KR20200049008A - Cosmetic composition with antioxidative and skin heat dreasing effect - Google Patents

Abstract

The present invention relates to a cosmetic composition in an aerosol formulation comprising an extract prepared by extracting Scutellaria baicalensis, Coptis japonica, Phellodendron amurense, and Houttuynia cordata with a solvent comprising 1,3 butylene glycol, and a method for manufacturing the same. Proposed is a cosmetic composition which exhibits antioxidant, antiaging, and immediate heat reducing effects.

Description

COSMETIC COMPOSITION WITH ANTIOXIDATIVE AND SKIN HEAT DREASING EFFECT}

The present invention relates to a cosmetic composition having anti-oxidation and heat-reducing efficacy and a method for manufacturing the same.

Skin is an important organ responsible for a part of the immune system. It is always exposed to physical stimuli from the outside, chemical stimuli caused by chemical substances, or environmental stimuli caused by sudden changes in humidity and temperature, light stimuli caused by ultraviolet rays, and irritation caused by inappropriate eating habits. It is. Therefore, in the case of cosmetics or lotions that need to soothe skin exposed to various external stimuli and apply directly to the skin, it is beneficial to use products that exclude artificial and chemical components as much as possible. There is a growing trend in interest.

In addition, even in the development of the formulation of the cosmetic composition, there is a need for the development of cosmetic materials based on research on extracting conditions that extract the components that are most beneficial to skin health and improve the functionality of the components beyond simple plant extracts. .

Under this background, the inventors of the present invention find beneficial effects on the skin of the mixed extracts of golden, hwangnyeon, hwangbaek and eoseongcho, find extraction conditions for optimal efficacy, and in addition, these extracts can be applied most effectively to the skin. As a result of repeated research on the development of formulations, the present invention has been completed.

In order to solve the above problems, one aspect of the present invention is a cosmetic composition having a sulfation and heat sensation efficacy, the cosmetic preparation comprises a mixed extract of a mixture of golden, yellow lotus, hwangbaek and eoseongcho, the mixed extract is 1,3 butylene glycol (1,3 butylene glycol) and ethanol ratio of 1: 1 is obtained by extraction of a solvent having a 1: 1 ratio or 1,3 butylene glycol and water ratio of 1: 1, and the mixed extract is The included cosmetic composition provides a cosmetic composition characterized by having a formulation in the form of an aerosol. The golden, hwangnyeon, hwangbaek and eoseongcho can be extracted by mixing in the same weight ratio. In addition, the mixed extract may be included in 0.001 to 10% by weight relative to the total weight of the cosmetic composition.

In another aspect of the present invention, the cosmetic composition provides a cosmetic composition further comprising a second mixed extract obtained by mixing each extract obtained by ethanol solvent extraction of golden, hwangnyeon, hwangbaek and eoseongcho. The second mixed extract is 90 to 95 mg / g for golden, 45 to 47 mg / g for hwangnyeon, 80 to 83 mg / g for hwangbaek, and 60 to 62 mg / g for polyphenols. Containing; Gold may contain 10 to 12 mg / g, hwangnyeon 23 to 27 mg / g, hwangbaek to 8 to 10 mg / g, and eoseongcho to 4 to 6 mg / g flavonoids, respectively.

Another aspect of the present invention, the first step of preparing a mixture by mixing golden, hwangnyeon, hwangbaek and Eoseongcho; The mixture is added 30 to 70 by adding at least one solvent selected from the group consisting of a solvent having a 1: 1 ratio of 1,3 butylene glycol and ethanol or a solvent having a 1: 1 ratio of 1,3 butylene glycol and water. A second step of extracting for 5-10 hours at ℃; A third step of filtering the extract obtained from the second step with a filtering device; And a fourth step of preparing the cosmetic composition containing the filtrate obtained from the third step in an aerosol form formulation. The extraction of the second step may be made under the condition of extracting the mixture at 60 ° C. for 6 hours.

The present invention is a cosmetic composition of an aerosol formulation containing an extract obtained by extracting golden, hwangnyeon, hwangbaek, and eoseongcho with a solvent containing 1,3 butylene glycol, which is extracted under the condition where the antioxidant and anti-aging effects are best exhibited, and Choose a formulation and show immediate heat sensation.

Figure 1 shows the content of polyphenols and flavonoids extracted by 1,3 butylene glycol, ethanol, water or mixed solvents of golden, hwangnyeon, hwangbaek, and eoseongcho.
Figure 2 shows the results of the electron donating ability of the mixed extract.
Figure 3 shows the ABTs radical cation scavenging ability of the mixed extract.
Figure 4 shows the results of measuring the cytotoxicity using the MTT assay of the mixed extract.

Golden (黃 芩; Scutellariae Radix) is a dicotyledonous plant that is cultivated in Korea, China, and eastern Siberia. It is a root of the perennial plant ( Scutellaria baicalensis) , belonging to the family Lamitae, and has been dysentery and pyrexia. ), Jaundice and carbuncle. The main components of gold include flavonoids such as baicalin, baicalein, wogonin, and wogonin glucuronide. Among them, baicalin has been reported to have various physiological and pharmacological effects such as anti-allergic, anti-inflammatory, and anti-cancer effects. It is known to have viruses and antioxidant effects.

The rhizome of the perennial plant of the Ranunculus family, the herbaceous plant, Coptis japonica , is called Coptidis Rhizoma in Chinese medicine and is cultivated in China, Korea, and Japan. It is used for the treatment of diarrhea or vomiting caused by stomach fever by making the stomach healthy and having sedative, anti-inflammatory, and antibacterial effects. One component of the extract, berberine, has a strong inhibitory effect on various bacteria, and has anti-inflammatory, hemostatic, anti-hypertensive and anti-cancer effects.

Hwangbaek (黃 hello; Phellodendri Cortex) is a dried bark of the bark of the Phellodendron amurense belonging to the Ruthaceae family, and is distributed in all regions except Jeju and Jeonnam regions in Korea. It is distributed in Edo and grows in hybrid forests or mountain valleys. In oriental medicine, inflammation, pneumonia, bone tuberculosis, colds, antiseptics, and anti-inflammatory drugs have been used. In addition, several studies have reported that the skin melanin biosynthesis inhibition, lipolysis effect, etc. of hwangbaek tree are studied, and various pharmacological effects of berberine, which is a representative main component, have been studied.

Houttuynia cordata (魚腥草; Houttuyniae Herba) is a perennial herbaceous smells as outposts of Houttuynia Cordata (Houttuynia cordata) belonging to saururaceae (Saururaceae) peculiar smell of fish from the leaves and stems. Since ancient times, it has been used as a therapeutic agent for fever, boils, bronchitis, detoxification, lipid peroxidation inhibitory activity, antibacterial activity, antimutation, antiviral, upper limiting activity, intermediate rapid toxicity suppression effect and liver protection effect.

Samhwang and Eoseongcho, which are the experimental materials, are classified as blue fever medicines that give heat in herbal medicine, and the main components such as baicalin, Bicalein, Berberine, and Quercetin have a wide range of antibacterial effects against Gram-positive and negative bacteria and fungi. Inflammation, hemostasis, lowering blood pressure, lowering blood sugar, immune activation, and anti-cancer effects.

Example 1-Extraction conditions and extraction method

Gold (Scutellariae Radix, abbreviated as SR for convenience in the following experiments), Hwangnyeon (Coptidis Rhizoma, abbreviated as CR), Hwangbaek (Phellodendri Cortex, abbreviated as PC) and Eoseongcho (Houttuyniae Herba, HH) for carrying out this research project (Abbreviated as)) was purchased from “Omni Herb”, a specialized herbal medicine company, and was used as an extraction sample. Distilled water corresponding to 10 times the weight (w / w) of Samwha and Eoseongcho was placed in a round flask attached with a reflux cooling tube vertically, and water was extracted using Reflux extractor (Changshin, Korea) for 6 hours at 60 ℃. It was referred to as an extract (W; Water extract), and ethanol at 70 ° C was refluxed in the same manner as above under the conditions of 60 ° C as a solvent, respectively, to make an ethanol extract (E; Ethanol extract). In addition, extracting 1,3 Butylene glycol equivalent to 10 times (w / w) of Samhwang and Eoseongcho as a solvent, BG100 (1,3 Butylene glycol 100%), water and 1,3 Butylene glycol were 1: 1. Extracted by mixing at a ratio of WBG (1,3 Butylene glycol in water) and ethanol and 1,3 Butylene glycol were each diluted 1: 1 and the extract used as a solvent was EtBG (1,3 Butylene glycol in ethanol). Each extract was filtered with filter paper (Whatman No 2) and used as a sample to measure useful components and physiological activities of the extracts of Samhwang and Eoseongcho under each condition.

Example 2-Polyphenol and flavonoid compound content measurement

1) Total polyphenol compound content

The content of the polyphenol compound in the extract of Samhwang and Eoseongcho extracted with different solvents was measured by the Folin-Denis (AOAC, 2005) method. To 0.2 mL of the sample, 1.8 mL of distilled water and 0.2 mL of Folin-Ciocalteu's phenol reagent (Junsei Chemical Co. Tokyo, Japan) were added, mixed and left at room temperature for 3 minutes, and then 0.4 mL of saturated Na ₂ CO ₃ solution was added and mixed. After adding 1.4 mL of distilled water and reacting for 1 hour at room temperature, the absorbance was measured at 725 nm using a spectrophotometer (Mecasys Optizen POP, Korea). The total polyphenol compound was taken at 725 nm in the same manner as above by taking the final concentrations of 0, 31.25, 62.5, 125, 250, 500 μg / mL using tannic acid (Sigma-Aldrich Co., St Louis MO, USA). The absorbance was measured to obtain the content of polyphenol compounds for golden, hwangnyeon, hwangbaek and eoseongcho extracts from the standard curve.

2) Total flavonoid compound content

To quantify the total flavonoid, modify the method (2000) of Nieva Moreno et al., Add 0.1 mL of 10% aluminum nitrate, 0.1 mL of 1M potassium acetate, and 4.7 mL of 80% ethanol to 0.1 mL of each concentration extract. After reacting at 25 ° C for 40 minutes, absorbance was measured at 415 nm. Total flavonoids were quantified using quercetin (Sigma Chemical Co.) to have final concentrations of 0, 15.625, 31.25, 62.5, 125, 250, 500 μg / mL, and extracted under different conditions from the standard curve measured in the same manner as above. The total flavonoid compound content of Samhwang and Eoseongcho extracts was determined.

3) Measurement result

The golden extract was analyzed to contain polyphenols of 92.00 mg / g in ethanol extract (SR-EtOH) and 69.01 mg / g in water extract (SR-W), with 100% 1,3 Butylene glycol and water and ethanol. Diluted WBG and EtBG were found to contain 1.77 mg / g, 0.09 mg / g, and 1.96 mg / g. In the Hwangnyeon extract, CR-W extracted with water contained the most polyphenol at 85.20 mg / g, and then CR-EtOH using ethanol as a solvent contained 46.69 mg / g. Hwangbaek contained 82.29 mg / g and 82.80 mg / g CP-W and CP-EtOH, respectively, and Eoseongcho contained HH-W 82.80 mg / g and HH-EtOH 61.67 mg / g. It was analyzed that the 1,3 butylene glycol extract contained much lower polyphenols when water and ethanol were used as solvents.

The content of the flavonoid compound was that the golden extract contained the most flavonoids with SR-EtOH 10.80 mg / g, followed by SR-W 4.96 mg / g and SR-WBG 2.43 mg / g. The extract of Hwangnyeon was found to contain the most flavonoids among the total extracts of CR-EtOH 25.69 mg / g, and CR-W was found to contain 15.64 mg / g. The Hwangbaek extract also contained 4.89 mg / g and 9.43 mg / g in water and ethanol extracts CP-W and CP-EtOH, respectively, and extracts containing 1,3 butylene glycol showed low flavonoid content. Eoseongcho extract was analyzed to contain 10.93 mg / g HH-W and 4.74 mg / g HH-EtOH. Each value is expressed as mean ± SD. The results of these experiments are shown in Table 1 and FIG. 1 below.

Extraction solvent (Extract) Compound (mg / g) Polyphenols Flavonoids SR-W 69.01 ± 0.49 ^{2) b3)} 4.96 ± 0.09 ^b SR-EtOH 92.00 ± 0.34 ^a 10.80 ± 0.31 ^a SR-BG100 1.77 ± 0.02 ^d 0.56 ± 0.00 ^e SR-WBG 0.09 ± 0.01 ^e 2.43 ± 0.06 ^c SR-EtBG 1.96 ± 0.02 ^c 0.93 ± 0.01 ^d CR-W 85.20 ± 1.01 ^a 15.64 ± 0.57 ^b CR-EtOH 46.69 ± 0.64 ^b 25.69 ± 0.11 ^a CR-BG100 0.97 ± 0.01 ^d 0.94 ± 0.01 ^d CR-WBG 0.07 ± 0.01 ^e 1.34 ± 0.03 ^c CR-EtBG 1.38 ± 0.01 ^c 1.40 ± 0.02 ^c CP-W 82.29 ± 0.93 ^a 4.89 ± 0.03 ^b CP-EtOH 82.80 ± 1.13 ^a 9.43 ± 0.15 ^a CP-BG100 0.42 ± 0.04 ^d 0.15 ± 0.00 ^e CP-WBG 2.14 ± 0.02 ^b 0.50 ± 0.01 ^c CP-EtBG 1.27 ± 0.01 ^c 0.37 ± 0.00 ^d HH-W 52.80 ± 0.49 ^b 10.93 ± 0.10 ^a HH-EtOH 61.67 ± 0.75 ^a 4.74 ± 0.08 ^b HH-EtBG 1.17 ± 0.03 ^d 0.79 ± 0.02 ^c

Example 3-Electron donating ability

The electron donating ability of Samhwang and Eoseongcho extracts was measured by the effect of hydrogen donating to DPPH (1,1-diphenyl-2-picryl hydrazyl) according to Blois' method (1958). 1 mL of DPPH diluted in absolute ethanol at a concentration of 0.2 mM was added to 2 mL of a sample of Samsul and Eoseongcho extracted at a certain concentration, mixed, and reacted at 37 ° C. for 30 minutes. The absorbance of this reaction solution was measured at 517 nm, and the electron donating ability was expressed as a percentage (%) of the difference in absorbance before and after adding the sample.

Table 2 shows the electron donating ability of the extracts according to the solvents of Samhwang and Eoseongcho. As a result of measuring the electron donating ability for each golden solvent, SR-EtOH at the concentration of 3% showed the highest electron donating ability at 92.12%, and showed electron donating activity of SR-WBG 91.44% and SR-WBG 91.44%, the control Vit The results were similar to C. In particular, in the case of SR-WBG, even at 0.5% concentration, it showed 83.66% higher electron donating ability than Hwangnyeon, Hwangbaek and Eoseongcho. In the case of Hwangnyeon, the highest electron donating activity was found in CR-BG100 and CR-EtOH at 86.04% and 85.37%, respectively. Unlike the results of polyphenols and flavonoids, it was slightly lower at 68% in CR-W and CR-E. Showed activity. Electron donating ability by sulfur white solvent was found in the order of CP-EtBG 89.70%> CP-EtOH 89.12%> CP-WBG 88.55%> CP-W 57.40%> CP-BG100 52.36%, the highest in the solvent containing ethanol. Showed electron donating activity. In the case of Eoseongcho extract, HH-EtOH showed the highest electron donating effect with 90.22%, followed by HH-WBG (82.03%) and HH-BG100 (74.65%). The results are shown in Table 2 and Figure 2 below.

Extraction solvent Concentration (%) ^a 0.5 1.0 2.0 3.0 SR-W 48.18 ± 1.93 ^{2) d3)} 69.95 ± 0.74 ^c 84.79 ± 1.70 ^c 80.68 ± 4.15 ^b SR-EtOH 41.81 ± 2.23 ^e 66.23 ± 1.14 ^d 80.86 ± 1.84 ^d 92.12 ± 1.52 ^a SR-BG100 72.60 ± 3.68 ^b 78.54 ± 3.42 ^b 84.09 ± 3.65 ^bc 90.53 ± 1.52 ^a SR-WBG 83.66 ± 1.40 ^a 86.90 ± 0.98 ^a 89.23 ± 0.59 ^a 91.44 ± 1.14 ^a SR-EtBG 61.06 ± 0.19 ^c 75.19 ± 1.78 ^b 86.41 ± 0.86 ^b 90.51 ± 1.14 ^a CR-W 19.27 ± 2.76 ^e 29.46 ± 1.71 ^e 61.01 ± 2.52 ^c 68.72 ± 3.49 ^c CR-EtOH 29.33 ± 1.34 ^d 42.47 ± 2.05 ^d 56.79 ± 0.50 ^d 68.85 ± 0.95 ^c CR-BG100 51.67 ± 2.94 ^b 65.86 ± 1.40 ^b 83.85 ± 4.01 86.04 ± 3.29 ^a CR-WBG 45.80 ± 0.60 ^c 61.42 ± 1.72 ^c 74.93 ± 1.20 ^b 81.89 ± 3.61 ^b CR-EtBG 78.34 ± 1.93 ^a 80.39 ± 0.7 ^a 1 83.56 ± 1.71 ^a 85.37 ± 0.90 ^a CP-W 20.11 ± 3.01 ^d 32.30 ± 2.39 ^d 48.71 ± 1.55 ^b 57.40 ± 2.15 ^b CP-EtOH 48.99 ± 2.54 ^b 56.76 ± 1.81 ^b 85.90 ± 0.99 ^a 89.12 ± 1.60 ^a CP-BG100 26.38 ± 1.36 ^c 28.87 ± 1.98 ^e 43.44 ± 2.53 ^d 52.36 ± 2.19 ^c CP-WBG 54.20 ± 1.33 ^a 71.01 ± 2.80 ^a 85.80 ± 1.33 ^a 88.55 ± 0.91 ^a CP-EtBG 49.83 ± 2.18 ^b 65.34 ± 3.28 ^b 81.28 ± 1.95 ^b 89.70 ± 0.66 ^a HH-W 8.42 ± 1.30 ^d 17.15 ± 1.10 ^e 32.60 ± 0.99 ^e 49.07 ± 1.32 ^e HH-EtOH 9.90 ± 1.08 ^d 23.00 ± 1.08 ^d 37.01 ± 1.59 ^d 60.61 ± 2.48 ^d HH-BG100 38.15 ± 2.67 ^c 52.58 ± 2.15 ^c 67.84 ± 2.64 ^c 74.65 ± 1.06 ^c HH-WBG 50.89 ± 0.65 ^b 58.31 ± 2.87 ^b 73.04 ± 2.71 ^b 82.03 ± 2.04 ^b HH-EtBG 59.73 ± 2.93 ^a 78.34 ± 2.41 ^a 85.70 ± 0.36 90.22 ± 0.83 ^a Vit C (Comparative Example) 88.15 ± 2.00 90.52 ± 0.68 93.48 ± 0.93 94.37 ± 1.80

Example 4-ABTs radical cation scavenging ability

The antioxidant power of the extract using ABTs radical was measured by ABTs + ㆍ cation decolorization assay method (Roberta et al., 1999). 7mM 2,2-azino-bis (3-ethyl benzthiazoline-6-sulfonic acid) and 2.4 mM potassium persulfate were mixed and left in a cow at room temperature for 24 hours to form ABTs + ㆍ, diluted with ethanol, and then diluted to ABTs + ㆍ 100uL. After 100 μL of sample was added and left for 1 minute, absorbance was measured at 732 nm using a UV / VIS spectrophotometer to indicate the difference in absorbance between the sample treated and untreated groups as a percentage (%). It was expressed as the ability to scavenge ABTs radical cation for the extract.

Table 3 shows the results of measuring the radical scavenging ability of ABTs of golden, hwangnyeon, hwangbaek and eoseongcho. Golden BG100 and WBG showed the highest ABTs radical scavenging activity at concentrations of 0.5% at 97.40% and 97.44%, especially SR-WBG showed 92% scavenging activity at 0.05% concentration, the highest scavenging activity than other extracts at the same concentration. Showed. The extract of hwangnyeon was 98.98% in CR-WBG, showing similar activity to vitamin C as a control, and showed scavenging activity in the order of CR-EtBG 87.55% and CR-BG100 83.50%. As a result of measuring the radical scavenging ability of ABTs by extracts of each yellow-white solvent, CP-WBG showed the highest activity at 91.96%, and CP-EtBG had an effect of 87.62%. In Eoseongcho extract, HH-WBG showed high scavenging activity of 95.27% and HH-EtOH 90.38%, and water extract HH-W showed less than 50% activity at a concentration of 0.5%. The experimental results are shown in Table 3 and FIG. 3 below.

Extracts Concentration (%) 0.05 0.1 0.25 0.5 SR-W 9.33 ± 0.37 ^{2) c3)} 31.75 ± 0.91 ^d 44.34 ± 1.77 ^c 61.23 ± 0.77 ^c SR-EtOH 1.56 ± 0.83 ^d 11.76 ± 0.52 ^e 30.27 ± 1.67 ^d 62.34 ± 1.11 ^c SR-BG100 57.60 ± 0.15 ^b 78.82 ± 1.21 ^b 96.74 ± 0.13 ^a 97.40 ± 1.30 ^a SR-WBG 92.32 ± 1.14 ^a 94.83 ± 0.74 ^a 96.95 ± 0.66 ^a 97.44 ± 0.25 ^a SR-EtBG 56.45 ± 1.46 ^b 69.43 ± 1.27 ^c 85.98 ± 1.30 ^b 89.83 ± 0.45 ^b CR-W 10.82 ± 0.27 ^c 19.62 ± 1.12 ^d 37.32 ± 0.99 ^d 61.32 ± 0.87 ^d CR-EtOH 6.25 ± 0.58 ^d 18.37 ± 1.89 ^d 38.28 ± 0.93 ^d 61.03 ± 0.90 ^d CR-BG100 32.05 ± 2.51 ^b 51.53 ± 2.61 ^c 72.04 ± 0.48 ^c 83.50 ± 0.90 ^c CR-WBG 48.52 ± 1.68 ^a 69.25 ± 1.00 ^a 90.12 ± 3.09 ^a 98.98 ± 1.64 ^a CR-EtBG 48.29 ± 1.64 ^a 63.04 ± 1.04 ^b 79.07 ± 1.67 ^b 87.55 ± 1.15 ^b CP-W 4.98 ± 1.14 ^e 15.49 ± 0.83 ^e 42.37 ± 0.76 ^e 64.09 ± 1.09 ^d CP-EtOH 11.73 ± 0.73 ^d 22.49 ± 0.61 ^d 51.19 ± 0.35 ^c 69.58 ± 0.79 ^c CP-BG100 21.40 ± 2.15 ^c 27.43 ± 1.63 ^c 39.89 ± 1.49 ^d 50.37 ± 2.11 ^e CP-WBG 52.73 ± 0.62 ^a 68.85 ± 1.20 ^a 81.40 ± 0.54 ^a 91.96 ± 1.07 ^a CP-EtBG 40.97 ± 1.77 ^b 60.20 ± 1.05 ^b 78.45 ± 1.74 ^b 87.62 ± 1.98 ^b HH-W 9.11 ± 1.81 ^d 14.18 ± 2.18 ^e 31.08 ± 0.16 ^e 48.17 ± 0.61 ^e HH-EtOH 5.90 ± 0.31 ^e 22.31 ± 0.83 ^d 40.59 ± 0.64 ^d 67.62 ± 0.24 ^d HH-BG100 32.05 ± 0.73 ^c 38.11 ± 1.26 ^c 54.77 ± 1.27 ^c 65.92 ± 0.74 ^c HH-WBG 65.80 ± 1.38 ^a 81.24 ± 1.16 ^a 92.12 ± 1.73 ^a 95.72 ± 0.21 ^a HH-EtBG 47.01 ± 1.93 ^b 62.09 ± 1.50 ^b 79.59 ± 2.12 ^b 90.38 ± 1.77 ^b Vit C (Comparative Example) 97.47 ± 0.71 98.03 ± 0.14 99.25 ± 0.22 99.01 ± 0.37

Example 5 -  Cytotoxicity measurement using MTT assay

Cytotoxicity of Samhwang and Eoseongcho with different extracts was investigated using the MTT assay for cytotoxicity of macrophage RAW 264.7 and HaCaT cells (Mosmann et al., 1983). RAW 264.7 and HaCaT cells were diluted with Dulbecco's modifide Eagle's medium (DMEM Gibco BRL., USA) medium at a concentration of 1 × 10 ⁵ cells / mL, dispensed 200 μL in 96 well plates (Falcon Co., USA), and 37 C. Incubator in 5% CO ₂ incubator for about 24 hours to adsorb RAW 264.7 and HaCaT cells to the bottom of the culture plate. Then, after removing all the supernatant by inhalation, the golden, hwangnyeon, hwangbaek, and eoseongcho extracts were diluted with DMEM medium by concentration (25, 50, 100, 200 ug / mL), and then treated with 200 uL in cells for 24 hours. Cultured for a while. After that, 100 µL of 5 ug / mL MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide, sigma, USA) solution was added to each well, 37 ℃, 5% Incubate for 3 hours in a CO ₂ incubator. After incubation, the supernatant was removed and 100 uL of DMSO (dimethyl sulfoxide) was dissolved in purple formazan crystals by reduction of MTT, dissolved and stirred for 10 minutes to measure absorbance at 570 nm using an ELISA reader. Cytotoxicity after sample treatment was compared to the negative control using the medium instead of the sample, and the survival rate (% control) was calculated.

As a result of confirming the survival rate of Raw 264.7 macrophages and HaCaT cells for the extracts of golden, hwangnyeon, hwangbaek and Eoseongcho extracted with different solvents by MTT assay, CR-E and HH-BG100 and HH-WBG were slightly different in Raw 264.7 macrophages. Although it exhibited toxicity, other extracts exhibited a survival rate of 85% or higher at a concentration of 200 ug / mL, thereby confirming cell viability. HaCaT cells, keratinocytes, showed a cell viability of 85% or more in all extracts, and thus no toxicity was observed affecting the cells. 4 shows this result.

Example 6- Temporary skin surface temperature reduction effectiveness evaluation Human application test

In August 2018, a human body application test was conducted at the Clinical Research Support Center for Cosmetics in Chungcheongbuk-do to evaluate the effect of temporarily reducing the skin surface temperature of "AtoBlue Ice Cooling" of L & T Nature Co., Ltd. of aerosol formulations. As a result, it was found that after 1 minute of application, a temporary skin surface temperature reduction effect of 27.770% was obtained.

Example 7- First skin irritation safety human application test

In August 2018, the Chungcheongbuk-do Cosmetics Clinical Research Support Center evaluated the skin irritation level of LTO2 Nature's "AtoBlue Ice Cooling" of the aerosol formulation and conducted a test to predict the degree of skin safety through this. As a result, evaluation results were found to be safe products that can be used in real life.

Claims (7)

As a cosmetic composition having sulfuric acid and heat efficacy,
The cosmetic preparation includes a mixed extract extracting a mixture of golden, hwangnyeon, hwangbaek and eoseongcho,
The mixed extract is obtained by a solvent in which the ratio of 1,3 butylene glycol and ethanol is 1: 1 or an extraction solvent in which the ratio of 1,3 butylene glycol and water is 1: 1,
A cosmetic composition comprising a cosmetic composition containing the mixed extract in the form of an aerosol. The cosmetic composition according to claim 1, wherein the golden, hwangnyeon, hwangbaek and eoseongcho are mixed and extracted in the same weight ratio. The cosmetic composition according to claim 1, wherein the mixed extract contains 0.001 to 10% by weight based on the total weight of the cosmetic composition. The cosmetic composition according to claim 1, wherein the cosmetic composition further comprises a second mixed extract obtained by mixing each extract obtained by ethanol solvent extraction of golden, yellow lotus, yellow white and eoseongcho. The method according to claim 4, wherein the second mixed extract is 90-95 mg / g for golden, 45-47 mg / g for hwangnyeon, 80-83 mg / g for hwangbaek, and 60-62 mg / g for eoseongcho. Each containing phenol (polyphenol);
Cosmetic composition characterized by containing 10 to 12 mg / g of golden, 23 to 27 mg / g of hwangnyeon, 8 to 10 mg / g of hwangbaek, and 4 to 6 mg / g of flavonoids, respectively. . A first step of preparing a mixture by mixing golden, hwangnyeon, hwangbaek and eoseongcho;
The mixture is added 30 to 70 by adding at least one solvent selected from the group consisting of a solvent having a 1: 1 ratio of 1,3 butylene glycol and ethanol or a solvent having a 1: 1 ratio of 1,3 butylene glycol and water. A second step of extracting for 5-10 hours at ℃;
A third step of filtering the extract obtained from the second step with a filtering device; And
Method for producing a cosmetic composition having a sulfated and heat sensation efficacy comprising a fourth step of preparing a cosmetic composition containing the filtrate obtained in the third step in an aerosol form. The method of claim 6, wherein the extraction of the second step extracts the mixture at 60 ° C. for 6 hours.

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