KR20190006888A

KR20190006888A - Composition comprising extracts, fractions and the isolated compounds of hibiscus hamabo siebold and zucc. for skin anti-pollution

Info

Publication number: KR20190006888A
Application number: KR1020170163326A
Authority: KR
Inventors: 박귀교; 장욱주; 김보윤; 김유아; 박병준; 김수영; 권혁준
Original assignee: 한국콜마주식회사; 대한민국(환경부 국립생물자원관장)
Priority date: 2017-07-11
Filing date: 2017-11-30
Publication date: 2019-01-21
Also published as: KR102018533B1

Abstract

The present invention relates to a composition for skin antipollution, and in the present invention, it is possible to provide a composition for skin antipollution, which comprises a compound separated from a component of a fungal extract, a fraction of the fungal extract and a fraction of the fungal extract, The present invention provides a composition for skin antipolution having an excellent antipollution effect.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a composition for skin antipollution comprising a compound separated from a fragrant extract, a fragrant extract, a fragrant extract and a fraction from a fragrant extract. FOR SKIN ANTI-POLLUTION}

The present invention relates to a composition for skin antipollution comprising a compound separated from the fractions of a fungus extract, a fungus extract and a fungus extract.

Skin is a thin film that surrounds the outside of the body. It functions to protect the human body from physical and chemical stimuli of the external environment. It prevents evaporation of moisture in the body and prevents excessive penetration of pollutants from the outside. Recently, as the use of chemicals has increased, the skin has been exposed to environmental pollution sources such as polycyclic aromatic hydrocarbons, aldehydes, nitrogen oxides, nitrates, and fine dusts generated during its manufacture, use and disposal, Itching, urticaria and inflammation, and skin diseases such as atopy, psoriasis and skin cancer are increasing.

Accordingly, the development of cosmetic compositions, cleaning agents, and external preparations having anti-pollution effects that prevent the environmental pollutants from adhering to the surface of the skin or penetrating the inside of the body through the skin is actively developed have. However, most of the products currently developed are merely removing the contaminants that are adhered to or penetrated into the skin and merely removing contaminants from the skin. In addition to not fundamentally protecting the skin from environmental pollutants, There is a problem in that the ability to protect the skin is insignificant.

In particular, since it contains a chemical substance as a main component, there is a problem that it acts as a cause of skin troubles and skin diseases. Accordingly, there is a need for the development of a composition having a low irritation effect on the skin and excellent antipollution effect using a natural substance.

On the other hand, Korean Patent No. 10-1571323 discloses a herb complex extract obtained by mixing peppermint, chamomile, rosemary, lemon grass, marigold, tea tree and lavender at the same weight ratio, Korean Patent Laid-Open No. 10-2017-0053388 discloses a cosmetic composition having an antipollution effect by containing an extract of Ulgum, apple, and grape. However, In the case of the above patent, the use of several extracts resulted in a high cost, but a small anti-pollution effect.

Korean Patent No. 10-1571323 Korean Patent Publication No. 10-2017-0053388

It is an object of the present invention to provide a skin anti-pilling agent which is excellent in antipollution effect that protects skin from environmental pollutants while having a low stimulus to the skin, by containing a compound separated from a natural substance, an extract of Huangguang, a fraction of a Huanggeong extract, And to provide a composition for a polyurethane.

The object of the present invention is not limited to the above-described technical problems, and another technical problem can be derived from the following description.

In order to accomplish the above object, the present invention provides a composition for skin antipollution comprising an extract of Huangguang as an active ingredient.

The extraction solvent of the above-described water-soluble extract may contain at least one selected from the group consisting of water, ethanol, methanol, acetone, and 1,3-butylene glycol.

In order to accomplish the above object, the present invention provides a skin anti-pollution composition comprising a fraction of the extract of Huangguang as an active ingredient.

The fractions of the above-described extracts may contain at least one selected from the group consisting of Ethyl acetate fraction, Methylene Chloride fraction and Hexane fraction of the above extract.

In order to achieve the above object, the present invention provides a composition for skin antipollution comprising tiliroside as an active ingredient.

The tiluroside may be isolated from the fraction of the Huangguang extract.

The composition may be to improve skin fouling, skin cell death or inhibition of skin cell proliferation by chemicals.

The chemical may include at least one selected from the group consisting of polycyclic aromatic hydrocarbons, aldehydes, nitrogen oxides, and nitrates.

The composition may be a cosmetic composition.

The composition for skin antipollution according to the present invention contains a compound separated from a natural substance such as a flaxseed extract, a flaxseed extract fraction, and a flaxseed extract fraction, and thus can be used as an anti-pollution agent that protects skin from environmental pollution sources Anti-pollution) effect can be excellent.

In particular, the composition for skin antipollution of the present invention may have excellent antipollution effect for protecting skin from environmental pollution sources such as polycyclic aromatic hydrocarbons, aldehydes, nitrogen oxides, nitrates, and fine dusts.

1 is a diagram showing a ¹ H NMR spectrum using nuclear magnetic resonance of a compound (Example 3) isolated from the ethyl acetate fraction of the crude extract prepared in Example 2. Fig.
2 is a diagram showing a ¹³ C NMR spectrum using nuclear magnetic resonance of a compound (Example 3) isolated from the ethyl acetate fraction of the crude extract prepared in Example 2. Fig.
FIG. 3 is a graph showing the results of evaluating the cytotoxicity of the extract of the yellow squash of Example 1, the fraction of Example 2, and the tiluroside of Example 3. FIG.
FIG. 4 is a graph showing cell proliferation rate when the extract of Example 1 was treated with ammonium nitrate. FIG.
Figure 5 is a plot showing the cell proliferation rate when the ethyl acetate fraction (EA fraction) of Example 2 was treated with benzopyrene.
Figure 6 is a plot showing the cell proliferation rate when the ethyl acetate fraction (EA fraction) of Example 2 was treated with formaldehyde.
Figure 7 is a plot showing the cell proliferation rate when the ethyl acetate fraction (EA fraction) of Example 2 was treated with ammonium nitrate.
Figure 8 is a plot showing the cell proliferation rate when the methylene chloride fraction (MC fraction) of Example 2 was treated with ammonium nitrate.
Figure 9 is a chart showing the cell proliferation rate when the hexane fraction (Hex fraction) of Example 2 was treated with ammonium nitrate.
10 shows the cell proliferation rate when the tiluroside of Example 3 was treated with ammonium nitrate.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings, which will be readily apparent to those skilled in the art. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. In order to clearly explain the present invention in the drawings, parts not related to the description are omitted.

It is to be understood that the terms or words used in the specification and claims are not to be construed in a conventional or dictionary sense and that the inventor may properly define the concept of a term in order to best describe its invention And should be construed in accordance with the principles and meanings and concepts consistent with the technical idea of the present invention.

In the specification of the present invention, when a component is referred to as " comprising ", it means that it can include other components as well as other components, .

Throughout the specification of the present invention, " A and / or B " means A or B, or A and B.

Throughout the specification of the present invention, " anti-pollution " may mean improving or preventing skin contamination, skin cell death or inhibition of skin cell proliferation by chemicals.

Throughout the specification of the present invention, the term " extract " includes all of the substances obtained by extracting the components of the natural substance, regardless of the extraction method, the extraction solvent, the extracted component or the form of the extract, It is a broad concept that includes all substances that can be obtained by extracting and then processing or treating the material obtained.

In the specification of the present invention, " no cytotoxicity " or " does not show cytotoxicity " may mean that the cell viability is 80% or more, but is not limited thereto.

The present invention has been specifically described below, but the present invention is not limited thereto.

SUMMARY OF THE INVENTION [ In the embodiment Skin containing Huang Geun Extract as an active ingredient For anti-pollution Lt; / RTI >

Hibiscus hamabo (Hibiscus hamabo or Hibiscus hamabo Siebold & Zucc.) Corresponds to the mulberry octopus of the dicotyledonous plant. It is about 1m high, and the bark is light gray brown and the young twigs have hairy densities. Leaves are alternate length 3 ~ 6 cm, width 3 ~ 7 cm, with hair on the front, grayish hairs on the back, pointed end, The flower has a 5 cm diameter yellow flower bloomed on the axilla of the end of the branch, and the center is dark purple.

The above-described extract of the present invention can be obtained by extracting an extract obtained by extraction treatment of a root-like plant such as root, stem, and leaf, a diluted solution or concentrate of the extract, a dried product obtained by drying the extract, Or a mixture thereof, extracts of all the formulations which can be formed using the extract itself and the extract. The extract may be preferably prepared in the form of a dry powder after extraction, and the above-described fimbriae outpost may be dried.

The extract of the present invention comprises (a) washing and drying the yellow hair; (b) mixing and extracting the dried yellow squid with the extraction solvent in step (a); And (c) filtration of the yolk in the mixture through step (b), and may be in a liquid form, which can be obtained by concentrating and drying to obtain an extract in powder form, Do not.

The extraction solvent of the above-mentioned water-soluble extract may contain at least one selected from the group consisting of water, ethanol, methanol, acetone and 1,3-butylene glycol, but not limited thereto, can do.

The composition for skin antipollution according to the present invention may have excellent antipollution effect with low irritation to the skin by containing the extract of Huangguang as an active ingredient. In particular, the composition for skin antipollution containing the above-described fungus extract can achieve an anti-pollution effect by preventing or ameliorating skin pollution, skin cell death or skin cell proliferation inhibition caused by environmental pollutants such as chemical substances.

The chemical substance may include at least one selected from the group consisting of polycyclic aromatic hydrocarbons, aldehydes, nitrogen oxides, and nitrates, but is not limited thereto.

The polycyclic aromatic hydrocarbons may include benzoprene, and the aldehydes may include formaldehyde. The nitrogen oxides may be applicable to all those that can be represented by the NOx formula, for _{example, NO, NO 2, N 2} O, N 4 O, N 2 O 3, N 2 O 4, N 2 O ₅ , N ₄ O ₆ , and the like. The nitrate salts include a salt consisting of a metal or a basic group and a nitrate group, preferably ammonium nitrate and an ionic salt thereof.

Preferably, in this embodiment, the extract may contain a methanol extract of methanol, and in particular, an ethanol extract of methanol extract of 25 to 100 μg / ml may have an anti-pollution effect on ammonium nitrate , But is not limited thereto.

The composition for skin antipollution according to the present invention contains 0.01-30% by weight, preferably 0.1-20% by weight, more preferably 1-10% by weight, most preferably 1-5% by weight . If the content of the extract is less than 0.01% by weight based on the total weight of the composition for skin antipollution, the antipollution effect is insignificant. If the extract contains more than 30% by weight of the extract, There may be a problem.

In addition, In the embodiment Of Huang Geun Extract The fraction Skin as active ingredient For anti-pollution Lt; / RTI >

The fraction of the extract of the present invention is obtained by fractionation of the extract of the present invention. The fraction can be subjected to chromatography, electrophoresis, principle separation or the like. The fractions of the extract of the present invention may contain at least one selected from the group consisting of ethyl acetate fraction, methylene chloride fraction and hexane fraction of the extract of Huangguang.

In addition, fractions of the above-described extracts of the above-described fungus extract may be used as they are, or may be used in the form of a dried product obtained by concentration after filtration and then extracting or concentrating and lyophilizing.

The composition for skin antipollution according to the present invention contains fractions of the extract of Huangguang as an active ingredient, so that it is possible to have an excellent antipollution effect while having low irritation to the skin. Particularly, the skin anti-pollution composition containing the fractions of the above-described extract of Huang Gui extract can prevent or improve skin pollution caused by environmental pollutants such as chemical substances, skin cell death or inhibition of skin cell proliferation, thereby realizing an anti-pollution effect.

Preferably, the composition for skin antipollution according to the present invention may comprise an ethyl acetate fraction of a water extract of the group A, and more preferably a water extract of a water extract of a group having a concentration of 10 to 75 μg / , The antipollution effect on benzopyrene, formaldehyde and ammonium nitrate may be excellent when the concentration of the ethyl acetate fraction is 18.75 to 75 μg / ml, but the present invention is not limited thereto.

The composition for skin antipollution according to the present invention contains 0.01 to 30% by weight, preferably 0.1 to 20% by weight, more preferably 1 to 10% by weight, and most preferably 1 to 5% by weight of the fractions of the above- %. If the fraction of the extract of the present invention is less than 0.01% by weight based on the total weight of the composition for skin antipollution, the antipollution effect is insignificant. When the fractions of the extract are contained in an amount of more than 30% There may be a problem that is not economical.

In addition, In the embodiment Tiliroside Skin as active ingredient For anti-pollution Lt; / RTI >

The tiluroside of the present invention may be obtained by separating the active fraction by performing a medium pressure liquid chromatography (MPLC) or a preparative liquid chromatography (prep-LC) on the fraction of the above-described extract of the above-described fungus extract, But is not limited thereto. In addition, in the present embodiment, tyllioside and isomers thereof may all be included.

[Chemical Formula 1]

The composition for skin antipollution according to the present invention contains tilirroside as an active ingredient, so that it is possible to provide a low-irritation effect on the skin and an excellent antipollution effect. In particular, the composition for skin antipollution including tiliroside of the present invention can prevent or improve skin pollution, skin cell death or inhibition of skin cell proliferation due to environmental pollutants such as chemical substances, thereby realizing an anti-pollution effect.

The chemical may be a nitrate salt, but is not limited thereto. The nitrate salts include a salt consisting of a metal or a basic group and a nitrate group, preferably ammonium nitrate and an ionic salt thereof.

Preferably, the tiluroside may be isolated from the ethyl acetate fraction of the extract of Huangguang in this example, and thus the antipollution effect against benzopyrene may be excellent, but the present invention is not limited thereto. Particularly, in this embodiment, tyllioside is separated from the ethyl acetate fraction of the extract of Huang Geun Extract. When the concentration of tiliroside is 0.01-5 μg / ml, preferably 2.5-5 μg / ml , It may have an anti-pollution effect on ammonium nitrate, but is not limited thereto.

The composition for skin antipollution according to the present embodiment contains 0.01 to 30% by weight, preferably 0.1 to 20% by weight, more preferably 1 to 10% by weight, and most preferably 1 to 5% by weight of the tilirroside. As shown in FIG. If the tilylside is contained in an amount of less than 0.01% by weight based on the total weight of the composition for skin antipollution, the antipollution effect is insignificant. If the tililoside is contained in an amount exceeding 30% by weight, There may be some problems.

The skin anti-pollution composition comprising the composition for skin antipollution comprising the above-described fungal extract, the composition for skin antipollution comprising the fraction of the fungal extract or the fraction isolated from the fraction of the fungal extract is preferably a cosmetic Composition, but is not limited thereto.

When the composition for skin anti-pollution is a cosmetic composition, it may contain softening agents, nutritional lotion, skin, mist, lotion, cream, essence, serum, pack, cleanser (cleansing cream, cleansing foam, Body lotion, body spray, body spray, etc.), powder, foundation, lipstick, mascara, makeup base, eyeliner, eye shadow, blusher, shampoo, rinse, conditioner, hair gel, hair serum, hair essence, body lotion . &Lt; / RTI >

The excipient may include a coloring agent, a fragrance, an emulsifier, a thickener, a pigment, a fragrance, an antioxidant, an antiseptic, a bactericide, a solvent, a thickener, an inorganic salt and a synthetic polymer material. Antioxidants, wrinkle removers, moisturizers, and the like.

Hereinafter, the composition for skin antipollution comprising as an active ingredient the compound isolated from the fractions of the present invention, the fractions of the above-described fractions of the above-described fragrance of the present invention, . These embodiments are only for illustrating the present invention, and thus the scope of the present invention is not construed as being limited by these embodiments.

[ Example ]

Example 1: Preparation of Huang Geun Extract

15 g of methanol was added to 700 g of the dried yellowfin tuna plant and extracted three times at room temperature for 72 hours. The extracted extract was filtered, the solvent was removed using a vacuum concentrator, and the resulting extract was lyophilized to prepare a methanol extract of yellow roots.

Example 2: The Hexane Fraction , Methylene chloride Fraction , Ethyl acetate Fraction Produce

The same procedure as in Example 1 was repeated by suspending the extract in methanol (methanol extract) in 1.5 L of distilled water, adding the same amount of hexane and fractionating with a fractionating funnel, repeating the procedure three times, extracting the fractionated hexane layer, Was removed and lyophilized to prepare a hexane fraction (9.21 g).

Then, the separated hexane layer was separated, and the remaining distilled water layer was fractionated according to the order of methylene chloride and ethyl acetate polarity to prepare methylene chloride fraction (1.34 g) and ethyl acetate fraction (1.16 g).

Example 3: Ethyl acetate of the Huang Geun Extract From the fraction Tillyloside Separation and structure identification

The ethyl acetate fraction (1.16 g) of the crude extract of Example 2 was subjected to MPLC under reversed phase silica gel and acetonitrile (ACN) -Water solvent gradient conditions to give a total of five fractions. Among them, the third fraction (fraction-3) was subjected to prep-LC with an ACN-Water solvent to separate the compounds. The obtained compound was confirmed by NMR to be purely isolated. A ¹ H, ¹³ C NMR spectrum (see FIGS. 1 and 2 and the following description) was obtained using a nuclear magnetic resonance machine (Bruker Co. 400 MHz, NMR), and these spectra were analyzed synthetically to determine the structure

^{1 H-NMR (400 MHz,} MeOD): δ 7.97 (2H, d, J = 8.4 Hz, H-2 ', 6'), 7.39 (1H, d, J = 16.0 Hz, H-7 ''') (2H, d, J = 9.2 Hz, H-3 ', 5'), 7.27 (2H, d, J = 8.4 Hz, H-2 ' , J = 8.8 Hz, H- 3 '''.5'''), 6.28 (1H, s, H-8), 6.11 (1H, s, H-6), 6.06 (1H, d, J = 16.0 Hz, H-8 ''' ), 5.24 (1H, m, H -1''), 4.31 (1H, d, J = 10.0 Hz, H-6β''), 4.19 (1H, dd, J = 11.6 , 6.4 Hz, H-6.alpha.), 3.52 (1H, m, H-5 ''), 3.48 3.46 (1H, m, 4 ")

^{13 C-NMR (400 MHz,} MeOD): δ 179.4 (C-4), 168.9 (C-9 '''), 165.9 (C-9), 163.0 (C-5), 161.6 (C-4') , 161.2 (C-4 '''), 159.4 (C-7), 158.4 (C-2), 146.6 '), 131.3 (C-2''', 6 '''), 127.1 (C-1'''), 122.8 (C- , 116.1 (C-3 ', 5'), 114.8 (C-8 '''), 105.7 ), 78.1 (C-3 ''), 75.9 (C-2 ''), 75.8 (C-5 '

As a result, the compound (light yellow powder) was identified as tiliroside of the above formula (1).

[ Experimental Example ]

Experimental Example 1: Evaluation of Huangguang extract

Experimental Example 1 - 1: MTT Cells using analytical methods Survival evaluation

The cells were cultured in a CO ₂ incubator at 37 ° C for 24 hours at a concentration of 1 × 10 ⁴ cells / ml in a 96-well plate. At this time, the medium was an IMDM medium containing 10% bovine serum and 1% antibiotics.

After the culture, the medium was replaced with an IMDM medium in which bovine serum and antibiotics were excluded. Then, the extract of the extract of Example 1 (methanol extract) was added at a concentration of 0.01 to 100 μg / ml for 21 hours in the same culture conditions.

Then, 5 mg / ml MTT solution was added and incubated for 3 hours. Then, the medium was removed and 150 μl of dimethyl sulfoxide was added to each well, shaking for 10 minutes, and the absorbance was measured at 570 nm. The results are shown in Fig.

In FIG. 3, the methanol is the yellowish methanol extract of Example 1, the n-hexane is the hexane fraction of Example 2, the methylene chloride is the methylene chloride fraction of Example 2, the ethyl acetate fraction is the ethyl acetate fraction of Example 2, Quot; means tiluroside of Example 3.

As shown in FIG. 3, it can be confirmed that the extract of Huanggeong showed no cytotoxicity at a concentration of 0.01 to 100 μg / ml.

Experimental Example 1 - 2: Anti-pollution Activity evaluation

In general, the degree of cell damage by ammonium nitrate (500 ppm) was measured using dermal keratinocytes that directly contacted with pollutants. The extract of the fermented milk of Example 1 (25 μg / ml, 50 μg / ml, 100 μg / ml) to determine the degree of cell damage by ammonium nitrate.

Human keratinocyte HaCaT was aliquoted at a concentration of 1 × 10 ⁴ cells / ml in a 96-well plate and incubated at 37 ° C. in a CO ₂ incubator for 24 hours. At this time, the medium was an IMDM medium containing 10% bovine serum and 1% antibiotics.

After the culture, the medium was replaced with an IMDM medium in which bovine serum and antibiotics were excluded. Then, the medium was treated with ammonium nitrate and further treated with the fermented methanol extract of Example 1 for 21 hours. After incubation for 3 hours with 5 mg / ml MTT solution, media was removed and 150 μl of dimethyl sulfoxide was added to each well. After shaking for 10 minutes, absorbance was measured at 570 nm. The results are shown in FIG. 4 and Table 1.

Huang Geun Extract (Concentration) Blank (-) Blank (+) 100 μg / ml 50 μg / ml 25 μg / ml Cell proliferation (%) 100 45 66 84 71

4 and Table 1, it was confirmed that the effect of ammonium nitrate on the cell damage was suppressed when the extract was treated with the extract. More specifically, it can be confirmed that when the concentration of the Huang Guan extract is 25-100 μg / ml, the cell proliferation rate is superior to that of ammonium nitrate-treated group (blank (+)).

Taken together, the results of Table 1 and Figs. 3 to 4 show that the extract of the present invention has an antipollution effect against ammonium nitrate without exhibiting cytotoxicity when the concentration of the extract is 0.01 to 100 μg / ml have.

Experimental Example 2: Evaluation of fractions of the Huangguang extract

Experimental Example 2-1: Evaluation of cell viability using MTT assay

After the culture, the medium was replaced with the IMDM medium in which bovine serum and antibiotics were excluded, and then the ethyl acetate fraction, methylene chloride fraction, and hexane fraction of Example 2 were treated with each concentration (0.01 to 100 μg / ml) Lt; / RTI > for 21 hours.

3, it can be confirmed that the ethyl acetate fraction does not show cytotoxicity at a concentration of 0.01 to 75 μg / ml.

3, it can be confirmed that the methylene chloride fraction does not show cytotoxicity at a concentration of 0.01 to 10 μg / ml.

3, it can be confirmed that the hexane fraction does not show cytotoxicity at a concentration of 0.01 to 10 μg / ml.

Experimental Example 2-2: Antipollution Evaluation of Ethyl Acetate Fraction (EA Fraction)

In general, the degree of cell damage by each of benzopyran (2.5 ppm), formaldehyde (1.5 ppm), and ammonium nitrate (500 ppm) was measured using keratinocytes that directly contacted with pollutants. (2.5 ppm), formaldehyde (1.5 ppm) and ammonium nitrate (500 ppm) were treated with ethyl acetate fractions of methanol extracts at different concentrations (18.75 μg / ml, 37.5 μg / ml and 75 μg / ml) The degree of damage was measured, and the effect of inhibiting skin cell damage was confirmed.

After the culture, the medium was replaced with an IMDM medium in which bovine serum and antibiotics were excluded. Then, each of benzo-pyrene, formaldehyde and ammonium nitrate was treated with the ethyl acetate fraction of Example 2 and further cultured for 21 hours. After incubation for 3 hours with 5 mg / ml MTT solution, media was removed and 150 μl of dimethyl sulfoxide was added to each well. After shaking for 10 minutes, absorbance was measured at 570 nm. The results are shown in Figs. 5 to 7 and Tables 2 to 4.

Table 2 and FIG. 5 show the results of treatment with benzopyrene. Table 3 and FIG. 6 show the results of treatment with formaldehyde, and Table 4 and FIG. 7 show the results of treatment with ammonium nitrate.

EA fraction (concentration) Blank (-) Blank (+) 75 μg / ml 37.5 μg / ml 18.75 μg / ml Cell proliferation (%) 100 50 102 109 77

EA fraction (concentration) Blank (-) Blank (+) 75 μg / ml 37.5 μg / ml 18.75 μg / ml Cell proliferation (%) 100 57 108 114 90

EA fraction (concentration) Blank (-) Blank (+) 75 μg / ml 37.5 μg / ml 18.75 μg / ml Cell proliferation (%) 100 51 80 103 84

5 to 7 and Tables 2 to 4, it was confirmed that the treatment with ethyl acetate fraction of the extract of Huang Geun Extract inhibited cytotoxicity against benzopyran, formaldehyde and ammonium nitrate.

More specifically, when the concentration of the ethyl acetate fraction is 18.75 to 75 μg / ml, it can be confirmed that the cell proliferation rate is superior to that of the group treated with only benzo-pyrene, formaldehyde and ammonium nitrate (Blank (+)). In particular, it was confirmed that the treatment with benzo-pyrene, formaldehyde, and ammonium nitrate showed the best cell proliferation rate when the ethyl acetate fraction treatment concentration was 37.5 μg / ml.

As a result of the above Experimental Examples 2-1 and 2-2, the ethyl acetate fraction of the Huang Geun Extract did not exhibit cytotoxicity when the concentration was 0.01 to 75 μg / ml, but the antipollution effect against benzopyrene, formaldehyde and ammonium nitrate As shown in Fig.

Experimental Example 2 - 3: Antipollution Evaluation of Methylene Chloride Fraction (MC fraction)

In general, the degree of cell damage by ammonium nitrate (500 ppm) was measured using keratinocytes that directly contacted with pollutants. The methylene chloride fraction of the extract of the fermented methanol extract of Example 2 (2.5 μg / ml , 5 μg / ml, and 10 μg / ml) to determine the degree of cell damage by ammonium nitrate (500 ppm).

After the culture, the medium was replaced with an IMDM medium in which bovine serum and antibiotics were excluded. After treatment with ammonium nitrate, the methylene chloride fraction was treated in Example 2 and further cultured for 21 hours. After incubation for 3 hours with 5 mg / ml MTT solution, media was removed and 150 μl of dimethyl sulfoxide was added to each well. After shaking for 10 minutes, absorbance was measured at 570 nm. The results are shown in FIG. 8 and Table 5.

MC fraction (concentration) Blank (-) Blank (+) 10 μg / ml 5 μg / ml 2.5 μg / ml Cell proliferation (%) 100 46 65 63 46

In Table 5 and FIG. 8, it was confirmed that the methylene chloride fraction of the extract of Huanggeong extract inhibited cell damage to ammonium nitrate. More specifically, it can be confirmed that when the treatment concentration of the methylene chloride fraction is 5 to 10 μg / ml, the cell proliferation rate is superior to that of the control (Blank (+)) treated with ammonium nitrate only.

Taken together, Experimental Examples 2 - 1 and 2 - 3 show that the methylene chloride fraction of the Huang Geun Extract has an antipolution effect against ammonium nitrate even without cytotoxicity when the concentration is 0.01 to 10 μg / ml .

Experimental Example 2 - 4: Evaluation of antipollution of hexane fraction

In general, the degree of cell damage by ammonium nitrate (500 ppm) was measured using dermal keratinocytes, which were directly contacted with pollutants. The hexane fraction of the fermented methanol extract of Example 2 (2.5 μg / ml, 5 μg / ml, and 10 μg / ml) to determine the degree of cell damage by ammonium nitrate (500 ppm).

After incubation, the medium was replaced with bovine serum and antibiotic-free IMDM medium, treated with ammonium nitrate, treated with the hexane fraction of Example 2, and further cultured for 21 hours. After incubation for 3 hours with 5 mg / ml MTT solution, media was removed and 150 μl of dimethyl sulfoxide was added to each well. After shaking for 10 minutes, absorbance was measured at 570 nm. The results are shown in FIG. 9 and Table 6.

Hexane fraction (concentration) Blank (-) Blank (+) 10 μg / ml 5 μg / ml 2.5 μg / ml Cell proliferation (%) 100 46 85 66 52

In Table 6 and FIG. 9, it was confirmed that the hexane fraction of the extract of Huanggeong extract inhibited cell damage to ammonium nitrate. More specifically, when the treatment concentration of the hexane fraction is 2.5 to 10 μg / ml, it can be confirmed that the cell growth rate is superior to that of the control (Blank (+)) treated with ammonium nitrate only.

Taken together with Experimental Examples 2 - 1 and 2 - 4, it can be confirmed that the hexane fraction of the Huang Guan extract has an antipolution effect on ammonium nitrate even when the concentration is 0.01 to 10 μg / ml, even without cytotoxicity.

Experimental Example 3: Evaluation of tiluroside

Experimental Example 3-1: Evaluation of cell viability using MTT assay

After the culture, the medium was replaced with the IMDM medium in which bovine serum and antibiotics were excluded. Then, the tiluroside of Example 3 was treated at different concentrations (0.01 to 100 μg / ml) and further cultured for 21 hours under the same culture conditions.

3, it can be confirmed that the tiliroside of Example 3 does not show cytotoxicity at a concentration of 0.01 to 5 μg / ml.

EXPERIMENTAL EXAMPLE 3-2 Evaluation of Antipollution Activity

In general, the degree of cell damage by ammonium nitrate (500 ppm) was measured using dermal keratinocytes that directly contacted with pollutants. The tiliroside of Example 3 (1.25 μg / ml, 2.5 μg / ml, 5 μg / ml) to determine the degree of cell damage by ammonium nitrate.

After the culture, the medium was replaced with an IMDM medium in which bovine serum and antibiotics were excluded. After treatment with ammonium nitrate, the tiliroside of Example 3 was further treated for 21 hours. After incubation for 3 hours with 5 mg / ml MTT solution, media was removed and 150 μl of dimethyl sulfoxide was added to each well. After shaking for 10 minutes, absorbance was measured at 570 nm. The results are shown in Fig. 10 and Table 7.

Tilyloside (concentration) Blank (-) Blank (+) 5 μg / ml 2.5 μg / ml 1.25 μg / ml Cell proliferation (%) 100 46 59 54 43

In Table 7 and FIG. 10, when the tiluroside of Example 3 was treated, the effect of inhibiting cell damage on ammonium nitrate could be confirmed. More specifically, it can be confirmed that when the treatment concentration of tiliroside is 1.25-5 μg / ml, the cell growth rate is higher than that of the control group treated with ammonium nitrate alone (Blank (+)).

Taken together, Experimental Examples 3 - 1 and 3 - 2 show that when the concentration of tiliroside of Example 3 is 0.01 to 5 μg / ml, it has anti-pollution effect against ammonium nitrate even without cytotoxicity have.

Formulation Example

Formulation Example 1: Manufacture of softening longevity

The composition of the softening water containing the above Examples 1, 2 or 3 as an active ingredient is shown in Table 8 below.

Specifically, sodium hyaluronate was dispersed in purified water with a propeller mixer (3000 rpm) to prepare a 1% solution. Ingredients 1 to 8, except for sodium hyaluronate, were homogenized at 500 rpm in a water-dissolving tank using a propeller mixer, completely dissolved by heating at 75 ° C, and then cooled to room temperature. Thereafter, the raw materials 9 to 11 were completely dissolved in a separate dissolution tank, and the mixture was added to the above-mentioned water dissolution tank and mixed with stirring. Example 1, Example 2 or Example 3 were added thereto, and the mixture was thoroughly stirred and mixed to prepare a softening agent. On the other hand, in addition to the components described in the table for the above-mentioned preparation, it is possible to further include known components necessary for production.

ingredient Content (% by weight) One Example 1, Example 2, or Example 3 0.01 2 Polyoxyethylene hardened castor oil 0.5 3 Glycine 3.3 4 Dipotassium glycyrrhizate 0.1 5 1,3-butylene glycol 3.0 6 Sodium hyaruronate 0.1 7 ethanol 5.0 8 Antioxidant 0.1 9 Triethanolamine 0.1 10 EDTA 0.1 11 antiseptic Suitable amount 12 Purified water Balance

Formulation Example 2. Manufacture of nutrition lotion

The composition of the nutritional lotion containing the above Example 1, Example 2 or Example 3 as an active ingredient is shown in Table 9 below.

Specifically, the carbomer was dispersed at 4000 rpm using a propeller mixer to prepare a 2% solution. Raw materials 1 to 6 were fed into an aqua yard, stirred and dispersed with a homomixer (2000 rpm), and then heated to 75 캜. The raw material 7-14 was added to the oily melting tank and dissolved by heating to 75 ° C. Then, the oil phase dissolved in the water dissolution tank was charged and emulsified (3000 rpm / 5 minutes) and then cooled to room temperature. Example 1, Example 2 or Example 3 was added thereto, and the mixture was thoroughly stirred and mixed to prepare a nutrition lotion. On the other hand, in addition to the components described in the table for the above-mentioned preparation, it is possible to further include known components necessary for production.

Nutritional lotion manufacturing ingredient Content (% by weight) One Example 1, Example 2, or Example 3 0.01 2 glycerin 7.0 3 Sorbitan stearate sucrose cocoate 2.0 4 Mineral oil 4.0 5 Trioctanoin 1.0 6 Stearic Acid 1.0 7 Glyceryl stearate 0.5 8 Sorbitan monostearate 1.0 9 Dimethicone 0.5 10 Antioxidant 0.3 11 Triethanolamine 0.1 12 Carbomer 0.2 13 EDTA 0.1 14 antiseptic Suitable amount 15 Purified water Balance

Formulation Example 3. Manufacture of Essence

The composition of the essence containing the above Example 1, Example 2 or Example 3 as an active ingredient is shown in Table 10 below.

Specifically, sodium hyaluronate and hydroxyethylcellulose were each dispersed in purified water in a propeller mixer (2000 rpm) to prepare a solution containing 1% by weight. The carbomer was also dispersed in purified water with a propeller mixer (4000 rpm) to prepare a solution containing 2% by weight. On the other hand, raw materials 1 to 12 were added to an aqueous dissolution tank, stirred and dispersed with a homomixer (2000 rpm), and then heated to 75 캜, and the warmed water phase was further cooled to room temperature. The ingredients 13 to 15 were completely dissolved in a separate dissolution tank, and the mixture was added to the above-mentioned water dissolution tank and mixed by stirring. Thus, an essence containing Example 1, Example 2 or Example 3 as an active ingredient was prepared. On the other hand, in addition to the components described in the table for the above-mentioned preparation, it is possible to further include known components necessary for production.

Essence manufacturing ingredient Content (% by weight) One Example 1, Example 2, or Example 3 0.01 2 glycerin 5.0 3 1,3-butylene glycol 2.0 4 Polyethylene glycol 2.0 5 Carbomer 1.0 6 Sodium hyaruronate 0.1 7 Glycine 3.0 8 Polyacrylamide 2.0 9 Hydroxyethylcellulose 0.2 10 ethanol 3.0 11 Antioxidant 0.3 12 Triethanolamine 0.1 13 Polyoxyethylene hardened castor oil 1.0 14 EDTA 0.1 15 antiseptic Suitable amount 16 Purified water Balance

Formulation Example 4. Manufacture of cream

The composition of the cream containing the above Example 1, Example 2 or Example 3 as an active ingredient is shown in Table 11 below.

Specifically, raw materials 1 to 8 were fed into an aeration dissolving tank, stirred and dispersed with a homomixer (2000 rpm), and then heated to 75 캜. The raw materials 9 to 16 were added to a separate oily melting tank and heated to 80 DEG C to dissolve the oil. The oil phase dissolved in the water-dissolving tank was introduced to emulsify (3000 rpm / 10 minutes) and then cooled to room temperature. Example 1, Example 2 or Example 3 were added thereto, and the mixture was thoroughly stirred and mixed to prepare a cream. On the other hand, in addition to the components described in the table for the above-mentioned preparation, it is possible to further include known components necessary for production.

Cream composition ingredient Content (% by weight) One Example 1, Example 2, or Example 3 0.01 2 1,3-butylene glycol 3.0 3 glycerin 3.0 4 Hydrogenated lecithin 1.0 5 Octyldodecanol 3.0 6 Trioctanoin 2.0 7 Stearic Acid 1.5 8 Cetostearyl alcohol 2.0 9 Polysorbate 60 1.5 10 Sorbitan sesquioleate 2.0 11 Dimethicone 3.0 12 Antioxidant 0.3 13 Xanthan gum 0.2 14 Triethanolamine 0.1 15 EDTA 0.1 16 antiseptic Suitable amount 17 Purified water Balance

Formulation Example 5. Manufacture of pack

The composition of the pack containing the above Example 1, Example 2 or Example 3 as an active ingredient is shown in Table 12 below. Specifically, raw materials 1 to 8 were completely dissolved in a separate dissolution tank and stirred and mixed. After Example 1, Example 2 or Example 3 was added, the mixture was thoroughly stirred and mixed to prepare a pack. On the other hand, in addition to the components described in the table for the above-mentioned preparation, it is possible to further include known components necessary for production.

Composition of pack ingredient Content (% by weight) One Example 1, Example 2, or Example 3 0.01 2 glycerin 7.0 3 1,3-butylene glycol 3.0 4 Squalene 3.0 5 Dimethicone 3.0 6 Sodium hyaluronate 0.1 7 Glycine 2.0 8 Polyacrylamide 5.0 9 Antioxidant 0.3 10 Triethanolamine 0.1 11 EDTA 0.1 12 antiseptic Suitable amount 13 Purified water Balance

Formulation Example 6. Manufacture of Cleansing Foam

The composition of the cleansing foam containing the above Example 1, Example 2 or Example 3 as an active ingredient is shown in Table 13 below. The water phase and the oil phase were dispersed and dissolved, respectively, followed by cooling to room temperature to prepare a cleansing foam. On the other hand, in addition to the components described in the table for the above-mentioned preparation, it is possible to further include known components necessary for production.

Composition of Cleansing Foam ingredient Content (% by weight) One Example 1, Example 2, or Example 3 0.01 2 Stearic acid 6.5 3 Myristic acid 28.0 4 Self-emulsifying glycerin monostearate 3.0 5 Propylene glycol 5.0 6 Concentrated glycerin 10.0 7 Sodium hydroxide 7.0 8 Sodium ethylenediaminetetraacetate 0.1 9 Spices Suitable amount 10 Purified water Balance

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the present invention. You will understand. It is therefore to be understood that the embodiments described above are illustrative in all aspects and not restrictive. For example, each component described as a single entity may be distributed and implemented, and components described as being distributed may also be implemented in a combined form.

The scope of the present invention is defined by the appended claims rather than the detailed description and all changes or modifications derived from the meaning and scope of the claims and their equivalents are to be construed as being included within the scope of the present invention do.

Claims

A composition for skin antipollution comprising an extract of Hibiscus hamabo Siebold & Zucc. As an active ingredient.

The method according to claim 1,
The extraction solvent of the above-
And at least one member selected from the group consisting of water, ethanol, methanol, acetone, and 1,3-butylene glycol.

A composition for skin antipollution characterized by comprising a fraction of a fungus extract as an active ingredient.

The method of claim 3,
The fractions of the above-
Wherein the composition comprises at least one selected from the group consisting of Ethyl acetate fraction, Methylene Chloride fraction, and Hexane fraction.

A composition for skin antipollution, which comprises tiliroside as an active ingredient.

6. The method of claim 5,
Wherein the tiliroside is separated from the fraction of the extract of the Huang Geun Extract.

7. The method according to any one of claims 1 to 6,
The composition
Wherein the composition for skin antipollution is characterized in that it improves skin pollution caused by chemicals, skin cell death or inhibition of skin cell proliferation.

8. The method of claim 7,
The chemical
Wherein the composition comprises at least one selected from the group consisting of aliphatic hydrocarbons, polycyclic aromatic hydrocarbons, aldehydes, nitrogen oxides, and nitrates.

7. The method according to any one of claims 1 to 6,
Wherein the composition is a cosmetic composition.