KR20180018055A - Anti-wrinkle Cosmetic Composition Comprising Functional Components from Sturgeon, Method for Obtaining the Functional Components and Method for Producing Anti-wrinkle Cosmetic Using the Functional Components - Google Patents

Abstract

The present invention relates to a technology for effectively obtaining useful components from sturgeons and using the same as a raw material for anti-wrinkle cosmetics. An anti-wrinkle cosmetic composition according to the present invention comprises: collagen peptide derived from sturgeon cartilage; and a caviar extract derived from caviar which is roe of sturgeons. The collagen peptide is obtained by hydrolyzing collagen protein, which is extracted by hydrolyzing sturgeon cartilage with pepsin, with alcalase. The caviar extract is obtained by adding caviar in an organic solvent and extracting the same at 60-80°C.

Description

TECHNICAL FIELD The present invention relates to an anti-wrinkle cosmetic composition containing a useful substance derived from sturm shark, a method for obtaining the useful substance, and a method for manufacturing a wrinkle-reducing cosmetic product using the useful substance. for Producing Anti-wrinkle Cosmetic Using the Functional Components}

More particularly, the present invention relates to a cosmetic composition for improving wrinkles containing a useful substance derived from sturgeon, a useful substance derived from the sturgeon, and a method for producing the same. The present invention relates to a technique for effectively obtaining a useful substance from sturgeon, And a method for producing a wrinkle-reducing cosmetic product using the sturgeon-derived useful substance.

In Hamyang-gun and other provinces in Gyeongsangnam-do, local sturgeon fishery industry is being developed as a specialized industry, and it takes longer than 5 ~ 6 years to produce fish and caviar. However, the technology to utilize the product derived from sturgeon is less than foreign countries.

More than 70% of sturgeon is composed of cartilage and is characterized by having more eggs than other fish. The sturgeon egg is called Caviar, and the caviar salted food is famous as one of the three gourmet foods in the world. Sturgeon caviar is similar to human skin protein structure, so it is known that it not only has quicker absorption of the skin than other ingredients, but also shows a good effect of improving the wrinkles of the skin by supplying the continuous nutrition to the skin after being absorbed. Sturgeon cartilage contains abundant calcium and calcium to strengthen the skeleton of children and women, and is also effective in preventing osteoporosis.

On the other hand, functional cosmetics is a cosmetic product with special functions other than cleaning and cosmetic purposes. It is an intermediate product between cosmetics and medicines, and has improved efficacy and safety. It is limited to products that protects skin from wrinkles, whitening, and ultraviolet rays. have. Among the functional cosmetics, the development of anti-aging cosmetics including wrinkle improvement is a major concern of the cosmetics industry due to an increase in the population of aging population, expansion of various consumer groups, development of beauty and medical technology, . In recent years, the demand for cosmetics has increased due to the increase in income due to the advancement of women and the increase in the educational attainment of women, as well as the so-called "early wrinkle improvement" which controls wrinkles in the late 20s or early 30s due to skin aging caused by stress, environmental pollution, Products' market is growing.

The most common symptom of skin aging is wrinkles, and the precise mechanism of its development has not been elucidated yet. Damage to matrix proteins such as collagen and elastic fibers present in the skin is the main cause of wrinkles. In addition, skin is one of the most outer skin layers of the body and is one of the most exposed to oxidative stress due to external stimuli. Various external factors such as ultraviolet rays, drugs, environmental pollutants increase the generation of reactive oxygen species Oxidative stress, which is caused by oxidative damage caused by binding with unsaturated fatty acids in cell membranes, is known to be a major factor of wrinkles. Among the wrinkles that occur as skin aging, small wrinkles are related to the shape of the epidermis and upper dermis of the skin, and large and deep wrinkles and sagging are basically caused by the qualitative and quantitative changes of the dermis, namely, collagen and elastin.

Retinol and adenosine, which are known to help improve wrinkles, have been used as raw materials for wrinkle-improving functional cosmetics, but they mainly focus on specific antioxidant ingredients. They are not only capable of causing skin troubles It is difficult to expect an effective effect. Currently, most of the physiologically active materials used as the main raw materials for functional cosmetics depend on imports. Therefore, it is necessary to study new materials for physiologically active materials using domestic technology and domestic natural materials.

Korean Patent No. 10-1191372 Korean Patent No. 10-1517950

The present invention has been developed to provide a new material for functional cosmetics using natural materials derived from sturgeon, and it is a technical problem to provide a technique for effectively obtaining a beneficial substance from sturgeon and utilizing it as a raw material for cosmetics for improving wrinkles .

In order to solve the above-mentioned technical problems, the present invention provides a wrinkle-improving cosmetic composition comprising a useful substance derived from sturgeon, a method for obtaining the useful substance, and a method for producing a wrinkle-reducing cosmetic product using the useful substance.

The cosmetic composition for improving wrinkles derived from sturgeon according to the present invention comprises a collagen peptide derived from sturgeon cartilage and a caviar extract derived from sturgeon alin caviar, wherein the collagen peptide is produced by adding sturgeon cartilage to pepsin And extracting the collagen protein extracted by digestion with an alcalase. The caviar extract is obtained by adding caviar to an organic solvent and extracting at 60 to 80 ° C.

The method for obtaining the sturdy shark-derived useful substance according to the present invention comprises: (S1) preparing a collagen peptide; And a step S2 of preparing a caviar extract. In step S1, the sturgeon cartilage is added to an alkali solution, stirred, and then centrifuged to wash the flask until the pH becomes neutral. Then, pepsin Stirring the mixture, and centrifuging the mixture to recover the supernatant; Dialyzing the recovered supernatant, and lyophilizing it to obtain a collagen protein; Lyophilized collagen protein is added with proteolytic enzyme in water, stirred, and centrifuged to separate the residue and the filtrate; And separating the collagen peptide by passing the separated filtrate through the ultrafiltration membrane. In the step 2, the caviar is added to 8 to 12 times by weight of ethanol and extracted at 65 to 75 ° C. .

The method for manufacturing a wrinkle-reducing cosmetic product using the useful substance derived from sturgeon in accordance with the present invention comprises: a first step of obtaining a sturdy shark-derived useful substance; A cosmetic composition comprising a collagen peptide and a caviar extract obtained in the first step, a first component including purified water, adenosine, a second component including an oil and an emulsifier, a third component including a lubricant, A second step of preparing the first step; A third step of heating the purified water to 70 to 80 캜; A fourth step of adding collagen peptide and first component to heated purified water and stirring at 60 to 70 캜; A fifth step of mixing and stirring the second component and the third component at 60 to 70 캜; A sixth step of mixing and stirring the agitated product of the fourth step and the agitated product of the fifth step at 55 to 65 ° C; A seventh step of cooling the stirred product of step 6 to 40 to 50 캜; Adding caviar extract to the cooled product of step 7, stirring the mixture at 40 to 50 ° C, and adding a pH adjusting agent to adjust the pH.

According to the present invention, the following effects can be expected.

First, since the present invention proposes extraction conditions optimized for extracting and separating useful substances from sturgeon cartilage and caviar, the useful components of sturgeon cartilage and caviar can be maximized.

Second, since the present invention proposes a material technology and a commercialization technique for wrinkle-improving functional cosmetics (composite product utilizing a conspicuous raw material) utilizing the useful materials of the sturgeon cartilage and caviar, It will contribute greatly to the development of the local economy through income increase, and also to the enhancement of national competitiveness.

1 is a photograph of collagen peptide derived from cartilaginous cartilage isolated according to [Example 4] of the present invention.
FIG. 2 is a photograph of a sturgeon-derived caviar extract extracted according to [Example 7] of the present invention.
Figure 3 shows the results of the cytotoxicity test conducted in Example 8 of the present invention
Fig. 4 and Fig. 5 show the results of intracellular collagen production test conducted in [Example 8] of the present invention.
Figs. 6 to 8 are examples of the cosmetic composition according to the present invention and are examples of formulations or cosmetic compositions for use.
FIG. 9 shows an eye cream production process at a blending ratio according to FIG.
10 is a photograph of a prototype manufactured by the method for manufacturing a cosmetic product according to the present invention.

More particularly, the present invention relates to a cosmetic composition for improving wrinkles containing a useful substance derived from sturgeon, a useful substance for cosmetics thereof, and the like. And a method for producing a cosmetic product for improving wrinkles using the cosmetic composition for improving wrinkles.

1. Improve wrinkles containing useful materials derived from sturgeon Cosmetics  Composition

A collagen peptide derived from sturgeon cartilage, and a caviar extract derived from sturgeon alin caviar. Here, the collagen peptide is obtained by hydrolyzing collagen protein extracted by hydrolysis of sturgeon cartilage with pepsin into alcalase, and the caviar extract is extracted at 60 to 80 ° C by adding caviar to organic solvent . It is preferable that the collagen peptide is contained in the total cosmetic composition in an amount of 0.1 to 0.6% by weight, and the caviar extract is contained in the total cosmetic composition in an amount of 0.1 to 0.6% by weight in view of economic efficiency and wrinkle-improving functional performance.

2. Wrinkle improvement from sturgeon For cosmetics  Of useful material Method of obtaining

The present invention establishes optimal conditions for effectively extracting and separating collagen peptides from sturgeon cartilage and optimum conditions for obtaining extracts from sturgeon caviar.

First, the optimal conditions for extracting and separating collagen peptide from the cartilage of sturgeon are examined step by step. Sturgeon cartilage is added to an alkaline solution, stirred, washed with water until the pH becomes neutral, centrifuged after adding pepsin in an acidic solution, and centrifuged to recover the supernatant; Dialyzing the recovered supernatant, and lyophilizing it to obtain a collagen protein; Lyophilized collagen protein is added with proteolytic enzyme in water, stirred, and centrifuged to separate the residue and the filtrate; And separating the collagen peptide by passing the separated filtrate through the ultrafiltration membrane.

Step S11 is a step of treating the sturgeon cartilage with an alkali solution to remove the insoluble portion, neutralize with water, and then hydrolyze with pepsin to obtain a collagen protein. In step S11, the sturgeon cartilage is pulverized to a size of 5 to 20 mm and added to a 0.1 N sodium hydroxide solution, which is 10 times by weight, stirred for 24 hours, centrifuged to neutralize the pH , And then 0.1% (w / v) pepsin was added to a 0.5 M acetic acid solution, which was 10 times by weight, and hydrolyzed by stirring for 2 days, followed by centrifugation at 8,000 rpm for 30 minutes to recover the supernatant.

The step S12 it is preferred that after the process to remove impurities to give the collagen protein, the supernatant was dialyzed in 20mM Na ₂ HPO ₄ proceeds in such a way that the freeze-dried.

Step S13 is a process of obtaining a collagen peptide by hydrolyzing a collagen protein with a protease. When step S13 is optimized, 10 times as much water as water is added to the lyophilized collagen protein, 2% (w / v) of a protease, alcalase, is hydrolyzed at 60 ° C for 6 hours, After the heat treatment, the residue is removed by centrifugation and the filtrate is separated.

Step S14 is a process of obtaining a collagen peptide having an average molecular weight of 800 to 1,200 Da by removing calcite residues and impurities using an ultrafiltration membrane. The collagen peptide is separated by filtration through an ultrafiltration membrane, followed by lyophilization.

Next, the optimal conditions for extracting caviar extract from sturgeon caviar are examined. According to the following examples, the extraction yield can be increased when caviar is added to ethanol at 8 to 12 times by weight and extracted at 65 to 75 ° C.

The characteristics of collagen peptide and caviar extracts from the sturgeon cartilage produced by the above method were investigated as raw materials for cosmetics (characteristics, heavy metals, etc.).

3. Manufacturing method of wrinkle-improving cosmetics originating from sturgeon shark

The present invention proposes a cosmetic preparation method using a sturdy cartilage and a caviar-derived useful substance as a wrinkle-improving functional cosmetic raw material. The oil-in-water-derived useful substances (collagen peptide and caviar extract) are basically emulsified because oil and water are present at the same time. Hereinafter, the method of manufacturing cosmetics according to the present invention will be described step by step.

First, a sturdy shark-derived useful substance is obtained (step 1), which is obtained according to the above-mentioned optimum conditions. A second component comprising an oil and an emulsifier; a third component comprising a lubricant; a second component comprising an oil and an emulsifier; a third component comprising a lubricant; (Step 2). The collagen peptide and the caviar extract obtained in the first step are suitably used in an amount of 0.1 to 0.6% by weight based on the entire cosmetic composition for cosmetics. Further, a stabilizer, a wetting agent, an emulsifier, a moisturizer, a coloring agent, a flavoring agent and the like can be further prepared to improve stability and usability.

Next, the purified water is heated to 70 to 80 캜 (step 3), the collagen peptide and the first component are added to the heated purified water, and the mixture is stirred at 60 to 70 캜 (step 4) The components are mixed and stirred at 60 to 70 ° C (Step 5). Then, the agitated product of the fourth step and the agitated product of the fifth step are mixed and emulsified at 55 to 65 ° C (step 6), and the agitated product of step 6 is cooled to 40 to 50 ° C Step 8) The caviar extract is added to the cooled product of Step 7, stirred at 40 to 50 ° C, and the pH is adjusted by adding a pH adjusting agent (Step 8). This completes the cosmetic product.

[ Example 1 ] Collagen from sturgeon cartilage Peptides  Separation manufacturing

1. Sturgeon cartilage preparation

After shaved cartilage, it was crushed to a size of 5 ~ 20 mm. The crushed cartilage was stored frozen (-20 ℃) and used as a material for extraction and separation of collagen protein.

2. Collagen Peptides  Separation manufacturing

Sturgeon cartilage was added to 0.1 N NaOH solution, and the mixture was firstly stirred. The mixture was centrifuged and washed with water until the pH became neutral. Then, pepsin was added in 0.5 M acetic acid solution and the mixture was stirred for 2 hours. (Step S11). The recovered supernatant was dialyzed against 20 mM Na ₂ HPO ₄ (pH 7.2) and lyophilized to obtain collagen protein (Step S12). The lyophilized collagen protein was mixed with water to prepare a substrate. Alcalase, a protease, was added to the substrate, followed by stirring. After centrifugation, the residue was removed to separate the filtrate (Step S13). Finally, the separated filtrate was subjected to ultrafiltration membrane to remove calcite residues and impurities, and then lyophilized to finally produce a collagen peptide (Step S14).

[ Example 2 ] Collagen protein extraction conditions from sturgeon cartilage

One. 0.1N  The optimum amount of NaOH solution added

In order to confirm the optimal addition amount of 0.1N NaOH solution, a test was conducted to extract collagen protein with different amounts of 0.1 N NaOH solution. The procedure of step S11 of [Example 1] was repeated while 0.1 N NaOH solution was added at 5, 10, and 15 times the weight of sturgeon cartilage. In step S11, NaOH solution was added thereto, The mixture was firstly stirred at a stirring time of 24 hours, then added with 0.1% (w / v) pepsin, and stirred at 50 rpm for 24 hours. Then, step S12 was performed to confirm the extraction yield.

As a result of the test, it was confirmed that removal of foreign matters and collagen extraction were facilitated as the amount of 0.1 N NaOH solution was increased. However, there was no significant difference between 10 and 15 times of addition, and the addition of 0.1 N NaOH solution 10 times as much as the weight of sturgeon cartilage was the optimum condition, and the extraction yield of collagen protein was 45.6% (w / w) .

2. Optimal Stirring speed  And Stirring time

In order to confirm the optimal agitation time, a test was conducted to extract collagen protein with different agitation times. (W / v) pepsin addition, 50 rpm agitation speed, 24 rpm, 50 rpm agitation speed, 12, 24, 36 and 48 hours agitation time, And the second stirring step was performed under the condition of time agitation time. Then, step S12 was performed to confirm the extraction yield.

As the agitation time increased, the removal of foreign matters and collagen extraction were facilitated. However, no significant difference was observed at 24, 36 and 48 hours, and the agitation for 24 hours under optimum conditions (extraction yield 46.8 ± 0.6% w)).

3. Optimum amount of pepsin added

In order to determine the optimal amount of pepsin, pepsin was added at a concentration of 0.05, 0.1, 0.2, 0.5% (w) in 0.1 N NaOH solution at 10 times the weight of the sturgeon cartilage and at a stirring speed of 50 rpm for 24 hours. / v), stirring was carried out at a stirring rate of 50 rpm for 24 hours, and then the step S11 was carried out. Then, step S12 was performed to confirm the extraction yield.

The extraction yield was increased with increasing pepsin concentration, but the addition of 0.1% pepsin was considered to be the optimum condition considering extraction cost and extraction efficiency (extraction yield 46.8 ± 0.6%).

4. Optimum after pepsin addition Stirring time

In order to confirm the optimal agitation time after the addition of pepsin, 0.1% (w / v) of pepsin was added after the primary stirring at 10 times of 0.1N NaOH solution, the stirring speed of 50 rpm, and the stirring time of 24 hours, The step S11 was carried out while stirring at 50 rpm under agitation time of 12, 24, 36, 48 and 60 hours. Then, step S12 was performed to confirm the extraction yield.

As the agitation time increased, the yield of collagen extraction increased. However, the yield at 48 hours of stirring time was almost the same as that of 0.2% (w / v) addition of pepsin (52.6 ± 0.9% at 96% level). Considering the extraction cost and extraction efficiency It can be seen as an optimal condition.

[ Example 3 ] Sturgeon cartilage-derived collagen Enzyme hydrolysis  Condition

1. Collagen protein extraction

Stirring was carried out at 10 rpm in 0.1N NaOH solution, 50 rpm stirring speed and 24 hours stirring time, and then added with pepsin 0.1% (w / v) Step S11 was performed while stirring the tea. Then, step S12 was carried out to obtain a lyophilized product of collagen protein, wherein the extraction yield was 52.6 ± 0.9%. Enzymatic hydrolysis conditions of the lysate of the collagen protein thus obtained were tested.

2. Enzymatic hydrolysis  Optimal for Substrate concentration

To determine the optimum substrate concentration for enzyme hydrolysis, 1% Alcalase (1%) was added at a substrate concentration of 10, 15, 20, 25 and 30% (w / w) And step S13 was performed. Here, the reaction conditions are a reaction temperature of 50 ° C, a reaction pH of 8.0 (20 mM Tris-HCl buffer), a reaction time of 3 hours, and a stirring speed of 50 rpm. As a result, the enzyme hydrolysis was markedly increased up to 20% substrate concentration, but the enzyme hydrolysis was moderately increased.

3. Enzymatic hydrolysis  Optimal Enzyme Concentration

To determine the optimal enzyme concentration for enzyme hydrolysis, the Alcalase concentration was adjusted to 0.% at a substrate concentration of 20%, a reaction temperature of 50 ° C, a reaction pH of 8.0 (20 mM Tris-HCl buffer), a reaction time of 2 days, , 1%, 2%, 3%, and 5% (w / v), respectively. As a result, the hydrolysis of collagen protein increased rapidly up to 2% (w / v) Alcalase concentration.

4. Enzymatic hydrolysis  Optimum reaction temperature for

45 ° C, 50 ° C, 55 ° C, 60 ° C, and 60 ° C, respectively, under conditions of a substrate concentration of 20%, an Alcalase concentration of 2%, a reaction pH of 8.0 (20 mM Tris-HCl buffer), a reaction time of 3 hours, 65 < 0 > C and 70 < 0 > C, respectively. As a result of the test, optimum activity was shown at a reaction temperature of 60 ° C.

5. Enzymatic hydrolysis  Optimal for Reaction pH

S13 was carried out at pH 7.0, 7.5, 8.0, and 8.5 (20 mM Tris-HCl buffer) under conditions of a substrate concentration of 20%, an Alcalase concentration of 2%, a reaction temperature of 60 ° C, a reaction time of 3 hours and a stirring speed of 50 rpm. The highest activity was observed at pH 8.0.

6. Enzymatic hydrolysis  Optimum Reaction Time for

The reaction was carried out at 3, 6, 9 and 12 hours under conditions of a substrate concentration of 20%, an Alcalase concentration of 2%, a reaction temperature of 50 ° C, a reaction pH of 8.0 (20 mM Tris-HCl buffer) and a stirring speed of 50 rpm Respectively. As a result of the test, the activity was the highest at the reaction condition for 6 hours.

7. Enzyme hydrolyzate Granulation ( Powdered )

Based on the above test results, step S13 was performed under conditions of a substrate concentration of 20%, an Alcalase concentration of 2%, a reaction temperature of 60 ° C, a reaction pH of 8.0 (20 mM Tris-HCl buffer), a stirring speed of 50 rpm, Then, step S14 was performed to obtain a powder. As a result, a collagen peptide having an average molecular weight of 800 to 1,200 Da was obtained with an extraction yield of 87.4 ± 0.6% (w / w).

[ Example 4 ] Collagen derived from sturgeon cartilage Of peptide  Character rating

The collagen peptide obtained according to [Example 3] above is shown in Fig. 1, and the characteristics thereof were evaluated. The evaluation results are shown in Table 1 below.

Characterization of collagen peptides Item standard Analytical value method of inspection Remarks Appearance It is hygroscopic with an off-white powder fitness Sensory test fitness Average molecular weight (Av. MW) 1,000 800 ~ 1,200 Da GPC fitness Crude protein 93.0% or more 95.54% Food circulation
General Test Methods fitness moisture Less than 7.0% 4.2% fitness Ash Not more than 1.0% 0.26% fitness pH (1% solution) 5.5 ± 1.0 6.38 pH meter fitness heavy metal lead 20 ppm or less Non-detection Food circulation
General Test Methods fitness arsenic 10 ppm or less Non-detection fitness Mercury  1 ppm or less Non-detection fitness Total aerobic bacteria count Not more than 1,000 CFU / ml (g) 60 fitness Coliform group voice voice fitness

[ Example 5 ] Caviar  Extract preparation

One. Caviar  Ready

The frozen caviar (2 kg) was thawed at room temperature and then pulverized by a crusher. The crushed caviar crushed material was subdivided and stored frozen (-20 ° C).

2. Ethanol Extraction

The supernatant was recovered by ethanol centrifugation (8,000 rpm, 10 minutes), and then evaporated in vacuo and the solids content was measured.

[ Example 6 ] Caviar  Extract extraction condition

1. Optimum amount of ethanol added

The supernatant was recovered by centrifugation (8,000 rpm, 10 minutes) at a stirring speed of 50 rpm at room temperature, and the supernatant was recovered by evaporation at a rate of 5, 7.5, 10, 12.5 and 15 times the weight of the caviar crushed material. The yield was measured. The results are shown in Table 2 below. As can be seen, the extraction yield was increased up to the 10-fold ethanol addition amount, but there was no significant difference in the extraction yield.

Extraction yield according to the amount of ethanol added Ethanol amount (weight ratio) The extraction yield (%, w / w) 5 times 8.6 ± 0.2% 7.5 times 10.1 ± 0.3% 10 times 10.8 ± 0.3% 12.5 times 10.8 ± 0.2% 15 times 10.9 ± 0.3%

2. Ethanol Extraction Time optimal  Addition amount

The supernatant was recovered by centrifugation (8,000 rpm, 10 minutes) at a stirring speed of 50 rpm, extraction time of 30 minutes, 60 minutes, 90 minutes and 120 minutes at a heating mantle of 70 ° C with a 10-fold increase in the amount of ethanol relative to the weight of the caviar crushed material. and the extraction yield was measured by evaporation. During the concentration process of caviar extract, it was extracted at 70 ℃ in Heating Mantle considering the process of raising the temperature to about 70 ℃ in order to remove moisture contained in the caviar. The measurement results are shown in Table 3 below. As can be seen, the difference in the extraction yields was not great at over 60 minutes. However, the extraction yield was 13.5 ± 0.3% at 60 min, which was 1.25 times higher than the extraction yield of 10.8 ± 0.3% extracted at room temperature for 24 hours.

Extraction yield according to extraction time Extraction time The extraction yield (%, w / w) 30 minutes 8.4 ± 0.2% 60 minutes 13.5 ± 0.3% 90 minutes 13.3 ± 0.2% 120 minutes 13.5 ± 0.2%

[ Example 7 ] Sturgeon originated Caviar  Characterization of extracts

The caviar extract (60 min, extraction yield: 13.5 ± 0.3%) extracted according to the above [Example 6] is shown in FIG. 2, and the results of the evaluation are shown in Table 4 below.

Characterization of caviar extract Item
standard
Analytical value
method of inspection
Remarks
Appearance Colorless ~ clear solution of light yellow color, slight special odor fitness Sensory test fitness pH
(Primary extract solution) 6.5 ± 1.0 7.02 pH meter fitness importance 0.95 to 1.06 1.02 hydrometer fitness heavy metal lead 20 ppm or less Non-detection Food circulation
General Test Methods fitness arsenic 10 ppm or less Non-detection fitness Mercury  1 ppm or less Non-detection fitness Total aerobic bacteria count Not more than 1,000 CFU / ml (g) Non-detection fitness Fungus number Non-detection fitness

[ Example 8 ] Wrinkle improvement effect test

One. MTT  assay for cytotoxicity

The cytotoxicity test for the wrinkle-improving effect test of the collagen peptide according to [Example 4] and the caviar extract according to [Example 7] through MTT assay was performed. The test results are shown in Fig. As shown, both caviar extract and collagen peptide showed almost no cytotoxicity up to a concentration of 1% (cell survival rate: 97.7 ± 0.6%).

2. Intracellular  Collagen Synthesis Assay

To compare the degree of cellular collagen production increase in fibroblast cultures with those of blank test, the effectiveness of functional cosmetics to help improve wrinkles among the guidelines for evaluating the effectiveness of functional cosmetics presented by Korea Food and Drug Administration (Guideline for Efficacy Evaluation of Functional Cosmetics). Here, HaCaT, a primary cell line, was used as a fibroblast. Fibroblasts were seeded in 48-well plates at a density of 5 × 10 ⁴ cells per well. Cells were cultured for 24 h, then washed with 10% PBS (phosphate buffered saline) After culturing for a time, the amount of collagen was measured by taking the culture solution. The amount of collagen measured was corrected by the amount of total collagen obtained by the Lowry assay.

Test results are shown in Figs. 4 and 5. FIG. 4 shows the collagen production ability according to the addition concentration. As can be seen, the caviar extract showed an excellent collagen production ability at a concentration of 0.2%, more than 3 times that of the control and 2 times of the collagen peptide. FIG. 5 shows a comparison of cell morphology of human fibroblast (HaCaT) when 0.2% of caviar extract and collagen peptide were added and cultured.

[Example 9] Production of wrinkle-improving cosmetics

1. Establishing the mixing ratio

Based on the results of Example 8, the ratio of functional cosmetic ingredients is checked using the collagen peptide derived from sturgeon cartilage (0.2%, caviar extract (0.2%).) At this time, Adenosine (0.04%) is used. The ratio of the cosmetic composition to the raw material of the cosmetic composition is appropriately determined according to the formulation or use (essence, eye cream, nutritional cream, etc.) As shown in the figure, raw materials for cosmetics include collagen peptide derived from cartilaginous cartilage and caviar extract prepared according to the present invention, purified water, a first component including adenosine, a composition containing an oil and an emulsifier A second component including a lubricant, and a pH adjuster.

2. Manufacturing process

FIG. 9 illustrates a process of manufacturing an eye cream with a composition according to the blending ratio of FIG. 6, and a process of manufacturing a cosmetic according to FIG. 9 will be described. First, a collagen peptide derived from cartilaginous cartilage and a caviar extract are first prepared (step 1), and a cosmetic raw material is prepared by mixing purified water, a first ingredient including adenosine, a second ingredient including an oil and an emulsifying agent, , a pH adjusting agent and the like are prepared (step 2). Then, the purified water is heated to 70 to 80 ° C (Step 3), and the collagen peptide and the first component are added to the heated purified water, followed by stirring at 60 to 70 ° C (Step 4). Next, the second component and the third component are mixed and stirred at 60 to 70 ° C (Step 5), and the stirred mixture of Step 4 and Step 5 is mixed and stirred at 55 to 65 ° C Step 6), and the stirring of the sixth step is cooled to 40 to 50 캜 (step 7). Finally, the caviar extract is added to the cooling water of step 7, stirred at 40 to 50 ° C, and the pH is adjusted by adding a pH adjusting agent (step 8). Between steps 8 and 8, a perfume or the like may be further added.

As a result, a wrinkle-improving cosmetic product as shown in Fig. 10 is completed. The stability and heavy metal testing of the prototype products were commissioned by KB Cosmetics and Korea Food Research Institute. The tests were carried out according to functional cosmetics standards and test methods for quality cosmetics manufacturing and quality control (KB-501-03, revised 2015.06.30) and the Food and Drug Administration (No. 2015-15, 2015.03.25). As a result, it was confirmed that the stability of the prototype (pH, total aerobic bacteria count, fungus count) meets the criteria of the phytosanitary standard and all the heavy metals (lead, arsenic, mercury) are not detected.

Claims (5)

A collagen peptide derived from sturgeon cartilage;
A caviar extract derived from sturgeon alin caviar,
The collagen peptide is obtained by hydrolyzing collagen protein extracted by hydrolysis of sturgeon cartilage with pepsin into an alcalase,
Wherein the caviar extract is obtained by adding caviar to an organic solvent and extracting at 60 to 80 ° C. The method of claim 1,
The collagen peptide comprises 0.1 to 0.6% by weight in the total cosmetic composition,
Wherein the caviar extract is composed of 0.1 to 0.6% by weight of the total cosmetic composition. A step S1 of producing a collagen peptide;
And a step (S2) of producing a caviar extract,
The step (S1)
Sturgeon cartilage is added to an alkaline solution, stirred, washed with water until the pH becomes neutral, centrifuged after adding pepsin in an acidic solution, and centrifuged to recover the supernatant; Dialyzing the recovered supernatant, and lyophilizing it to obtain a collagen protein; Lyophilized collagen protein is added with proteolytic enzyme in water, stirred, and centrifuged to separate the residue and the filtrate; And separating the collagen peptide by passing the separated filtrate through the ultrafiltration membrane,
The step (S2)
Caviar is added to 8 to 12 times by weight of ethanol and extracted at 65 to 75 ° C. Wherein the method comprises the steps of: 4. The method of claim 3,
The step (S1)
Sturgeon cartilage was added to 10 times 0.1 N sodium hydroxide solution by weight, stirred for 24 hours, and centrifuged to wash the pH until neutrality. Then, 0.1 M (w / v) pepsin, stirring for 2 days, hydrolyzing and centrifuging to recover the supernatant;
The recovered supernatant was dialyzed in 20mM Na ₂ HPO ₄ and then the step S12 to obtain a collagen protein and lyophilized;
10% by weight of water was added to the lyophilized collagen protein and 2% (w / v) of protease, alcalase, was hydrolyzed at 60 ° C for 6 hours and then heat-treated at 80 ° C for 30 minutes. Step S13 of removing the residue and separating the filtrate;
Separating the collagen peptide, and lyophilizing the filtered collagen peptide to obtain a useful substance for improving cosmetic wrinkles derived from sturgeon sharks. As a method for manufacturing a cosmetic product using a useful substance derived from sturgeon,
A process for producing a sturdy shark-derived useful substance according to claim 3 or 4, comprising the steps of:
A cosmetic raw material comprising a first component comprising purified water, adenosine, a second component comprising an oil and an emulsifying agent, a third component comprising a lubricant, a second component comprising an oil and an emulsifier, A second step of preparing an adjusting agent;
A third step of heating the purified water to 70 to 80 캜;
A fourth step of adding collagen peptide and first component to heated purified water and stirring at 60 to 70 캜;
A fifth step of mixing and stirring the second component and the third component at 60 to 70 캜;
A sixth step of mixing and stirring the agitated product of the fourth step and the agitated product of the fifth step at 55 to 65 ° C;
A seventh step of cooling the stirred product of step 6 to 40 to 50 캜;
Adding caviar extract to the cooled product of step 7, stirring at 40 to 50 ° C, and adjusting pH by adding a pH adjusting agent;
Wherein the wrinkle-improving cosmetic material is a wrinkle-improving cosmetic product.

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