KR102120475B1 - Cosmetic composition comprising Hermetia illucens larva - Google Patents

Abstract

The present invention relates to a cosmetic composition using Dongae and the like, and according to an aspect of the present invention, a cosmetic composition using wrinkle improvement, antioxidant activity and antibacterial activity of the extract is provided to Dongae. In addition, Dongae et al. extract further includes an extract of baekbaekpi, centella extract, and Eoseongcho extract to further improve the functionality of the cosmetic composition, improve flavor and color, and reduce side effects by not using organic synthetic preservatives.

Description

Cosmetic composition containing extract in Dongae etc. {Cosmetic composition comprising Hermetia illucens larva}

The present invention relates to a cosmetic composition containing an extract in Dongae.

The Hermetiaillucens (black sodier fly) is a representative environmental cleansing insect of the family Diptera, known as the American Dongae. Hermetia illucens is an environmental cleansing insect. It is a powerful antibacterial substance that can quickly decompose these organic wastes while living in organic wastes (food waste, animal wastes, agricultural by-products, etc.) that contain large amounts of bacteria, fungi, and viruses. It possesses a mechanism for expressing. Therefore, it is expected to be used as a natural antibiotic because it is not only excellent as a development material for natural antibiotics that can replace chemical antibiotics, but also has a very fast growth rate compared to other insects and does not require a separate feed cost.

In order to apply Dongae lamp to the industry, mass production technology was developed and research is actively conducted in various fields such as animal feed manufactured with Dongae lamp and waste treatment technology using Dongae lamp. It is insufficient.

It is to provide an extract to Dongae, etc. as an active ingredient of a functional cosmetic composition.

It is to provide a cosmetic composition that does not use organic synthetic preservatives or reduces its content, but does not increase replacement cost.

The technical problem is not limited to the above, and another technical problem may be derived from the following description.

Provides a cosmetic composition containing an extract as an active ingredient in Dongae. It provides a cosmetic composition containing a mixed natural extract including an extract of baekbaekpi, centella extract, and Eoseongcho as an active ingredient.

Functional cosmetics containing the extract as an active ingredient for wrinkle improvement, antioxidant activity and antibacterial activity can be provided to Dongae.

By further including the extract of Dongae et al., baekbaekpi extract, centella extract, Eoseongcho extract, it is possible to provide a cosmetic composition that improves wrinkle improvement, antioxidant activity, antibacterial activity, and improves color and odor.

By using the extract as a natural antibiotic in Dongae and the like, it is possible to provide a cosmetic composition that limits the use of artificial synthetic synthetic preservatives or reduces its content.

1 is a graph showing the Elastase inhibitory activity of each of Examples 1-3 and Comparative Examples 1,2, and showing their values.
2 is a graph showing the values of Tyrosinase inhibitory activity of Examples 1-3 and Comparative Examples 1,2, respectively.
3 is a graph showing the DPPH free radical removal activity of each of Examples 1-3 and Comparative Examples 1,2, and showing the value.
FIG. 4 is a graph showing the activity of Hydroxyl radical scavenging activity of Examples 1-3 and Comparative Examples 1,2, respectively.
5 is a graph showing the values of Fe ²⁺ -chelating activity of Examples 1-3 and Comparative Examples 1,2, respectively.
6 is a graph showing the values of the ABTS radical scavenging activity of Examples 1-3 and Comparative Examples 1,2, respectively.

Hereinafter, the contents of the present invention will be described in more detail.

The present invention relates to a functional cosmetic composition using an extract in Dongae. It is known that a substance secreted by larvae for biodefense in Dongae and the like has resistance to proteolytic enzymes. Conventionally, it has been known that the extract has antimicrobial activity in Dongae, etc., but there is no known effect on wrinkle improvement and antioxidant activity.

In the present invention, as well as having a wrinkle improvement effect by confirming that the extract of Dongae et al. has Elastase inhibitory activity and Tyrosinase inhibitory activity, it also has an antioxidant effect by confirming that it has DPPH free radical scavenging activity, Hydroxyl radical scavenging activity, and ABTS radical scavenging activity. Said to have

Dongae et al extract used in the present invention can be prepared through drying, grinding, and extraction processes from larvae of Dongae et al.

Drying larvae in Dongae and the like; Crushing the larvae on the dried Dongae lamp to form a larva powder on the Dongae lamp; Forming an larva powder solution in Dongae lamp by adding an extraction solvent to the larva powder in Dongae lamp; Recovering the supernatant by centrifuging the larva powder solution on the heated Dongae etc.; And concentrating the supernatant.

In the present invention, the larvae of Dongae lamps are those of the American Dongae lamps ( Hermetia illucens ), which are stored at a low temperature of -80°C to 20°C.

In the step of drying the larvae to the Dongae, drying may be dried by irradiating with hot air, freezing or microwaves. In order to have a high yield and optimal activity of the extract in Dongae, etc., it is performed for 12 hours to 24 hours at 60°C to 80°C for hot air drying, and for 36 hours to 65 hours at -60°C to -40°C for freeze drying It is carried out for 10 minutes to 50 minutes when microwave irradiation.

In the step of crushing the larvae on the dried dongae lamp to form larva powder on the dongae lamp, the crushing speed is performed at 2500 rpm to 3700 rpm so that the larvae powder on the dried dongae lamp can be obtained from 100 to 1000 mesh sized larvae. .

In the step of forming and heating the larval powder solution in Dongae lamp by adding an extraction solvent to the larva powder in the Dongae lamp, the extraction solvent contains distilled water and ethanol, preferably distilled water and ethanol in a volume ratio of 60:40 to 80:20 Mix with. The heating condition is performed at a temperature of 80°C to 120°C for 10 minutes to 48 hours, and shows antibacterial activity optimized in the temperature range and heating time.

In the step of recovering the supernatant by centrifuging the larvae powder solution in the heated Dongae lamp, the centrifugation is performed for 10 minutes to 50 minutes at a speed of 2000 rpm to 6000 rpm at 1 °C to 10 °C.

In the step of concentrating the supernatant, the recovered supernatant is concentrated for 6 to 12 hours using a vacuum concentrated centrifuge.

The extract of Dongae etc. prepared in the above may be stored in distilled water, ethanol or a solvent mixed with distilled water and ethanol for preservation of safety and storage.

In the present invention, the mixed natural extracts further containing the extract of Dongae et al., the extract of baekpi, the extract of medicinal herb, and the extract of Eoseongcho can provide a cosmetic composition having improved wrinkle improvement effect and antioxidant activity as well as improved odor and color. Extracts from Dongae, etc., as animal raw materials, can be a factor in inhibiting product marketability when used in cosmetics with high content due to the smell and color. By further including the plant-derived natural ingredients, baekbaekpi extract, centella extract, and eoseongcho extract, it is possible to improve the flavor and color by enhancing the preference. In addition, each extract has Elastase inhibitory activity, Tyrosinase inhibitory activity, DPPH free radical scavenging activity, Hydroxyl radical scavenging activity, and ABTS radical scavenging activity. Antibacterial effect.

)의 뿌리껍질로서 주피를 제거한 것으로 뛰어난 미백 및 보습원료로 쓰이며 임상 미백원료 중에서 거의 트러블이 없는 원료로 입증되어 있으며 미백과 보습의 이중기능 원료로서의 효과가 뛰어나다. 상백피 추출물은 뽕나무 뿌리를 말려서 물이나 보습제하에서 끓이거나 추출해서 사용한다. Morus alba Linn

It is used as an excellent whitening and moisturizing ingredient as it removes the main skin as the root bark of) and is proven as a material that is almost trouble-free among clinical whitening raw materials, and has excellent effects as a dual function ingredient for whitening and moisturizing. Sangbaekpi extract is used by drying the root of a mulberry tree, boiling it under water or a moisturizer, or extracting it.

Centella asiatica L. Urban is a perennial plant belonging to the Apiaceae family, and is also called tiger grass because it sees wounded tigers rolling and healing in the place where the diseased grass grows. As the main ingredients, beta-carotene, beta-sitosterol, campesterol, camphor, camperol, saponin, stigmasterol, calcium, iron, magnesium, manganese, phosphorus, potassium, selenium, zinc, vitamins B1, B2, B3, C, etc. Contains. It has a collagen-producing effect, which has a skin regeneration effect and suppresses Bacillus bacteria to improve skin diseases such as acne. It has various effects such as sedation, free radical removal, antioxidant action, and strengthening of immunity. The centella extract is obtained by washing the leaves of centella, drying with hot air, then weighing in a blender, adding 10 times the extraction solution, ultrasonically extracting for 30 minutes, centrifuging at 3000 rpm for 10 minutes, and filtering and concentrating the supernatant.

Houttuynia cordata is a wild herb of the perennial herb belonging to the saururaceae family and is native to China, Japan, and Korea. Eoseongcho is also known as medicinal wheat, and has been reported to have a wide range of therapeutic effects such as edema, syphilis, cystitis, uteritis, pneumonia, and paralysis. The active substance, quercitrin, has been reported to have a variety of bioactive functions. Eoseongcho extract is obtained by reflux extraction of 60% methanol for 2 hours, filtered, and concentrated using a vacuum rotary concentrator.

In the present invention, the mixed natural extract containing the extract of Dongae et al. not only has wrinkle improvement and antioxidant activity but also has antibacterial activity.

In general, in order to increase the shelf life of the cosmetic composition, organic synthetic preservatives such as methylparaben, propylparaben, and butylparaben are used, or expensive natural antibiotics such as melittin and bee venom are used. Methylparaben, an organic synthetic preservative used in cosmetics, causes wrinkles and blotch when exposed to UV rays, promotes skin aging, and is absorbed into the human body through the skin, causing allergies, or causing confusion in the hormone secretion system. Has the side effect of increasing sperm count and reducing sperm count in men. Dongae has antibacterial activity and can replace organic synthetic preservatives. In addition, Dongae, etc., can be replaced with expensive natural antibiotics such as Merlittin and bee venom derived from bees because it is easier to breed than bees and has excellent breeding ability, so it can be supplied at low cost. In addition, the extract of baekbaekpi, centella extract, and eoseongcho extract also have antibacterial activity, and thus, synergistically with the extract of Dongae, show more enhanced antibacterial and anti-inflammatory effect, thus maintaining stability without using synthetic preservatives and having excellent preservative cosmetic Compositions can be provided.

In the present invention, butylene glycol, 1,2-hexanediol, and ethylhexylglycerin act as solvents in the cosmetic composition and help the extract and the mixed natural extract to penetrate the skin.

According to an example of the present invention, the cosmetic composition is 20-25 parts by weight of Dongae et al. extract, 20-25 parts by weight of Sangbaekpi extract, 20-25 parts by weight of Centella extract, 20-25 parts by weight of Eoseongcho extract, butylene glycol 3 -7 parts by weight, 1-5 parts by weight of 1,2-hexanediol, and 0.01-1 parts by weight of ethylhexylglycerin. If the content of the extract in Dongae etc. is less than 20 parts by weight, wrinkle improvement, antioxidant effect and antibacterial activity are not expressed from the extract in Dongae etc., and it is difficult to act as an active ingredient, and if it exceeds 25 parts by weight, the peculiar smell is strong and desired because of the slippery texture There is a difficulty in preparing the cosmetic composition of the formulation.

The cosmetic composition according to the present invention is used in formulations such as lotion, essence, foundation, powder, lotion, cream, pack, gel, ointment, patch, spray, mask sheet and detergent.

Hereinafter, the present invention will be described in more detail through examples, comparative examples, and test examples.

Example 1. Preparation of extracts from Dongae etc.

The larvae (H. illucens) were used by Dongae lamps raised in a breeding room (25℃ ~ 27℃, relative humidity 60) after being sold by the Ministry of Agriculture and Biology of the National Academy of Agricultural Science, Rural Development Administration. First, 1,000 g of larvae were dried for 45 minutes in a frozen lamp (-20°C to -80°C) to obtain about 340 g of dried copper lamp. Dry Dongae etc. are ground by a pulverizer and powdered, and then 3.4 L of extraction solvent (distilled water: ethanol = 70: 30) is added and heated at 100° C. for 30 minutes, followed by centrifugation (4° C., 4500 rpm, 30 min). Did. After centrifugation, the supernatant was recovered, and only the pure supernatant was recovered by filtering the recovered supernatant using a 0.45 μm filter. The pure supernatant was concentrated for 9 hours using a vacuum concentrated centrifuge (EYLYA Co., Tokyo, Japan) to prepare an extract in Dongae lamp.

Example 2. Preparation of mixed natural extract

A mixed natural extract was prepared by mixing 25 g of epithelium extract, 25 g of coriander extract, and 25 g of Eoseongcho extract against 25 g of extract from Dongae et al. prepared in Example 1.

Example 3. Preparation of cosmetic composition

With respect to 23 g of Dongae et al. prepared in Example 1, 23 g of epithelium extract, 23 g of Centella extract, 22.95 g of Eoseongcho extract, 5 g by weight of butylene glycol, 3 g of 1,2-hexanediol, ethyl hexylglycerin 0.05 A cosmetic composition in which g was mixed was prepared.

Comparative Example 1.

A cosmetic composition was prepared by mixing 33.3 g of lettuce extract, 33.3 g of Centella extract, and 33.4 g of Eoseongcho extract.

Comparative Example 2.

A cosmetic composition was prepared in which 30.65 g of lettuce extract, 30.65 g of Centella extract, 30.65 g of Eoseongcho extract, 5 g by weight of butylene glycol, 3 g of 1,2-hexanediol, and 0.05 g of ethylhexylglycerin were prepared.

Examples 1-3 prepared in the above, Comparative Examples 1,2 are shown in Table 1 below.

Ingredient name (%) Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 2 Extract of Dongae 100 25 23 - - Lettuce extract - 25 23 33.3 30.65 Centella extract - 25 23 33.3 30.65 Eoseongcho Extract - 25 22.95 33.4 30.65 Butylene Glycol - - 5 - 5 1,2-hexanediol - - 3 - 3 Ethylhexylglycerin - - 0.05 - 0.05 total 100 100 100 100 100

Test Example 1. Elastase inhibitory ability

Porcine pancreas elastase inhibitory activity of each of Examples 1-3 and Comparative Examples 1,2 prepared as described above was measured, and the values are shown in Table 2 below and the graphs are shown in FIG. 1.

Porcine pancreas elastase inhibitory activity was measured using N-succinyl-(L-Ala)3-p-nitoanilide as a substrate, and the amount of p-nitoanilide produced at 37°C for 30 minutes was measured (Cannell et al., 1988). Each test solution was prepared to a constant concentration, and 0.1 ml was taken into a test tube, and 0.05 ml of elastase, pancreatic solution, and Type I: From Porcine Pancreas (0.6 units/ml) in 50 mM Tris-HCl buffer (pH 8.6) was added. After adding 0.1 ml of N-succinyl-(L-Ala)3-p-nitoanilide (1 mg/ml) dissolved in 50 mM Tris-HCl buffer (pH 8.6) as a substrate, react for 30 minutes and absorbance at 410 nm. Was measured. Elastase inhibitory activity was expressed as the rate of decrease in absorbance of the sample solution with and without additives.

The results of the experiment were calculated as the mean±standard deviation (Mean±. Statistical analysis according to the statistical significance test between each group was significant when p<0.05 by performing a Duncan post-test comparison of ANOVA (one-way analysis of variance test). It was judged (SPSS V12., SPSS Inc, Chicago, IL, USA).

＼ Concentration (㎍/ml) 0 10 30 50 100 300 500 1000 3000 5000 10000 Example 1 0 0.702 0.853 1.096 4.259 5.978 6.288 10.96 13.096 14.785 16.936 Example 2 0 0.889 0.396 1.186 6.752 7.899 8.645 13.78 14.52 18.33 20.96 Example 3 0 0.808 0.288 1.16 5.171 6.446 7.935 12.164 12.86 13.122 15.907 Comparative Example 1 0 0.454 0.585 0.73 2.64 3.47 3.861 5.148 7.263 6.932 8.234 Comparative Example 2 0 0.212 0.114 1.02 2.24 2.867 3.942 5.961 8.126 7.865 9.259

As shown in Table 2, in Example 1 it can be confirmed that there is an Elastase inhibitory ability. From this, it can be seen that the extract of Dongae et al. has a wrinkle improvement effect. In detail, in all of Examples 1 to 3, the activity was insignificant until the concentration of 50 μg/ml, and the inhibitory activity began to gradually increase from the concentration of 100 μg/ml, and at the concentration of 10000 μg/ml, the highest processing concentration of the sample. Example 1 showed 16.93%, Example 2 20.96%, and Example 3 15.91%.

When comparing Example 2 and Comparative Example 1, it can be seen that Elastase inhibitory ability is higher when the extract is included in Dongae. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the wrinkle improvement effect of the mixed natural extract.

When comparing Example 3 and Comparative Example 2, it can be seen that Elastase inhibitory ability is higher when the extract is included in Dongae. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the wrinkle improvement effect of the cosmetic composition.

Test Example 2. Tyrosinase inhibitory ability

Tyrosinase activity of each of Examples 1-3 and Comparative Examples 1,2 prepared as described above was measured, and the values are shown in Table 3 below and the graphs are shown in FIG. 2.

Tyrosinase activity was measured by the method of Yagi et al. (1986). To a test tube, 0.2 ml of substrate solution in which 10 mM L-DOPA was dissolved in 0.5 ml of 1.15 M sodium phosphate buffer at pH 6.8 and 0.1 ml of sample solution was added to 0.2 ml of 110 Unit/ml mushroom tyrosinase. After reacting at 25° C. for 2 minutes, the absorbance of DOPA chrome generated in the reaction solution was measured at 475 nm using a microplate reader. Tyrosinase inhibitory activity was shown as the absorbance reduction rate of the sample solution added and non-added groups.

The results of the experiment were calculated as the mean±standard deviation (Mean±. Statistical analysis according to the statistical significance test between each group was significant when p<0.05 by performing a Duncan post-test comparison of ANOVA (one-way analysis of variance test). It was judged (SPSS V12., SPSS Inc, Chicago, IL, USA).

＼ Concentration (㎍/ml) 0 10 30 50 100 300 500 1000 3000 5000 10000 Example 1 0 0.36 0.3346 2.312 3.152 4.245 6.862 11.248 18.324 20.185 34.285 Example 2 0 0.26 0.458 2.045 2.685 3.264 7.358 12.364 19.248 22.34 36.47 Example 3 0 0.197 0.248 1.548 2.449 2.829 6.322 10.061 17.694 19.361 32.863 Comparative Example 1 0 0.124 0.262 1.295 2.245 2.744 5.645 7.89 13.479 16.25 24.586 Comparative Example 2 0 0.214 0.425 1.264 2.024 2.426 5.314 8.242 12.245 14.695 21.54

As shown in Table 3, it can be confirmed that in Example 1 there is Tyrosinase inhibitory ability. From this, it can be seen that the extract of Dongae et al. has a wrinkle improvement effect. In detail, in all of Examples 1 to 3, the activity was minimal until the concentration of 50 μg/ml, and the inhibitory activity gradually increased from the concentration of 100 μg/ml, and the concentration of 10000 μg/ml, the highest concentration of the sample, Example 1 showed 34.28%, Example 2 36.47%, and Example 3 32.863%.

When comparing Example 2 and Comparative Example 1, it can be seen that Tyrosinase inhibitory activity is higher when the extract is included in Dongae. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the wrinkle improvement effect of the mixed natural extract.

When comparing Example 3 and Comparative Example 2, it can be seen that Tyrosinase inhibitory ability is higher when the extract is included in Dongae. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the wrinkle improvement effect of the cosmetic composition.

Test Example 3. DPPH free radical removal ability

By measuring the DPPH inhibitory activity of each of Examples 1-3 and Comparative Examples 1,2 prepared as described above, the values are shown in Table 4 below and the graphs are shown in FIG. 3.

DPPH (1,1-diphenyl-2-picryldrazyl) radical removal activity was measured according to the method of Hsu et al. (2006). Samples are diluted by concentration using DMSO (dimethyl sulfoxide), mixed with 40 μl of sample and 760 μl of 300 mM DPPH, reacted at 37° C. for 30 minutes, and then 515 nm using Infinite 200pro (Tecan, PerkinElmer, USA) Absorbance was measured at.

The antioxidant activity of the extract according to the concentration was calculated by the following equation.

Scavenging capacity (%) = {1-(absorbance of the extract-added group/absorbance of the extract-free group)}×100

The results of the experiment were calculated as the mean±standard deviation (Mean±. Statistical analysis according to the statistical significance test between each group was significant when p<0.05 by performing a Duncan post-test comparison of ANOVA (one-way analysis of variance test). It was judged (SPSS V12., SPSS Inc, Chicago, IL, USA).

＼ Concentration (㎍/ml) 0 10 30 50 100 300 500 1000 3000 5000 10000 Example 1 0 1.62 1.961 3.041 4.142 4.781 4.912 5.071 6.51 7.621 8.712 Example 2 0 1.69 2.482 3.455 3.86 4.68 4.768 5.264 6.788 8.014 9.934 Example 3 0 0.92 1.255 1.367 1.272 1.894 1.528 2.006 3.618 5.35 8.23 Comparative Example 1 0 0.425 0.621 0.785 0.855 1.744 2.854 2.652 3.42 4.254 5.14 Comparative Example 2 0 0.29 0.496 0.899 0.69 1.001 1.21 1.894 2.024 2.65 3.78

As shown in Table 4, it can be seen that in Example 1 there is a DPPH free radical removal ability. From this, it can be seen that the extract of Dongae et al. has antioxidant activity. Specifically, in all of Examples 1 to 3, as the concentration increased, the inhibitory activity began to increase steadily, and at the concentration of 10000 μg/ml, the highest concentration of the sample, 8.712% in Example 1 and 9.934 in Example 2 %, Example 3 showed an activity of 8.23%.

When comparing Example 2 and Comparative Example 1, it can be seen that when the extract is included in Dongae, DPPH free radical removal ability is higher. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the antioxidant activity of the mixed natural extract.

When comparing Example 3 and Comparative Example 2, it can be seen that when the extract is included in Dongae, DPPH free radical removal ability is higher. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the antioxidant effect of the cosmetic composition.

Test Example 4. Hydroxyl radical removal ability

Hydroxyl radical inhibitory activity of each of Examples 1-3 and Comparative Examples 1,2 prepared as described above was measured, and the values are shown in Table 5 below and the graphs are shown in FIG. 4.

Hydroxyl radical removal activity was performed according to the method of Smirnoff and Cumbes (1989). After mixing 250 µl of FeSO4 and 175 µl of H2O2 with 75 µl concentration-specific samples and reacting for 30 minutes at 37°C, absorbance was measured at 560 nm using Infinite 200pro (Tecan, PerkinElmer, USA).

The antioxidant activity of the extract according to the concentration was calculated by the following equation.

Scavenging capacity (%) = {1-(absorbance of the extract-added group/absorbance of the extract-free group)}×100

The results of the experiment were calculated as the mean±standard deviation (Mean±. Statistical analysis according to the statistical significance test between each group was significant when p<0.05 by performing a Duncan post-test comparison of ANOVA (one-way analysis of variance test). It was judged (SPSS V12., SPSS Inc, Chicago, IL, USA).

＼ Concentration (㎍/ml) 0 10 30 50 100 300 500 1000 3000 5000 10000 Example 1 0 1.96 1.742 2.241 3.426 5.142 7.326 8.962 14.268 20.159 37.685 Example 2 0 1.86 1.957 1.678 2.044 2.867 7.795 9.0875 15.125 22.01 39.68 Example 3 0 1.597 0.088 1.166 0.157 1.387 6.527 9.755 13.821 19.039 28.085 Comparative Example 1 0 0.962 1.264 1.062 2.134 1.376 6.421 6.861 11.294 12.69 20.085 Comparative Example 2 0 0.622 0.062 0.851 1.264 0.965 3.162 5.264 9.162 13.246 18.325

As shown in Table 5, it can be seen that in Example 1, there is the ability to remove Hydroxyl radicals. From this, it can be seen that the extract of Dongae et al. has antioxidant activity. In detail, in all of Examples 1 to 3, the activity was insignificant until the concentration of 100 μg/ml, and the inhibitory activity gradually increased from the concentration of 300 μg/ml, and at the concentration of 10000 μg/ml, which is the highest processing concentration of the sample. In Example 1, 37.685%, Example 2 39.68%, and Example 3 28.085%.

When comparing Example 2 and Comparative Example 1, it can be confirmed that when the extract is included in Dongae, the ability to remove Hydroxyl radicals is higher. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the antioxidant activity of the mixed natural extract.

When comparing Example 3 and Comparative Example 2, it can be seen that when the extract is included in Dongae, the ability to remove Hydroxyl radicals is higher. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the antioxidant effect of the cosmetic composition.

Test Example 5. Fe ²⁺ -chelating activity

The Fe ²⁺ -chelating activity of each of Examples 1-3 and Comparative Examples 1,2 prepared as described above was measured, and the values are shown in Table 6 below and the graphs are shown in FIG. 5.

The Fe ²⁺ -chelating activity assay is performed by modifying the method of Hsu et al. (2006). 15 μl of 2 mM FeCl 2 and 150 μl of sample, 30 μl of 5 mM ferrozine, and 605 μl of distilled water were mixed and reacted at room temperature for 15 minutes, followed by Fe ^2+-at 562 nm using Infinite 200pro (Tecan, PerkinElmer, USA) The absorbance of the ferrozine mixed solution was measured.

The antioxidant activity of the extract according to the concentration was calculated by the following equation.

Scavenging capacity (%) = {1-(absorbance of the extract-added group/absorbance of the extract-free group)}×100

The results of the experiment were calculated as the mean±standard deviation (Mean±. Statistical analysis according to the statistical significance test between each group was significant when p<0.05 by performing a Duncan post-test comparison of ANOVA (one-way analysis of variance test). It was judged (SPSS V12., SPSS Inc, Chicago, IL, USA).

＼ Concentration (㎍/ml) 0 10 30 50 100 300 500 1000 3000 5000 10000 Example 1 0 5.032 8.675 11.285 15.324 18.24 23.485 38.24 57.425 77.241 86.34 Example 2 0 4.682 9.264 12.245 14.261 17.154 25.124 37.127 60.142 75.142 92.17 Example 3 0 3.042 8.235 8.707 9.315 10.138 14.164 19.83 48.811 64.317 81.576 Comparative Example 1 0 3.152 6.578 7.265 7.942 8.695 11.264 13.482 32.547 48.48 56.248 Comparative Example 2 0 2.861 6.485 7.594 8.124 9.348 12.067 15.751 37.152 58.142 68.325

As shown in Table 6, it can be confirmed that in Example 1, there is Fe ²⁺ -chelating activity. From this, it can be seen that the extract of Dongae et al. has antioxidant activity. Specifically, in all of Examples 1 to 3, as the concentration increased, the inhibitory activity began to increase steadily, and the concentration of 10000 μg/ml, the highest concentration of the sample, was 86.34% in Example 1 and 92.17 in Example 2. %, Example 3 showed an activity of 81.576%.

When comparing Example 2 and Comparative Example 1, it can be confirmed that when the extract is included in Dongae, Fe2+-chelating activity is higher. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the antioxidant activity of the mixed natural extract.

Comparing Example 3 and Comparative Example 2, it can be seen that Fe2+-chelating activity is higher when the extract is included in Dongae. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the antioxidant effect of the cosmetic composition.

Test Example 6. ABTS removal ability

Example 1-3 and Comparative Examples 1,2 prepared as described above, ABTS radical scavenging activity was measured, and the values are shown in Table 7 below and the graphs are shown in FIG. 6.

The ABTS radical scavenging activity is measured by the ABTS antioxidant assay method. 7 mM 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (Sigma, A1888) and 2.45 mM potassum persulfate were mixed 1:1 to react for 16 hours to form ABTS ⁺ Dilute with distilled water to use the absorbance at 734 nm, resulting in a value of 0.7. Absorbance was measured at 734 nm after 15 minutes by mixing 190 μl of diluted ABTS ⁺ solution and 10 μl of sample.

＼ Concentration (㎍/ml) 0 10 30 50 100 300 500 1000 3000 5000 10000 Example 1 0 1.042 1.34 2.421 2.832 3.247 4.352 5.265 16.75 20.24 33.254 Example 2 0 1.24 1.379 2.245 2.0175 3.479 4.685 4.873 16.25 18.48 36.85 Example 3 0 0.2 One 0.745 -0.051 1.216 1.539 2.987 10.878 15.466 27.89 Comparative Example 1 0 0.26 0.853 0.762 1.212 0.862 1.537 2.42 10.075 11.86 20.14 Comparative Example 2 0 0.145 0.751 0.753 0.921 1.004 1.402 1.945 8.312 11.24 18.24

As shown in Table 7, it can be confirmed that in Example 1 there is ABTS radical scavenging activity. From this, it can be seen that the extract of Dongae et al. has antioxidant activity. In detail, in all of Examples 1 to 3, the activity was insignificant until the concentration of 100 μg/ml, and the inhibitory activity gradually increased from the concentration of 300 μg/ml, and at the concentration of 10000 μg/ml, which is the highest processing concentration of the sample. Example 1 showed 33.254%, Example 2 36.85%, and Example 3 27.89%.

When comparing Example 2 and Comparative Example 1, it can be confirmed that when the extract was included in Dongae, ABTS radical scavenging activity was higher. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the antioxidant activity of the mixed natural extract.

Comparing Example 3 and Comparative Example 2, it can be confirmed that when the extract is included in Dongae, ABTS radical scavenging activity is higher. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the antioxidant effect of the cosmetic composition.

Test Example 7. Antibacterial activity

The antimicrobial activity of each of Examples 1-3 and Comparative Examples 1 and 2 prepared as described above and methyl paraben, Merlittin, and bee venom were measured and the values are shown in Table 8 below.

In order to confirm the antibacterial activity of the perfume composition according to the present invention, antibacterial experiments were performed on Examples 1-3, Comparative Examples 1,2 and methylparaben. The specific experimental method is as follows. Staphylococcus aureus , Escherichia coli , and Pseudomonas aeruginosa strains used in the experiment were from Tryptic Soy Broth and Candida Albicans. ( Candida albicans ) and Aspergillus niger strains were cultured in Sabouraud Dextrose Broth. The diluted solution diluted 1/100 (1/10 for the Candida albicans strain) in the culture medium was used as the test solution. Aspergillus niger used a spore suspension prepared to be 2 x 10 ⁸ cfu/ml as a test solution. 0.15 ml of the test solution was added to 15 ml of each medium, and well mixed was used as a dilution solution. In the 96-well plate (96 well plate) row 1, 16 µl of the sample was added, and 184 µl of the diluted solution was added. 100 µl of the diluted solution was added to the remaining wells. After mixing well in the mixed solution of the 1st row, take 100 µl, put it in the 2nd row, mix it well, and then take 100 µl and dilute it twice by adding it to the 3rd row.

Staphylococcus aureus , Escherichia coli , and Pseudomonas aeruginosa in a 32 °C thermostatic bath, Candida albicans , Aspergillus niger niger ) was incubated in a 25°C thermostat. After 48 hours, the microbial growth was checked by suspension and microscope to determine the minimum inhibitory concentration (MIC) value.

Substance name Minimum inhibitory concentration (MIC), unit% Staphylococcus aureus Escherichia coli Pseudomonas aeruginosa Candida albicans Aspergillus niger Methylparaben 0.15~0.2 0.2 0.2~0.25 0.1 0.1 Meryltin > 0.1 0.35 0.2 > 0.1 > 0.1 Bee venom 0.2 0.5 0.5 0.37 > 0.1 Example 1 0.08 0.125 0.18 0.075 0.068 Example 2 0.12 0.18 0.08 0.12 0.078 Example 3 0.48 0.375 0.668 0.12 0.064 Comparative Example 1 0.88 0.54 1.02 0.08 0.08 Comparative Example 2 0.69 0.55 0.86 0.08 0.07

As shown in Table 2, it can be confirmed that in Example 1, the antibacterial activity was higher than that of methylparaben. From this, it can be seen that the extract of Dongae et al. has antibacterial activity.

When comparing Example 2 and Comparative Example 1, it can be seen that the antibacterial activity is higher when the extract is included in Dongae. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the antibacterial activity of the mixed natural extract.

When comparing Example 3 and Comparative Example 2, it can be confirmed that when the extract was included in Dongae, the antibacterial activity was higher. From this, it can be seen that the extract of Dongae et al acts as an active ingredient in the antibacterial activity of the cosmetic composition.

So far, preferred examples, comparative examples, and test examples for the present invention have been described. Although specifically described through Examples, Comparative Examples and Test Examples of the present invention, this is not intended to limit the present invention, but merely to demonstrate the present invention. Therefore, it will be apparent to those skilled in the art to which the present invention pertains that the scope of the present invention is not limited by these examples according to the gist of the present invention. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent range should be interpreted as being included in the present invention.

Claims (9)

delete delete delete delete Dongae et al. extract 20-25 parts by weight, baekbaekpi extract 20-25 parts by weight, centella extract 20-25 parts by weight, Eoseongcho extract 20-25 parts by weight, butylene glycol 3-7 parts by weight, 1,2-hexanediol 1 A cosmetic composition having antibacterial activity containing as an active ingredient a mixed natural extract containing -5 parts by weight and 0.01-1 part by weight of ethylhexylglycerin. delete delete delete The method of claim 5, wherein the cosmetic composition is a cosmetic lotion, essence, foundation, powder, lotion, cream, pack, gel, ointment, patch, spray, mask sheet and cleaning agent having any one of the formulation characterized in that it has an antibacterial activity Cosmetic composition.

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