KR102454297B1 - Cosmetic composition for anti-oxidation, anti-inflammation or skin improvement containing propolis fermented extracts - Google Patents

Abstract

The present invention includes a propolis fermented extract obtained by fermenting and extracting a propolis complex including gold propolis, green propolis and red propolis as an active ingredient, thereby achieving significantly excellent antioxidant, anti-inflammatory or skin improvement effects. It relates to a cosmetic composition and a method for manufacturing the same.

Description

COSMETIC COMPOSITION FOR ANTI-OXIDATION, ANTI-INFLAMMATION OR SKIN IMPROVEMENT CONTAINING PROPOLIS FERMENTED EXTRACTS comprising propolis fermentation extract as an active ingredient

The present invention relates to a cosmetic composition for antioxidant, anti-inflammatory or skin improvement comprising a propolis fermented extract as an active ingredient.

Propolis is a substance made by bees by mixing their own saliva and enzymes with resin-like substances extracted from various plants for their survival and reproduction. It is also called Russian penicillin or natural penicillin. There are many differences in the composition and content of propolis depending on what plant the bees use to make it. .

In general, propolis contains a large amount of active ingredients such as flavonoids, amino acids, and minerals, and thus can implement effects such as antioxidant, anti-inflammatory, immune activity, antibacterial, and anticancer. Accordingly, as disclosed in Korean Patent Registration No. 10-1247375, it is mainly used as a material for foods and health foods that activate the functions of the human body through ingestion.

Recently, various attempts have been made to apply such propolis to cosmetic compositions.

For example, in Korean Patent Registration No. 10-1884660, a mixed extract of hydrolytic enzyme-treated propolis, royal jelly and honey and an enzyme extract of kava kava, turmeric and cacao seeds are used as active ingredients, 0.1 to 10 based on the total weight of the cosmetic composition. Disclosed is a cosmetic composition containing % by weight. In addition, Republic of Korea Patent No. 10-1835799 discloses a first formulation containing propolis extract, glycerin, butylene glycol, etc., propolis extract, adenosine, beta-glucan, Heotgae tree extract, green tea extract, black bean extract, etc. Disclosed is a cosmetic composition in which a second agent containing

In the prior art, including the above patent, most of them only consider the fact that a fermented product or extract of one type of propolis is mixed with other active ingredients to realize a skin beauty effect, but different types of propolis are mixed and used. The point is not taken into consideration. In other words, propolis has significant differences in composition and content depending on the type of plant and climate, but until now, only a technology using only one type of propolis has been mainly utilized, and a plurality of propolis with different constituents and content have been used. Attempts to realize various effects by mixing were insufficient.

Accordingly, the present inventor completed the present invention by developing a fermented propolis extract with significantly improved antioxidant, anti-inflammatory or skin improvement effects by constructing a plurality of propolis with different characteristics, fermenting and extracting them. did.

Republic of Korea Patent No. 10-1247375 Republic of Korea Patent No. 10-1884660 Republic of Korea Patent No. 10-1835799

A first object of the present invention is to provide a cosmetic composition that is remarkably excellent in antioxidant, anti-inflammatory or skin improvement effects by including a fermented propolis extract obtained by mixing and fermenting different types of propolis.

A second object of the present invention is to provide a method for preparing a cosmetic composition having excellent antioxidant, anti-inflammatory or skin improvement effects, including the fermented propolis extract.

The object of the present invention is not limited to the technical problem as described above, and another technical problem may be derived from the following description.

In order to achieve the first object, the present invention provides an antioxidant, anti-inflammatory or A cosmetic composition for improving skin is provided.

The propolis complex may include the gold propolis, green propolis and red propolis in a weight ratio of 1: 0.1-5: 0.1-5.

The fermentation may be fermenting the propolis complex into a Lactobacillus strain.

The Lactobacillus strain may include Lactobacillus plantarum ( Lactobacillus plantarum ).

The skin improvement may be skin moisturizing or skin softness improvement.

The cosmetic composition may include 0.01 to 30% by weight of the fermented propolis extract based on the total weight.

The cosmetic composition is from the group consisting of softening lotion, nourishing lotion, astringent lotion, skin, lotion, essence, serum, cream, massage cream, pack, makeup base, BB cream, foundation, powder, cleansing foam, cleansing cream and cleansing water. It may be one formulation selected.

In order to achieve the second object, the present invention is a preparation step of preparing a propolis complex comprising gold propolis, green propolis and red propolis; A primary fermentation step of aging the propolis complex in a jar; a secondary fermentation step of inoculating the first fermented fermented product with a strain and fermenting it; and an extraction step of extracting the second fermented fermented product to form a propolis fermented extract; A manufacturing method is provided.

The cosmetic composition of the present invention contains, as an active ingredient, a fermented propolis extract obtained by fermenting and extracting a propolis complex containing gold propolis, green propolis and red propolis, thereby exhibiting remarkably excellent antioxidant, anti-inflammatory or skin improvement effects. can be implemented

In particular, in the present invention, by mixing and using different types of propolis, better antioxidant, anti-inflammatory or skin improvement effects can be realized than when one type of propolis is used.

In addition, according to the manufacturing method of the cosmetic composition of the present invention, a propolis complex comprising gold propolis, green propolis and red propolis is double fermented according to two steps to obtain a fermented propolis extract, thereby providing remarkably excellent antioxidant, It is possible to prepare a cosmetic composition that implements an anti-inflammatory or skin improvement effect.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art can easily carry out the present invention. However, the present invention may be embodied in several different forms and is not limited to the embodiments described herein. And in order to clearly explain the present invention in the drawings, parts irrelevant to the description are omitted.

The terms or words used in the specification and claims of the present invention are not to be construed as limited in their ordinary or dictionary meanings, and the inventor may appropriately define the concept of the term in order to best describe his invention. Based on the principle, it should be interpreted as meaning and concept consistent with the technical idea of the present invention.

In the entire specification of the present invention, when a part "includes" a certain component, it means that other components may be further included, rather than excluding other components unless otherwise stated. .

Throughout the specification of the present invention, "A and/or B" means A or B, or A and B.

Although the present invention has been specifically described below, the present invention is not limited thereto.

The present invention provides a cosmetic composition for antioxidant, anti-inflammatory or skin improvement comprising a propolis fermented extract as an active ingredient.

In one embodiment of the present invention, by including a fermented propolis extract obtained by fermenting and extracting a propolis complex containing gold propolis, green propolis and red propolis as an active ingredient, significantly excellent antioxidant, anti-inflammatory or skin improvement effect It provides a cosmetic composition having.

Specifically, in this embodiment, a propolis complex containing gold propolis, green propolis and red propolis is fermented and extracted, and a propolis fermented extract with a high content of active ingredients such as polyphenol and artepillin C By including, it is possible to provide a cosmetic composition having a remarkably excellent antioxidant, anti-inflammatory or skin improvement effect. In particular, in this embodiment, by mixing and using different types of propolis, a significantly superior antioxidant, anti-inflammatory or skin improvement effect can be realized than when one type of propolis is used.

Implementing the skin improvement effect may mean implementing the skin moisturizing improvement effect and the skin softness improvement effect, but is not limited thereto. Specifically, the fermented propolis extract of this embodiment and the cosmetic composition comprising the same have antioxidant and anti-inflammatory effects, thereby restoring the function of the skin dermis, thereby improving skin moisture, skin complexion, and skin softness.

The propolis complex includes gold propolis, green propolis and red propolis.

Gold propolis is also called brown propolis or yellow propolis, and refers to propolis having a color of gold, brown, or yellow. The gold propolis is produced in various countries such as Brazil, New Zealand, North America, and China. Specifically, the gold propolis of this embodiment may be from Australia, but is not limited thereto.

Green propolis is mainly manufactured using the resin of the Baccharis plant, and it is a green propolis, and contains a high content of flavonoids and Artepillin C, It can have antibacterial, anticancer, and antioxidant effects by itself. Specifically, the green propolis of this embodiment may be from Brazil, but is not limited thereto.

Red propolis is a red propolis, and it may mean that it is made with the red sap of a tree that grows only in the clean areas of northeastern Brazil. In particular, red propolis has a higher polyphenol content than gold propolis and green propolis, and contains isoflavones, an antioxidant component, so it has effects such as antioxidant and skin immunity enhancement. Specifically, the red propolis of this embodiment may be from Brazil, but is not limited thereto.

In this embodiment, gold propolis, green propolis and red propolis are not included alone, but all three types of propolis having different colors are included, so that the fermented propolis extract of this embodiment is polyphenol, artephyllin It may contain an active ingredient such as C in a high content. Accordingly, the cosmetic composition of the present embodiment can implement excellent antioxidant, anti-inflammatory or skin improvement effects.

In particular, the propolis complex of this embodiment includes the three types of propolis described above, and may include the gold propolis, green propolis and red propolis in a weight ratio of 1: 0.1 to 5: 0.1 to 5. According to the above, when the propolis complex of this embodiment contains the gold propolis, green propolis and red propolis in a weight ratio of 1: 0.1 to 5: 0.1 to 5, the fermented propolis extract of this embodiment and its containing The antioxidant, anti-inflammatory or skin improvement effect of the cosmetic composition may be improved.

Specifically, the propolis complex of this embodiment may include the gold propolis, green propolis and red propolis in a weight ratio of 1: 2 to 4: 1. According to the above, when the propolis complex of this embodiment contains the gold propolis, green propolis and red propolis in a weight ratio of 1: 2 to 4: 1, the fermented propolis extract of this embodiment and a cosmetic comprising the same The antioxidant, anti-inflammatory or skin improving effect of the composition may be further enhanced. In particular, in this embodiment, by including a higher content of green propolis than gold propolis and red propolis, the above effects can be further improved.

More specifically, the propolis composite of this embodiment may include the gold propolis, green propolis and red propolis in a weight ratio of 1: 4: 1. According to the above, when the propolis complex of this embodiment contains the gold propolis, green propolis and red propolis in a weight ratio of 1: 4: 1, the fermented propolis extract of this embodiment and the cosmetic composition comprising the same Antioxidant, anti-inflammatory or skin-improving effects can be significantly improved.

The fermented propolis extract is fermented and extracted from the propolis complex.

The fermented propolis extract of this embodiment may be obtained by fermenting and then extracting the propolis complex.

Specifically, the fermentation may be carried out by inoculating and fermenting the propolis complex with a strain applicable to fermentation, but is not limited thereto.

More specifically, the fermentation may be carried out by secondary fermentation in which the propolis complex is put in a jar and aged for primary fermentation, and then the primary fermented ferment is inoculated with a strain applicable to fermentation and fermented. not limited In particular, the fermented propolis extract of this embodiment may contain a higher content of active ingredients such as polyphenols and artepylline C by double fermenting a propolis complex mixed with three types of propolis. Accordingly, the antioxidant, anti-inflammatory or skin improvement effect of the cosmetic composition comprising the fermented propolis extract can be significantly improved.

In this case, the strain applicable to the fermentation may be a Lactobacillus strain, but is not limited thereto.

Specifically, the strain applicable to the fermentation may include, but is not limited to, Lactobacillus plantarum .

More specifically, the strain applicable to the fermentation may include Lactobacillus plantarum KCC-32 (Accession No.: KACC92135P), but is not limited thereto. In particular, in this embodiment, as the strain includes Lactobacillus plantarum KCC-32 (accession number: KACC92135P), polyphenol, artephilin C, etc. A police fermented extract may be provided. Accordingly, the antioxidant, anti-inflammatory or skin improvement effect of the cosmetic composition comprising the fermented propolis extract can be significantly improved.

The extraction may be carried out by any known extraction method, for example, the extraction is one or more extraction methods selected from the group consisting of hot water extraction method, room temperature stirring extraction method, reflux cooling extraction method, ultrasonic extraction method, high temperature pressure extraction method, and low temperature high pressure extraction method It may be carried out by, but is not limited to.

The cosmetic composition of this embodiment may contain 0.01 to 30% by weight of the fermented propolis extract, specifically 0.1 to 20% by weight, more specifically 1 to 10% by weight based on the total weight of the cosmetic composition. In particular, when the cosmetic composition of this embodiment contains 0.01 to 30% by weight of the fermented propolis extract based on the total weight, it is possible to provide a cosmetic composition having a stable formulation while excellent in antioxidant, anti-inflammatory or skin improvement effect. .

The cosmetic composition of this embodiment consists of softening lotion, nutrient lotion, astringent lotion, skin, lotion, essence, serum, cream, massage cream, pack, makeup base, BB cream, foundation, powder, cleansing foam, cleansing cream and cleansing water. It may be one formulation selected from the group, but is not limited thereto, and may be implemented as a formulation of all known cosmetic compositions.

The cosmetic composition of this embodiment may include excipients or additives necessary for the formulation of each formulation in addition to the fermentation extract of propolis. For example, the excipients or additives may include surfactants, emulsifiers, emulsifying aids, silicone polymers, pigments, pigments, fragrances, preservatives, bactericides, oxidation stabilizers, organic solvents, thickeners, emollients, antioxidants, free radical scavengers, opacity Topical, stabilizer, emollient, silicone, antifoam, humectant, vitamin, insect repellent, preservative, anti-inflammatory, filler, polymer, propellant, basifying agent, acidifying agent, colorant, thickening agent, skin conditioning agent, whitening agent, antioxidant, anti-wrinkle agent and the like, but is not limited thereto.

In addition, the present invention provides a method for preparing a cosmetic composition for anti-oxidation, anti-inflammatory or skin improvement comprising the fermented propolis extract as an active ingredient.

In another embodiment of the present invention, a preparation step of preparing a propolis complex comprising gold propolis, green propolis and red propolis; A primary fermentation step of aging the propolis complex in a jar; a secondary fermentation step of inoculating the first fermented fermented product with a strain and fermenting it; and an extraction step of extracting the secondary fermented fermented product to form a propolis fermented extract do.

In particular, in this embodiment, a propolis complex containing gold propolis, green propolis and red propolis is fermented and extracted, and a propolis fermented extract with a high content of active ingredients such as polyphenol and artepillin C is used. can be manufactured. Accordingly, it is possible to provide a cosmetic composition having a remarkably excellent antioxidant, anti-inflammatory or skin improvement effect.

In particular, in this embodiment, by extracting and fermenting a propolis complex mixed with different types of propolis to prepare a propolis fermented extract, significantly better antioxidant, anti-inflammatory or skin improvement than when one type of propolis is used effect can be implemented.

Hereinafter, the manufacturing method of the cosmetic composition has been described in detail.

(1) a preparation step of preparing a propolis complex containing gold propolis, green propolis and red propolis;

In the preparation step of this embodiment, a propolis complex including gold propolis, green propolis and red propolis is prepared. At this time, the preparation step of this embodiment may be carried out at room temperature conditions, but is not limited thereto.

Specifically, in the preparation step of this embodiment, a propolis complex can be prepared by mixing and pre-treating propolis including gold propolis, green propolis and red propolis in a resin state.

More specifically, in the preparation step of this embodiment, gold propolis, green propolis and red propolis in a resin state are mixed, immersed in an organic solvent, and separated into a lower layer and an upper layer containing wax, and the upper layer is filtered and By concentrating under reduced pressure and freeze-drying, the propolis complex in powder form can be prepared.

The organic solvent may include any known organic solvent, for example, may include ethanol, but is not limited thereto.

(2) a primary fermentation step of aging the propolis complex in a jar;

In the primary fermentation step of this embodiment, the primary fermentation is carried out in which the propolis complex prepared according to the above preparation step is put in a jar and aged. In particular, in the case of using a jar in the primary fermentation, sufficient fermentation may occur due to the activity of enzymes without artificial inoculation of microorganisms due to the unique air permeability of the jar. At this time, the primary fermentation step of this embodiment may be carried out at room temperature conditions, but is not limited thereto.

Specifically, in the primary fermentation step of this embodiment, the propolis complex is mixed with an organic solvent and water, and then put in a jar and aged for a certain period of time, thereby performing primary fermentation. The organic solvent may include all known organic solvents, for example, 1,3-butylene glycol, but is not limited thereto.

The primary fermentation of this embodiment may be carried out for 5 to 10 days, specifically 6 to 8 days. If the primary fermentation is carried out for less than 5 days or more than 10 days, the antioxidant of the fermented propolis extract and the cosmetic composition comprising the same than when the primary fermentation is carried out for 5 to 10 days , the anti-inflammatory or skin improvement effect may be insignificant.

(3) a secondary fermentation step of inoculating the first fermented fermented product with a strain and fermenting;

In the second fermentation step of this embodiment, the second fermentation is performed by inoculating the fermented product that is first fermented according to the first fermentation step with a strain applicable to fermentation.

Specifically, in the secondary fermentation step of this embodiment, the secondary fermentation may be performed by inoculating the Lactobacillus strain into the primary fermented ferment. In particular, by using a Lactobacillus strain for fermentation, the fermented propolis extract prepared according to this example contains an active ingredient in a high content, and its antioxidant, anti-inflammatory or skin improvement effect can be excellent.

More specifically, in the secondary fermentation step of this embodiment, the secondary fermentation may be performed by inoculating the first fermented fermented product with Lactobacillus plantarum . In particular, by using Lactobacillus plantarum as a fermentation strain, the fermented propolis extract prepared according to this example contains an active ingredient in a high content, and its antioxidant, anti-inflammatory or skin improvement effect is more can be excellent

Most specifically, in the secondary fermentation step of this embodiment, the secondary fermentation may be performed by inoculating the fermented product with Lactobacillus plantarum KCC-32 (Accession No.: KACC92135P). In particular, by using Lactobacillus plantarum KCC-32 (Accession No.: KACC92135P) as a fermentation strain, the content of active ingredients in the fermented propolis extract prepared according to this example is significantly increased, and its antioxidant, anti-inflammatory, or skin The improvement effect may be remarkably excellent.

The secondary fermentation of this embodiment may be carried out at a temperature of 28 to 40 ℃, specifically 30 to 35 ℃, but is not limited thereto. If, the secondary fermentation of this embodiment is carried out at less than 28 ℃ or is carried out at more than 40 ℃, than when the secondary fermentation is carried out at 28 ~ 40 ℃, the activity of microorganisms is inhibited, propolis fermentation extract and The improvement of the antioxidant, anti-inflammatory, or skin improvement effect of the cosmetic composition comprising the same may be insignificant.

The secondary fermentation of this example may be carried out at a pH condition of pH 5.5 to 6.8, specifically pH 6 to 6.5, but is not limited thereto. If, if the secondary fermentation of this example is carried out at less than pH 5.5 or at more than pH 6.8, the activity of microorganisms is inhibited than in the case of pH 5.5 to 6.8, and the fermentation extract of propolis and the cosmetic composition comprising the same The improvement in antioxidant, anti-inflammatory or skin-improving effects may be negligible.

The secondary fermentation of this embodiment may be carried out for 5 to 10 days, specifically 6 to 8 days. If the secondary fermentation is carried out for less than 5 days or for more than 10 days, the antioxidant of the fermented propolis extract and the cosmetic composition comprising the same than when the secondary fermentation is carried out for 5 to 10 days , the anti-inflammatory or skin improvement effect may be insignificant.

(4) an extraction step of extracting the second fermented fermented product to form a propolis fermented extract;

In the extraction step of this embodiment, a fermented propolis extract is formed by extracting the fermented product secondary to the secondary fermentation step.

Specifically, in the extraction step of this embodiment, the fermented product secondary fermented according to the second fermentation step is selected from the group consisting of hot water extraction method, room temperature stirring extraction method, reflux cooling extraction method, ultrasonic extraction method, high temperature pressure extraction method, and low temperature and high pressure extraction method After extraction by one or more extraction methods, the liquid component is filtered and removed to form a propolis fermentation extract.

More specifically, in the extraction step of this embodiment, the fermented product fermented secondarily according to the second fermentation step is extracted by ultrasonic extraction, and then the liquid component is filtered and removed to form a propolis fermentation extract, but limited thereto. doesn't happen In this case, the ultrasonic extraction may be performed at a temperature of 15 to 20 ℃ at a frequency of 20 to 40 kHz, but is not limited thereto.

The propolis fermentation extract obtained according to the above may be further subjected to a known post-treatment, but is not limited thereto. For example, the post-treatment may include freeze-drying, high-temperature drying, pulverization, etc., but is not limited thereto.

Thereafter, the cosmetic composition can be prepared by mixing the excipients or additives necessary for formulation of the formulation with the fermentation extract of propolis obtained according to the above.

For example, the excipients or additives may include surfactants, emulsifiers, emulsifying aids, silicone polymers, pigments, pigments, fragrances, preservatives, bactericides, oxidation stabilizers, organic solvents, thickeners, emollients, antioxidants, free radical scavengers, opacity Topical, stabilizer, emollient, silicone, antifoam, humectant, vitamin, insect repellent, preservative, anti-inflammatory, filler, polymer, propellant, basifying agent, acidifying agent, colorant, thickening agent, skin conditioning agent, whitening agent, antioxidant, anti-wrinkle agent and the like, but is not limited thereto.

The manufacturing method of the cosmetic composition of this embodiment includes all of the above 'cosmetic composition for anti-oxidation, anti-inflammatory or skin improvement containing fermented propolis extract as an active ingredient'.

Hereinafter, through Examples, Comparative Examples, and Experimental Examples, a cosmetic composition for antioxidant, anti-inflammatory or skin improvement containing the fermented propolis extract of the present invention as an active ingredient, and a method for preparing the same will be described in detail. These examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention by these examples.

[Preparation Examples 1 to 9: Preparation of Propolis Complex]

(1) Preparation of propolis complex

According to Table 1 [unit: weight ratio], three types of propolis were mixed. Thereafter, the mixture was immersed in ethanol, centrifuged to remove the lower wax phase, and the upper layer was filtered to obtain a filtrate. At this time, the total amount of the three types of propolis was the same.

The preparation of the propolis complex according to the above was carried out under the following conditions.

-Temperature: 40~50℃

- Stirring speed: 50 ~ 100 rpm

- Vacuum degree: 750 mmHg

Then, the filtrate was concentrated under reduced pressure to volatilize the solvent, and then lyophilized to prepare a propolis complex (sample).

[Examples 1 to 9: Preparation of fermented propolis extract using Lactobacillus plantarum strain]

By using the propolis complex prepared according to Preparation Examples 1 to 9 and fermenting as follows, a fermented propolis extract and a cosmetic composition comprising the same were prepared.

Example 1

(2) Primary fermentation

The propolis complex of Preparation Example 1 prepared according to the above was placed in each jar and aged for 7 days, while primary fermentation was performed. At this time, a mixture of the propolis complex, an organic solvent and water was used, and based on the total amount, 1 wt% of the propolis complex (powder), 10 wt% of 1,3-butylene glycol, and 89 wt% of water were used. It was mixed and fermented.

(3) Secondary fermentation

Lactobacillus plantarum was inoculated into each of the primary fermentation products of Preparation Example 1 formed according to the above, and secondary fermentation was performed for 7 days. At this time, the secondary fermentation was carried out while 55 g of MRS Broth medium (Difco) was added per 1 L, and left for 7 days at about 30 to 35° C. and pH 6 to 6.5 conditions.

The Lactobacillus plantarum used for the secondary fermentation was the following, and 0.5 to 1% of Lactobacillus plantarum for the primary fermentation (concentration: 2 × 10 ⁵ cfu/ml) was inoculated with

- Sales place: KCTC (Korea Biological Resources Center)

-Strain number: 3108

(4) Extraction

After ultrasonic extraction of each of the secondary fermentation products of Preparation Example 1 formed as described above, and filtering them, the propolis fermentation extract of Example 1 was finally obtained. The ultrasonic extraction was carried out for 2 hours at a temperature of 15 to 20 ℃ under a frequency condition of about 20 to 40 kHz.

The filtration was performed by ultrasonically extracting each of the secondary fermented products, and then filtering them with GFC (1-3 μm filter paper), 0.8 μm filter paper, and 0.2 μm filter paper in this order.

Examples 2 to 9

In the same manner as above, the propolis ferment extract of Examples 2 to 9 was prepared using the propolis complex of Preparation Examples 2 to 9.

- Example 2: Manufactured using the propolis composite of Preparation Example 2

- Example 3: Manufactured using the propolis composite of Preparation Example 3

- Example 4: Manufactured using the propolis composite of Preparation Example 4

- Example 5: Manufactured using the propolis composite of Preparation Example 5

- Example 6: Manufactured using the propolis composite of Preparation Example 6

- Example 7: Manufactured using the propolis composite of Preparation Example 7

- Example 8: Prepared using the propolis composite of Preparation Example 8

- Example 9: Manufactured using the propolis composite of Preparation Example 9

[Examples 10 to 18: Preparation of fermented propolis extract using Lactobacillus plantarum KCC-32 (accession number: KACC92135P)]

Example 10

Using Preparation Example 1, except that Lactobacillus plantarum KCC-32 (Accession No.: KACC92135P) was used as a fermentation strain, a propolis fermentation extract was prepared in the same manner as in Example 1.

The depository institutions of Lactobacillus plantarum KCC-32 (accession number: KACC92135P) used for the fermentation are as follows.

- Deposited institution: Agricultural Gene Resource Center, National Academy of Agricultural Sciences

Examples 11 to 18

Using Preparation Examples 2 to 9, a fermented propolis extract was prepared in the same manner as in Example 10.

- Example 11: Manufactured using the propolis composite of Preparation Example 2

- Example 12: Prepared using the propolis composite of Preparation Example 3

- Example 13: Preparation using the propolis composite of Preparation Example 4

- Example 14: Prepared using the propolis composite of Preparation Example 5

- Example 15: Prepared using the propolis composite of Preparation Example 6

- Example 16: Prepared using the propolis composite of Preparation Example 7

- Example 17: Prepared using the propolis composite of Preparation Example 8

- Example 18: Prepared using the propolis composite of Preparation Example 9

[Comparative Preparation Examples 1-3: Preparation of Propolis Complex]

(1) Preparation of propolis

It was carried out in the same manner as in Preparation Example 1, except that propolis was prepared according to Table 2 [unit: weight ratio].

Comparative Examples 1 to 3

It was carried out in the same manner as in Example 1, except that each of the propolis of Comparative Preparation Examples 1 to 3 was used.

- Comparative Example 1: Manufactured using the propolis composite of Comparative Preparation Example 1

- Comparative Example 2: Manufactured using the propolis composite of Comparative Preparation Example 2

- Comparative Example 3: Manufactured using the propolis composite of Comparative Preparation Example 3

Comparative Examples 4 to 6

It was carried out in the same manner as in Example 10, except that each of the propolis of Comparative Preparation Examples 1 to 3 was used.

- Comparative Example 4: Manufactured using the propolis composite of Comparative Preparation Example 1

- Comparative Example 5: Manufactured using the propolis composite of Comparative Preparation Example 2

- Comparative Example 6: Manufactured using the propolis composite of Comparative Preparation Example 3

[Experimental example]

Experimental Example 1: DPPH radical scavenging activity (antioxidant)

The DPPH radical scavenging ability of the propolis complex (propolis) of the Preparation Examples and Comparative Preparation Examples and the fermented propolis extract of Examples and Comparative Examples was evaluated as follows, and the results are shown in Table 3 [unit: %].

At this time, the DPPH radical scavenging ability was evaluated as follows.

First, 50 μl of a 50 mM DPPH solution was added to 100 μl of a sample, and then reacted at room temperature for about 30 minutes to measure absorbance at 517 nm. As a control, L-ascorbic acid (500 μg/ml) was used, and the DPPH radical scavenging ability of each sample was calculated by the following equation.

In the case of the control, L-ascorbic acid, DPPH radical scavenging ability of about 87.26% was shown.

DPPH radical scavenging ability (%) = (1-absorbance of no sample addition group / absorbance of no treatment group) * 100

Referring to Table 3, it can be seen that the DPPH radical scavenging ability is further improved in Examples 1 to 18 in which fermentation was performed, than in Preparation Examples 1 to 9, which is the propolis complex before fermentation. In particular, when comparing Examples 1 to 9 and Examples 10 to 18, it can be seen that when Lactobacillus plantarum KCC-32 (Accession No.: KACC92135P) is used as a fermentation strain, the effect is significantly improved.

In addition, when comparing Examples 1 to 18 and Comparative Examples 1 to 6, it can be seen that the DPPH radical scavenging ability is more excellent when three types of propolis are mixed and used. In particular, referring to Examples 1 to 18, it can be seen that the effect is further improved when the three components are included, but gold propolis, green propolis and red propolis are included in a weight ratio of 1: 2 to 4: 1. have.

Experimental Example 2: Analysis of total polyphenol content (antioxidant/anti-inflammatory)

50 μl of Folin-ciocalteu reagent was mixed with 50 μl of the sample and reacted for about 5 minutes in the dark and at room temperature, and then 50 μl of 10% Na ₂ CO ₃ solution was treated thereto. Then, after reacting at room temperature for about 1 hour, absorbance was measured at 640 nm to measure the total polyphenol content, and the results are shown in Table 4 [unit: mg/g].

At this time, Tannic acid (sigma chemical co.) was used as a standard material.

Referring to Table 4, it can be seen that the total polyphenol content is increased in Examples 1 to 18 in which the fermentation is performed, compared to Preparation Examples 1 to 9, which are the propolis complexes before fermentation. In particular, when comparing Examples 1 to 9 and Examples 10 to 18, when Lactobacillus plantarum KCC-32 (Accession No.: KACC92135P) was used as the fermentation strain, it was found that the total polyphenol content was significantly improved. have.

In addition, when comparing Examples 1 to 18 and Comparative Examples 1 to 6, it can be seen that the total polyphenol content is high when the three types of propolis are mixed and used even though the total amount of the total propolis is the same. In particular, referring to Examples 1 to 18, when the three components are included, but gold propolis, green propolis and red propolis are included in a weight ratio of 1: 2 to 4: 1, the total polyphenol content is significantly increased. can be found

Experimental Example 3: Analysis of Artepylline C Content

The sample was analyzed by high performance liquid chromatography (HPLC, YL9100 HPLC system; Younglin Instruments CO., Ltd., Gyeonggi, Korea) to measure the content of artephyllin C in the sample, and the results are shown in Table 5 [unit: ppm] shown in At this time, artepylline C was detected at a retention time of 37 minutes.

HPLC analysis conditions are as follows.

- Analytical instrument: YL 9100 HPLC

- Analysis column: YL C18-4E (4.6 x 250mm) 5μm

- Flow rate: 1.0 ml/min (injection amount: 20 μl)

- Injection concentration: 1000, 500, 250, 125, 62.5 ppm in Ethanol

- Detection conditions: UV, 314 nm

- Column temperature: 40℃

- Mobile phase: 2% Acetic acid in Acetonitrile solvent

Referring to Table 5, it can be seen that the artephyllin C content was higher in Examples 8 and 17 in which the fermentation was performed than in Preparation Example 8, which is the propolis complex before fermentation. In particular, when comparing Examples 8 and 17, it can be seen that when Lactobacillus plantarum KCC-32 (Accession No.: KACC92135P) was used as the fermentation strain, the artephilin C content was significantly increased.

In addition, when comparing Examples 8 and 17 with Comparative Examples 1 to 6, even though the total amount of propolis is the same, it can be seen that the content of artepylline C is high when three types of propolis are mixed and used. .

Experimental Example 4: NO production inhibitory ability

Nitric oxide (NO) content was measured by the amount of nitric oxide (NO) produced in the supernatant of the cell. The nitrite reduced with nitrate was used as a griess reagent (Sigam, USA), and 2 X 10 ⁶ cells in a 6 well plate were washed twice with PBS when the confluence was 80%, and then using a serum-free medium. Incubated for more than 12 hours, and reacted by treatment with LPS (lipopolysaccharide) 10 ㎍ / ml. Then, after about 2 hours, a sample (treatment concentration: 1%) was treated and tested.

After 24 hours, the supernatants were collected and reacted with griess reagent for 30 minutes, and then the absorbance was measured at 540 nm.

Referring to Table 6, it can be seen that the NO production inhibitory ability is improved in the cases of Examples 1 to 18, which were fermented, than in Preparation Examples 1 to 9, which are the propolis complexes before fermentation. In particular, when comparing Examples 1 to 9 and Examples 10 to 18, when Lactobacillus plantarum KCC-32 (Accession No.: KACC92135P) was used as a fermentation strain, it can be seen that the NO production inhibitory effect was significantly improved. have.

In addition, when comparing Examples 1 to 18 and Comparative Examples 1 to 6, it can be seen that the NO production inhibitory ability is improved when the three types of propolis are mixed and used even though the total amount of the total propolis is the same. In particular, referring to Examples 1 to 18, it can be seen that, when the three components are included, but gold propolis, green propolis and red propolis are included in a weight ratio of 1: 2 to 4: 1, the NO production inhibitory ability is significantly improved. can

Experimental Example 5: TNF-α activity inhibitory ability

In a 96 well plate, 1 X 10 ⁴ cells/well of fibroblasts (Fibroblast HS86) were cultured for 24 hours. Thereafter, it was replaced with a serum-free medium and treated with 100 mM H ₂ O ₂ for 30 minutes, and then the samples were treated by concentration and cultured for about 24 hours.

At this time, as a positive control, dexamethasone was treated at concentrations of 0.1%, 0.5% and 1.0%. The concentration (%) of TNF-α in the culture solution treated with the sample (fermented propolis extract) was analyzed compared to the untreated comparative culture solution.

At this time, the TNF-α was measured by ELISA method, and the results for the TNF-α inhibition rate (%) are shown in Table 7 [unit: %].

Referring to Table 7, it can be seen that, according to this example, the TNF-α inhibitory effect is remarkably excellent.

Experimental Example 6: Evaluation of cell protection

The protective effect on the cells of the sample by oxidation was analyzed by applying the MTT assay method.

This is a method of measuring the conversion to formazan by mitochondrial dehydrogenases, and 1 X 10 ⁴ cells/well of fibroblasts (Fibroblast HS86) were cultured for 24 hours in a 96 well plate. Thereafter, it was replaced with a serum-free medium and cultured for 24 hours, followed by reaction with 100 mM H ₂ O ₂ for 30 minutes. Then, the sample was treated by concentration and treated with 20 μl of MTT solution per well, and then reacted for 4 hours at 37 ° C under 5% CO ₂ conditions, and then the change in absorbance at 490 nm using a microplate reader was measured, and the control Cell viability was expressed as a percentage, and the results are shown in Table 8 [unit: %].

Referring to Table 8, when comparing Example 17 with Comparative Examples 4 to 6, it can be seen that the cytoprotective effect is improved when the three types of propolis are mixed and used even though the total amount of the total propolis is the same.

[Formulation Example]

Formulation Example 1: Cream

According to Table 9 [unit: wt%], a cosmetic composition comprising the fermented propolis extract of Examples and Comparative Examples was prepared.

Formulation Example 2: Serum

According to Table 10 [unit: wt%], a cosmetic composition comprising the fermented propolis extract of Examples and Comparative Examples was prepared.

Experimental Example 7: Formulation stability evaluation of cosmetic compositions

The stability of the cosmetic composition (cream formulation) prepared according to the above was evaluated, and the results are shown in Table 11.

The stability was evaluated by visually observing the formulation stability of the cosmetic composition after leaving it at each temperature for 80 days, and the results are shown in Table 11.

Specifically, if there was no layer separation of the formulation and there was no discoloration or discoloration, it was marked as 'none', and if there was layer separation, discoloration or discoloration of the formulation, it was marked as 'change'.

Referring to Table 11, it can be seen that the formulation is stable according to this example.

Experimental Example 8: Evaluation of the skin improvement effect of the cosmetic composition

The cosmetic composition (cream formulation) prepared according to the above was directly applied to the skin, and the degree of skin condition improvement was evaluated according to the following, and the results are shown in Tables 12 [unit: score] and 13.

In the experiment, each group (composed of 20 women) was applied immediately after face-washing, twice a day, for about 2 weeks, and the skin moisturizing improvement effect was evaluated on a scale of 1 to 5, and the average value is shown in Table 12. It was. At this time, the closer to 5 points, the better the skin moisturizing effect.

In addition, the skin softness improvement effect was evaluated on a scale of 1 to 5 in the same manner as above, and the average value thereof is shown in Table 13. At this time, the closer to 5 points, the better the skin softness improvement effect.

Referring to Tables 12 and 13, it can be seen that, according to this example, the skin moisturizing improvement effect and the skin softness improvement effect are excellent. That is, according to this embodiment, it means that the skin condition is improved by providing moisture to the skin and softening the skin surface.

Referring to Experimental Examples 1 to 8, it can be seen that the fermented propolis extract according to this example has excellent DPPH radical scavenging ability and has an antioxidant effect as it contains polyphenol and artephyllin C in high content.

In addition, it can be seen that the fermentation extract of propolis according to this example has an anti-inflammatory effect as it contains polyphenols and artefylline C in high content while inhibiting NO production and TNF-α activity.

In addition, the fermented propolis extract according to this example has excellent DPPH radical scavenging ability, NO production inhibitory ability, and TNF-α activity inhibitory activity, and contains active ingredients such as polyphenol and artephyllin C in a high content, improving skin condition it can be seen that

That is, when the above contents are put together, it can be seen that the fermented propolis extract according to this embodiment has excellent antioxidant, anti-inflammatory and skin improvement effects without cell damage.

As described above, the description of the present invention is for illustration, and those of ordinary skill in the art to which the present invention pertains can easily transform into other specific forms without changing the technical spirit or essential features of the present invention. You will understand. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. For example, each component described as a single type may be implemented in a distributed manner, and likewise components described as distributed may also be implemented in a combined form.

The scope of the present invention is indicated by the following claims rather than the above detailed description, and all changes or modifications derived from the meaning and scope of the claims and their equivalent concepts should be interpreted as being included in the scope of the present invention. do.

Claims (8)

A cosmetic composition for antioxidant, anti-inflammatory or skin improvement, comprising as an active ingredient a fermented propolis extract obtained by fermenting and extracting a propolis complex containing gold propolis, green propolis and red propolis,
The propolis complex is
The cosmetic composition comprising the gold propolis, green propolis and red propolis in a weight ratio of 1: 2 to 4: 0.5 to 2.
delete The method of claim 1,
The fermentation is
A cosmetic composition for antioxidant, anti-inflammatory or skin improvement, characterized in that the propolis complex is fermented with a Lactobacillus strain.
4. The method of claim 3,
The Lactobacillus strain is
Lactobacillus plantarum ( Lactobacillus plantarum ) Antioxidant, anti-inflammatory or cosmetic composition for skin improvement, characterized in that it contains.
The method of claim 1,
The skin improvement
A cosmetic composition for antioxidation, anti-inflammatory or skin improvement, characterized in that it is skin moisturizing or skin softness improvement.
The method of claim 1,
The cosmetic composition is
A cosmetic composition for antioxidant, anti-inflammatory or skin improvement, comprising 0.01 to 30% by weight of the fermented propolis extract based on the total weight.
The method of claim 1,
The cosmetic composition is
One formulation selected from the group consisting of softening lotion, nourishing lotion, astringent lotion, skin, lotion, essence, serum, cream, massage cream, pack, makeup base, BB cream, foundation, powder, cleansing foam, cleansing cream and cleansing water A cosmetic composition for antioxidant, anti-inflammatory or skin improvement, characterized in that
A preparatory step of preparing a propolis complex containing gold propolis, green propolis and red propolis;
A primary fermentation step of aging the propolis complex in a jar;
a secondary fermentation step of inoculating the first fermented fermented product with a strain and fermenting it; and
An extraction step of extracting the secondary fermented fermented product to form a propolis fermented extract As a method,
The propolis complex is
The method comprising the gold propolis, green propolis and red propolis in a weight ratio of 1: 2 to 4: 0.5 to 2.