KR101500316B1 - Cosmetic composition for improving skin aging and wrinkle comprising extract of Ishige okamurae by n-butanol - Google Patents

Abstract

The present invention provides a method for extracting an effective ingredient for prevention of skin aging and wrinkles from Ishige okamurae . The present invention also provides a cosmetic composition for preventing skin aging and improving wrinkles, which contains an butanol extract of Laurus as an active ingredient and has antioxidative activity and inhibits activity of elastin degrading enzyme and collagenase. The cosmetic composition is expected to inhibit destruction of skin tissue and contribute to maintaining healthy skin.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a cosmetic composition for preventing skin aging and improving wrinkles containing an extract of n-butanol,

The present invention relates to a cosmetic composition containing an extract of Ishige okamurae as an active ingredient and more particularly to a cosmetic composition containing an extract of butanol as an active ingredient to prevent aging of the skin, ≪ / RTI >

The lobsters are sea-dominated arboreous plants, which are Ishige okamurae Yendo and Ishige sinicola .

Ishige okamurae is distributed all over the southern coast, Jeju Island, Japan. Morphological features are like stiff tree branches, dark brown color, dry black, and the inner structure of the body is filamentous, The outer layer is a small cell that is perpendicular to the surface of the body and has heat.

Ishige sinicola is distributed in all parts of the southern coast, Jeju island, Japan and China. Morphological characteristics are leafy, blackish brown, bubble-like part is yellowish brown, dry and black, It lives in a community on the central rock, and is used for food.

In recent years, studies on functional compositions using extracts of such lavandula have been actively conducted. Korean Patent Laid-Open Publication No. 10-2003-0015536 (published on Feb. 25, 2003) discloses a method of adding an extracting solvent to dried and crushed wortlets, obtaining an extract by immersion, , And a whitening cosmetic composition which is excellent in the inhibitory effect of tyrosinase activity containing a ginseng extract obtained by concentrating the ginseng extract of the present invention, and Korean Patent Publication No. 10-0981184 (published on September 10, 2010, A composition for natural antioxidation and a method for producing the same), an extract containing diphlorethohydroxycarmalol, which is a substance exhibiting antioxidative activity from a disc, has excellent antioxidative activities such as radical scavenging activity and cell damaging activity Respectively.

One of the areas of greatest interest in cosmetics is wrinkles associated with aging, reduced elasticity, dryness and lack of gloss. The skin is subject to a variety of changes through aging. The epidermis, the dermis, and the subcutaneous tissue of the skin become thinner and the ECM component, which gives the skin elasticity, changes. Collagen, which accounts for 70 ~ 80% of the extracellular matrix, is abruptly decreased in production with age, and has a close relationship with wrinkle formation. Collagen, elastin, proteoglycan, glucosaminoglycan, laminin, fibronectin, etc., which form the skin connective tissues, are oxidized and lose their function, Loss of elasticity and wrinkles are formed excessively, and the skin is changed into senile skin.

As you get older, the collagen content in the connective tissue of the skin decreases, which is due to degradation of collagen synthesis and promotion of degradation. Therefore, degradation of collagen biosynthesis and promoting collagen degradation are the major causes of wrinkles in the skin. Collagen biosynthesis is regulated by factors involved in transcription and post-translational levels, and collagen degradation is regulated by collagenase (collagenase), which degrades collagen by ultraviolet light Promoting the expression of matrix metalloproteases (MMPs), such as the same, promotes collagen degradation and reduces collagen content. In addition, as the collagen's deformation accelerates due to the external environment, the skin becomes wrinkled and deepened.

As a result, skin aging is caused by non-homogenization of cells, loss of elastin, destruction of collagen, reduction and delay of collagen synthesis, and the like. This occurs in the epidermis of the skin but can be seen as a phenomenon that occurs in the dermis.

Various cosmetic compositions have been studied for solving the problem of skin aging, and the wrinkle improvement effect of the skin is visible in some parts. There have been various reports on retinoids widely used for improvement of skin wrinkles, spots and pigmentation, among which retinol has been widely reported to remove skin wrinkles. Various retinol-containing cosmetics have been reported to be used for skin wrinkles, And the like. U.S. Patent No. 4,603,146 (Jun. 29, 1986) discloses an example of wrinkle improvement using retinol (vitamin A), which is effective in inhibiting collagen degradation due to inhibition of collagenase activity.

In recent years, interest in skin and cosmetics has been rapidly increasing, and research on cosmetic compositions containing natural extracts or containing natural extracts has been actively conducted.

Korean Patent Publication No. 10-2003-0015536, Feb. 25, 2003. Korean Patent Registration No. 10-0981184, Sep. 10, 2010. U.S. Patent No. 4,603,146, July 29, 1986.

The present invention has antioxidative activity and is excellent in inhibiting activity of elastase and collagenase in a solvent extract of a tiger plover, especially butanol extract, And to apply it to a cosmetic composition for improving wrinkles.

Accordingly, it is an object of the present invention to provide a method for efficiently extracting an effective ingredient for preventing skin aging and improving wrinkles, which is excellent in antioxidative activity from a hand.

It is another object of the present invention to provide a cosmetic composition which is effective for preventing skin aging and improving wrinkles, which comprises butanol extract of pa as an active ingredient.

To achieve the above object, there is provided a method for extracting an effective ingredient for preventing skin aging and improving wrinkles from a hand. Preferably, the extract is obtained by removing the n-hexane extract and the chloroform extract from the extract obtained by extracting the dough with methanol, dividing the extract into n-butanol and concentrating it under reduced pressure to obtain an extract of the active ingredient.

In addition, the present invention provides a cosmetic composition for preventing skin aging and improving wrinkles, which contains butanol extract of pa as an active ingredient and has an antioxidative activity and inhibits the activity of elastase and collagenase.

The butanol extract of P obtained by the extraction method of the present invention has an excellent antioxidative activity and an excellent effect of inhibiting the activity of elastase and collagenase. Accordingly, the present invention can provide a cosmetic composition which can prevent aging of the skin and elastically improve the skin to effectively relieve wrinkles.

FIG. 1 shows a flow chart of the separation process of butanol extract inhibiting the activity of elastase and collagenase from Lard.
FIG. 2 shows the TLC pattern of the solvent extract obtained from the disc. Lane 1: Hexane, Lane 2: CHCl ₃ , Lane 3: BuOH, Lane 4: H ₂ O
Figure 3 shows the TLC pattern of the butanol extract obtained from the lard. A) CHCl ₃ : MeOH: H ₂ O = 7: 3: 0.5, B) CHCl ₃ : MeOH = 1: 1

The present invention provides a method for extracting an effective ingredient which is excellent in antioxidative activity from a test piece and inhibits elastase and collagenase activity. Preferably, the n-hexane extract and the chloroform extract are removed from the extract obtained by extracting the plaque with methanol, and the extract is separated into n-butanol and concentrated under reduced pressure to extract the active ingredient.

The present invention provides a cosmetic composition containing as an active ingredient a butanol extract of a pear to prevent skin aging and improve wrinkles.

The cosmetic composition of the present invention contains the butanol extract of the present invention in an amount of 0.1 to 10.0% by weight based on the total weight of the composition. If the content is less than 0.1% by weight, the effect is insufficient. If the content is more than 10.0% by weight, cytotoxicity may be exhibited.

The composition of the present invention is not particularly limited in its formulation depending on the body part to be applied. Specifically, it may be, for example, a cosmetic composition having a formulation of a softening agent, a nutritional lotion, a massage cream, a nutritional cream, an eye cream, a pack, a gel or a skin adhesive type cosmetic composition, And may be a transdermal dosage form such as a spray.

Hereinafter, the present invention will be described in more detail with reference to specific examples. However, it should be understood that the present invention is not limited to the following embodiments, and various modifications and changes may be made without departing from the spirit and scope of the present invention.

Unless otherwise defined in the technical and scientific terms used herein, the meaning is generally understood by those of ordinary skill in the art to which this invention belongs. Repeated descriptions of the same technical constitution and operation as those of the conventional art will be omitted.

(Example 1) Production of solvent extract of Ishige okamurae

The extracts were separated from the extracts using various solvents and concentrated under reduced pressure.

1) First wash the pellet with purified water, dry it, and crush it with a mixer.

2) Add about 1.5 kg of crumbled lysate into 10 L of methanol (MeOH) and shake for 12 to 36 hours at 100 to 150 rpm, preferably for 24 hours at 130 rpm, and then filter to obtain MeOH extract. After the MeOH extraction is repeated two more times, the MeOH solution is dried under reduced pressure.

3) After dissolving MeOH extract in 3L of 80% MeOH, solvent fraction is applied using 3L of n-Hexane. This is repeated 3 times to obtain n-hexane solution, which is then dried under reduced pressure to obtain n-hexane solvent extract .

4) Remove n-Hexane and concentrate again. Make 3L of 30% MeOH solution. Repeat 3 times with 3L chloroform to obtain chloroform solvent extract.

5) After removing the chloroform, concentrate the remaining solution again and distribute to 2L of n-butanol and distilled water. Separate each solution and concentrate under reduced pressure to obtain an extract.

Through the above process, various solvent extracts as shown in [Table 1] were obtained and used in future experiments. FIG. 1 is a schematic diagram of the extraction process.

Table 1. Solvent extracts obtained through the extraction process of this Example

(Example 2) The TLC pattern of the solvent extract obtained from the beaker

As shown in Table 1, in Example 1, various solvents were used to obtain extracts from the tiles. The TLC pattern on the silica gel of the extract was examined.

First, TLC was performed on a silica gel mixed with CHCl ₃ : MeOH = 1: 1 as a mobile phase on TLC patterns of n-Hexane, CHCl ₃ , BuOH and Water extracts.

Figure 2 shows the TLC pattern on silica gel of various solvent extracts of tars. Each of the solvent extracts was well separated without overlapping components. It can be seen that the active ingredients of the paddle were well extracted and separated by solvent extraction.

(Example 3) Antioxidant activity of the solvent extract obtained from Lard

Antioxidant activity was determined by the Blois method to detect DPPH radical scavenging activity. Specific experimental methods are as follows.

First, 1200 μL of DPPH stock solution was added to 2 mg of DPPH (2,2-diphenyl-1-picrylhydrazyl) and 15 mL of ethanol was added to the solution. Then, 500 μL of DMSO and 3000 μL of ethanol were mixed and finally DPPH solution . Place 450 μL of the prepared DPPH solution in a quartz cuvette and measure the initial absorbance (B) at 518 nm.

At this time, the initial absorbance should be adjusted so as to fall within the range of 0.94 to 0.97. If the absorbance deviates from 0.94 to 0.97, that is, when the initial absorbance is 0.97 or more, ethanol is added to dilute the solution. If the absorbance is lower than 0.94, an additional DPPH solution is added to obtain an initial absorbance value of 0.94 to 0.97 Respectively. When the initial absorbance is properly adjusted, 50 μL of the sample dissolved in methanol is added to the cuvette containing the DPPH solution adjusted for absorbance, and the sample is measured at 518 nm for 10 minutes at intervals of 10 seconds. The antioxidant activity at this time is denoted as reduction (%) and is determined by the following method.

Antioxidant activity (ΔA) = B (initial absorbance) - C (absorbance of sample solution)

Where B is the initial absorbance of 0.94 to 0.97 and C is the lowest absorbance measured for 10 minutes. The ΔA thus obtained is calculated again by reduction (%) by the following equation.

Reduction (%) = (A / B) x 100

The antioxidative activity of various solvent extracts obtained from Lard was measured at a concentration of 0.1 mg / ml and the results are shown in Table 2 below. As shown in Table 2, the butanol extract showed the highest antioxidant activity.

Table 2. Antioxidant activity of solvent extracts obtained from L.

(Example 4) Elastase and Collagenase Inhibitory Activity of Solvent Extracts Derived from Lard

In order to confirm the effect of various solvent extracts obtained from the extracts on the prevention of skin aging and wrinkles, the ability to inhibit the activity of enzymes degrading elastin or collagen was measured.

1. Elastase activity inhibitory activity measurement

EnzCheck Elastase assay kit (Invitrogen E12056) was used to measure elastinase activity, and DQTM elastin (from bovine neck ligament, flourescein conjugate) was used as a substrate. That is, 110 μl of reaction buffer, 20 μl of sample, 50 μl of elastase (1 units / ml) and 30 μl of elastin (100 μg / ml) were added to a 96-well plate and mixed well at 37 ° C. for 1 hour And the results were measured at 505/515 nm using a fluorescence microplate reader. N-Methoxysuccinyl-Ala-Ala-Pro-Val-chloromethylketone was used as a positive control.

2. Measurement of inhibitory activity of collagenase activity

EnzCheck Gelatinase / Collagenase assay kit (Invitrogen E12055) was used to measure collagenolytic enzyme activity, and DQTM collagen type 1 (from bovine skin, flourescein conjugate) was used as a substrate. To the 96-well plate, 150 μl of reaction buffer, 20 μl of sample, 20 μl of collagenase (10 units / ml) and 10 μl of collagen type 1 (100 μg / ml) Lt; / RTI > Experimental results were measured at 495/515 nm using a fluorescence microplate reader. As a positive control, 1,10-phenanthroline and monohydrate were used.

The inhibitory activities of elastase and collagenase, enzymes related to aging of skin, were measured on various solvent extracts obtained from Lard, and the results are shown in Table 3 below. As shown in Table 3, the n-butanol extract shows a high inhibitory activity against both elastase and collagenase.

Table 3. Elastase and Collagenase Inhibitory Activity of Solvent Extracts Derived from Lard

(Example 5) TLC pattern of n-butanol extract

As shown in [Table 2] and [Table 3], among the various solvent extracts obtained from Lard, extracts extracted with n-butanol exhibited the best antioxidative activity and enzymes such as elastase and collagenase . Therefore, the TLC pattern of the n-butanol extract obtained from the truffle was investigated on silica gel.

(B) mixed with CHCl ₃ : MeOH: H ₂ O = 7: 3: 0.5 mixed solvent (A) and CHCl ₃ : MeOH = 1: 1 as a mobile phase on silica gel to examine the TLC pattern of n-butanol extract, Were used to perform TLC, respectively.

Figure 3 shows the TLC pattern on silica gel of the n-butanol extract of Lys. It can be seen that the n-butanol extract in the A and B mobile phase solvents contains almost no other impurities in the form of a mixture of two components.

The butanol extract obtained from the tongue by the extraction method of the present invention is excellent in antioxidant activity and exhibits an effect of inhibiting elastin degrading enzyme and collagen degrading enzyme. Therefore, it is possible to prevent aging of the skin which contributes to maintenance of healthy skin by inhibiting destruction of skin tissue. And is contained as an effective ingredient in a cosmetic composition for improving wrinkles.

The cosmetic composition of the present invention contains the butanol extract of the present invention in an amount of 0.1 to 10.0% by weight based on the total weight of the composition. The cosmetic composition is not particularly limited in its formulation depending on the body part to be applied. For example, it may be a cosmetic composition having a formulation of a softening agent, a nutritional lotion, a massage cream, a nutritional cream, an eye cream, a pack, a gel or a skin adhesive type cosmetic composition and may also contain a lotion, ointment, The same transdermal dosage form can also be used.

Claims (7)

Ishige okamurae is washed with purified water, dried and pulverized, extracted with methanol at 100 to 150 rpm for 12 to 36 hours, and the methanol residue obtained by further extracting the remaining residue with methanol twice is dried under reduced pressure to prepare a methanol extract step; Dissolving the methanol extract in 80% methanol, performing a solvent fraction using n-hexane, and repeating this three times to remove n-hexane extract; Removing the n-hexane extract, concentrating the remaining solution, dissolving in 30% methanol, performing the solvent fraction using the same amount of chloroform, repeating this three times to remove the chloroform extract; And removing the chloroform, concentrating the remaining solution, and distributing the same to an equal amount of n-butanol and distilled water, separating each solution, and concentrating under reduced pressure to prepare an n-butanol extract. -Butanol < / RTI >
Ishige okamurae is washed with purified water, dried and pulverized, extracted with methanol at 100 to 150 rpm for 12 to 36 hours, and the methanol residue obtained by further extracting the remaining residue with methanol twice is dried under reduced pressure to prepare a methanol extract step; Dissolving the methanol extract in 80% methanol, performing a solvent fraction using n-hexane, and repeating this three times to remove n-hexane extract; Removing the n-hexane extract, concentrating the remaining solution, dissolving in 30% methanol, performing the solvent fraction using the same amount of chloroform, repeating this three times to remove the chloroform extract; And removing the chloroform to concentrate the remaining solution, and then distributing the same to an equal amount of n-butanol and distilled water, separating each solution, and concentrating under reduced pressure to prepare an n-butanol extract. Butanol extract as an active ingredient.
3. The method of manufacturing a cosmetic composition according to claim 2, wherein the composition has an antioxidative activity.
3. The method for producing a cosmetic composition according to claim 2, wherein the composition inhibits the activity of elastin degrading enzyme and collagenase.
[Claim 3] The method according to claim 2, wherein the composition contains an n-butanol extract prepared from tallow in an amount of 0.1-10.0 wt% based on the total weight of the composition.
6. The method for producing a cosmetic composition according to any one of claims 2 to 5, wherein the composition is used for skin aging prevention and wrinkle improvement.
The composition according to claim 6, wherein the composition has any one of the formulations selected from the group consisting of softening agents, nutritional lotion, massage cream, nutritional cream, eye cream, lotion, ointment, pack, gel, patch, Gt;

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