WO2022119074A1 - Method for preparing cosmetic composition comprising fermented fig alcohol extract, and cosmetic composition prepared thereby - Google Patents

Abstract

The present invention relates to a method for preparing a cosmetic composition comprising a fermented fig alcohol extract, and a cosmetic composition prepared thereby, the method comprising: a first step of obtaining an extract from fig leaves and figs by means of 70% alcohol; and a second step of obtaining a fermented product by fermenting the extract by means of a nine-type lactobacillus mixture or one type of yeast. The 70% alcohol can preferably be 70% ethanol. By fermenting an extract which has been extracted from fig leaves and figs by means of alcohol, the anti-inflammatory activity and whitening activity of the extract can be enhanced.

Description

Method for manufacturing a cosmetic composition comprising a fermented product of fig alcohol extract and a cosmetic composition prepared therefrom

The present invention relates to a method for manufacturing a cosmetic composition and a cosmetic composition prepared therefrom, and to a method for manufacturing a cosmetic composition comprising a fermented product of an alcohol extract of figs and a cosmetic composition prepared therefrom.

Figs are known as non-flowering fruits. Since fig flowers bloom inside the actual fruit, it is only impossible to identify them from the outside, but perhaps for this reason, it is not easy to encounter cases where figs are found as an aesthetic object.

However, figs contain various components such as niacin, sodium, protein, carbohydrate, retinol, beta-carotene, vitamins A, B1, B2, B6, C, E, dietary fiber, zinc, folic acid, phosphorus, lipid, iron, potassium, and calcium. As it contains, it is easy to come across cases where you want to use the ingredients contained in figs nutritionally or pharmacologically.

Recently, there are also cases of extracting fig ingredients and applying them to cosmetics. Korean Patent Application Laid-Open No. 10-2020-0114810 discloses an invention related to a method for preparing a cosmetic composition comprising an alcohol extract of fig leaf, and the alcohol extract of fig leaf is disclosed to have antioxidant, anti-inflammatory and whitening activity.

The present invention aims to devise a method that can improve anti-inflammatory performance and whitening performance using fig alcohol extract.

The present invention, as a method for producing a cosmetic composition comprising a fermented product of a fig alcohol extract, the first step of obtaining an extract of fig leaves and fruits using 70% alcohol, and the extract using 9 types of mixed lactic acid bacteria and a second step of fermenting to obtain a fermented product. The 9 types of mixed lactic acid bacteria are composed of the following lactic acid bacteria, and 70% alcohol is 70% ethanol.

1) Lactobacillus plantarum ( Lactobacillus plantarum )

2) Lactobacillus rhamnosus ( Lactobacillus rhamnosus )

3) Lactobacillus casei ( Lactobacillus casei )

4) Lactobacillus paracasei ( Lactobacillus paracasei )

5) Lactobacillus fermentum ( Lactobacillus fermentum )

6) Lactobacillus acidophilus ( Lactobacillus acidophilus )

7) Streptococcus thermophiles ( Streptococcus thermophiles )

8) Bifidobacterium longum ( Bifidobacterium longum )

9) Bifidobacterium bifidum ( Bfidobacterium bifidum )

The fermented product includes rutin and quercetin, and the second step is a step of increasing the amount of quercetin included in the extract by decomposing rutin included in the extract.

The present invention, comprising a fermented product of an ethanol extract of fig leaves and fruits, wherein the rutin is contained in an amount less than the amount of rutin contained in the ethanol extract of fig leaves and fruits, and in the ethanol extract of fig leaves and fruits It is possible to provide a cosmetic composition comprising a greater amount of quercetin than the amount of quercetin included.

On the other hand, the present invention, the first step of obtaining an extract of fig leaves and fruits using 70% alcohol and Saccharomyces cerevisiae fermenting the extract using Saccharomyces cerevisiae to obtain a fermented product It involves two steps. Here, 70% alcohol is 70% ethanol.

The fermented product includes rutin and quercetin, and the second step is a step of decomposing rutin and quercetin contained in the extract.

The present invention can provide a cosmetic composition comprising a fermented product of an ethanol extract of fig leaves and fruits, and comprising rutin and quercetin in an amount smaller than the amount of rutin and quercetin contained in the ethanol extract of fig leaves and fruits. there is

The present invention improves anti-inflammatory and whitening performance by applying a fermented product of an alcoholic extract of fig leaves and fruits (hereinafter, “fig leaves and fruits” may be referred to as “fig leaves/fruits”) to a cosmetic composition can do it

1 is a flowchart of a method for manufacturing a cosmetic composition comprising a fermented product of an alcohol extract of figs according to the present invention.

Figure 2 shows the cell viability when the extraction medium for fig leaf/fruit extraction was hot water, 50% ethanol, 70% ethanol, and 1,3-butylene glycol (1,3-BG), respectively.

3 shows the anti-inflammatory performance when the extraction medium for fig leaf/fruit extraction was hot water, 50% ethanol, 70% ethanol, and 1,3-butylene glycol (1,3-BG), respectively.

Figure 4 shows the cell viability of the extract when the fig leaf / fruit is extracted using 70% ethanol and the fermented product when the extract is fermented with 9 types of mixed lactic acid bacteria.

5 shows the extract when fig leaves/fruits are extracted using 70% ethanol and the anti-inflammatory performance of the fermented product when the extract is fermented with 9 types of mixed lactic acid bacteria.

6 shows the extract when fig leaves/fruits are extracted using 70% ethanol and the whitening performance of the fermented product when the extract is fermented with 9 types of mixed lactic acid bacteria.

7 is a chromatogram of a standard product, FIG. 7(A) is a chromatogram of rutin, and FIG. 7(B) is a chromatogram of quercetin.

8 is a TIC (Total Ion Chromatogram) result of each sample, FIG. 8(A) is a TIC result of a 70% ethanol extract, FIG. 8(B) is a TIC result of a lactic acid bacteria fermented product of a 70% ethanol extract, FIG. 8( C) is a TIC result of a malt fermented product of 70% ethanol extract, and FIG. 8(D) is a TIC result of a yeast fermented product of 70% ethanol extract.

9 is an EIC (Extracted Ion Chromatogram) result of 70% ethanol extract, FIG. 9(A) is an EIC result of rutin, and FIG. 9(B) is an EIC result of quercetin.

Figure 10 is the EIC result of the lactic acid bacteria fermented product of 70% ethanol extract, Figure 10 (A) is the EIC result of rutin, Figure 10 (B) is the EIC result of quercetin.

11(A) is a mass spectrum of rutin ( m/z : 609.2 [MH] ^- ), and FIG. 11(B) is a mass spectrum of quercetin ( m/z : 301.1 [MH] ^- ).

Figure 12 shows the anti-inflammatory performance of 70% ethanol extract, fermented product and EGCG (Epigallocatechin gallate) when the extract is fermented with 9 types of mixed lactic acid bacteria.

13 shows the whitening performance of 70% ethanol extract, fermented product and arbutin when the extract is fermented with 9 types of mixed lactic acid bacteria.

14 shows the cell viability of the extract when fig leaves/fruits are extracted using 70% ethanol and the fermented product when the extract is fermented with one type of yeast.

15 shows the extract when fig leaves/fruits are extracted using 70% ethanol and the anti-inflammatory performance of the fermented product when the extract is fermented with one type of yeast.

Figure 16 shows the whitening performance of the extract when fig leaves/fruits are extracted using 70% ethanol and the fermented product when the extract is fermented with one type of yeast.

Fig. 17 is a chromatogram of a standard product, Fig. 17 (A) is a chromatogram of rutin, and Fig. 17 (B) is a chromatogram of quercetin.

18 is a chromatogram of a 70% ethanol extract, FIG. 18(A) is the TIC result of the ethanol extract, FIG. 18(B) is the EIC result of rutin, and FIG. 18(C) is the EIC result of quercetin.

19 is a chromatogram of a yeast fermented product of 70% ethanol extract, FIG. 19(A) is a TIC result of the yeast ferment, FIG. 19(B) is an EIC result of rutin, and FIG. 19(C) is an EIC result of quercetin to be.

20(A) is a mass spectrum of rutin ( m/z : 609.2 [MH] ⁻ ), and FIG. 20(B) is a mass spectrum of quercetin ( m/z : 301.1 [MH] ^- ).

21 shows the cell viability of the extract when fig leaves/fruits are extracted using 70% ethanol and the fermented product when the extract is fermented with one type of lactic acid bacteria and one type of yeast, respectively.

Figure 22 shows the anti-inflammatory performance of the extract when fig leaves/fruits are extracted using 70% ethanol and the fermented product when fermented with one type of lactic acid bacteria and one type of yeast, respectively.

23 shows the whitening performance of the extract when fig leaves/fruits are extracted using 70% ethanol and the fermented product when the extract is fermented with one type of lactic acid bacteria and one type of yeast, respectively.

[Explanation of code]

S100: alcohol extraction of fig leaves/fruits to obtain extraction

S200: lactic acid bacteria or yeast fermentation of the extract to obtain a fermented product

Hereinafter, the present invention will be described in detail. However, the present invention is not limited or limited by the exemplary embodiments. Objects and effects of the present invention can be naturally understood or made clearer by the following description, and the objects and effects of the present invention are not limited only by the following description. In addition, in the description of the present invention, if it is determined that a detailed description of a known technology related to the present invention may unnecessarily obscure the gist of the present invention, the detailed description thereof will be omitted.

1 is a flowchart of a method for manufacturing a cosmetic composition comprising a fermented product of an alcohol extract of figs according to the present invention. Referring to Figure 1, the present invention, the first step (S100) of obtaining an extract of fig leaf / fruit using 70% alcohol and fermenting the extract using 9 types of mixed lactic acid bacteria to obtain a fermented product A second step (S200) is included. The description of the 9 types of mixed lactic acid bacteria is as follows, and 70% alcohol is preferably 70% ethanol.

1) Lactobacillus plantarum ( Lactobacillus plantarum ): It is known as lactic acid bacteria that act in the late fermentation of kimchi. Recently, as well as intestinal health, it is also known as lactic acid bacteria for skin health such as skin itch improvement and skin moisturizing.

2) Lactobacillus rhamnosus ( Lactobacillus rhamnosus ): It helps the proliferation of lactic acid bacteria and suppresses harmful bacteria, and is known as a lactic acid bacteria that helps to facilitate bowel movements.

3) Lactobacillus casei ( Lactobacillus casei ): It is known as a lactic acid bacteria used in the production of cheese. Recently, it is also known as lactic acid bacteria that helps improve abdominal pain and bloating.

4) Lactobacillus paracasei ( Lactobacillus paracasei ): It is known as a lactic acid bacterium that helps to improve irritable bowel syndrome by inhibiting fungi, and is also known as an effective lactic acid bacterium for improving skin diseases such as atopy and psoriasis.

5) Lactobacillus fermentum ( Lactobacillus fermentum ): It has excellent acid resistance or intestinal epithelial cell adhesion, so most of them can reach the intestine when administered orally, and are known as lactic acid bacteria that can effectively inhibit the growth of harmful bacteria in the intestine.

6) Lactobacillus acidophilus ( Lactobacillus acidophilus ): It is known as a lactic acid bacterium that helps to strengthen immunity through intestinal health, and is also known as a lactic acid bacterium that can produce folic acid and lactase that digests milk, especially among B vitamins. .

7) Streptococcus thermophiles : It is known as a lactic acid bacterium that can promote intestinal peristalsis by generating lactic acid and prevent damage from toxins generated by harmful bacteria in the intestine.

8) Bifidobacterium longum ( Bifidobacterium longum ): It is known as a lactic acid bacterium that has a large inhibitory effect on harmful bacteria in the intestine, and is also known as a lactic acid bacterium that helps to improve diarrhea and enteritis.

9) Bfidobacterium bifidum ( Bfidobacterium bifidum ): It is known as a lactic acid bacterium that mainly inhabits the large intestine of breastfed babies, and is also known as a lactic acid bacterium with excellent ability to kill harmful bacteria by producing antibiotics.

The present invention, the first step (S100) of obtaining an extract of fig leaf / fruit using 70% alcohol and Saccharomyces cerevisiae fermenting the extract using Saccharomyces cerevisiae to obtain a fermented product A second step (S200) is included. Saccharomyces cerevisiae is known as brewer's yeast. Here, the 70% alcohol is preferably 70% ethanol.

The first step of the present invention is to obtain an extract from fig leaves/fruits. As the extraction medium, one of hot water, alcohol, 1,3-butylene glycol (1,3-BG), and ethyl acetate may be selected. However, since the purpose of extraction is to extract useful substances from fig leaves/fruits and use them as a cosmetic composition, an extraction medium may be selected in consideration of the use and effect of the cosmetic composition. When alcohol is selected as the extraction medium, 40 to 80% ethanol may be used, and preferably 70% ethanol (ethanol:water=7:3) may be used.

The second step of the present invention is a step of fermenting the fig leaf/fruit alcohol extract. Fermentation uses the above-mentioned 9 kinds of lactic acid bacteria or the above-mentioned one type of yeast. Fermentation refers to an action in which microorganisms such as bacteria or yeast decompose organic compounds to generate alcohols, organic acids, carbon dioxide, and the like. Fermentation in the present invention refers to a process of decomposing rutin contained in the fig leaf/fruit alcohol extract. Rutin is one of flavonol glycosides (glycosides) and refers to light yellow needle-shaped crystals, and glycosides contain aglycone, a non-saccharide component. In the present invention, aglycone refers to quercetin.

As a microorganism capable of fermenting the fig leaf/fruit alcohol extract, it may include Lactobacillus kimchicus in addition to the above-mentioned 9 types of lactic acid bacteria or the above-mentioned 1 type of yeast. Lactobacillus kimchicus is known as kimchi lactic acid bacteria. Microorganisms capable of fermenting fig leaf/fruit alcohol extract are not limited thereto. Any microorganism capable of degrading rutin may be included therein. Hereinafter, experimental examples of the present invention will be described.

2 and 3 show cell viability and anti-inflammatory performance when the extraction medium for fig leaf/fruit extraction was hot water, 50% ethanol, 70% ethanol, and 1,3-butylene glycol (1,3-BG), respectively. indicates

The process of obtaining a sample of the fig leaf/fruit to be extracted is as follows. After washing the fig leaves thoroughly, they were dried for 7 days in a well-ventilated place and out of sunlight. After washing the fig fruit, each fruit was divided into 4 parts and dried in a dryer for 5 days. The dried fig leaves/fruits were stored in a dry, cool and dark place, and powder samples were obtained by pulverizing them into 40 mesh particles with a grinder. The method of obtaining the fig leaf/fruit extract using each extraction medium is as follows.

In the case of hot water, 10 g of fig leaf/fruit powder and 100 g of water were mixed at a ratio of 1:10, incubated in a sonicator (42° C., 3 hours), and extracted using a filter with a pore size of 11.0 μm. The fig leaf/fruit extract ( hereinafter, Sample 1 ) was obtained by filtration.

In the case of 50% ethanol, 10 g of fig leaf/fruit powder and 100 g of 50% ethanol were mixed at a ratio of 1:10 and incubated in a sonicator (60° C., 1.5 hours) for extraction. The fig leaf/fruit extract was cooled to 30° C., filtered using a filter having a pore size of 11.0 μm, and concentrated under reduced pressure at 45° C. and 70 mbar, followed by centrifugation at 3,000 RPM for 30 minutes. An extract ( hereinafter, Sample 2 ) was obtained.

In the case of 70% ethanol, 10 g of fig leaf/fruit powder and 100 g of 70% ethanol were mixed at a ratio of 1:10 and incubated in a sonicator (60° C., 1.5 hours) for extraction. The fig leaf/fruit extract was cooled to 30° C., filtered using a filter with a pore size of 11.0 μm, and centrifuged at 3,000 RPM for 30 minutes after concentration under reduced pressure at 45° C. and 70 mbar, and the fig leaf/fruit extract ( hereinafter, Sample 3 ) was obtained.

In the case of 1,3-butylene glycol, 10 g of fig leaf/fruit powder and 100 g of 1,3-butylene glycol are mixed in a 1:10 ratio, incubated in a sonicator (42°C, 3 hours), and extracted. It was filtered using a filter having a pore size of 11.0 μm to obtain a fig leaf/fruit extract ( hereinafter, Sample 4 ).

The method of measuring cell viability (cytotoxicity test) is as follows. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to measure cell viability. 3Х10 ³ B16F10 mouse melanoma (whitening) and Raw264.7 (anti-inflammatory) cells were inoculated into a 96 well plate, and the extract was treated after 48 hours. The extract-treated cells were cultured for 48 hours, then treated with 0.5 mg/mL MTT (Sigma-Aldrich) and cultured at 37° C. for 1 hour. After 1 hour, the medium was removed and the resulting MTT formazan was dissolved in DMSO (dimethyl sulfoxide), and absorbance was measured at 595 nm using a microplate reader. Cell viability was shown by comparing the measured absorbance.

The anti-inflammatory performance measurement method is as follows. The anti-inflammatory performance can be expressed through whether the transcriptional ability of NF-kB, a transcription factor of cytokines that plays an important role in the immune response, is inhibited by fig leaf/fruit extract. Therefore, NF-kB reporter assay was performed to confirm the extent to which NF-kB transcriptional ability was suppressed when immune cells were treated with fig leaf/fruit extract.

The promoter region of a gene known to be transcribed by NF-kB was cloned into a vector containing a luciferase gene and inserted into immune cells. Then, NF-kB transcription-inducing lipopolysaccharide (lipopolysaccharide, hereinafter referred to as "LPS") was treated to activate NF-kB transcriptional ability, and then fig leaf/fruit extract was treated to induce luciferase ( luciferase) activity was measured.

For reference, luciferase activity is determined by the minimal IL-8 promoter (position A luciferase reporter gene construct constructed to be located upstream of -67 to +44) was used, and the gene was inserted into immune cells using lipofectamine plus (Gibco-BRL) according to the reagent manual.

The gene-inserted immune cells were treated with LPS to activate the transcriptional ability of NF-kB, and then cultured for 24 hours. Cultured immune cells were harvested by directly adding 0.1 ml of lysis buffer (0.1 M HEPES, pH 7.6, 1% Triton-X, 1 mM DTT and 2 mM EDTA) at the time of harvest to harvest the cell lysate at 4 °C. Cell proteins were obtained by centrifugation at 14,000 rpm for 10 minutes. After measuring the protein concentration using Bradford assay (Bio-Rad Laboratories, Hercules, CA), 20 μg of protein was added to luciferase assay mix (25mM glycylglycine, 15mM MgSO ₄ , 1mg/ml bovine serum albumin (BSA), 5mM ATP and After addition to 1 mM D-luciferin (Analytical Luminescence Laboratory, San Diego, CA)), the luminescence was measured three times for 20 seconds using Monolight 2010 (Analytical Luminescence Laboratory), and the average value was obtained and displayed.

Referring to FIG. 2 , in the case of Samples 1 to 3, it can be seen that the cell viability of 90% or more appears even at a sample concentration of 200 μg/ml. In the case of sample 4, it can be seen that a lower cell viability appears when compared to samples 1 to 3 based on a sample concentration of 50 μg/ml.

Referring to FIG. 3 , it can be seen that Sample 3 of Samples 1 to 3 exhibited the best anti-inflammatory performance. In the case of Sample 4, it can be seen that the anti-inflammatory performance similar to that of Sample 2 is shown based on the sample concentration of 50 μg/ml.

4 to 6 are extracts when fig leaves/fruits are extracted using 70% ethanol ( hereinafter, sample 5 ) and fermented products when the extracts are fermented with 9 types of mixed lactic acid bacteria ( hereinafter, sample 6 ) Cell viability, anti-inflammatory performance and whitening performance are shown. The types of 9 types of mixed lactic acid bacteria were as described above, and Lactomason Co., Ltd.'s 9 types of mixed lactic acid bacteria products (Lot no.: S-190916-1) were used.

The process of obtaining a sample of the fig leaf/fruit to be extracted and the method of obtaining the fig leaf/fruit extract using 70% ethanol as an extraction medium are the same as described above. Sample 6 was obtained by adding the 9 kinds of mixed lactic acid bacteria to sample 5 and fermenting at 37° C. for 5 days. The method of measuring cell viability and anti-inflammatory performance is the same as described above, and the measuring method of whitening performance is as follows.

For measurement of whitening performance, a method of extracting melanin in B16F10 mouse melanoma cells and measuring absorbance was used. 2Х10 ⁵ B16F10 mouse melanoma cells were inoculated in a 60-mm cell culture dish and cultured for 48 hours. After culturing, cells were treated with α-MSH, a substance that induces melanin production, and fig leaf/fruit extract, and cultured for 48 hours. After the culture was completed, the cells were harvested, washed once with PBS (Phosphate-buffered saline), and the number of cells was counted. 100 μL of 1 N aqueous sodium hydroxide solution (NaOH, Sigma-Aldrich) was added to each sample in which the same amount of cells were dispensed, and the cells were lysed at 95° C. for 15 minutes. Thereafter, the melanin content was confirmed by measuring the absorbance at a wavelength of 450 nm using a microplate reader.

4 to 6 , it can be seen that sample 5 and sample 6 exhibited a cell viability of 90% or more even at a sample concentration of 50 μg/ml, and that sample 6 showed better anti-inflammatory and whitening performance than sample 5 can be checked

Hereinafter, results of quantitative analysis of rutin and quercetin included in Sample 6 using liquid chromatography mass spectrometry (LC-MS/MS) will be described. However, in order to draw a clearer conclusion without limiting the sample used for quantitative analysis to sample 6, sample 5, malt fermented product of 70% ethanol extract ( hereinafter, sample 7 ), and yeast fermented product of 70% ethanol extract ( hereinafter referred to as sample 7) , and the results of quantitative analysis of sample 8 ) will be described together.

First, as a pretreatment process for each sample, samples 5 to 8 were treated with MeOH/D.W. After dissolving in the mixed solvent, filtration with filter paper was used as a sample solution, and 1 mg of rutin and quercetin standard was precisely measured, 1 mL of MeOH was added, followed by ultrasonic shaking for 20 minutes, and then filtered to obtain a standard stock solution. This standard stock solution was diluted by the serial dilution method to prepare standard solutions for calibration curves with concentrations of 0.5, 1, 5, 10, and 100 μg/mL.

Instrument	Agilent Technologies 6410 Triple Quad (LC-MS/MS)
Column	Chromasil C183.0 mm Х 150 mm, 3.0 μm
Solvent	A; 0.1% Formic acid in Water B; 0.1% Formic acid in Acetonitrile
flow rate	0.4 mL/min
Injection vol.	5 μL
gradient	(1) 0 min, 5% (2) 30 min, 100% (3) 32 min 100%
Ionization Mode	-ESI, scan mode
Gas temp.	350 ℃
capillary volt.	4000 V
Nebulizer	40 psig
Fragmentor	135 V

7(A) and 7(B) are chromatograms of rutin and quercetin of the standard. 8(A) to 8(D) are TIC results of Sample 5, Sample 6, Sample 7, and Sample 8, respectively. 9(A) and 9(B) are EIC results of rutin and quercetin in Sample 5. 10(A) and 10(B) are EIC results of rutin and quercetin in sample 6. 11(A) and 11(B) show the mass spectra of rutin ( m/z : 609.2 [MH] ^- ) and the mass spectrum of quercetin ( m/z : 301.1 [MH] ^- ).

Table 2 is information on a calibration curve using rutin and quercetin as target substances, and Table 3 is a table showing the amounts of rutin and quercetin included in samples 5 to 8. In Table 3, the unit of each value is μg/mg, and “N.D.” means Not Detected.

Compound	Regression equation	Linear range (μg/mL)	correlation coefficient (R 2 )
Routine	y = 62,420.8765x + 8,997.3845	0.5-10	0.9992
quercetin	y = 314,373.8219x + 39,651.7192	0.5-5	0.9989

sample	Routine	quercetin
sample 5	1.884	0.002
sample 6	1.002	0.132
sample 7	N.D.	N.D.
sample 8	N.D.	N.D.

7 to 11 and Tables 2 and 3, it can be seen that sample 6 contains a smaller amount of rutin than sample 5, and sample 6 includes a larger amount of quercetin than sample 5. Therefore, when fermenting the 9 types of mixed lactic acid bacteria of 70% ethanol extract, as the amount of quercetin, an aglycone, is increased by decomposition of rutin, a glycoside, that is, anti-inflammatory performance and whitening through biological conversion of substances through fermentation It can be seen that the performance is improved.

12 is a 70% ethanol extract ( hereinafter, sample 9 ), a fermented product ( hereinafter, sample 10 ) and EGCG (Epigallocatechin gallate) ( hereinafter, sample 11 ) when the extract is fermented with 9 types of mixed lactic acid bacteria. Anti-inflammatory performance indicates The types of 9 types of mixed lactic acid bacteria, the fermentation process, and the method for measuring anti-inflammatory performance are the same as described above.

Epigallocatechin gallate (EGCG, gallate-3-epigallocatechin) or 　catechin of FIG. 12 is a type of polyphenol that is an extract of green tea leaves and is known to have a strong antioxidant action.

Referring to FIG. 12 , it can be seen that the anti-inflammatory performance is improved as the concentrations of samples 9 and 10 are increased. In addition, it can be confirmed that the anti-inflammatory performance when the concentration of sample 10 is 50 μg/ml is close to the anti-inflammatory performance when the concentration of sample 11 is 10 μg/ml. For reference, the concentration μg/ml of sample 11 means (mass of EGCG) / (volume of medium containing EGCG), and the medium is RPMI medium (Raw264.7 cells).

Figure 13 shows the whitening performance of 70% ethanol extract ( hereinafter, sample 12 ), fermented product ( hereinafter, sample 13 ) and arbutin ( hereinafter, sample 14 ) when the extract is fermented with 9 types of mixed lactic acid bacteria indicates. The types of 9 kinds of mixed lactic acid bacteria, fermentation process, and methods for measuring whitening performance are the same as described above.

Arbutin of FIG. 13 is known to have physiological activity to inhibit melanin production by inhibiting tyrosinase activity, which is an enzyme that regulates melanin production, and thus is used as a skin whitening agent.

Referring to FIG. 13 , it can be seen that the whitening performance is improved as the concentrations of Samples 12 and 13 are increased, and the whitening performance when the concentration of Sample 13 is 50 μg/ml is the same as when the concentration of Sample 14 is 20 μg/ml. It can be seen that the whitening performance is superior to that of ml. For reference, the concentration μg/ml of sample 14 means (mass of arbutin) / (volume of medium containing arbutin), and the medium is DMEM medium (B16F10 cells).

14 to 16 show an extract ( hereinafter, Sample 15 ) when fig leaves/fruits are extracted using 70% ethanol and a fermented product ( hereinafter, Sample 16 ) when the extract is fermented with one type of yeast Cells It shows the survival rate, anti-inflammatory performance and whitening performance. One type of yeast is the aforementioned Saccharomyces cerevisiae .

The process of obtaining a sample of the fig leaf/fruit to be extracted and the method of obtaining the fig leaf/fruit extract using 70% ethanol as an extraction medium are the same as described above. Sample 16 was obtained by mixing sample 15 and a culture solution, inoculating brewer's yeast, and culturing at 25° C., 150 rpm, and 24 h. Methods for measuring cell viability, anti-inflammatory performance and whitening performance are the same as described above.

14 to 16 , it can be seen that both sample 15 and sample 16 show a cell viability of 90% or more even at a sample concentration of 20 μg/ml, and sample 16 has a sample concentration of 10 μg/ml and 20 μg/ml than sample 15. It can be seen that the anti-inflammatory performance is better in , and it can be seen that sample 16 has better whitening performance than sample 15 at all sample concentrations.

17(A) and 17(B) are chromatograms of rutin and quercetin of a standard. 18(A) is the TIC result of Sample 15, FIG. 18(B) is the EIC result of rutin of Sample 15, and FIG. 18(C) is the EIC result of quercetin of Sample 15. 19(A) is the TIC result of sample 16, FIG. 19(B) is the EIC result of rutin of sample 16, and FIG. 19(C) is the EIC result of quercetin of sample 16. 20(A) is a mass spectrum of rutin ( m/z : 609.2 [MH] ⁻ ), and FIG. 20(B) is a mass spectrum of quercetin ( m/z : 301.1 [MH] ^- ).

Compound	Regression equation	Linear range (μg/mL)	correlation coefficient (R 2 )
Routine	y = 72,770.1869x + 12,519.4791	0.5-10	0.9992
quercetin	y = 554,724.5714x - 9,711.0000	0.25-1	1.0000

sample	Routine	quercetin
sample 15	4.020	0.042
sample 16	0.960	0.016

Table 4 is information on a calibration curve using rutin and quercetin as target substances, and Table 5 is a table showing the results of quantitative analysis of rutin and quercetin included in samples 15 and 16. In Table 5, the unit of each numerical value is μg/mg. Quantitative analysis methods and processes for samples 15 and 16 were performed under the same conditions as those for quantitative analysis methods and processes for samples 5 to 8 described above.

17 to 20 and Tables 4 and 5, it can be seen that sample 16 contains less rutin and quercetin than sample 15. Compared with the results of Table 4, it is common that rutin is degraded by fermentation and the amount of rutin decreases, but it can be concluded that the amount of quercetin increases during lactic acid fermentation and decreases during yeast fermentation.

21 to 23 are extracts when fig leaves/fruits are extracted using 70% ethanol ( hereinafter, sample 17 ), fermented products when sample 17 is fermented with one type of lactic acid bacteria ( hereinafter, sample 18 ) and samples The cell viability, anti-inflammatory performance and whitening performance of the fermented product ( hereinafter, Sample 19 ) when 17 was fermented with one type of yeast are shown. One type of lactic acid bacteria is Lactobacillus kimchicus , also called kimchi lactic acid bacteria, and one type of yeast is the aforementioned Saccharomyces cerevisiae ( Saccharomyces cerevisiae ).

The process of securing a sample of the fig leaf/fruit to be extracted, the method of obtaining the fig leaf/fruit extract using 70% ethanol as an extraction medium, the lactic acid bacteria fermentation method, and the yeast fermentation method are as described above. However, unlike the above, each sample (sample) was prepared by varying the vol%, and the vol% is based on the volume of the fermentation product/[the volume of the fermentation product + the volume of the animal cell culture medium].

Referring to FIG. 21 , it can be seen that all of samples 17 to 19 exhibited a cell viability of 90% or more at 0.5 vol% or less. Referring to FIG. 22 , it can be seen that sample 18 exhibits superior anti-inflammatory performance than sample 17 at sample concentrations of 0.25 vol% and 0.5 vol%, and sample 19 exhibits superior anti-inflammatory performance than sample 17 at all sample concentrations. Referring to FIG. 23 , it can be seen that samples 18 and 19 exhibited superior whitening performance than sample 17 at all sample concentrations.

The cosmetic composition of the present invention has a fermented product of fig leaf/fruit alcohol extract as an essential component, and may include other components that are normally formulated in cosmetic compositions, and as a compounding component, an oil and fat component, a moisturizer, an emollient, and an interface activators, emulsifiers, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, fragrances, blood circulation promoters, cooling agents, limiting agents, purified water, and the like.

In addition, according to an embodiment of the present invention, it is possible to provide a formulation capable of caring for skin troubles, such as alleviating or treating acne, based on the anti-inflammatory performance of the fermented product of fig leaf/fruit alcohol extract.

In addition, according to another embodiment of the present invention, it is possible to provide a formulation capable of whitening care based on the whitening performance of the fermented fig leaf/fruit alcohol extract.

The above-mentioned formulations for skin trouble care and/or whitening care include skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, and essence. , nutritional essence, pack, soap, shampoo, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, press powder, and loose powder may be any one selected from the group consisting of.

Although the present invention has been described in detail through representative embodiments above, those of ordinary skill in the art to which the present invention pertains will understand that various modifications are possible within the limits without departing from the scope of the present invention with respect to the above-described embodiments. will be. Therefore, the scope of the present invention should not be limited to the described embodiments, but should be defined by all changes or modifications derived from the claims and equivalent concepts as well as the claims to be described later.

Claims (2)

1. A first step of obtaining an extract of fig leaves and fruits using 70% alcohol; and

   A second step of fermenting the extract using 9 types of mixed lactic acid bacteria to obtain a fermented product,

   The 9 kinds of mixed lactic acid bacteria,

   Lactobacillus plantarum ( Lactobacillus plantarum );

   Lactobacillus rhamnosus ( Lactobacillus rhamnosus );

   Lactobacillus casei ( Lactobacillus casei );

   Lactobacillus paracasei ( Lactobacillus paracasei );

   Lactobacillus fermentum ( Lactobacillus fermentum );

   Lactobacillus acidophilus ( Lactobacillus acidophilus );

   Streptococcus thermophiles ;

   Bifidobacterium longum ; and

   It is composed of Bfidobacterium bifidum ,

   The 70% alcohol is 70% ethanol,

   The second step is a step of increasing the amount of quercetin contained in the extract by decomposing the rutin contained in the extract,

   The fermented product is a method for producing a cosmetic composition comprising a fermented product of fig alcohol extract, characterized in that it contains rutin and quercetin.
2. A fermented product of an ethanolic extract of fig leaves and fruits,

   The fermented product of the ethanol extract of fig leaves and fruits contains rutin in an amount less than the amount of rutin contained in the ethanol extract of fig leaves and fruits,

   The fermented product of the ethanol extract of the fig leaf and fruit contains quercetin in an amount greater than the amount of quercetin contained in the ethanol extract of the fig leaf and fruit,

   The fermented product of the ethanol extract of the fig leaf and fruit is obtained by fermenting the ethanol extract of the fig leaf and fruit using 9 types of mixed lactic acid bacteria,

   The 9 kinds of mixed lactic acid bacteria,

   Lactobacillus plantarum ( Lactobacillus plantarum );

   Lactobacillus rhamnosus ( Lactobacillus rhamnosus );

   Lactobacillus casei ( Lactobacillus casei );

   Lactobacillus paracasei ( Lactobacillus paracasei );

   Lactobacillus fermentum ( Lactobacillus fermentum );

   Lactobacillus acidophilus ( Lactobacillus acidophilus );

   Streptococcus thermophiles ;

   Bifidobacterium longum ; and

   Bifidobacterium bifidum ( Bfidobacterium bifidum ) A cosmetic composition, characterized in that it consists of.

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