KR102232341B1 - Cosmetic composition for preventing or improving skin aging or skin inflammation comprising fermented product of Mentha piperascens leaf and ginger as an active ingredient - Google Patents

Abstract

The present invention relates to a cosmetic composition for preventing or improving skin aging or skin inflammation containing a fermented product of peppermint leaf and ginger as an active ingredient, and specifically, Lactobacillus planta in a liquid medium containing peppermint leaf and ginger powder. Rum ( Lactobacillus plantarum ) relates to a cosmetic composition for preventing or improving skin aging or skin inflammation containing a fermented product obtained by inoculation and fermentation as an active ingredient.
The cosmetic composition of the present invention contains fermented products of peppermint and ginger as active ingredients, so that it is less irritating and exhibits excellent antioxidant activity and anti-inflammatory activity, thereby exhibiting excellent effects in preventing or improving skin aging or skin inflammation.

Description

Cosmetic composition for preventing or improving skin aging or skin inflammation comprising fermented product of Mentha piperascens leaf and ginger as an active ingredient }

The present invention relates to a cosmetic composition for preventing or improving skin aging or skin inflammation containing a fermented product of peppermint leaf and ginger as an active ingredient, and specifically, Lactobacillus planta in a liquid medium containing peppermint leaf and ginger powder. Rum ( Lactobacillus plantarum ) relates to a cosmetic composition for preventing or improving skin aging or skin inflammation containing a fermented product obtained by inoculation and fermentation as an active ingredient.

China, Beijing, India and Korea are currently in a very serious stage of fine dust, and these fine dusts lead to skin diseases as well as respiratory diseases for all modern people, causing various types of skin diseases such as contact dermatitis and eczema, and health and life. It is showing serious damage.

In the case of the steadily growing overseas and domestic cosmetics industry and, in particular, in the'Derma Cosmetics' market, which combines skin science with cosmetics, the market size of soothing products for sensitive skin-related troubles caused by fine dust, etc. is 500 billion won, growing by more than 15% every year. have.

Fermentation refers to the process of decomposing organic matter using microbial enzymes, and the fermentation method, the oldest technology in history, is widely used in various fields such as food, pharmaceuticals, and cosmetics. Among them, food fermentation is not only in Korea but also in the world. It is the most widely used. In the case of food fermentation, it is known that it has the effect of improving the flavor, flavor, and texture of food through the enzyme action of microorganisms as a traditional method of processing refinement, destroying toxic substances through the fermentation process, and enhancing physiologically active substances. .

Mint and ginger are known for their physiological activities such as antibacterial and antioxidant effects, but due to the stimulation of their unique strong scent, consumers' preference is low, so they are used as spices in foods or only as some flavors of fragrance ingredients.

Therefore, by utilizing fermentation that can enhance physiologically active substances, the shortcomings of peppermint and ginger were supplemented, and the activation as a functional ingredient for flavoring was maximized.

Korean Patent Registration No. 10-1694229

The main object of the present invention is to provide a cosmetic composition that contains a fermented product obtained through a specific fermentation of peppermint and ginger, thereby reducing irritation and exhibiting an excellent effect on the skin.

According to one aspect of the present invention, the present invention comprises a fermented product obtained by inoculating and fermenting Lactobacillus plantarum in a liquid medium containing peppermint leaf and ginger powder as an active ingredient. It provides a cosmetic composition for preventing or improving skin inflammation.

In the cosmetic composition of the present invention, it is preferable that the liquid medium contains 10 to 30 g/L of the mint leaf powder, 10 to 30 g/L of the ginger powder, and further contains sugar.

In the cosmetic composition of the present invention, the fermentation product is preferably obtained by inoculating Lactobacillus plantarum in the liquid medium and shaking fermentation at 35 to 38° C. at 100 to 200 rpm for 2 to 4 days.

The cosmetic composition of the present invention contains fermented products of peppermint and ginger as active ingredients, so that it is less irritating and exhibits excellent antioxidant activity and anti-inflammatory activity, thereby exhibiting excellent effects in preventing or improving skin aging or skin inflammation.

1 shows DPPH radical scavenging activity of fermented peppermint leaves and ginger according to an embodiment of the present invention. vitamin C, control; Ginger & peppermint, fermented product treatment group of the present invention; 0d, before fermentation; 3d, 3 days after fermentation; 7d, 7 days after fermentation.
Figure 2 shows the total polyphenol content of peppermint leaf and ginger fermentation according to an embodiment of the present invention. control (DDW), control; Ginger & peppermint, fermented product treatment group of the present invention; 0d, before fermentation; 3d, 3 days after fermentation; 7d, 7 days after fermentation.
Figure 3 shows the total flavonoid content of peppermint leaves and ginger fermentation according to an embodiment of the present invention. Ginger & peppermint, fermented product treatment group of the present invention; 0d, before fermentation; 3d, 3 days after fermentation; 7d, 7 days after fermentation.
Figure 4 shows the results of the cytotoxicity test of peppermint leaves and ginger fermentation according to an embodiment of the present invention. control, control (no treatment); 10 ~ 2.5mg / ml, treatment group by concentration of the fermented product of the present invention; Before, before fermentation; 3 days, 3 days after fermentation; 7 days, 7 days after fermentation.
5 shows the experimental results of the NO production inhibitory effect of fermented peppermint leaves and ginger according to an embodiment of the present invention. control, control (no treatment); LPS, LPS (10 ng/ml) treatment group; Peppermint + ginger, fermented product treatment group of the present invention; 0d, before fermentation; 3d, 3 days after fermentation; 7d, 7 days after fermentation.
6 shows the experimental results of the NO production inhibitory effect of fermented peppermint leaves and ginger according to an embodiment of the present invention. control, control (no treatment); lps 10ng/ml, LPS (10ng/ml) treatment group; 0d, pre-fermentation mixture treatment group; 3d, the fermented product treatment group of the present invention fermented for 3 days; The fermented product treatment group of the present invention fermented on 7d and 7 days.
7 shows the manufacturing sequence of a cosmetic formulation according to an embodiment of the present invention.
8 shows a questionnaire for evaluating skin irritation of a cosmetic formulation according to an embodiment of the present invention.
9 shows the skin measurement results according to the use of the cosmetic formulation according to an embodiment of the present invention. Before, before use of the formulation; 1st after 15 minutes, 15 minutes after formulation use; Second time after 30 minutes, 30 minutes after use of the formulation.

The cosmetic composition of the present invention is characterized in that it contains a fermented product obtained by inoculating and fermenting Lactobacillus plantarum in a liquid medium containing peppermint leaf and ginger powder as an active ingredient.

In the present invention, the powder of mint leaves and ginger can be obtained through a conventional method of powdering plant materials, but it is preferable to use powdered mint leaves and ginger by lyophilizing them in order to increase the effect of the present invention. Do.

In the present invention, the liquid medium can be obtained by adding nutrients necessary for the survival of Lactobacillus plantarum to water together with the powder of mint leaves and ginger. It is preferable that the liquid medium contains 10 to 30 g/L of the mint leaf powder, 10 to 30 g/L of the ginger powder, and further contains sugar as an additional nutrient. At this time, it is preferable to use glucose and sucrose as the sugar, and it is preferable to contain 1 to 10 g/L of glucose and 1 to 10 g/L of sucrose in the liquid medium.

It is preferable to sterilize the liquid medium before inoculating the fermenting bacteria in order to prevent erroneous fermentation due to the growth of various bacteria that may occur during the fermentation process. At this time, sterilization may be performed using a conventional liquid sterilization method, but it is preferable to use a method of heating at high temperature and high pressure.

In the present invention, Lactobacillus plantarum is used as the fermenting bacteria, and in this case, it is preferable to use KCCM 11322, ATCC 8014, IFO 3070, DSM 20205 or NRRL B-531 as the Lactobacillus plantarum strain.

In the present invention, the inoculation may be performed by adding the pre-culture solution of the fermenting bacteria to the liquid medium. At this time, the pre-culture solution of the fermenting bacteria can be obtained by inoculating and culturing the fermenting bacteria in a liquid medium in which the fermenting bacteria can grow. At this time, it is preferable to use a method of shaking culture for 1 to 3 days at 35 to 39°C and 100 to 200 rpm in MRS liquid medium. In order to increase fermentation efficiency, it is preferable to inoculate by adding 0.5 to 2% (v/v) of the preculture thus obtained.

In the present invention, the fermentation may be performed by maintaining a liquid medium containing mint leaves and ginger powder inoculated with the fermenting bacteria at a temperature capable of growth and fermentation of the fermenting bacteria for an appropriate time. At this time, the temperature is preferably 35 to 38°C, and it is preferable to perform shaking fermentation at 100 to 200 rpm for 2 to 4 days.

In the present invention, the fermented product prepared through the above method can be used as it is, but it is preferable to use it after removing the solid material and lyophilizing it. At this time, the solids may be removed by conventional methods such as sedimentation of solids, filtration, and centrifugation.

The cosmetic composition of the present invention may be prepared in a form conventionally prepared in the art, and may be prepared by adding the fermented product during the manufacturing process of the cosmetic. At this time, the amount of fermented product added may be appropriately selected in consideration of the type, shape, purpose, and method of manufacture of the cosmetic, but it is preferable to contain the fermented product in an amount of 0.1 to 1% by weight based on the weight of the total cosmetic composition, More preferably, it is judged to be good to contain 0.1 to 0.5% by weight.

Hereinafter, the present invention will be described in more detail through examples. Since these examples are for illustrative purposes only, the scope of the present invention is not to be construed as being limited by these examples.

[Example]

1. Manufacture of peppermint leaves and ginger fermentation products

1-1. Preparation of fermentation microorganisms

Lactobacillus plantarum (KCCM 11322) was plated on MRS agar solid medium, cultured at 37°C for 3 days, and then cultured strain 1 platinum was inoculated on 100 ml of MRS liquid medium and inoculated at 36°C for 2 days. It was cultured with shaking at 160 rpm.

1-2. Fermentation production

Liquid medium per liter Last name content glucose 5g sucrose 5g Mint leaf freeze-dried powder 20g Ginger Freeze-Dried Powder 20g Distilled water up to 1000ml

Prepare a liquid medium as shown in Table 1, sterilize at 121°C for 30 minutes at 1.5psi, cool, add 10 ml of the liquid culture solution of Lactobacillus plantarum of 1-1 and inoculate, and inoculate at 36.5°C at 160 ~ 180 rpm. It was fermented with shaking for ~ 7 days.

Thereafter, the fermentation broth was settled at 4° C. for 24 hours at low temperature, and then centrifuged at 4° C. and 4,500 rpm to obtain a supernatant, which was filtered twice with a filter of 5 μm and 0.45 μm pore size, respectively, and the filtrate was freeze-dried.

2. Antioxidant effect

The antioxidant effect of the fermented product (lyophilized powder) sample of Example 1 by concentration (10 mg/ml, 5 mg/ml, 2.5 mg/ml) was investigated.

The antioxidant effect was investigated by the DPPH radical scavenging activity test method, and the DPPH radical scavenging activity test method modified from the method of Seon-hee you (2016) was used. After adding an equal amount of 0.1mM DPPH (2,2-diphenyl-1-picrylhydrazyl) solution to a 96-well plate in an equal amount of 180 µl each, 20 µl of samples of various concentrations dissolved in distilled water were added each and reacted at 37°C for 30 minutes. Absorbance was measured at 517 nm. At this time, ascorbic acid (vitamin C) was used as a control. DPPH radical scavenging activity was calculated by the following equation.

DPPH radical scavenging activity (%) = 100-{(absorbance in added group / absorbance in no added group) x 100}

As a result, as shown in FIG. 1, all of the fermented products of Example 1 exhibited excellent antioxidant activity, and particularly, when fermented for 3 days, it was found that the antioxidant activity was excellent.

3. Total polyphenol content

The total polyphenol content of the fermented product (lyophilized powder) sample of Example 1 was investigated.

The total polyphenol content was measured using the method of Na Hyun Lee et al. (2015). _{2% Na 2} CO ₃ solution in 100 μl of sample solution (diluted with water to a concentration of 10 mg/100 μl), DDW (negative control) or Gallic acid standard (0, 20, 40, 60, 80, 100 mg/L) 2 ml was added, stirred sufficiently, and allowed to stand for 3 minutes. After vortexing with 100 µl of 50% Folin &Ciocalteu's phenol reagents, the mixture was reacted in a dark room for 30 minutes, and the absorbance was measured at 750 nm using a blank as a control. To quantify the phenolic compound, it was calculated from a calibration curve obtained using gallic acid as a standard material.

As a result of this, as shown in FIG. 2, it was found that the total polyphenol content was high in all of the fermented products of Example 1, but the content was rather decreased due to fermentation. When these results and the results of the antioxidant activity test are combined, it can be predicted that components exhibiting antioxidant activity other than polyphenols were produced due to fermentation.

4. Total flavonoid content

The total flavonoid content of the fermented product (lyophilized powder) sample of Example 1 was investigated.

The total flavonoid content was measured by modifying the method of Hongmei Geng. (2008). 2 ml of sample solution (diluted with water to a concentration of 10 mg/100 µl), DDW (negative control) or Rutin hydrate standard (0, 10, 20, 30, 40, 50 mg/L) and 0.3 _{ml of 5% NaNO 2} After mixing, react at room temperature for 6 minutes _{, add 0.3 ml of 10% Al(NO 3} ) ₃ to react at room temperature for 6 minutes, add 4 ml of 10% NaOH solution, stir sufficiently, and then fill 10 ml with secondary distilled water and mix. After reacting for 15 minutes, the blank was controlled at 510 nm using a spectrophotometer (OPTIZEN POP Meccasys), and the absorbance of each solution was measured. The total flavonoid content was calculated from a calibration curve prepared using Rutin hydrate as a standard material.

As a result of this, as shown in FIG. 3, it was found that the total flavonoid content was high in all of the fermented products of Example 1, but the content was rather decreased due to fermentation. When these results and the results of the antioxidant activity experiment are combined, it can be predicted that components exhibiting antioxidant activity other than flavonoids were produced due to fermentation.

5. Cytotoxicity test

The cytotoxicity of the fermented product (lyophilized powder) sample of Example 1 was investigated.

5-1. Cell culture

RAW264.7 (TIB-71; ATCC, USA) macrophages used in the experiment were purchased from American type cell collection (ATCC, USA), and the medium used was 10% FBS in DMEM (Dulbecco's Modified Eagle's Medium; Gibco, USA) medium. (Fetal Bovine Serum; Gibco, USA) and 1% of Penicillin (100U/ml)/Streptocycin (100µg/ml)/Amphotericin B (0.25µg/ml) were added, and then under the conditions of _{37°C and 5% CO 2} It was used after culturing.

5-2. Cell Viability Assay (MTT assay)

The cell viability of the sample was measured by the EZ-Cytox (Enhanced Cell Viability Assay kit, DoGenBio Co., Ltd.) method. Cells were dispensed into a 96-well plate at a concentration of 5×10 ⁴ cells/well and cultured for 24 hours, and then the medium was removed. Add 20 µl of each sample diluted by concentration to 180 µl of new DMEM (10% FBS, 1% Penicillin (100U/ml)/Streptocycin (100µg/ml)/Amphotricin B (0.25µg/ml)). After incubation for time, 10 µl of EZ-Cytox reagent was added to each well and incubated for 1 hour at 37° C. and 5% CO ₂ . After reacting for 1 hour, absorbance was measured at 450 nm using an ELISA reader.

5-3. result

Experimental results As shown in Figure 4, the fermented product of Example 1 was found to have no significant cytotoxicity compared to the control at all concentrations.

6. NO production inhibitory effect

The effect of inhibiting NO generation of the fermented product (lyophilized powder) sample of Example 1 was investigated. The NO production inhibitory effect is used as an indicator of the anti-inflammatory effect.

RAW 264.7 cells ^{were dispensed into a 12-well plate at a concentration of 2.5×10 5} cells/ml, and cultured for 24 hours, and then the medium was removed. Here, a sample diluted to 1 mg/ml in a new DMEM (10% FBS, 1% Penicillin (100U/ml)/Streptocycin (100µg/ml) Amphotericin B (0.25µg/ml)) medium was pretreated for 1 hour. Then, LPS (10 ng/ml) was treated and incubated for 24 hours at 37° C. and 5% CO ₂ . After 24 hours of sample treatment, the cell culture solution was centrifuged, and then 100 μl of the supernatant and 50 μl of Griess reagent A solution (1% sulfanilamide) + 50 μl of the B solution (0.1% N-(1-naphtyl) ethylenediamine dihydrochloride) were mixed to 96 well plates After reacting at for 20 minutes, the absorbance was measured at 540 nm using an ELISA reader. As a result _{, a standard curve was drawn with 1M NaNO 2,} and the value of NO contained in the cell culture supernatant was calculated.

As a result, as shown in FIGS. 5 and 6, both of the fermented products of Example 1 were found to have an excellent inhibitory effect on NO generation, and in particular, when fermented for 3 days, the effect of inhibiting NO generation was excellent.

7. Formulation manufacturing using fermented products

In order to use the fermented product of Example 1 as a cosmetic additive, 0.2% (w/w) by mixing the fermented product (freeze-dried powder) 3 days after fermentation by applying the results of the evaluation of the efficacy of cosmetic functions such as antibacterial activity and antioxidant. An essence mask pack and a moisture cream were prepared with the content of (see Table 2). At this time, while maximizing the cosmetic and functional efficacy of the fermented product, the degree of preference for cytotoxicity and color was considered. The order of preparation of the formulation is shown in FIG. 7.

Prize Raw material name Content (% by weight) Awards D.W To 100 Glycerin 5.00 Sodium Hyaluronate (1%) 6.00 1,3-Butylene Glycol 6.50 Disodium EDTA 0.02 D-M 0.05 Paid Cetyl Ethylhexanoate 0.50 Xanthan gum 1.00 Dimethicone 1.00 Carbopol941 0.10 adding Allantoin 0.10 Dipropylene Glycol 0.15 TEA 0.10 Fermented product of Example 1 0.20 P.F 0.10

8. Evaluation of skin irritation of the formulation

A questionnaire survey on skin irritation after using the essence mask pack of Example 7 was conducted on 20 subjects using a questionnaire as shown in FIG. 8.

Very strong Strong weakness No irritation (%) Eradication 0 1(5) 2(10) 17(85) Itching (pruritus) 0 1(5) 1(5) 18(90) Stinging 1(5) 1(5) 1(5) 17(85) Burning (burning sensation) 1(5) 0 2(10) 17(85) Soreness 0 0 2(10) 18(90) ache 0 1(5) 0 19(95) Pull/Tighten 0 1(5) 1(5) 18(90) Snow 0 0 2(10) 18(90) Open eyes 0 0 1(5) 19(95) Eye pain (pain) 0 0 2(10) 18(90) Redness (erythema) 0 0 2(10) 18(90) Swelling (edema) 0 0 0 20(100) Millet 0 0 0 20(100) Hardening (hardening) 0 0 0 20(100) pimple 0 0 0 20(100) acne 0 0 0 20(100) Keratin 0 0 0 20(100) Blisters or blisters 0 0 0 20(100) Pigmentation 0 0 0 20(100) Eye congestion 0 0 0 20(100) Tears 0 0 0 20(100)

As a result of this, as shown in Table 3, 1 patient (5%), weak 2 (10%), strong itching, 1 (5%), weak 1 (5%) when the degree of itching is strong. ), soreness, very strong, strong, weak 1 person (5%) each, strong burning 1 person (5%), weak case 2 people (10%), soreness weak 2 people (10%), blindness Two patients (10%) were weak, 1 (5%) weakened eyes, and 2 patients (10%) had weak eye pain and redness, and most were evaluated as having no skin irritation.

9. Effects on the skin by using the formulation

Skin measurements were performed on the subject before and after using the essence mask pack of Example 7.

Skin moisture, skin melanin, skin elasticity, and sebum secretion were measured using a skin measuring device. The device used for skin measurement is the Multi skin test center MC 1000 (Courage-Khazaka Electronic Co., Germany). The implementation method was 5 minutes after cleansing, through a skin measuring device, skin moisture, skin melanin, and sebum secretion were measured using probes, respectively, and the first measurement 15 minutes after using the essence mask pack, and 30 minutes after use. The second measurement was made at one time point.

As a result of this, as shown in FIG. 9, in the case of skin moisture level, moisture increased from 51.3 to 55.7 in the first measurement after 15 minutes before applying the essence mask pack, and to 62 in the second measurement after 30 minutes, and in the case of melanin level, from 34.7 to 30 minutes before application. In the second measurement, it showed a slightly clearer skin tone at 32.3. The amount of sebum secretion increased from 2.7 before application to 4.7 in the first measurement 15 minutes after application, and 4.5 in the second measurement 30 minutes after application. According to these results, the use of the essence mask pack containing the fermented product of the present invention helps a lot in the oil-water balance that maintains the moisture and barrier of the skin, and maintains the skin elasticity to properly supply moisture and oil to the dry skin. It is believed that zooming can help calm and prevent skin troubles or fine dust.

Claims (3)

A cosmetic composition for preventing or improving skin inflammation, comprising a fermented product obtained by inoculating and fermenting Lactobacillus plantarum in a liquid medium consisting of mint leaf powder, ginger powder, sugar and water. The method of claim 1,
The liquid medium is a cosmetic composition, characterized in that the mint leaf powder is contained in an amount of 10 to 30g/L, and the ginger powder is contained in an amount of 10 to 30g/L. The method of claim 1,
The fermentation product is a cosmetic composition, characterized in that obtained by inoculating Lactobacillus plantarum in the liquid medium and shaking fermentation at 35 to 38° C. at 100 to 200 rpm for 2 to 4 days.

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