KR20160114980A

KR20160114980A - Cosmetic composition comprising protein-polysaccharide extracted from linum usitatissimum seed, protein-polysaccharide extracted from salvia hispanica seed and polyglutamic acid, and method of preparing the same

Info

Publication number: KR20160114980A
Application number: KR1020150041558A
Authority: KR
Inventors: 이근하; 한정희; 이지언; 이정민; 권순상
Original assignee: (주)모아캠
Priority date: 2015-03-25
Filing date: 2015-03-25
Publication date: 2016-10-06

Abstract

The present invention provides a cosmetic composition comprising protein-polysaccharide extracted from Linum usitatissimum seeds, protein-polysaccharide extracted from Salvia hispanica seeds, and polyglutamic acid as active ingredients, and to a producing method thereof. The cosmetic composition has an excellent skin moisturizing effect, an anti-aging effect, and a skin wrinkle inhibiting effect.

Description

FIELD OF THE INVENTION The present invention relates to a cosmetic composition containing a protein polysaccharide extracted from flaxseed, a protein polysaccharide extracted from a tooth, and polyglutamic acid as an active ingredient, and a method for producing the same. BACKGROUND ART ACID, AND METHOD OF PREPARING THE SAME}

The present invention relates to a cosmetic composition containing as an active ingredient a protein polysaccharide extracted from flaxseed, a protein polysaccharide extracted from tooth root, and polyglutamic acid, and a method for producing the same.

Flaxseed is the annual plant ahmagwa pool that is widely distributed from the Mediterranean to South Asia, India's Flax (Linum usitatissimum ) and long oval-shaped seeds. It has long been used in Australia as a traditional medicinal product for diseases such as respiratory, cold and cough, and has a lot of mucus in the cell wall. It contains a large amount of lignan, a dietary fiber and a plant estrogen component. In addition, it contains omega-3 fatty acid, linoleic acid and oleic acid, and is also used for edible and ointment.

Chi is a seed of Salvia hispanica which is originated from Mexico and is rich in nutrients such as magnesium, potassium, iron and calcium, and is rich in dietary fiber.

Nutrient-rich flaxseed and toothseed are used in a relatively diverse range for edible purposes, but various studies on the effect of skin on cosmetics have not been made in earnest.

Korean Patent Laid-Open Publication No. 2014-0131701 (published Nov. 14, 2014)

The present invention provides a cosmetic composition having excellent skin moisturizing effect, anti-aging effect, skin wrinkle inhibiting effect, skin wrinkle reducing effect, anti-inflammatory effect and antibacterial effect.

The present invention provides a cosmetic composition containing, as an active ingredient, a protein polysaccharide extracted from flaxseed, a protein polysaccharide extracted from tooth root, and polyglutamic acid.

(A) separating the protein polysaccharide layer from flaxseed and filtering the protein polysaccharide layer separated from flaxseed to obtain a protein polysaccharide extracted from flaxseed;

(B) separating the protein polysaccharide layer from the tooth tissue and filtering the protein polysaccharide layer separated from the tooth tissue to obtain a protein polysaccharide extracted from the tooth tissue;

(C) mixing the protein polysaccharide extracted from flaxseed with the protein polysaccharide extracted from toothseal; And

(D) adding polyglutamic acid to the protein polysaccharide extracted from mixed flaxseed and the proteinaceous polysaccharide extracted from toothseal.

The present invention provides a cosmetic composition having excellent skin moisturizing effect, anti-aging effect, skin wrinkle inhibiting effect, skin wrinkle improving effect, anti-inflammatory effect and antibacterial effect.

FIG. 1 shows the measurement result of the moisturizing effect according to the content ratio of Experimental Example 3. FIG.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is well known and commonly used in the art.

In the present invention, the linseed is of annual plant ahmagwa pool that is widely distributed from the Mediterranean to the West Asia, India in Flax (Linum usitatissimum ) seeds.

In one embodiment of the present invention, the content of the protein polysaccharide extracted from the flaxseed is 0.1 to 20% by weight based on the weight of the cosmetic composition.

In one embodiment of the present invention, the protein polysaccharide extracted from flaxseed can be any one or more than one selected from the group consisting of rhamnose, galactose, xylose, glucuronic acid, glucose, fucose, mannose and arabinose .

The main constituent sugar of the protein polysaccharide extracted from flaxseed may be mannose represented by the following formula (1) or glucose represented by the following formula (2).

[Chemical Formula 1]

(2)

In the present invention, tooth seed refers to seeds of Lamiaceae and Salvia hispanica .

It is rich in omega-3, which is antioxidant, and is rich in dietary fiber. It is rich in nutrients such as magnesium, potassium, iron, and calcium. Especially, it has a high mucus content due to the seed properties that form natural gel, and these components have a property of absorbing water more than 10 times the tooth volume.

In one embodiment of the present invention, the protein polysaccharide extracted from tooth root is selected from the group consisting of galactose, glucuronic acid, rhamnose, xylose, glucose, fucose, mannose, arabinose, &Lt; / RTI >

The major constituent sugar of the protein polysaccharide extracted from flaxseed can be galactose represented by the following formula (3) or glucuronic acid represented by the following formula (4).

(3)

[Chemical Formula 4]

In one embodiment of the present invention, the content of the protein polysaccharide extracted from the tooth root is 0.1 to 20% by weight based on the weight of the cosmetic composition.

In the present invention, the polyglutamic acid means a polymer in which D-glutamic acid and L-glutamic acid are glutamyl bonded, and specifically may be polygamma glutamic acid, and more specifically polygamargamic acid represented by the following formula .

[Chemical Formula 5]

In the poly (gamma) glutamic acid represented by Formula 5, n in Formula 5 may be 780 to 15,740.

In one embodiment of the present invention, the molecular weight of the poly-gamma glutamic acid is 1,000,000 to 2,000,000 Da.

The polyglutamic acid may be a mucous substance, and may be chemically synthesized by polymerizing poly-α-glutamic acid, N-carboxylic acid anhydride of glutamic acid γ-benzyl ester and then dehydrogenating it with hydrogenated hydrogen, or fermenting food such as rice straw Can be obtained by using a strain of Bacillus sp. Isolated from Korean traditional soybean fermented food, such as Chungkukjang, Japanese traditional soybean fermented food, Natto, and traditional soybean fermented food, Kinema.

In one embodiment of the present invention, the polyglutamic acid is 0.1 to 20% by weight based on the weight of the cosmetic composition.

In one embodiment of the present invention, the cosmetic composition is for skin moisturizing.

In the present invention, skin moisturization refers to the ability to prevent water loss and to supply moisture to the skin, which allows the skin to stay in the skin for a long time.

The cosmetic composition according to the present invention can significantly increase skin moisturizing activity as compared with the case where flaxseed, toothseed and polyglutamic acid are used alone.

In one embodiment of the present invention, the cosmetic composition is anti-aging.

The anti-aging cosmetic composition may be specifically for antioxidant.

In one embodiment of the present invention, the cosmetic composition is for suppressing skin wrinkles or improving skin wrinkles.

In one embodiment of the present invention, the cosmetic composition is anti-inflammatory.

In one embodiment of the present invention, the cosmetic composition is for antibacterial use.

The method for producing a cosmetic composition of the present invention comprises (a) separating a protein polysaccharide layer from flaxseed (hereinafter referred to as step (a)).

In one embodiment of the present invention, the step (a) further comprises a pretreatment step of adding flaxseed to distilled water and stirring the flaxseed.

The distilled water may be specifically the third distilled water.

In one embodiment of the present invention, the step (a) separates the protein polysaccharide layer from the amylase using a filter material having a mesh unit of 50 to 100. When the mesh unit of the filter body is less than 50, there is a problem that the impurities are mixed. When the mesh unit is more than 100, the filter efficiency is lowered in terms of filtration refining ability because the pore becomes smaller.

In the present specification, a mesh unit means the number of filtration holes entering a length of one inch.

In one embodiment of the present invention, the step of obtaining the protein polysaccharide extracted from flaxseed is carried out under a pressure of 10 to 100 hPa. When the pressure is less than 10 hPa, impurities are mixed. When the pressure is more than 100 hPa, the separation speed of the polysaccharide decreases due to the reduced pressure.

(B) obtaining a protein polysaccharide extracted from flaxseed (hereinafter referred to as step (b)) by filtering a protein polysaccharide layer separated from flaxseed.

In one embodiment of the present invention, step (b) comprises obtaining a protein polysaccharide extracted from flaxseed from a protein polysaccharide layer separated from flaxseed using a filter press.

In the present invention, a filter press means a process of pressurizing a raw liquid into a closed filter chamber and separating the solid and the liquid through the filter body.

It is not easy to separate impurities having a large particle size mixed with the polysaccharide by using a filter because of high viscosity. However, in the case where the step (b) is a protein polysaccharide using a filter press, Compressing and transporting the stock solution to the filter plate, the polysaccharide can be easily separated from the impurities in a short time. The feed pump may be a positive pressure type.

In one embodiment of the present invention, the filter press uses a filter body having 200 to 300 mesh units. When the mesh unit of the filter press is less than 200, there is a problem that the impurities are mixed. When the mesh unit is more than 300, the pore becomes smaller and the filter efficiency is lowered in terms of the filtration speed.

In one embodiment of the present invention, the filter press is performed under a pressure of 4.5 to 7 kgf / cm ² . When the pressure of the filter press is less than 4.5 kgf / cm ² , there is a problem that impurities are mixed, and when the pressure exceeds 7 kgf / cm ² , impurities due to excessive feed pressure applied to the filter plate are not filtered And a problem of leakage occurs.

The method for producing a cosmetic composition of the present invention comprises (c) separating a protein polysaccharide layer from a tooth tooth (hereinafter referred to as step (c)).

The above step (c) can be applied to the separation of the protein polysaccharide layer of step (a) described above, except that tooth seed is used instead of flaxseed.

The method for producing a cosmetic composition of the present invention comprises the steps of (d) obtaining a protein polysaccharide extracted from tooth root by filtering a protein polysaccharide layer separated from tooth root (hereinafter referred to as step (d)).

The above step (d) can be applied to obtaining the protein polysaccharide of step (b) described above, except that the protein polysaccharide layer separated from toothseal is used instead of the protein polysaccharide layer separated from flaxseed.

The method for producing a cosmetic composition of the present invention comprises (e) mixing a protein polysaccharide extracted from flaxseed with a protein polysaccharide extracted from a toothseal (hereinafter referred to as step (e)).

In one embodiment of the present invention, the step (e) is carried out after the steps (a) to (d) are carried out or simultaneously with the steps (a) to (d).

In one embodiment of the present invention, the mixing ratio of the protein polysaccharide extracted from the flaxseed extract to the protein polysaccharide extracted from the toothseal is 1: 1 to 2: 1 by weight, preferably 1: 1.

The method for producing a cosmetic composition of the present invention comprises the step of (f) adding polyglutamic acid to a protein polysaccharide extracted from a mixed polysaccharide and a proteinaceous polysaccharide extracted from a tooth.

In one embodiment of the present invention, the step (f) further comprises a step of pretreating flaxseed obtained by adding distilled water to polyglutamic acid and stirring.

In one embodiment of the present invention, the mixing ratio of the protein polysaccharide: polyglutamic acid extracted from the flaxseed is 4: 1 to 4: 2 by weight, preferably 2: 1.

In one embodiment of the present invention, the mixing ratio of the protein polysaccharide: polyglutamic acid extracted from the tooth root is 4: 1 to 4: 2 by weight, preferably 2: 1.

In the method for producing a cosmetic composition according to the present invention, the description of the above-mentioned cosmetic composition of the present invention can be applied unless otherwise specified.

The composition of the present invention may be formulated as an external preparation for skin, in particular as a cosmetic composition, and may be formulated containing a cosmetically or dermatologically acceptable medium or base. In addition, the composition of the present invention may be provided in any form suitable for topical application, for example, as a solution, an emulsion obtained by dispersing an oil phase in water phase, an emulsion obtained by dispersing water phase in water phase, a suspension, a solid, a gel, Pastes, foams, or aerosol compositions. Compositions of such formulations may be prepared according to conventional methods in the art.

In addition, the composition according to the present invention may contain, in addition to the above-mentioned substances, other ingredients which can give a synergistic effect to the main effect, to the extent that the main effect is not impaired. In addition, the composition according to the present invention may further comprise a moisturizing agent, an emollient agent, an ultraviolet absorber, an antiseptic, a bactericide, an antioxidant, a pH adjuster, an organic and inorganic pigment, a fragrance, a cold agent or a limiting agent. The compounding amount of the above components can be easily selected by those skilled in the art within a range not to impair the objects and effects of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

< Reference example 1>

1,000 mL of third distilled water was added to 1 to 5 wt% of flax seed oil and 10 to 50 g of water and the resulting mixture was stirred for 2 hours at 400 to 800 rpm at 25 ° C using a propeller mixer (BL3000, Heidon, Japan) , Primix, Model 2.5) at 2000 to 5000 rpm at 20 to 30 ° C for 30 minutes to separate the shell and the protein polysaccharide layer. Then, the inner seeds and the protein polysaccharide layers were separated under a reduced pressure of 10-100 hPa using a 50-100 mesh unit of the dispersion in which the inner flax seed layer and the protein polysaccharide layer were separated. The separated solution was filtered using a filter press equipped with a filter body of 200 mesh or more to remove impurities and to obtain a gel containing flaxseed protein polysaccharide. The thus obtained product was referred to as Comparative Example 1 and used in the Experimental Example.

< Reference example 2>

1,000 mL of tertiary distilled water was added to 1 to 3 wt% of tooth seed and 10 to 30 g of water, and the mixture was stirred for 2 hours using a propeller mixer (BL3000, Heidon, Japan) at 400 to 800 rpm at 20 to 30 DEG C, The shell and the protein polysaccharide layer were separated by dispersing using a dispersing machine (TKHomodisper, Primix, Model 2.5) at 2,500 rpm and 20 to 30 ° C for 30 minutes. Thereafter, the inner seed layer and the protein polysaccharide layer were separated from the inner tooth layer and the protein polysaccharide layer under a reduced pressure of 10 to 100 hPa using a 50-100 mesh unit filter. The separated solution was filtered using a filter press equipped with a filter body of 200 mesh or more to remove impurities and to obtain a gel containing flaxseed protein polysaccharide. The thus obtained product was referred to as Comparative Example 2 and used in the Experimental Example.

< Reference example 3>

1,000 mL of tertiary distilled water was added to polyglutamic acid (trade name: r-poly glutamic acid, manufactured by Vedan Co., Ltd., 1 to 5% by weight and 10 to 50 g) and 100 to 500 and dispersed and dissolved for 1 hour at 20 to 30 DEG C. The thus prepared product was referred to as Comparative Example 3 and used in the Experimental Example.

< Example 1>

(1,000 mL) was added to 1 to 10 wt% of flaxseed, 10 to 100 g, and 1 to 10 wt% of tooth seed and 10 to 100 g of the mixture. The mixture was then stirred for 2 hours using a propeller mixer (BL3000, Heidon, Japan) , And the mixture was firstly stirred at 20 to 30 ° C and then dispersed at 2000 to 5000 rpm at 25 ° C for 30 minutes using a dispersing machine (TKHomodisper, Primix, Model 2.5) to separate the shell and the protein polysaccharide layer. Thereafter, the inner seed layer and the protein polysaccharide layer were separated from the inner seed layer and the protein polysaccharide layer under a reduced pressure using a 50-100 mesh unit filter. The thus separated liquid was filtered using a filter press equipped with a filter body of 200 mesh or more to remove impurities and to obtain a gel containing the seed protein polysaccharide.

1,000 mL of third distilled water was added to 10 to 50 g of polyglutamic acid and dispersed and dissolved in a disperser (TKHomodisper, Primix, Model 2.5) at 200 to 500 rpm at 25 ° C for 1 hour, .

< Experimental Example 1> From flaxseed Extracted protein polysaccharide and From Mr. Chi Analysis of components of extracted protein polysaccharide

i) Molecular weight analysis

The molecular weights of the protein polysaccharides extracted from flaxseed and the protein polysaccharides extracted from the teeth were analyzed as follows.

Gel permeation chromatography was performed on the samples obtained in Reference Examples 1 and 2, respectively. (ELSD 3300, Alltech, USA) using a pL AquaGel OH-30 (7.5X 300 mm, VARIAN, USA) column and high pressure liquid chromatography (Agilent Technologies 1200 Series HPLC / GPC, Agilent, ) Was used to detect ionized water in mobile phase, a column temperature of 25 캜, a flow rate of 0.7 ml per minute, a detector temperature of 50 캜 and a nitrogen gas flow rate of 1.4 ℓ / min. PEG (PolyEthyleneGlycol) The molecular weight distribution of the protein polysaccharide extracted from flaxseed and the molecular weight distribution of the protein polysaccharide extracted from the tooth root according to Reference Example 2 according to Reference Example 1 were confirmed by preparing a calibration curve of the molecular weight according to the delay time in the column of the sample, Are shown in Table 1 below.

sample Calibration curve Delay time (min) Mn (Da) Reference Example 1 Y = -0.2322x + 7.0908 5.043 / 11.987 1,277,800 / 71,186 Reference Example 2 Y = -0.2322x + 7.0908 5.051 / 11.725 1,276,600 / 83,288

ii) Content of protein components

The protein contents of the protein polysaccharides extracted from flaxseed and the protein polysaccharides extracted from the teeth were analyzed as follows.

BCA (bicinchoninic acid) was measured for the samples obtained in Reference Examples 1 and 2, respectively. BCA assay uses the property that protein can reduce copper ions (Cu ² ⁺ , Cu ⁺ ). Copper ions reduced by protein react with BCA solution to produce violet compound (Cu ⁺ / BCA chromophore) This violet compound shows the maximum absorbance at a specific wavelength of 562 nm. Therefore, the amount of protein was measured using the increase in absorbance at 562 nm as the amount of protein increased and the amount of the purple compound increased. The experimental method is as follows. First, 150 μl of each sample was added to each well of 96 microplate using a micro BCA protein measurement reagent kit (Pierce, Cat. No. 23227), and then the samples obtained in Referential Examples 1 and 2 were added to each well. Then, BCA (bicinchoninic acid) kit WR Working Reagent reagent was added and mixed vigorously for 30 seconds. Then, the reaction was carried out at 37 ° C for 2 hours, and a 540 nm absorbance was measured with a microplate reader (UVT-06685, Thermo max, USA) to prepare a calibration curve and quantify the protein. The results are shown in Table 2.

sample Protein content (mg / g) Reference Example 1 77 mg / g Reference Example 2 28 mg / g

As shown in Table 2, the protein content of the protein polysaccharide extracted from flaxseed according to Reference Example 1 was found to be 77 mg / g, and the protein content of the protein polysaccharide extracted from tooth root according to Reference Example 2 was found to be 28 mg / g.

iii) Type and content of sugar ingredient

The analysis of the sugar components of the protein polysaccharides extracted from flaxseed and the protein polysaccharides extracted from the teeth were carried out in the following manner.

2.5 mg of each of the samples obtained in Reference Examples 1 and 2 were dissolved in 1 ml of tertiary distilled water and hydrolyzed with TFA (trifluoroacetic acid). The mixture was then derivatized using PMP (1-phenyl-3-methyl-5-pyrazolone) and purified by Shim-pack (6.0x150mm, CLC-ODS) and high pressure liquid chromatography (Agilent Technologies 1200 Series HPLC, Agilent, USA) at UV 245 nm and 1 mM mixture (mannose, glucuronic acid, The sugar content of the protein polysaccharide extracted from flaxseed extract according to Reference Example 1 and the amount of the protein polysaccharide extracted from tooth root according to Reference Example 2 were measured by using a calibration curve After confirming the sugar components, the results are shown in Table 3 below.

sample Mol (%) Mannos Glucuronic acid Rams North Glucose Galactose Xylose Arabinose Fucose Reference Example 1 3.5 18.7 6.6 8.6 7.4 3.6 1.2 3.0 Reference Example 2 1.5 23.6 4.6 5.1 39.5 7.1 0.5 2.9

As a result, it was confirmed that the sugar of the protein polysaccharide extracted from flaxseed according to Reference Example 1 was mainly composed of rhamnose, galactose, xylose, glucuronic acid, glucose, fucose, mannose and arabinose.

It was confirmed that the sugar of the protein polysaccharide extracted from tooth seed according to Reference Example 2 contained mainly galactose and glucuronic acid and also contained a small amount of rhamnose, xylose, glucose, fucose, mannose and arabinose, The polysaccharide sugars were mainly xylose, rhamnose, and galactose, glucuronic acid, glucose, fucose, mannose, and arabinose.

iv) Type and content of sugar ingredient

0.2 g of each of the samples obtained in Reference Examples 1 and 2 was hydrolyzed in 20 ml of 6N HCl at 130 ° C for 24 hours to break the peptide bond of the protein to make a free amino acid. Then, free amino acids were separated by using OPA (o-phthalaldehyde, Agilent) and FMOC (9-fluorenylmethylchloroformate) derivatives to form fluorescence isoindole, followed by high performance liquid chromatography (Agilent Technologies 1200 Series HPLC, Agilent, USA (FLD, Agilent Technologies, USA). Calibration curves were prepared using a standard 250 pM amino acid analysis standard (AAS, Agilent, USA), and the amino acid content of the protein polysaccharide extracted from flaxseed extract according to Reference Example 1 and the amino acid content of the protein polysaccharide extracted from toothseal according to Reference Example 2 The results are shown in Table 4 below.

sample Reference Example 1 Reference Example 2 Aspartic acid 3.23 1.85 Glutamic acid 22.4 3.24 Serine 2.07 0.82 Histidine 0.48 - Glycine 6.42 3.58 Threonine 1.53 0.98 Arginine 6.42 0.91 Alanine 2 2.25 Tyrosine 0.81 0.49 Cysteine 27.6 3.97 Balin 1.99 1.21 Methionine 0.47 - Phenylalanine 1.3 0.59 Isoleucine 1.34 0.62 Leucine 1.8 0.84 Lysine 3.96 - Proline 2.29 - Total amino acid content 86.11 21.35

As a result, the amino acid of the flaxseed polysaccharide was mainly composed of glutamic acid, glycine, arginine, cysteine, and lysine. The total content of 17 kinds was found to be 86.11 mg / g, and the amino acid of the tooth seed protein polysaccharide was mainly cysteine, glutamic acid, Glycine, total 17 kinds and the content was 21.35 mg / g.

< Experimental Example 2> Measurement of moisturizing effect

In order to evaluate the moisturizing effect of the cosmetic composition according to the present invention, skin moisture loss and skin moisture content were measured. Each sample and a control group of hyaluronic acid solution were applied directly to the skin to evaluate changes in water content and moisture content of skin with time under certain conditions. Experimental methods and results were evaluated as follows.

The moisture content of the skin was measured by using a Tewameter (TM300, MPA 6). The moisture content of the skin area (g / m ² h) was measured. Moisture content was measured by using a Coreneometer (CM825, MPA6). Based on the electronic capacitance of the skin surface, the moisture content was measured and the capacitance value was set to an arbitrary capacitance units (AU) value between 0 and 120 Respectively.

Subjects were kept in a constant temperature and humidity condition (relative humidity: 45 ± 2%, temperature: 25 ± 2 ℃) 1 hour before starting the test for 10 persons in their 20s and 40s. Then, the initial moisture content of each sample application site was measured, and then the sample was applied. 20 μL / cm ² was applied to the inside of the left forearm. In order to minimize the error according to the area, each sample was applied with different application areas of samples 1 to 4. Skin loss and skin moisture content were measured according to a certain period of time (before application, after application, 10 minutes, 20 minutes, 30 minutes, 60 minutes, 120 minutes, 180 minutes after application). Each measurement site was repeated 3 times and the average value was calculated for the amount of water loss and skin moisture content.

The samples used in the tests are Comparative Examples 1 to 3 and Example 1, and as a control group, a 1% aqueous solution of hyaluronic acid (product name: sodium hyaluonate, manufactured by China Spechem Co., Ltd.), which is known to have excellent skin moisturizing effect, The results of the evaluation are shown in Table 5 (unit: g / m ² h).

Time elapsed (min) Martial arts Control group Comparative Example 1 Comparative Example 2 Comparative Example 3 Example 1 0 (before coating) 42 43 43 42 41 45 10 (after application) 52 83 84 82 86 90 20 52 76 77 74 76 86 30 46 72 71 69 72 78 60 44 58 57 54 58 65 120 41 47 46 44 45 57 180 40 44 43 40 43 53

* Control group: 1% aqueous solution of hyaluronic acid

As shown in Table 1, the water content of Example 1 increased up to 41% compared with the non-mud bath at 10 to 30 minutes after application, and the moisture content was maintained at about 21% higher than the mud bath after 3 hours . In addition, the comparative group has a skin moisture content of about 17% higher than that of hyaluronic acid, which is known to have excellent moisturizing effect. In addition, it has an excellent skin moisture content as compared with Comparative Examples 1 to 3.

Next, skin moisture loss was measured using a Tewameter under the same experimental conditions. The evaluation results of the amount of water loss of the hard skin are shown in Table 6 (unit: g / m ² h).

Time elapsed (min) Martial arts Control group Comparative Example 1 Comparative Example 2 Comparative Example 3 Example 1 0 (before coating) 12.5 13.5 12.6 13.0 12.6 12.5 10 (after application) 11.3 10.5 9.5 10.6 10.6 7.7 20 10.0 8.0 8.6 9.6 8.2 6.9 30 10.2 8.3 8.7 9.4 8.3 6.3 60 10.1 8.5 8.7 8.9 8.4 6.5 120 10.2 8.7 9.0 9.1 8.6 7.6 180 11.0 8.9 9.1 9.2 9.0 7.9

* Control group: 1% aqueous solution of hyaluronic acid

As shown in Table 6, after 3 hours from the application of the sample 4, the amount of water vapor evaporation in the non-molluscum group was 11 g / m ² h, 8.9 g / m ² h in the control group and 7.9 g / m ² h in the sample 4, (30 min) and a maximum of about 27% (10 min) compared to hyaluronic acid, compared to the control group. Also, after 3 hours, the moisture content of the epidermis is about 28% lower than that of the non-mud bath, and the moisture retention is maintained. In addition, it was found that the water loss of the hard surface was further reduced compared to Comparative Examples 1 to 3.

< Example 2 to 5>

(1,000 mL) was added to 10-100 g of flaxseed and the mixture was stirred for 2 hours using a propeller mixer (BL3000, Heidon, Japan) at 400 to 800 rpm at 20 to 30 ° C, and then dispersed in a disperser (TKHomodisper, Primix, Model 2.5) at 2000 ~ 5000 rpm at 25 ℃ for 30 minutes to separate the shell and the protein polysaccharide layer.

Thereafter, the inner seed layer and the protein polysaccharide layer were separated from the inner seed layer and the protein polysaccharide layer under a reduced pressure using a 50-100 mesh unit filter. The thus separated liquid was filtered using a filter press equipped with a filter body having 200 mesh units or more to obtain a gel containing the protein polysaccharide extracted from flaxseed (hereinafter referred to as Sample 1).

This procedure was repeated for 10 to 100 g of toothseal to obtain a gel (hereinafter referred to as Sample 2) containing the protein polysaccharide extracted from toothseed.

1000 mL of distilled water was added to 10 to 50 g of polyglutamic acid and dispersed and dissolved in a disperser (T. K. Homodisper, Primix, Model 2.5) at 200 to 500 rpm at 25 ° C for 1 hour to obtain Sample 3.

Samples 1 to 3 were mixed in the proportions shown in Table 7 below to obtain samples of Examples 2 to 5.

mix Mixed amount (g) Example 2 Example 3 Example 4 Example 5 Sample 1 40 g 40 g 40 g 40 g Sample 2 10 g 20 g 30 g 40 g Sample 3 50 g 40 g 30 g 20 g

< Experimental Example 3> Measurement of moisturizing effect by content ratio

In order to evaluate the moisturizing effect according to the ingredient content ratio of the cosmetic composition according to the present invention, the weights of the samples obtained in the above-mentioned Examples 2 to 5 were applied to the mandarin shells, respectively, and the weights were measured. The post weight was measured. In addition, as the respective control groups for the samples of Examples 2 to 5, the same weight of orange peel was allowed to stand at room temperature for 24 hours without any treatment, and then weighed.

The results of measurement of the weight change of the tangerine peel for the samples of Examples 2 to 5 were calculated by the following equation (1), and the results are shown in FIG. 1 and Table 8 below.

Example 2 Example 3 Example 4 Example 5 Water loss rate (%) -19 -16 -12 -10

As shown in Table 8, the water loss rate of Example 5, in which the weight ratio of the protein polysaccharide extracted from flaxseed: the protein polysaccharide extracted from tooth root: poly glutamic acid was 4: 4: 2, was much better than that of each of Examples 2 to 4 can confirm.

Therefore, the protein polysaccharide extracted from flaxseed: the mixing ratio of the protein polysaccharide extracted from the tooth root is 1: 1 by weight, the ratio of the protein polysaccharide: polyglutamic acid extracted from flaxseed is 4: 2 by weight, When the mixing ratio of polyglutamic acid is 4: 2 by weight, it can be seen that the moisturizing effect is the most excellent.

< Experimental Example 4> Measurement of antioxidant effect

Free radical scavenging activity was measured using DPPH (2,2-diphenyl-1-picrylhydrazyl) to evaluate the antioxidative ability of the cosmetic composition according to the present invention. Free radicals are considered to be the causative agent of aging, especially skin aging.

DPPH, a reagent used for the free radical scavenging activity of this experiment, is a very stable free radical, a violet compound that exhibits characteristic absorption at 520 nm. This substance is decolorized by proton-radical scavenger among various antioxidant mechanisms, so it has an advantage of being able to easily observe the antioxidant activity visually. Thus, the DPPH solution and the sample dissolved in the solvent were mixed and reacted for 30 minutes. After 30 minutes, the absorbance at 520 nm was measured to measure the FSC50 value, which is the concentration of the sample required to reduce the concentration of DPPH by 50% The free radical scavenging rate was then calculated.

&Quot; (2) "

free radical scavenging rate (%) = {1- absorbance of experiment group-absorbance of blank test solution / absorbance of control group} × 100

The active oxygen scavenging activity was also evaluated using xanthine oxidase. Xanthine (oxygen oxidoreductase) is an enzyme that acts as an oxidizing agent such as hydrogeneperoxide or produces superoxide radicals in the process of producing uric acid with xanthine as substrate.

Xanthine oxidase inhibitor has been used as a therapeutic agent for urethral acidosis because it can reduce the amount of uric acid in the blood without significantly affecting the purine metabolism. Allopuriol (hypoxanthine derivative) alloxanthine, probenecid, colchicine and the like are known as a market antiepileptic agent. Gout is generally known to be caused by the accumulation of uric acid in the xanthine-producing urinary tract by xanthine oxidase in purine metabolites.

In this experiment, 0.2 mL of a solution of xanthine (2 mM) dissolved in 0.1 mL of each sample solution and 0.6 mL of 0.1 M potassium phosphate (pH 7.5) was added, and 0.1 mL of xanthine oxidase (0.2 U / mL) The reaction was terminated by adding 1 mL of 1N HCl, and the amount of uric acid produced in the reaction solution was measured at 292 nm to calculate an active oxygen scavenging rate according to the following equation (3).

&Quot; (3) "

Active oxygen scavenging rate (%) = (1-generation amount of uric acid in the sample addition group / amount of uric acid production in the no-addition group) × 100

sample Free radical scavenging rate IC ₅₀ (%) Active oxygen scavenging rate IC ₅₀ (%) Comparative Example 1 3.65 4.84 Comparative Example 2 4.47 4.65 Comparative Example 3 9.65 6.87 Example 1 1.85 2.09

As shown in Table 9, the mixture of Example 1 exhibited superior antioxidative effects compared to the flaxseed extract, tooth extract, and polyglutamic acid solutions of Comparative Examples 1 to 3, which are the sole materials.

< Experimental Example 5> Ella Stays Active inhibition measure

In order to evaluate the wrinkle formation inhibitory ability of the cosmetic composition according to the present invention, an inhibition of ellastase activity was carried out.

Skin aging, especially wrinkle formation, is thought to be caused mainly by action of active oxygen and destruction of extracellular matrix by matrix metalloproteinases (MMPs: collagenase, elastase, etc.). Therefore, measurement of inhibitory activity of MMPs is very important for inhibiting skin aging. 7.5 μL of the sample solution and 92.5 μL of the buffer solution were added to 0.13 M of Tris-Cl (pH 8.0) and 1.0 mM of N-succinyl- (Ala) 3-p-nitroanilide, For 10 min, 100 μL of ELASTASE solution was added thereto, and the mixture was incubated at 25 ° C. for 10 minutes in a water bath. Absorbance was measured at 410 nm, and the inhibition rate of ELASTASE was calculated according to the following equation (4).

&Quot; (4) "

Elastase inhibition rate (%) = 100 - ((absorbance of sample) / absorbance of control group) × 100)

As a control, 0.12 mg / mL of PMSF (phenylmethylsulfonyl fluoride, Sigma), a specific inhibitor of Elastase, was used.

sample Elastase inhibitory activity IC ₅₀ (%) Comparative Example 1 4.68 Comparative Example 2 8.96 Comparative Example 3 12.65 Example 1 3.64 PMSF 0.12 mg / mL

As shown in Table 10, Example 1, which is a mixture, exhibited significantly better elastase inhibitory activity than the flaxseed extract, tooth seed extract, and polyglutamic acid solutions of Comparative Examples 1 to 3 as sols.

< Experimental Example 6> Measurement of collagen synthesis effect

In order to examine the effect of the cosmetic composition according to the present invention on improving the skin wrinkles, the collagen aggregation performance was measured. Collagen is a constituent component of the skin dermis matrix layer together with the elastin, and the skin slowly ages, and the constituents constituting the matrix layer in the dermis are decomposed to form wrinkles. The collagen is a component of the dermis matrix layer The effect of improving the skin wrinkles can be confirmed. The test method proceeds as follows.

HDF-N cells (ATCC) were seeded at 1.4 × 10 ⁴ cells / well in a 48-well plate and cultured for 24 h in cell culture conditions. Thereafter, the medium was removed, and the starvation state of the cells was maintained for 24 hours. After that, the test substance was diluted in the cell culture medium, FBM (without media supplement) The culture medium was harvested 24 hours later and the procollagen type I c-peptide (PIP) EIA kit (TAKARA, MK101) was used for positive procollagen Were measured. Cells attached to the bottom were washed with PBS, lysed with 1N NaOH, and the total amount of protein was measured to determine the amount of procollagen synthesis per protein. The amount of each procollagen and the amount of protein were calculated according to the calibration curve.

Collagen
Synthesis effect (%) sample Concentration (%) 0 One 2 5 Comparative Example 1 100.00 100.6 105.3 107.2 Comparative Example 2 100.00 94.6 96.8 99.6 Comparative Example 3 100.00 101.5 104.8 105.9 Example 1 100.00 103.8 112.8 125.6 Ascorbic acid (vitamic-c) 100.00 105.6 120.6 132.6

As shown in Table 11 above, Example 1, which is a mixture, exhibited significantly higher collagen synthesis effect than the flaxseed extract, toothseed extract, and polyglutamic acid solution of Comparative Examples 1 to 3 as sols.

< Experimental Example 7> MMP -1 expression inhibition activity measurement

Enzyme immunoassay (ELISA) was performed to measure the concentration of MMP (matrix metalloproteinase) -1 after UV irradiation and sample addition of the cosmetic composition according to the present invention. The experimental method is as follows.

This test was conducted to compare the inhibition of collagenase production in the fibroblasts by the test substance. HDF-N cells (ATCC) were seeded at a density of 1 × 10 4 cells / well in a 24-well plate and cultured for 24 hours under cell culture conditions. After 24 hours, the medium was discarded and washed with DPBS, followed by the addition of 200 μl of DPBS and irradiation with UV-A 5 J / cm 2. Samples were diluted in the medium to an appropriate concentration, and then treated with cells and cultured for 48 hours under cell culture conditions. The positive control group was treated with retinoic acid and the concentration was 10 μM. The amount of collagenase was measured using a human total MMP-1 ELISA kit (R & D system, DY901, USA). The amount of collagenase measured was corrected to the total amount of protein.

sample Treatment concentration (%) MMP-1 inhibitory activity (%) Comparative Example 1 One 16 2 26 Comparative Example 2 One 4 2 9 Comparative Example 3 One 21 2 32 Example 1 One 25 2 42 Retinol 0.1 20

As shown in Table 12, the sample of Example 1, which was a mixture of 1% to 2% of each treatment concentration, was superior to the flaxseed extract, toothseed extract and polyglutamic acid solution of Comparative Examples 1 to 3, 1 inhibitory activity.

< Experimental Example 8> NO inhibitory activity measurement

In order to test the anti-inflammatory effect of the cosmetic composition according to the present invention, NO inhibitory activity was evaluated. Inflammation is the mechanism by which the wound is protected by a protective mechanism when physical stimulation, bacterial infection, or penetration by chemical substances occurs in the wound or tissue and regenerates the damaged area. Excessive inflammation, however, can damage surrounding tissues, worsening the condition of the disease or inducing new diseases. The inhibitory effect of LPS (lipopolysaccharide) - induced cytokine production on the NO inhibitory activity of the samples was measured. The experimental method is as follows.

RAW 264.7 cells (ATCC) were cultured for 18 hours, and then treated with LPS to induce inflammation. At the same time, the amount of NO produced was measured after 24 hours. Compared with LPS alone, Samples 1 to 4 inhibited NO production in a concentration-dependent manner. In particular, Sample 1 inhibited NO production by 80% or more at a treatment concentration of 100 μg / ml. In the case of Sample 1, NO generation inhibitory effect was more than twice as high as that in Samples 2 to 3. On the other hand, no cytotoxicity was observed in the treatment concentrations of all the sample groups

sample Treatment concentration (%) NO inhibitory activity (%) Comparative Example 1 0.5 35 1.0 40 Comparative Example 2 0.5 34 1.0 42 Comparative Example 3 0.5 25 1.0 26 Example 1 0.5 56 1.0 62

As shown in Table 13, Example 1, which was a mixture of 0.5 to 1.0% of each treatment concentration, exhibited superior NO inhibitory activity to the sols of Comparative Examples 1 to 3 than the flaxseed extract, toothseed extract and polyglutamic acid solution .

< Experimental Example 9> Measurement of preservation activity

Experiments were conducted to measure the preservative activity of the cosmetic composition according to the present invention. It used a circular filter paper method (paper disc method) as the test method, using Staphylococcus aureus to Gram-positive bacteria is a strain (Staphylococcusaureus, ATCC 6538P), E. coli gram-negative bacteria (Escherichia coli , ATCC 8739), Pseudomonas aeruginosa, ATCC 9027), a yeast Candida yeast (Candida albicans , ATCC 10231), and Aspergillus niger (ATCC 22343) was used as a fungus .

Approximately 50 μl of each of the cultivated strains was added to a solidified plate medium, and the mixture was dispensed into a plate culture medium and uniformly smoothed using a spreader. About 40 μl of the test sample solution was absorbed into a paper disk having a diameter of 10 mm, the solvent was dried, and the strain was closely adhered to the surface of the smear medium. After culturing for more than 48 hours in an incubator at 25 to 35 ° C., The size of the clear zone formed around the disk was measured to confirm the antibacterial effect. Since microorganisms can not grow near the paper disk, the larger the average diameter value, the better the antimicrobial activity. The test sample solutions were tested as Comparative Examples 1 to 3 and Example 1, and as a result, it was confirmed that Example 1 exhibited excellent antimicrobial activity because the average diameter value was largest (Table 14).

sample Diameter of transparent circle per microbial strain (mm) Average diameter of transparent ring (mm) S. aureus E. coli P. aeruginosa C. albicans A. niger Comparative Example 1 9 10 10 9 9 9.4 Comparative Example 2 10 9 10 10 9 9.6 Comparative Example 3 9 10 11 10 10 10 Example 1 12 11 11 11 12 11.4

Claims

A cosmetic composition comprising, as an active ingredient, a protein polysaccharide extracted from flax seed, a protein polysaccharide extracted from a tooth root, and polyglutamic acid.

The cosmetic composition according to claim 1, wherein the content of the protein polysaccharide extracted from the flaxseed is 0.1 to 20% by weight based on the weight of the cosmetic composition.

The cosmetic composition according to claim 1, wherein the content of the protein polysaccharide extracted from the tooth root is 0.1 to 20% by weight based on the weight of the cosmetic composition.

The cosmetic composition according to claim 1, wherein the content of the polyglutamic acid is 0.1 to 20% by weight based on the weight of the cosmetic composition.

[Claim 3] The cosmetic composition according to claim 1, wherein the mixing ratio of the protein polysaccharide extracted from the flaxseed extract to the protein polysaccharide extracted from the toothseal is 1: 1 to 2: 1 by weight.

The cosmetic composition according to claim 1, wherein the mixing ratio of the protein polysaccharide: polyglutamic acid extracted from the flaxseed is 4: 1 to 4: 2 by weight.

The cosmetic composition according to claim 1, wherein the mixing ratio of the protein polysaccharide: polyglutamic acid extracted from the teeth is 4: 1 to 4: 2 by weight.

The cosmetic composition according to claim 1, wherein the cosmetic composition is for moisturizing the skin.

The cosmetic composition according to claim 1, wherein the cosmetic composition is anti-aging.

The cosmetic composition according to claim 1, wherein the cosmetic composition is for skin wrinkle suppression or skin wrinkle improvement.

The cosmetic composition according to claim 1, wherein the cosmetic composition is anti-inflammatory.

The cosmetic composition according to claim 1, wherein the cosmetic composition is antibacterial.

(A) separating the protein polysaccharide layer from flaxseed and filtering the protein polysaccharide layer separated from flaxseed to obtain a protein polysaccharide extracted from flaxseed;
(B) separating the protein polysaccharide layer from the tooth tissue and filtering the protein polysaccharide layer separated from the tooth tissue to obtain a protein polysaccharide extracted from the tooth tissue;
(C) mixing the protein polysaccharide extracted from flaxseed with the protein polysaccharide extracted from toothseal; And
(D) adding polyglutamic acid to the protein polysaccharide extracted from mixed flaxseed and the protein polysaccharide extracted from toothseed.