KR102018533B1

KR102018533B1 - Composition comprising extracts, fractions and the isolated compounds of hibiscus hamabo siebold and zucc. for skin anti-pollution

Info

Publication number: KR102018533B1
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: 2019-09-05
Also published as: KR20190006888A

Abstract

The present invention relates to a composition for skin antipollution, and in the present invention, as it comprises a compound isolated from the root extract, the fraction of the root extract, and the fraction of the root extract, the skin is hypoallergenic and the skin from an environmental pollutant. It provides a composition for skin antipollution excellent protective antipollution effect.

Description

COMPOSITION COMPRISING EXTRACTS, FRACTIONS AND THE ISOLATED COMPOUNDS OF HIBISCUS HAMABO SIEBOLD AND ZUCC. FOR SKIN ANTI-POLLUTION}

The present invention relates to a composition for skin antipollution, comprising a compound isolated from the extract of Rhus, Rhus, and Rhus.

The skin is a thin film that surrounds the outside of the body and protects the body from physical and chemical stimuli of the external environment. It also prevents excessive penetration of pollutants from the outside while suppressing evaporation of moisture in the body. With the recent increase in the use of chemicals, the skin is exposed to environmental pollutants such as polycyclic aromatic hydrocarbons, aldehydes, nitrogen oxides, nitrates, and fine dust generated during the manufacture, use, and disposal of erythema, edema, Mild skin problems such as itching, hives and inflammation, and skin diseases such as atopy, psoriasis and skin cancer are increasing.

For this reason, the development of cosmetic compositions, cleaning agents, and external preparations having an anti-pollution effect that prevents the above-mentioned environmental pollutants from adhering to the surface of the skin or penetrating into the body through the skin is actively progressed. have. However, most of the products currently being developed merely serve to remove contaminants attached or penetrated to the skin, and simply remove contaminants from the skin, and do not fundamentally protect the skin from environmental contaminants. There was a slight problem with the ability to protect the skin from.

In particular, because it contains a chemical as a main component there was a problem that acts as a cause of skin problems and skin diseases. Accordingly, it is necessary to develop a composition having excellent antipollution effect while being hypoallergenic on the skin using natural materials.

On the other hand, as a natural material, Republic of Korea Patent No. 10-1571323, peppermint, chamomile, rosemary, lemongrass, wild chrysanthemum, tea tree and lavender are mixed with the same weight ratio, respectively, by using an anti-poll extract Disclosed is a cosmetic composition for improving skin having a lution effect, and Korean Patent Publication No. 10-2017-0053388 discloses a cosmetic composition having an anti-pollution effect as it contains turmeric, apple and grape extracts. In the case of the patent, the use of a plurality of extracts, the cost was high, but there was a problem that the antipollution effect is insignificant.

Republic of Korea Patent No. 10-1571323 Republic of Korea Patent Publication No. 10-2017-0053388

It is an object of the present invention to include a compound isolated from the extract of the Rheum undulatum, the extract of the Rheum undulatum and the extract of the Rheum undulatum, which is a natural substance, the skin anti-hypoallergenic and excellent anti-pollution effect to protect the skin from environmental pollution sources It is to provide a composition for a solution.

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

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

The extract solvent of the root extract may include one or more selected from the group consisting of water, ethanol, methanol, acetone and 1,3-butylene glycol.

In addition, in order to achieve the above object, the present invention provides a composition for skin antipollution comprising a fraction of the root extract as an active ingredient.

The fraction of the root extract may include one or more selected from the group consisting of ethyl acetate fraction, methylene chloride fraction and hexane fraction of the root extract.

In addition, in order to achieve the above object, the present invention provides a composition for skin antipollution comprising tirilloroside (Tiliroside) as an active ingredient.

The tirilloroside may be isolated from the fraction of the root extract.

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

The chemical substance may include one or more 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 of the present invention includes a compound isolated from a natural substance of Rheum undulatum extract, a fraction of Rheum ulus extract and a fraction of Rhizome extract, which is hypoallergenic to the skin and protects the skin from environmental pollutants ( Anti-pollution) effect can be excellent.

In particular, the composition for skin antipollution of the present invention may be excellent in the antipollution effect of protecting the skin from environmental pollutants such as polycyclic aromatic hydrocarbons, aldehydes, nitrogen oxides, nitrates, fine dust.

FIG. 1 is a diagram showing ¹ H NMR spectra using nuclear magnetic resonance of a compound (Example 3) isolated from an ethyl acetate fraction of Rhus root extract prepared in Example 2. FIG.
FIG. 2 is a diagram showing ¹³ C NMR spectra using nuclear magnetic resonance of a compound (Example 3) isolated from an ethyl acetate fraction of Rhubarb extract prepared in Example 2. FIG.
Figure 3 is a view showing the results of evaluating the cytotoxicity of the Rhizome extract of Example 1, the fraction of Example 2, the tyriloside of Example 3.
Figure 4 is a diagram showing the cell proliferation rate when treated with the root extract of Example 1 together with ammonium nitrate.
FIG. 5 is a diagram showing the cell proliferation rate when the ethyl acetate fraction (EA fraction) of Example 2 was treated with benzopyrene.
FIG. 6 is a diagram showing the cell proliferation rate when the ethyl acetate fraction (EA fraction) of Example 2 was treated with formaldehyde.
7 is a diagram showing the cell proliferation rate when the ethyl acetate fraction (EA fraction) of Example 2 was treated with ammonium nitrate.
8 is a diagram showing the cell proliferation rate when the methylene chloride fraction (MC fraction) of Example 2 was treated with ammonium nitrate.
9 is a diagram showing the cell proliferation rate when the hexane fraction (Hex fraction) of Example 2 was treated with ammonium nitrate.
FIG. 10 is a diagram showing the cell proliferation rate when the tiloriside of Example 3 was treated with ammonium nitrate.

DETAILED DESCRIPTION Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings so that those skilled in the art may easily implement the present invention. As those skilled in the art would realize, the described embodiments may be modified in various different ways, all without departing from the spirit or scope of the present invention. In the drawings, parts irrelevant to the description are omitted in order to clearly describe the present invention.

The terms or words used in the specification and claims of the present invention are not to be construed in a conventional or dictionary sense, and the inventors may appropriately define the concept of terms in order to best explain their invention in the best way. Based on the principle, it should be interpreted as meaning and concept corresponding to the technical idea of the present invention.

Throughout the specification of the present invention, when a part is said to "include" a certain component, it means that it can further include other components, without excluding the other components unless otherwise stated. .

In 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 to improve or prevent skin contamination, skin cell death or inhibition of skin cell proliferation by chemicals.

Throughout the specification of the present invention, the "extract" includes all 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, and also extracts the components of the natural substance It is a broad concept that includes all of the materials that can be obtained by extracting the material obtained and then processing or processing it in another way.

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

Hereinafter, the present invention has been described in detail, but the present invention is not limited thereto.

The present invention is In the embodiment Skin containing yellow root extract as an active ingredient For anti-pollution To provide a composition.

Rhizome (Hibiscus Hamabo or Hibiscus hamabo Siebold & Zucc.) Is a dicotyledonous mallow, mallow, about 1m high, and the bark is light grayish brown with hairs on young twigs. Leaves are alternate, 3-6 cm long, 3-7 cm wide, with hairs on the front, gray-white hairs on the back, pointed ends, and cupped on the edges. Flowers bloom yellow flowers 5 cm in diameter in the leaf axes at the end of branches, dark purple in the middle.

The root extract of the present embodiment is an extract obtained by the extraction process of root roots, including roots, stems, leaves, etc., diluents or concentrates of the extracts, dried products obtained by drying the extracts, crudes or purified products of the extracts, Or extracts of all formulations that can be formed using the extract itself and the extract, such as mixtures thereof. The extract may be prepared and preferably used in the form of a dry powder after extraction, the root may be dried.

Rhubarb extract of this embodiment comprises the steps of (a) washing and drying the rhubarb; (b) extracting the yellow root dried in step (a) with an extraction solvent; And (c) may be obtained according to the step of filtering the yellow root in the mixture passed through step (b), may be in liquid form, it can be obtained by extracting the powder form by concentrating and drying it, but not limited thereto. Do not.

The extraction solvent of the root extract may include one or more selected from the group consisting of water, ethanol, methanol, acetone and 1,3-butylene glycol, but is not limited thereto, and further includes all known types of extraction solvents. can do.

The composition for skin antipollution of the present embodiment may include an anti-pollution effect as well as a hypoallergenic effect on the skin as the root extract is an active ingredient. In particular, the composition for skin antipollution comprising the root extract may implement an antipollution effect by preventing or improving skin contamination, skin cell death or inhibition of skin cell proliferation by environmental pollutants such as chemicals.

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

The polycyclic aromatic hydrocarbon may include benzopyrene, and the aldehydes may include formaldehyde. The nitrogen oxides may be anything that can be represented by the formula of NOx, 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 It} may include, but is not limited to. The nitrates include salts composed of metal or positive basic groups and nitrate groups, preferably ammonium nitrate and ionization thereof.

Preferably, the present invention may include a methanol extract of sulfur root using methanol as an extraction solvent, and among them, may have an antipollution effect on ammonium nitrate when the concentration of the methanol extract is 25 to 100 μg / ml. This is not restrictive.

Skin antipollution composition according to the present embodiment is 0.01 to 30% by weight, preferably 0.1 to 20% by weight, more preferably 1 to 10% by weight, most preferably 1 to 5% by weight of the yellow root extract It may include. If, based on the total weight of the composition for skin antipollution, including the root extract less than 0.01% by weight, the anti-pollution effect is insignificant, and if it contains more than 30% by weight of the root extract is not economical There may be a problem.

In addition, the present invention In the embodiment Of rhubarb extract Fractions Skin to include as active ingredient For anti-pollution To provide a composition.

The fraction of the root extract of the present embodiment is obtained through the fractionation operation of the extract of the root of this embodiment, the fraction may be carried out by chromatography, electrophoresis, principle separation and the like. The fraction of the root extract of the present embodiment may include one or more selected from the group consisting of ethyl acetate fraction, methylene chloride fraction and hexane fraction of the root extract.

In addition, the fraction of the rhubarb extract may be used as it is, or may be used in the form of an extract obtained by filtration and concentrated or in the form of a dried product obtained by freeze drying after concentration.

As the composition for skin antipollution of the present embodiment includes a fraction of the root extract as an active ingredient, the antipollution effect may be excellent while being hypoallergenic on the skin. In particular, the composition for skin antipollution comprising a fraction of the rhubarb extract can implement an antipollution effect by preventing or improving skin contamination, skin cell death or inhibition of skin cell proliferation by environmental pollutants such as chemicals.

Preferably, the composition for skin antipollution of the present embodiment may include an ethyl acetate fraction of rhubarb extract, especially, when the concentration of the ethyl acetate fraction of rhubarb extract is 10 to 75 μg / ml, more preferably rhubarb extract When the concentration of the ethyl acetate fraction of 18.75 ~ 75 μg / ml, the antipollution effect on benzopyrene, formaldehyde and ammonium nitrate may be excellent, but is not limited thereto.

Skin antipollution composition of the present embodiment is 0.01 to 30% by weight, preferably 0.1 to 20% by weight, more preferably 1 to 10% by weight, most preferably 1 to 5% by weight of the fraction of the root extract May contain%. If the anti-pollution extract contains less than 0.01% by weight, based on the total weight of the composition for skin antipollution, the antipollution effect is insignificant, and the fraction of the root extract is included in excess of 30% by weight. There can be problems that are not economic.

In addition, the present invention In the embodiment Tiliroside Skin to include as active ingredient For anti-pollution To provide a composition.

Tiliroside of the present embodiment may be obtained by separating the active fraction by performing medium pressure liquid chromatography (MPLC) or preparative liquid chromatography (prep-LC) to the fraction of the sulfur root extract, as shown in Formula 1 It is possible but not limited to this. In addition, the present embodiment may include both the tyriloside and an isomer thereof.

[Formula 1]

As the composition for skin antipollution of the present embodiment includes tirilloroside as an active ingredient, it may have an excellent antipollution effect while being hypoallergenic on the skin. In particular, the composition for skin antipollution comprising a tyloside of the present embodiment can implement an antipollution effect by preventing or improving skin contamination, skin cell death or inhibition of skin cell proliferation by environmental pollutants such as chemicals.

The chemical may be nitrate, but is not limited thereto. The nitrates include salts composed of metal or positive basic groups and nitrate groups, preferably ammonium nitrate and ionization thereof.

Preferably, in the present embodiment it may include a tirilloroside separated from the ethyl acetate fraction of the Rhizome extract, and thus may have an excellent antipollution effect on benzopyrene, but is not limited thereto. In particular, the present embodiment includes a tirilloroside separated from the ethyl acetate fraction of the Rheum undulatum extract, among these, when the concentration of the tiloriside is 0.01 ~ 5 μg / ml, preferably 2.5 ~ 5 μg / ml At this time, it may have an antipollution effect on ammonium nitrate, but is not limited thereto.

Skin antipollution composition of the present embodiment is 0.01 to 30% by weight, preferably 0.1 to 20% by weight, more preferably 1 to 10% by weight, most preferably 1 to 5% by weight of the tyloside It can be included as. If the anti-pollution composition contains less than 0.01% by weight of the anti-pollution composition based on the total weight of the skin antipollution, the antipollution effect is insignificant. There may be a problem.

The composition for skin antipollution comprising the root extract, the composition for skin antipollution comprising a fraction of the root extract, or the composition for skin antipollution comprising a compound separated from the fraction of the root extract is preferably a cosmetic It may be a composition, but is not limited thereto.

When the composition for skin antipollution is a cosmetic composition, along with a dermatologically acceptable excipient, softening longevity, nourishing longevity, skin, mist, lotion, cream, essence, serum, pack, face wash (cleansing cream, cleansing foam, Cleansing Water, Cleansing Powder), Powder, Foundation, Lipstick, Mascara, Makeup Base, Eyeliner, Eyeshadow, Blusher, Shampoo, Conditioner, Conditioner, Hair Gel, Hair Serum, Hair Essence, Body Lotion, Body Wash, Body Spray It may be prepared in the form of.

The excipients may include colorants, fragrances, emulsifiers, thickeners, pigments, fragrances, antioxidants, preservatives, fungicides, solvents, thickeners, inorganic salts, synthetic polymer materials, and as a functional ingredient skin conditioning agents, whitening agents, Antioxidants, anti-wrinkle agents, moisturizers and the like.

Through the following Examples, Comparative Examples, and Experimental Examples in detail the composition for skin antipollution comprising a compound isolated from the extract of the present invention, the fraction of the root extract and the fraction of the root extract as an active ingredient I will. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

[ Example ]

Example 1: Preparation of Rhubarb Extract

15 L of methanol was added to 700 g of dried humus root, and extracted three times at room temperature for 72 hours. The extracted extract was filtered and then the solvent was removed using a vacuum concentrator, and freeze-dried to prepare a methanol extract of yellow root.

Example 2: of rhubarb extract Hexane Fraction Methylene chloride Fraction Ethyl acetate Fraction Produce

The root extract (methanol extract) of Example 1 was suspended in 1.5 L of distilled water, and the same amount of hexane was added to fractionating with a fraction funnel three times. The fractionated hexane layer was extracted, and then the solvent was removed using a vacuum concentrator. Hexane fraction (9.21 g) was prepared by removing and freeze drying it.

Subsequently, the methylene chloride fraction (1.34 g) and the ethyl acetate fraction (1.16 g) were prepared by partitioning the fractionated hexane layer and the remaining distilled water layer in the order of methylene chloride and ethyl acetate polarity.

Example 3: ethyl acetate of rhubarb extract From fractions Tillyroside Separation and Structure Identification

The ethyl acetate fraction (1.16 g) of the rhubarb extract of Example 2 was subjected to MPLC using reverse phase silica gel and acetonitrile (ACN) -Water solvent gradient to obtain a total of five fractions. Among them, the third fraction (fraction-3) was subjected to prep-LC with ACN-Water solvent to separate compounds. The obtained compound was confirmed to be pure separation as confirmed by NMR. ¹ H, ¹³ C NMR spectra (see Fig. 1, Fig. 2 and the following description) were obtained using a nuclear magnetic resonance apparatus (Bruker Co. 400 MHz, NMR), and the structures were determined by comprehensive analysis of these spectra.

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 ''') , 7.28 (2H, d, J = 8.4 Hz, H-2 ''',6'''), 6.80 (2H, d, J = 9.2 Hz, H-3 ', 5'), 6.77 (2H, d , 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-6α ''), 3.52 (1H, m, H-5 ''), 3.48 (1H, m, H-3 ''), 3.47 (1H, m, H-2 ''), 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 (C-7'''), 135.3 (C-3), 132.3 (C-2 ', 6 '), 131.3 (C-2''', 6 '''), 127.1 (C-1'''), 122.8 (C-1 '), 116.8 (C-3''', 5 ''') , 116.1 (C-3 ', 5'), 114.8 (C-8`` '), 105.7 (C-10), 104.1 (C-1''), 100.0 (C-6), 94.9 (C-8 ), 78.1 (C-3``), 75.9 (C-2 ''), 75.8 (C-5 ''), 71.8 (C-4 ''), 64.4 (C-6 '')

As a result, the compound (light yellow powder) was identified as Tiliroside of Chemical Formula 1.

[ Experimental Example ]

Experimental Example 1: Evaluation of Yellow Root Extract

Experimental Example 1-1: MTT Cell using analytical methods Viability evaluation

Skin keratinocytes (HaCaT) were aliquoted in 96-well plates at a concentration of 1 × 10 ⁴ cells / ml and incubated in a 37 ° C. CO ₂ incubator for 24 hours. At this time, the medium used was IMDM medium containing 10% bovine serum and 1% antibiotics.

After culturing, it was replaced with IMDM medium without bovine serum and antibiotics, and then, the root extract (methanol extract) of Example 1 was treated by concentration (0.01 to 100 μg / ml) and further incubated for 21 hours under the same culture conditions.

Thereafter, 5 mg / ml concentration of MTT solution was added thereto, followed by incubation for 3 hours, and then the medium was removed. 150 μl of dimethyl sulfoxide was added to each well, shaking for 10 minutes, and absorbance was measured at 570 nm. The results are shown in FIG.

On the other hand, in Figure 3 methanol is the sulfuric methanol extract of Example 1, n-hexane is the hexane fraction of Example 2, Methylene chloride is the methylene chloride fraction of Example 2, Ethyl acetate is the ethyl acetate fraction of Example 2, Tiliroside is It means the tyriloside of Example 3.

Referring to FIG. 3, the rhubarb extract may be confirmed to have no cytotoxicity at a concentration of 0.01 to 100 μg / ml.

Experimental Example 1-2: Anti-pollution Active evaluation

In general, the extent of cellular damage caused by ammonium nitrate (500 ppm) was measured using skin keratinocytes that are in direct contact with the pollutant, and the concentration of sulfuric acid methanol extract of Example 1 (25 μg / ml, 50 μg / ml, 100 μg / ml) was measured for the degree of cell damage by ammonium nitrate, and compared to this to determine the effect of inhibiting skin cell damage.

Human keratinocytes HaCaT were dispensed in 200 μl aliquots in 96-well plates at 1 × 10 ⁴ cells / ml and incubated in 37 ° C., CO ₂ incubator for 24 hours. At this time, the medium used was IMDM medium containing 10% bovine serum and 1% antibiotics.

After incubation was replaced with IMDM medium without bovine serum and antibiotics, and then treated with ammonium nitrate, and then incubated for 21 hours by treatment with sulfuric methanol extract of Example 1. After that, MTT solution of 5 mg / ml concentration was incubated for 3 hours, the medium was removed, dimethyl sulfoxide was added to each well of 150 ul, shaking for 10 minutes, and absorbance was measured at 570 nm. The results are shown in Figure 4 and Table 1.

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

Looking at Figure 4 and Table 1, it was confirmed that the treatment of the root extract extracts the effect of inhibiting cell damage by ammonium nitrate. In more detail, when the concentration of the rhubarb extract is 25 ~ 100 μg / ml, it can be seen that it has an excellent cell proliferation rate compared to the group treated with ammonium nitrate only (Blank (+)).

By combining the results of Table 1 and Figures 3 to 4, it can be confirmed that the rhubarb extract of this example has an antipollution effect against ammonium nitrate without showing cytotoxicity when the concentration of the extract is 0.01 to 100 μg / ml. have.

Experimental Example 2 Evaluation of Fractions of Rhizome Extract

Experimental Example 2-1: Cell viability evaluation using MTT assay method

After the culture, the serum and the antibiotic-free IMDM medium were replaced, and the ethyl acetate fraction, the methylene chloride fraction, and the hexane fraction of Example 2 were each treated in different concentrations (0.01 to 100 μg / ml), respectively. Incubated for 21 h at.

3, it can be seen that the ethyl acetate fraction does not exhibit cytotoxicity at a concentration of 0.01-75 μg / ml.

3, it can be seen that the methylene chloride fraction shows no cytotoxicity at a concentration of 0.01 to 10 μg / ml.

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

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

Generally, skin keratinocytes in direct contact with the pollutants were used to measure the extent of cell damage caused by benzopyrene (2.5 ppm), formaldehyde (1.5 ppm), and ammonium nitrate (500 ppm), respectively. Ethyl acetate fractions of methanol extracts were treated by concentration (18.75 μg / ml, 37.5 μg / ml, 75 μg / ml) to treat cells with benzopyrene (2.5 ppm), formaldehyde (1.5 ppm) and ammonium nitrate (500 ppm). The extent of the damage was measured and compared to confirm the effect of inhibiting skin cell damage.

After incubation was replaced with IMDM medium without bovine serum and antibiotics, and then treated with benzopyrene, formaldehyde, ammonium nitrate, respectively, and further incubated for 21 hours by treating the ethyl acetate fraction of Example 2. After that, MTT solution of 5 mg / ml concentration was incubated for 3 hours, the medium was removed, dimethyl sulfoxide was added to each well of 150 ul, shaking for 10 minutes, and absorbance was measured at 570 nm. The results are shown in FIGS. 5 to 7 and Tables 2 to 4.

Tables 2 and 5 are the results of treatment with benzopyrene, Tables 3 and 6 are the results of treatment with formaldehyde, and Tables 4 and 7 are 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 rate (%) 100 50 102 109 77

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

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

5 to 7 and Tables 2 to 4, the treatment of the ethyl acetate fraction of the root extract was confirmed the effect of inhibiting cell damage to benzopyrene, formaldehyde and ammonium nitrate.

More specifically, when the concentration of the ethyl acetate fraction is 18.75 ~ 75 μg / ml, it can be seen that it has an excellent cell proliferation rate compared to the group treated with only benzopyrene, formaldehyde and ammonium nitrate (Blank (+)). In particular, when treated with benzopyrene, treated with formaldehyde, treated with ammonium nitrate, it can be seen that the best cell proliferation rate when the treated concentration of the ethyl acetate fraction is 37.5 μg / ml.

Combining the Experimental Examples 2-1 and 2-2, the ethyl acetate fraction of the Rhizome extract showed antipollution effect against benzopyrene, formaldehyde and ammonium nitrate without showing cytotoxicity when the concentration was 0.01-75 μg / ml. It can be confirmed that has.

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

In general, the extent of cellular damage caused by ammonium nitrate (500 ppm) was measured using skin keratinocytes that are in direct contact with the pollutants. The methylene chloride fraction of the methanol extract of Example 2 was determined by concentration (2.5 μg / ml). , 5 μg / ml, 10 μg / ml) was used to measure the degree of cell damage by ammonium nitrate (500 ppm), and compared to this to confirm the effect of inhibiting skin cell damage.

After incubation was replaced with IMDM medium without bovine serum and antibiotics, and then treated with ammonium nitrate and treated with methylene chloride fraction in Example 2 and further incubated for 21 hours. After that, MTT solution of 5 mg / ml concentration was incubated for 3 hours, the medium was removed, dimethyl sulfoxide was added to each well of 150 ul, shaking for 10 minutes, and 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 rate (%) 100 46 65 63 46

Looking at Table 5 and Figure 8, when the methylene chloride fraction of the root extract was treated it was confirmed the effect of inhibiting cell damage to ammonium nitrate. In more detail, when the treatment concentration of the methylene chloride fraction is 5 ~ 10 μg / ml, it can be seen that it has an excellent cell proliferation rate compared to the control group (Blank (+)) treated only with ammonium nitrate.

Combining the Experimental Examples 2-1 and 2-3, it can be seen that the methylene chloride fraction of the Rhizome extract has an antipollution effect against ammonium nitrate without the cytotoxicity when the concentration is 0.01 to 10 μg / ml. .

Experimental Examples 2-4: Antipollution Evaluation of Hexane Fraction

In general, the extent of cellular damage caused by ammonium nitrate (500 ppm) was measured using skin keratinocytes that are in direct contact with the pollutants. The concentration of hexane fractions of the methanol extract of Example 2 was determined by concentration (2.5 μg / ml, 5 μg / ml, 10 μg / ml) was used to measure the degree of cell damage caused by ammonium nitrate (500 ppm), and the skin cell damage inhibition effect was confirmed by comparison.

After incubation was replaced with IMDM medium without bovine serum and antibiotics, and then treated with ammonium nitrate, the hexane fraction of Example 2 was further incubated for 21 hours. After that, MTT solution of 5 mg / ml concentration was incubated for 3 hours, the medium was removed, dimethyl sulfoxide was added to each well of 150 ul, shaking for 10 minutes, and absorbance was measured at 570 nm. The results are shown in Figure 9 and Table 6.

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

Looking at Table 6 and Figure 9, the treatment of the hexane fraction of the root extract was confirmed the effect of inhibiting cell damage to ammonium nitrate. In more detail, when the treatment concentration of the hexane fraction is 2.5 ~ 10 μg / ml, it can be seen that it has an excellent cell proliferation rate compared to the control group (Blank (+)) treated only with ammonium nitrate.

Integrating the Experimental Examples 2-1 and 2-4, it can be seen that the hexane fraction of the root extract has an antipollution effect against ammonium nitrate without the cytotoxicity when the concentration is 0.01 to 10 μg / ml.

Experimental Example 3: Evaluation of Tiliroside

Experimental Example 3-1: Cell viability evaluation using MTT assay method

After incubation was replaced with IMDM medium excluding the bovine serum and antibiotics, and then treated with the tyriloside of Example 3 by concentration (0.01 ~ 100 μg / ml) and further incubated for 21 hours in the same culture conditions.

Looking at Figure 3, it can be seen that the tyloridoside of Example 3 does not exhibit cytotoxicity at a concentration of 0.01 ~ 5 μg / ml.

Experimental Example 3-2 anti-pollution activity evaluation

In general, the extent of cellular damage caused by ammonium nitrate (500 ppm) was measured using the skin keratinocytes in direct contact with the pollutants, and the tyriloside (1.25 μg / ml, 2.5 μg / ml, 5) of Example 3 was measured. μg / ml) was used to measure the degree of cell damage caused by ammonium nitrate, and the results were compared to confirm the skin cell damage inhibition effect.

After incubation was replaced with IMDM medium without bovine serum and antibiotics, and then treated with ammonium nitrate, followed by incubation for an additional 21 hours by treating the tiloriside of Example 3. After that, MTT solution of 5 mg / ml concentration was incubated for 3 hours, the medium was removed, dimethyl sulfoxide was added to each well of 150 ul, shaking for 10 minutes, and absorbance was measured at 570 nm. The results are shown in FIG. 10 and Table 7.

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

Looking at Table 7 and Figure 10, it was confirmed that the cell damage inhibition effect on ammonium nitrate when the tiloriside treatment of Example 3. More specifically, it can be seen that when the treatment concentration of tyrrilloside is 1.25-5 μg / ml, the cell proliferation rate was higher than that of the control treated with ammonium nitrate only (Blank (+)).

Combining the Experimental Examples 3-1 and 3-2, it can be confirmed that the concentration of the tyloridoside of Example 3 has an antipollution effect against ammonium nitrate without cytotoxicity when 0.01 ~ 5 μg / ml. have.

Formulation example

Formulation example 1: Preparation of Flexible Cosmetics

The composition of the flexible longevity containing Example 1, Example 2 or Example 3 as an active ingredient is shown in Table 8.

Specifically, sodium hyaluronate was prepared in a 1% solution by dispersing in purified water with a propeller mixer (3000 rpm). Raw materials 1 to 8, except for sodium hyaluronate, were homogenized at 500 rpm in an aqueous phase dissolution tank using a propeller mixer, warmed completely at 75 ° C, and then cooled to room temperature. Then, after completely dissolving the raw materials 9 to 11 in a separate dissolution tank, it was added to the aqueous phase dissolution tank and stirred and mixed. Example 1, Example 2 or Example 3 was put here, and it fully stirred and mixed, and manufactured flexible softening water. On the other hand, in addition to the components disclosed in the table for the preparation may further include known components required for the preparation.

ingredient Content (% by weight) One Example 1, Example 2 or Example 3 0.01 2 Polyoxyethylene Cured Castor Oil 0.5 3 Glycine 3.3 4 Dipotassium glycylizate 0.1 5 1,3-butylene glycol 3.0 6 Sodium hyaluronate 0.1 7 ethanol 5.0 8 Antioxidant 0.1 9 Triethanolamine 0.1 10 EDTA 0.1 11 antiseptic Quantity 12 Purified water Remaining amount

Formulation example 2. Preparation of Nutrients

The composition of nutrient cosmetics containing 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. The raw materials 1-6 were thrown into the water phase dissolution tank, it stirred with the homomixer (2000 rpm), and dispersed, and it heated up to 75 degreeC. The raw materials 7-14 were added to the oil phase dissolution tank, and it melt | dissolved by heating to 75 degreeC. Then, the oil phase dissolved in the aqueous phase dissolution tank was introduced 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 sufficiently stirred and mixed to prepare nutrient cosmetic water. On the other hand, in addition to the components disclosed in the table for the preparation may further include known components required for the preparation.

Nutritional Cosmetics 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 Trioctanoine 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 Quantity 15 Purified water Remaining amount

Formulation example 3. Preparation of Essence

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

Specifically, sodium hyaluronate and hydroxyethyl cellulose were each prepared by dispersing with a propeller mixer (2000 rpm) in purified water as a 1 wt% solution. 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 1-12 were thrown into the water phase dissolution tank, and it stirred and dispersed by the homomixer (2000 rpm), heated to 75 degreeC, and the warmed water phase was cooled to room temperature again. After completely dissolving the raw materials 13 to 15 in a separate dissolution tank, the mixture was added to the aqueous phase dissolution tank and stirred and mixed. 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 disclosed in the table for the preparation may further include known components required for the preparation.

Essence manufacturer 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 hyaluronate 0.1 7 Glycine 3.0 8 Polyacrylamide 2.0 9 Hydroxyethyl cellulose 0.2 10 ethanol 3.0 11 Antioxidant 0.3 12 Triethanolamine 0.1 13 Polyoxyethylene Cured Castor Oil 1.0 14 EDTA 0.1 15 antiseptic Quantity 16 Purified water Remaining amount

Formulation example 4. Preparation of Cream

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

Specifically, the raw materials 1 to 8 were introduced into the aqueous phase dissolution tank, stirred and dispersed with a homomixer (2000 rpm), and then heated to 75 ° C. Raw materials 9 to 16 were added to a separate oil phase dissolution tank to dissolve it at 80 ° C., and the oil phase dissolved in the aqueous phase dissolution tank was added to emulsify (3000 rpm / 10 minutes), and then cooled to room temperature. Example 1, Example 2 or Example 3 was put here, it stirred and mixed sufficiently, and the cream was prepared. On the other hand, in addition to the components disclosed in the table for the preparation may further include known components required for the preparation.

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 Trioctanoine 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 Quantity 17 Purified water Remaining amount

Formulation example 5. Manufacture of pack

The composition of the pack containing Example 1, Example 2 or Example 3 as an active ingredient is shown in Table 12 below. Specifically, the raw materials 1 to 8 were completely dissolved in a separate dissolution tank, and stirred and mixed. Example 1, Example 2 or Example 3 was added thereto, followed by sufficiently stirring and mixing to prepare a pack. On the other hand, in addition to the components disclosed in the table for the preparation may further include known components required for the preparation.

Composition of the 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 Quantity 13 Purified water Remaining amount

Formulation example 6. Preparation of Cleansing Foam

The composition of the cleansing foam containing Example 1, Example 2 or Example 3 as an active ingredient is shown in Table 13. The aqueous phase and the oil phase were each dissolved and dissolved, mixed and saponified, and then cooled to room temperature to prepare a cleansing foam. On the other hand, in addition to the components disclosed in the table for the preparation may further include known components required for the preparation.

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 Quantity 10 Purified water Remaining amount

As described above, the description of the present invention is for the purpose of illustration, and those skilled in the art to which the present invention belongs may easily change to other specific forms without changing the technical spirit or essential features of the present invention. I can understand. Therefore, it is to be understood that the embodiments described above are exemplary in all respects and not restrictive. For example, each component described as a single type may be implemented in a distributed manner, and similarly, components described as distributed may be implemented in a combined form.

The scope of the present invention is shown by the following claims rather than the above description, and all changes or modifications derived from the meaning and scope of the claims and their equivalents should be construed as being included in the scope of the present invention. do.

Claims

Skin antipollution composition comprising the extract of Hibiscus hamabo Siebold & Zucc. As an active ingredient.

The method of claim 1,
Extraction solvent of the yellow root extract
A composition for skin antipollution comprising at least one selected from the group consisting of water, ethanol, methanol, acetone and 1,3-butylene glycol.

Skin antipollution composition characterized in that it comprises a fraction of the root extract as an active ingredient.

The method of claim 3, wherein
The fraction of the rhubarb extract is
Ethyl acetate (Ethyl acetate) fraction, methylene chloride (Methylene Chloride) fraction and the hexane (Hexane) fraction of the root extract extract for skin antipollution composition characterized in that it comprises at least one.

delete

The method according to any one of claims 1 to 4,
The composition is
A composition for skin antipollution, characterized by improving skin contamination, skin cell death or inhibition of skin cell proliferation by chemicals.

The method of claim 7, wherein
The chemical
A composition for skin antipollution comprising at least one selected from the group consisting of polycyclic aromatic hydrocarbons, aldehydes, nitrogen oxides and nitrates.

The method according to any one of claims 1 to 4,
The composition is a composition for skin antipollution, characterized in that the cosmetic composition.