KR102334193B1 - Composition for skin cleansing comprising fermented extract of soapberry and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a composition for cleaning skin or hair containing a fermented product of a soapberry extract as an active ingredient, and a manufacturing method thereof, and more specifically, to a cosmetic composition for cleaning skin or hair containing a fermented product of a soapberry extract as an active ingredient; and a method for manufacturing a fermented product of a soapberry extract for cleaning skin or hair comprising: a step of manufacturing the soapberry extract, and a step of fermenting the manufactured soapberry extract. The composition is provided to ferment the soapberry extract through microorganisms, thereby providing excellent cleaning power by enhancing a content of glycolipid which is a biological surfactant and being used as a safe and effective skin or hair cleaner.

Description

Composition for cleaning skin containing fermented product of soapberry extract as an active ingredient and method for manufacturing the same

The present invention relates to a composition for cleaning skin or hair containing a fermented product of soapberry extract as an active ingredient and a method for preparing the same.

Soapberry ( Sapindus mukurossi), also known as the sick-free tree, is found in Korea, China, Taiwan, Japan, and India. Soapberry reaches 20m in height, branches are hairless and brown, and the leaves are alternate phyllotaxis and pinnate compound leaves once. Small leaves are 9-13, long oval-shaped, egg-shaped or long oval-shaped lancets, with flat edges, with wrinkles on the back and no hairs. The small petiole is 2-6mm, and the flowers are monovalent, light yellow-green, bloom in June, have a diameter of 4-5mm, and hang on the panicle. The bark of soapberry fruit contains a lot of saponin, and in folklore, the bark was used as an expectorant.

Regarding surfactants used as detergents, attempts to use biosurfactants instead of synthetic surfactants, which have harmful or side effects, are increasing. Biosurfactants have been defined as surface-active biomolecules produced by microorganisms with a wide range of applications, but have recently been defined to include all surfactants made using renewable natural plant materials.

Biomolecules produced by microorganisms include glycolipids, phospholipids, fatty acids, lipopeptides, or lipoproteins. As a linkage, it maintains the stability of cell membranes and plays a role in promoting immune responses, cell connections, and cell recognition. Research on the production of such glycolipids through microorganisms is in progress, and the glycolipids produced accordingly have advantages of low toxicity and excellent biodegradability.

Enterococcus faecalis (E. faecalis ) is a gram-positive cocci belonging to the genus Enterococcus, which resides mainly in the digestive tract of humans or mammals. It is also a hygienic factor that affects the shelf life of food. E. faecalis is a nonmotile microorganism that ferments glucose without gas production and does not undergo catalase reaction with hydrogen peroxide. This shows consistent growth on nutrient media consistent with being facultative anaerobic. Metabolizes a variety of energy sources, including glycerol, lactic acid, malic acid, citric acid, arginine, agmatine and many keto acids. In particular, E. faecalis possesses glycosyltransferases (GTs) enzymes capable of producing glycolipids, which link saccharides with lipid molecules and cells that respond to the presence of glycolipids on the cell surface. It is responsible for assembling the correct oligosaccharides so that the right receptor can be activated.

There is a need for a technology that can use surfactants that are used in various ways in our daily life in a safer and environmentally friendly way by utilizing these natural substances.

Republic of Korea Patent Publication No. 10-2013-0078929 (published on July 10, 2013)

An object of the present invention is to provide a composition for cleaning the skin using a fermented product of a natural product that is safe and has excellent cleaning power.

Another object of the present invention is to provide a method for producing a fermented product of a natural product that can be used for washing.

In order to achieve the above object, the present invention provides a cosmetic composition for cleaning skin or hair containing a fermented product of soapberry extract as an active ingredient.

In addition, the present invention comprises the steps of preparing a soapberry extract; and fermenting the prepared soapberry extract.

The composition for cleaning skin or hair according to the present invention can be used as a more effective skin or hair cleanser by fermenting the extract of soapberry through microorganisms to increase the content of glycolipids, which is a biosurfactant, and thereby exhibit excellent cleaning power.

The composition is prepared by using natural products and microorganisms, so it is safe to the human body and has less toxicity and side effects, and an environmentally friendly detergent can be prepared.

1 is a graph showing the cytotoxicity test results of Sopberry Fermented Examples and Comparative Examples according to an experimental example of the present invention.
Figure 2 shows the results of the detergency test of the fermented soapberry according to an experimental example of the present invention.

Hereinafter, the present invention will be described in detail.

The present invention provides a cosmetic composition for cleaning skin or hair containing a fermented product of soapberry extract as an active ingredient.

In the present specification, "soapberry (soapberry, Sapindus mukurossi)", as described above, is a fruit opened from a soapberry tree growing in the Himalayas wild, also called a sick-free tree, also called a soapnut or soapnut. It has a slightly acidic pH (pH 5.5-6.0) similar to the pH of our skin, and is rich in saponin components.

As used herein, the term "extract" refers to a substance obtained by extracting a component of a natural product regardless of an extraction method, an extraction solvent, an extracted component, or the form of an extract. It may include any material obtainable by processing or processing by the method.

In the present invention, the soapberry extract may be a soapberry extracted with water, C1 to C4 alcohol or a mixed solvent thereof, and preferably may be extracted with water, ethanol, or an aqueous ethanol solution, but is not limited thereto. .

The soapberry extract may be extracted according to a method commonly used in the art, for example, hot water extraction, ultrasonic extraction, filtration, reflux extraction, etc., which are performed alone or in combination of two or more methods can be performed.

Preferably, the soapberry extract may be a mixed extract of a soapberry ethanol extract and a soapberry hot water extract.

More specifically, the soapberry ethanol extract is mixed with soapberry and a 50% by weight aqueous ethanol solution in a weight ratio of 1: (8 to 12), more preferably 1:10, 40 to 60° C., more preferably may be extracted at 50° C. for 30 minutes to 2 hours, more preferably for 1 hour.

The soapberry hot water extract is obtained by mixing soapberry and purified water in a weight ratio of 1: (3 to 7), more preferably 1:5, at 90 to 110° C., more preferably at 100° C., 5 to 30 min, more preferably, it may be extracted for 15 min.

The mixed extract is obtained by mixing the soapberry ethanol extract and hot water extract in a weight ratio of 1: (8 to 12), more preferably 1:10 by weight, at 50 to 70° C., more preferably at 60° C., for 30 minutes. to 2 hours, more preferably, it may be extracted for 1 hour.

More details will be described later in the following experimental examples.

In the present invention, the fermented product of the soapberry extract may be a fermented product of the soapberry extract with Enterococcus faecalis NCK03 (Accession No.: KFCC11876P) strain.

The strain has a glycosyltransferases (glycosyltransferases, GTs) enzyme, it is possible to produce glycolipids (glycolipids), and thus produced glycolipids have the advantage of low toxicity and excellent biodegradability.

The fermented product fermented with the strain may be further fermented with glycolipid synthetase, but is not limited thereto.

Accordingly, the fermented product of the fermented soapberry extract has a high glycolipid content and has excellent cleaning power, so it can be used as a cosmetic composition for cleaning skin or hair including hands, body, and scalp.

As used herein, the term "cosmetics" is used in the human body to clean and beautify the human body to add attractiveness, to brighten the appearance, or to maintain or promote skin and hair health, and according to the present invention, infants and young children (only It refers to cosmetics emphasizing the effects of bathing and washing (for children under 3 years of age), bathing, body washing, and hair washing.

The cosmetic composition may further include one or more of the ingredients listed in the list of cosmetic raw materials registered with the Ministry of Food and Drug Safety and ICID (International Cosmetic Ingredients). For example, the cosmetic composition may include an organic solvent, a solubilizer, a thickener, a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a fragrance, a surfactant, an emulsifier, a filler, a sequestering agent, a preservative, a vitamin, a blocking agent, It may further comprise one or more adjuvants commonly used in the cosmetic field, such as wetting agents, dyes and pigments, hydrophilic or lipophilic actives, or any other ingredients commonly used in cosmetics.

The vitamin may include a water-soluble or oil-soluble vitamin, and the water-soluble vitamin is a water-soluble vitamin that can be formulated in cosmetics, preferably vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, and nicotinic acid amide, folic acid, vitamin C, vitamin H, and the like, and more preferably vitamin C, which helps collagen synthesis. In addition, salts thereof (thiamine hydrochloride, sodium ascorbate salt, etc.) or derivatives (ascorbic acid-2-phosphate sodium salt, ascorbic acid-2-phosphate magnesium salt, etc.) may be included, and the water-soluble vitamin is a microorganism transformation method , can be obtained by a conventional method such as a purification method from a culture of a microorganism, an enzymatic method or a chemical synthesis method.

The cosmetic composition may further include a skin, scalp or hair protectant, an active agent, and the like in order to enhance the cleaning effect and minimize side effects.

The cosmetic composition may be prepared in any formulation conventionally prepared in the art to which the present invention pertains. For example, it may be formulated as a lotion, emulsion, lotion, cream, paste, gel, solution, suspension, oil, wax, pack, powder, foundation, spray, surfactant-containing cleansing products, etc., but is limited thereto. no.

Preferably, it may be prepared in formulations such as cleansing cream, cleansing lotion, cleansing foam, cleansing water, solid soap, liquid soap, stick-type product, balm-type product, cleansing tissue, spray, etc., but is limited thereto no.

When the cosmetic composition is in a cream or gel formulation, animal oil, vegetable oil, wax, paraffin, starch, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be further included as a carrier component.

When the cosmetic composition is a solution or emulsion formulation, a solvent, solvating agent or emulsifying agent, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, propylene glycol, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid It may further include esters and the like.

When the cosmetic composition is a suspension formulation, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be further included.

When the cosmetic composition is in a powder or spray formulation, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be further included as a carrier component, and in particular, in the case of a spray formulation, additional chlorofluorohydrocarbon , may further include a propellant such as propane/butane or dimethyl ether.

When the cosmetic composition is a surfactant-containing cleansing formulation, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glycerides, fatty acid diethanolamide, vegetable oil, linoline derivatives or ethoxylated glycerol fatty acid esters may be further included.

The cosmetic composition may be used alone or by being applied in duplicate, or may be used by overlapping with other cosmetic compositions other than the present invention. In addition, the cosmetic composition may be used according to a conventional method of use, and the number of times of use may be varied according to the skin condition or taste of the user.

In addition, the present invention provides a method for preparing a fermented product of soapberry extract for cleaning skin or hair.

The manufacturing method comprises the steps of preparing a soapberry extract; and fermenting the prepared soapberry extract.

In the preparation method according to the present invention, the step of preparing the soapberry extract may be performed by extracting the soapberry with water, C1 to C4 alcohol or a mixed solvent thereof, but is not limited thereto.

Preferably, the preparing of the soapberry extract comprises the steps of preparing an ethanol extract of soapberry; preparing a soapberry hot water extract; and preparing a mixed extract by mixing the ethanol extract and the hot water extract.

In the step of preparing the soapberry ethanol extract, soapberry and 50% by weight of an aqueous ethanol solution are mixed in a weight ratio of 1: (8 to 12), more preferably 1:10, 40 to 60° C., more preferably may be carried out by extracting at 50 ° C. for 30 minutes to 2 hours, more preferably for 1 hour, and then cooling at 0 to 10 ° C., more preferably at 4 ° C.

In the step of preparing the hot water extract of soapberry, soapberry and purified water are mixed in a weight ratio of 1: (3 to 7), more preferably 1:5, at 90 to 110°C, more preferably at 100°C In, after extraction for 5 to 30 minutes, more preferably for 15 minutes, cooling at 0 to 10 °C, more preferably at 4 °C, for 20 to 40 minutes, more preferably for 30 minutes.

In the step of preparing the mixed extract, the prepared soapberry ethanol extract and hot water extract are mixed in a weight ratio of 1: (8 to 12), more preferably 1:10, 50 to 70° C., more preferably is carried out by extracting at 60° C. for 30 minutes to 2 hours, more preferably for 1 hour, and then cooling at 0 to 10° C., more preferably at 4° C., for 30 minutes to 2 hours, more preferably for 1 hour. can

Each of the extract steps may be repeated once or twice or more, but is not limited thereto.

In the manufacturing method according to the present invention, the step of fermenting the prepared soapberry extract includes: primary fermentation of the soapberry extract with Enterococcus faecalis NCK03 (Accession No.: KFCC11876P) strain; and secondary fermentation with glycolipid synthetase after the primary fermentation.

Preferably, in the step of primary fermentation, the prepared soapberry extract is inoculated with the strain and fermented at 30 to 40° C., more preferably at 37° C., for 3 to 7 days, more preferably for 5 days. can be performed.

The second fermentation step may be performed by adding glycolipid synthase to the first fermented product and fermenting at 30 to 40° C., more preferably at 37° C., for 5 to 10 days, more preferably for 7 days. .

More details will be described later in the following experimental examples.

Hereinafter, examples will be described in detail to help the understanding of the present invention. However, the following examples are merely illustrative of the content of the present invention, and the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art.

<Experimental Example 1> Enterococcus faecalis NCK03 ( Enterococcus faecalis Isolation of NCK03)

The strain is isolated from cheese, and in order to isolate bacteria with efficacy, the cheese is stationary cultured in RC (reinforced clostridial) medium (BD), and then 10 ^-2 , 10 ^-3 , 10 ^{-4 by a decimal dilution method.} , ^{was diluted to 10 -5} to isolate the strain. The isolated strain was peptone 10.0g/L, sodium chloride 5.0g/L, beef extract 10.0g/L, yeast extract 3.0g/L, dextrose ( dextrose) 5.0 g/L, starch 1.0 g/L, L-cysteine hydrochloride 0.5 g/L, sodium acetate 3.0 g/L, and culture medium for 3 days did

Strains grown in RC medium were subjected to rRNA sequence analysis to analyze the base sequence. From gDNA obtained by chromosomal DNA extraction, PCR was performed using universal primers 785F (5' GGA TTA GAT ACC CTG GTA 3') and 907R (5' CCG TCA ATT CMT TTR AGT TT 3') to perform nucleotide sequence was analyzed. The homology test was performed through BLAST search provided by NCBI. Was identified with the base sequence strains are shown to Enterococcus faecalis (Enterococcus faecalis) and 99.36% similarity belonging to Enterococcus (Enterococcus) genus (genus).

The sequence of the strain is as follows.

Enterococcus faecalis NCK03 (Accession No.: KFCC11876P):

<Experimental Example 2> Preparation of soapberry extract

Soapberries were washed and dried. In one embodiment, the drying was carried out by hot air drying so that the mass of moisture compared to soapberry was within 5%, and in the shredding step, it was shredded to a size of 0.1 mm. Thereafter, the soapberry was extracted in order to use the soapberry extract as a substitute for purified water.

First, soapberry and 50% ethanol were mixed in a ratio of 1:10, extracted at 50°C for 1 hour, and cooled at 4°C (step 1). In addition, soapberry was extracted with purified water at a ratio of 1:5 at 100° C. for 15 minutes, followed by cooling at 4° C. for 30 minutes, repeated three times (step 2). Finally, the extracts of Step 1 and Step 2 were mixed in a ratio of 1:10, extracted at 60° C. for 60 minutes, and cooled at 4° C. for 60 minutes, followed by extraction twice.

<Experimental Example 3> Preparation of Soapberry Fermented Product

Enterococcus faecalis NCK03 (Enterococcus faecalis NCK03) (Accession No.: KFCC11876P) in 7% of glucose and 1% of yeast extract in the soapberry extract obtained in Experimental Example 2 ^{3 × 10 5} to fit the number After inoculation, primary fermentation was performed at 37°C for 5 days. After 5 days of fermentation, 1 μg/L of glycolipid synthetase was added, and secondary fermentation was performed at 37° C. for 7 days. Glycolipid synthase was purchased from Merck (SAE0093-50UG).

After the second fermentation was completed, the strain was disrupted by ultrasonication and the strain was removed by a centrifuge, and then the first filter was performed using a 5 μm filter. After that, the second filter was performed once more with a 0.45 μm filter to proceed with sterilization.

Table 1 below shows Examples and Comparative Examples under various conditions.

In the following experimental examples, cytotoxicity analysis, glycolipid quantitative analysis, and detergency evaluation of Examples and Comparative Examples according to the conditions of Table 1 were performed.

Extraction method 1st fermentation 2nd fermentation Example 1 ethanol extraction ○ ○ Example 2 hot water extraction ○ ○ Example 3 Ethanol extraction + hot water extraction ○ ○ Example 4 ethanol extraction ○ × Example 5 hot water extraction ○ × Example 6 Ethanol extraction + hot water extraction ○ × Comparative Example 1 ethanol extraction × × Comparative Example 2 hot water extraction × × Comparative Example 3 Ethanol extraction + hot water extraction × ×

<Experimental Example 4> Efficacy test of fermented soapberry

4-1. Cytotoxicity assay

Keratinocytes (ATCC) were counted using a hemocytometer, and then dispensed in a 96-well plate at a ^{concentration of 1×10 4} cells/well. After culturing for 24 hours, the test substance or comparative substance of Example 1 was added to each well at a total concentration of 100 μg/mL to keratinocytes, and 100 μL of DMEM medium containing 5% penicillin/streptomycin was used at 37° C. , 5% CO ₂ Incubated for 24 hours in an incubator. Thereafter, the supernatant was removed and a mixture of DMEM medium and WST solution (EZ-cytox) in a weight ratio of 9:1 was added, followed by _{reaction in an incubator at 37° C., 5% CO 2} for 2 hours, and absorbance at 450 nm measured. The degree of cytotoxicity (cell viability) was expressed as a percentage (%) based on the absorbance intensity of the control group using the same solvent as the test substance and the comparative substance.

[Formula 1]

Cell viability (%) = Absorbance of sample material addition group / Absorbance of control group × 100

As a result, as shown in FIG. 1 , Examples 1 to 6 had no cytotoxicity compared to the untreated group, but Comparative Examples 1 to 3 had cytotoxicity compared to the untreated group.

4-2. Quantitative analysis of glycolipids

A glycolipid (glycolipid) was dissolved in chloroform (chloroform, 0.2 mg/mL), and 20 μL was injected into normal-phase HPLC (YL9100 plus) and analyzed. The same HPLC and column as for phospholipid analysis were used, and the mobile phase was gradient elution with solvent (A): chloroform and solvent (B): methanol: water (95:5, v/v) of 99:1 (v/v). After flowing in the ratio for 15 minutes, the ratio of 75:25 (v/v) for 5 minutes, the ratio of 10:90 (v/v) for 5 minutes, and the last 10 at the ratio of 99:1 (v/v) Minutes were flowed for analysis. A standard material, digalactosyl diacylglycerol, was purchased from Sigma-Aldrich Co. (St. Louis, MO, USA) and used. Analysis software was analyzed using YL-Clatiry.

As a result, as shown in Table 2 below, in Example 3, the glycolipid content was measured to be the highest at 31.27 ppm.

Glycolipid content (ppm) Example 1 26.18 Example 2 22.95 Example 3 31.27 Example 4 15.49 Example 5 12.82 Example 6 17.41 Comparative Example 1 6.58 Comparative Example 2 2.61 Comparative Example 3 7.98

4-3. cleaning power evaluation

20 mg each of abietic acid (Sigma-Aldrich) used as a contaminant was applied to one side of the slide glass. After that, 300 μL of the detergency test material and the comparative material were placed on a slide glass coated with contaminants, and then the detergency was evaluated.

As a positive control, 70% ethanol was used. For evaluation of cleaning power, the change in the number of particles remaining on the slide glass before and after cleaning was confirmed with an optical microscope (KI400). Using Optiview software, particles with a diameter of 100 to 300 μm were selected and the number was measured.

As a result, as shown in FIG. 2 and Table 3, Example 3 had the smallest number of residual particles.

Residual particle count Before cleaning (Control) TNTC Example 1 92 Example 2 107 Example 3 84 Example 4 203 Example 5 227 Example 6 209 Comparative Example 1 343 Comparative Example 2 382 Comparative Example 3 309 Positive control (70% ethanol) 27

As described above in detail a specific part of the content of the present invention, for those of ordinary skill in the art, it is clear that this specific description is only a preferred embodiment, and the scope of the present invention is not limited thereby. do. That is, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Korea Microorganism Conservation Center (domestic) KFCC11876P 202110105

Claims (13)

Contains fermented product of soapberry extract as an active ingredient,
The fermented product is
A cosmetic composition for cleaning skin or hair, characterized in that the soapberry extract is fermented with Enterococcus faecalis NCK03 (Accession No.: KFCC11876P) strain. The method of claim 1,
The soapberry extract is
A cosmetic composition for cleaning skin or hair, characterized in that soapberry is extracted with water, C1 to C4 alcohol, or a mixed solvent thereof. The method of claim 1,
The soapberry extract is
A cosmetic composition for cleaning skin or hair, characterized in that it is a mixed extract of soapberry ethanol extract and soapberry hot water extract. delete The method of claim 1,
The fermented product is
After fermentation of the strain, a cosmetic composition for cleaning skin or hair, characterized in that it is further fermented with glycolipid synthetase. preparing a soapberry extract; and
Including; fermenting the prepared soapberry extract;
The step of fermenting the prepared soapberry extract,
First fermentation of the soapberry extract with Enterococcus faecalis NCK03 (Accession No.: KFCC11876P) strain; and
After the first fermentation, a second fermentation with glycolipid synthase; 7. The method of claim 6,
The step of preparing the soapberry extract,
preparing an ethanol extract of soapberry;
preparing a soapberry hot water extract; and
Preparing a mixed extract by mixing the ethanol extract and the hot water extract; 8. The method of claim 7,
The soapberry ethanol extract,
Skin or hair washing, characterized in that it is prepared by mixing soapberry and 50 wt% ethanol aqueous solution in a weight ratio of 1: (8 to 12) and extracting at 40 to 60 ° C for 30 minutes to 2 hours and then cooling at 0 to 10 ° C. A method for producing a fermented product of dragon soapberry extract. 8. The method of claim 7,
The soapberry hot water extract,
Skin or hair washing, characterized in that it is prepared by mixing soapberry and purified water in a weight ratio of 1: (3 to 7) and extracting it at 90 to 110° C. for 5 to 30 minutes and then cooling it at 0 to 10° C. for 20 to 40 minutes. A method for producing a fermented product of dragon soapberry extract. 8. The method of claim 7,
The mixed extract is
The ethanol extract and the hot water extract prepared above are mixed in a weight ratio of 1: (8 to 12), extracted at 50 to 70° C. for 30 minutes to 2 hours, and then cooled at 0 to 10° C. for 30 minutes to 2 hours, characterized in that , A method for preparing a fermented product of soapberry extract for cleaning skin or hair. delete 7. The method of claim 6,
The first fermentation step is,
A method for producing a fermented product of a soapberry extract for skin or hair washing, characterized in that the prepared soapberry extract is inoculated with the strain and fermented at 30 to 40° C. for 3 to 7 days. 7. The method of claim 6,
The second fermentation step is,
A method for producing a fermented product of soapberry extract for skin or hair washing, characterized in that the glycolipid synthetase is added to the primary fermented product and fermentation is performed at 30 to 40° C. for 5 to 10 days.

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