pulverized product; And a step of reacting the kelp pulverized product and water at 3 MPa and a temperature of 180 ° C to 240 ° C to prepare a kelp kidney extract extract, wherein the kelp extract has a wrinkle preventing or improving ability.

The step of preparing the crushed tangle may be performed by using a conventional seaweed crusher, and further drying of the tangle may be performed prior to the crushed product. In this regard, the step of preparing the tuna corn crushed material may be performed by a method including drying the kelp and pulverizing the dried kelp.

The kelp drying can be carried out by conventional methods such as a seaweed drying method, a natural drying method, a hot air drying method, a freeze drying method, and the like. The natural drying method may be carried out at a temperature of 20 ° C to 30 ° C and a ventilating condition .

In the step of adding water to the tumbled ground water, the amount of water to be added may be 500 parts by weight to 1,500 parts by weight, or 700 parts by weight to 1300 parts by weight or 900 parts by weight to 1100 parts by weight, based on 100 parts by weight of the sea tangle ground powder.

The water may be general water or distilled water, and the distilled water may be primary distilled water or secondary distilled water or tertiary distilled water.

The step of preparing the kelp starch extract may be carried out by reacting the kelp powder and water at a constant temperature and constant pressure for a predetermined time.

More specifically, the pressure condition In detail, the pressure condition of the subcritical extraction may be 1 MPa to 5 MPa or 2 MPa to 4 MPa or 2.5 MPa to 3.5 MPa or 3 MPa, the temperature conditions The temperature conditions of the critical extraction can be 180 ° C to 240 ° C or 190 ° C to 230 ° C or 200 ° C to 220 ° C or 210 ° C. In addition, the reaction time, specifically the time to perform the subcritical extraction method for preparing the subcritical water extract is 5 seconds to 10 minutes, or 10 seconds to 5 minutes, or 30 seconds to 3 minutes or 45 seconds to 2 minutes or It may be one minute.

Specifically, the reaction conditions of the step of preparing the kelp starch extract may be performed at 3 MPa and 200 ° C to 220 ° C or 3 MPa and 210 ° C for 1 minute.

In the subcritical extraction, when a carrier is required, the carrier may be nitrogen gas. More specifically, the subcritical extraction method may use subcritical water as the subcritical fluid and nitrogen gas as the carrier.

In the case of the subcritical hydrothermal extraction method, not only solves the problem that excessive costs and processes are required for the removal of the organic solvent of the conventional extraction method and may cause environmental problems, and the extraction is performed under the optimum conditions of the present invention. In the case of performing, especially at the conditions of 3 MPa and 210 ℃ of the optimum conditions, the improved yield compared to the conventional solvent extraction method, specifically the increase in extraction efficiency of about 1,300% to 200% was confirmed, significantly improved It was evaluated that a yield can be obtained.

As described above, the method of producing the sea tangle extract having anti-wrinkle activity using the sub-coagulation of the present invention has safety and eco-friendliness as compared with the conventional method, and can be performed in a relatively simple process, The wrinkle activity is improved and the yield is also remarkably increased.

The kelp extract of the present invention can improve the esterase inhibitory activity and the collagen aggregation performance, that is, the anti-wrinkle activity, as well as the extraction yield, according to the conditions of the subcritical extraction, It is possible to have remarkably improved anti-wrinkle activity and yield as compared with the extraction method used. By using seaweed used as food and water solvent which is the safest solvent, safety against side effects is guaranteed, and as a result, And can be manufactured by a simple method using seaweeds produced abundantly in the west coast of Korea. Therefore, the cosmetic composition of the present invention can be economically advantageous and can contribute to the revitalization of the local economy and the income of the local farmers' In terms of the effect, it will be very large.

FIG. 1 is a graph showing collagen aggregation performance according to the kind of each extract according to an embodiment of the present invention, wherein the abscissa represents the temperature during the subcritical extraction condition, and the letter represents the type of solvent or the comparative group, ascorbic acid And the vertical axis indicates collagen aggregation performance relative to the control group.

Hereinafter, examples of the present invention will be described. The following examples are only for illustrating the present invention and the scope of the present invention is not limited by the following examples. However, the following Preparation Examples and Examples are merely illustrative examples for the purpose of facilitating understanding of the present invention, and can be modified into various other forms, and the scope of the present invention is not limited to the following Examples.

< Example  1> Preparation of kelp extract

1-1. Kelp Subcriticality  Preparation of extract

Seaweeds, that is, seaweeds, were prepared by crushing kelp collected from Jangheung area, Jeollanam - do with naturally dried kelp for 3 days under the temperature condition of 20 ℃ ~ 30 ℃ and ventilating condition.

The kelp crassiforme extract was prepared by adding 20 g of the above tangle crushed product and 200 mL of third distilled water into a subcritical extraction device and adjusting the temperature to 90 ° C., 150 ° C., 210 ° C. and 270 ° C. under a pressure of 3 MPa, The reaction was carried out under the conditions of pressure and temperature for 1 min. The prepared extract was filtered under reduced pressure using a paper filter paper (Whatman, No. 2), concentrated using a rotary vacuum evaporator after the filtration, and dried to dry using a freeze dryer after concentration. To prepare an extract. The final extract was stored at -20 ℃ was used in the experiment.

The kelp solvent extract was prepared by solvent extraction using water, methanol and ethanol. Specifically, the kelp hot-water extract was prepared by immersing 20 g of the above tangle kernels in 200 mL of tertiary distilled water and then extracting at 100 DEG C for 15 minutes. The prepared extract was filtered under reduced pressure using a paper filter paper (Whatman, No. 2), concentrated using a rotary vacuum evaporator after the filtration, and dried to dry using a freeze dryer after concentration. To prepare an extract. The final extract was stored at -20 ℃ was used in the experiment.

The extract of the kelp solvent using the organic solvent was prepared by dipping 20 g of the above kelp crushed product in 200 mL of methanol or ethanol for 24 hours. The prepared extract was filtered under reduced pressure using a paper filter paper (Whatman, No. 2), concentrated using a rotary vacuum evaporator after the filtration, and dried to dry using a freeze dryer after concentration. To prepare an extract. The final extract was stored at -20 ℃ was used in the experiment.

The extraction yield of the extract prepared by each of the above methods was confirmed by comparing the weight of the final extract obtained with respect to the first used tuna sample. The results are shown in Table 1 below.

Extract Type Subcritical Extraction (3M Pa) Solvent extraction 90 ° C 150 ℃ 210 ℃ 270 ℃ Heat number Methanol ethanol Extraction yield (%) 46.93 51.59 65.67 59.60 33.53 5.28 15.32

As shown in Table 1, it was confirmed that the extraction yield is relatively higher than the solvent extraction method by the subcritical extraction method. In the subcritical extraction method, the extraction yield was increased with increasing temperature up to 210 ℃, but the extraction yield decreased at 270 ℃ and 210 ℃ was most suitable. Particularly, it was confirmed that the subcritical extraction at 210 ° C improved the yield by about 200% or more and the yield was improved by about 1,200% as compared with the solvent extraction method using methanol, compared with the hot water extraction method in which the extraction yield was the best in the solvent extraction method.

Therefore, in terms of extraction yield, it was confirmed that it is most preferable to prepare the extract by performing subcritical extraction under the conditions of 3 MPa and 210 ° C.

< Example  2> Extraction method and extraction conditions of kelp extract Anti-wrinkle activity  Confirm

The kelp extract prepared in Example 1 was confirmed to have anti-wrinkle activity according to extraction method, extraction solvent and extraction conditions, and further cytotoxicity was confirmed.

2-1. Cytotoxicity measurement

In order to measure the cytotoxicity of the sea tangle extract prepared in Example 1, normal human dermal fibroblast, specifically, CCD-1064 Sk (normal human dermal fibroblast, ATCC CRL-2076, USA) MTT colometric assay. The CCD-1064Sk cells were cultured in Iscove's Modified Dulbecco's Medium (IMDM) containing 10% fetal bovine serum (FBS) at 37 ° C and 5% CO ₂ . Cell line) was used after 3 to 7 times subculture after initial thawing.

First, CCD-1064Sk was dispensed into a 96-well plate at a concentration of 1 × 10 ⁵ cells / mL at 200 μL / well, and cultured at 37 ° C and 5% CO ₂ for 24 hours. The final concentrations of each of the seaweed extracts prepared in Example 1 were adjusted to 0.1 mg / ml, 1 mg / ml and 2 mg / ml, respectively, and cultured for 24 hours. After the cultivation, 20 μL of MTT preparation was added to the culture solution at a concentration of 2 mg / mL and regenerated for 4 hours to form formazan. The formazan formed on the bottom of the 96-well plate was dissolved by adding 200 μL of dimethylsulfoxide (DMSO, Sigma, USA), centrifuged at 5000 rpm for 1 minute, and absorbance was measured at 540 nm using an ELISA reader. Relative toxicity was measured by the method of calculating the cell survival rate when the absorbance of the control was 100%, and the results are shown in Table 2 below.

sample Cell survival rate (%) Name of sample Extraction condition Sample concentration (mg / mL) 0.1 One 2 Subcritical extraction
(3M Pa) 90 ° C 97.72 ± 0.3 94.42 ± 1.8 91.93 ± 0.2 150 ℃ 105.64 ± 0.8 95.71 ± 1.2 92.99 ± 0.3 210 ℃ 99.32 ± 1.6 94.24 0.8 101.54 ± 0.8 270 ℃ 96.85 ± 2.1 94.33 + - 0.6 94.35 ± 0.5 Solvent extraction Heat number 97.723 ± 0.8 96.24 ± 0.5 96.31 + - 1.4 Methanol 103.54 ± 0.8 95.63 ± 0.7 99.56 + - 0.4 ethanol 98.45 ± 0.8 96.23 ± 0.7 108.87 ± 0.3

As shown in Table 2, all of the kelp extracts showed almost no toxicity, and in particular, when treated with 2 mg / mL, the kelp extract and kelp extract extracts which had undergone subcritical extraction at 3 MPa and 210 캜 and ethanol The cytotoxicity of the organic solvent seaweed extract using the solvent was found to be lowest.

2-2. Elastase  Through inhibitory effect Anti-wrinkle activity  Measure

The elastase inhibitory activity of the kelp extract prepared in Example 1 was measured in order to measure the anti-wrinkle activity of the kelp kidney extract extract through inhibitory activity against elastase known to be one of the causes of skin wrinkles.

More specifically, experiments were performed using elastase and substrate N-succinyl-l- (Ala) 3-ρ-nitroanilide derived from pigs. First, 60 μL of 50 mM Tri-HCl buffer (pH 8.0) was added to each well of a 96-well plate. Then, 10 μL of 10 mM N-succinyl-1- (Ala) 3-ρ-nitroanilide and 10 μL of each of sea tangle extract and distilled water (10 μL) was added to each well, pre-incubated at 37 ° C. for 5 minutes, and then 10 μL of the elastase prepared at a concentration of 1 unit / mL was added to each well. After 15 minutes at 37 ° C. Lt; / RTI &gt; After the reaction, the inhibitory activity was measured by measuring the absorbance at 415 nm. The IC ₅₀ values determined from the above-mentioned measurements are shown in Table 3 below. As a positive control, Ursolic acid, well known as an elastase inhibitor, was used. As a result of confirming the above uricolic acid by the same method, it was confirmed that the IC ₅₀ value was 12.9 ug / mL.

Extract Type Subcritical Extraction (3M Pa) Solvent extraction 90 ° C 150 ℃ 210 ℃ 270 ℃ Heat number Methanol ethanol Elastase inhibitory activity
(IC ₅₀ (mg / mL)) 46.93 51.59 65.67 59.60 33.53 5.28 15.32

As shown in Table 3, the inhibitory activity against elastase was found to be superior to the subcritical extraction method compared to the solvent extraction method. In addition, in relation to the subcritical extraction method, the extraction yield increased with increasing temperature up to 210 ° C., but the extraction yield was decreased at 270 ° C., so that the temperature of 210 ° C. was most suitable.

Therefore, it has been confirmed that it is most preferable to perform the subcritical extraction at the above-mentioned 3 MPa and 210 캜 in order to produce the extract, in relation to the anti-wrinkle activity through the elastase inhibitory activity.

Table 4 shows the results of measuring the amount of kelp powder used for the preparation of the extract on the basis of the subcritical extract under the conditions of 3 MPa and 210 캜 at which the elastase inhibitory activity was the most excellent.

Extract Type Subcritical Extraction (3M Pa) Solvent extraction 90 ° C 150 ℃ 210 ℃ 270 ℃ Heat number Methanol ethanol Kelp dry powder usage (mg) 2.94 1.53 1.00 1.48 4.94 16.42 6.53

As shown in Table 3, in order to exhibit the elastase inhibitory activity when the subcritical extract was prepared under the conditions of 3 MPa and 210 DEG C using 1 mg of the kelp dried powder sample, the sample required for each extract Is 1.48 mg to 2.94 mg in the case of the subcritical extraction method and 4.94 mg to 16.42 mg in the case of the solvent extraction method and that the subcritical extract is produced at the above-mentioned 3 MPa and 210 캜 under the conditions of the most economical Respectively.

From the above results, it was confirmed that the subcritical extraction was most preferably performed under the conditions of 3 MPa and 210 ° C in terms of the extraction yield and the cost, that is, the requirement of using the tallow sample as a raw material.

2-3. Collagen Generation ability  through Anti-wrinkle activity  Measure

In order to confirm the anti-wrinkle activity of the prepared sea tangle extract prepared in Example 1, the collagen production ability was confirmed by measuring the content of collagen released from the cells in a normal human dermal fibroblast.

Specifically, Sirius red dye binding assay was performed using the CCD-1064Sk (normal human dermal fibroblast, ATCC CRL-2079, USA). First, the CCD-1064Sk was dispensed into a 96-well plate at a concentration of 2 × 10 ⁵ cells / mL in an amount of 200 μL / well and cultured for 24 hours at 37 ° C and 5% CO ₂ . 1 was cultured at 37 ° C and 5% CO ₂ for 24 hours. The cells were treated with different concentrations of cadmium iodine extract at concentrations not showing cytotoxicity.

To 100 μL of the medium, 1 mL of 50 μM sirius red solution (0.5 M acetic acid) was added, followed by reaction at room temperature for 20 minutes. After the reaction, centrifugation was carried out at 5000 x g for 20 minutes, and then 200 μL of 0.5 M NaOH was added to the precipitate, and the collagen was eluted for 5 minutes. The eluted collagen dye was transferred to a 96-well plate, and the absorbance value at 540 nm was measured to determine the total collagen content released from the cell. The results are shown in FIG. Ascorbic acid was used as a positive control.

As shown in Fig. 1, the collagen aggregation performance was confirmed to be comparatively improved when compared with the solvent extraction method by the subcritical extraction method. In the subcritical extraction method, the extraction yield was increased with increasing temperature up to 210 ℃, but the extraction yield decreased at 270 ℃ and 210 ℃ was most suitable. Particularly, in the case of 210 ° C, an improvement effect of collagen production ability was confirmed to be about 15% compared to 150 ° C and about 14% compared to 270 ° C. In order to produce an extract having anti-wrinkle effect, The conditions were confirmed to be 210 占 폚.

Therefore, it was confirmed that, in terms of anti-wrinkle effect including collagen production ability, subcritical extraction was performed for 1 minute under the conditions of 3 MPa and 210 캜 to produce an extract, which is the most preferable method for producing sea tangle extract.

Claims (7)

Wrinkle prevention or improvement cosmetic composition comprising a kelp ( Laminaria japonica ) subcritical water extract extracted with a subcritical water of 3 MPa and 180 ℃ to 240 ℃ as an active ingredient. The method of claim 1,
The kelp subcritical water extract is a cosmetic composition for preventing or improving wrinkles that is extracted with a subcritical water of 3 MPa and 200 ℃ to 220 ℃. The method of claim 2,
The kelp subcritical water extract is a cosmetic composition for preventing or improving wrinkles that are extracted with subcritical water of 3 MPa and 210 ℃. Kelp ( Laminaria) japonica ) Wrinkle prevention or improvement cosmetic composition comprising the kelp subliminal water extract as an active ingredient, characterized in that the water is added to the pulverized product and extracted under conditions of 3 MPa and 180 ℃ to 240 ℃. Kelp ( Laminaria) japonica ) to prepare a kelp crushed powder; Adding water to the kelp pulverized product; And preparing kelp sublime extract by reacting the kelp pulverized product and water at 3 MPa and 180 ° C. to 240 ° C.
Wherein the kelp extract has a wrinkle preventing or improving ability. The method of claim 5,
The amount of water added to the kelp crushed powder is 500 parts by weight to 1,500 parts by weight based on 100 parts by weight of the kelp crushed, the reaction conditions of preparing the kelp sublime water extract is a condition of 3 MPa and 200 ℃ to 220 ℃ Kelp subcritical water extract manufacturing method having the ability to prevent or improve wrinkles. The method according to claim 6,
The step of preparing the kelp pulverization is the kelp ( Laminaria japonica ) is dried, and then pulverized, and the amount of water added to the kelp pulverized product is 900 parts by weight to 1100 parts by weight based on 100 parts by weight of kelp pulverized, and the step of preparing the kelp sublime water extract is Method for preparing kelp subcritical water extract having anti-wrinkle or improvement ability to be performed for 1 minute under conditions of 3 MPa and 210 ° C.

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