KR101824687B1 - Antimicrobial composition comprising natural plant extract - Google Patents

Abstract

The present invention relates to an antimicrobial composition comprising a natural plant extract, which has no toxicity, exhibits excellent antimicrobial effect and can be substituted for a conventional synthetic preservative, and an antimicrobial composition comprising an extract of Carbacol and Gadja as an active ingredient and a cosmetic composition .

Description

An antimicrobial composition comprising natural plant extract,

The present invention relates to an antimicrobial composition comprising a natural plant extract, and more particularly to an antimicrobial composition comprising carbacol and / or oregano extract and Gadja extract as an active ingredient and a cosmetic composition containing the same.

Conventional cosmetic compositions contain various components including water, and are likely to be contaminated by microorganisms such as bacteria or fungi. The negative aspects of contamination of cosmetic compositions can be summarized in two ways: the first is the risk to the consumer's health, and the second is the decline in the value of the product. Most cosmetic compositions are applied to the skin. Therefore, when a product contaminated with microorganisms is applied to skin, inflammation or skin trouble may occur due to microorganisms introduced through the wound. In particular, contamination with pathogenic microorganisms can cause serious health problems. Other products such as fungi and germs that are propagated cause changes in the product properties due to destruction of emulsion and emulsification, changes in pH of the product, and result in a drop in the value of the product. For this reason, the cosmetic composition should have a suitable level of preservative system depending on the characteristics of the formulation. Preservatives that can be used in cosmetic compositions vary from country to country. In Korea, the type of preservative and the compositional limit that can be used in cosmetics are specified by the Korea Food & Drug Administration (Cosmetic Act, Articles 4, 3, 9 and Article 13 (6), Korea Food & Drug Administration Notice 2000-27).

Preservatives used in conventional cosmetic compositions generally include chemical preservatives (synthetic preservatives) which are easy to manufacture, cheap in cost, and easy to mass-produce. Examples of such preservatives include Metyl Paraben, Ethyl Paraben, Propyl paraben, butyl paraben and the like and quaternium-15, imidazolidinyl urea diazolidinyl urea, phenoxyethanol, Benzalkonium Chloride, chlorohexidine gluconic acid and Trichlosan, and the like. However, such a chemical preservative inhibits the growth of microorganisms, thereby imparting stability to the cosmetic composition, but at the same time, it may cause irritation to the user's skin. Typical examples are formalin-free preservatives, including diazolidinyl urea and imidazolidinyl urea. These have recently been reviewed for safety worldwide and their use is on the decline and their use is limited. In addition, parabens such as Metyl Paraben, Ethyl Paraben, Propyl Paraben and Butyl Paraben, which are most safe as conventional chemical preservatives and commonly used in cosmetics and pharmaceuticals, Even preservatives are disbelief in the use of the criteria as permitted by skin allergies, the possibility of environmental hormones and the generation of resistant bacteria. Accordingly, there is a need for a natural preservative that can reduce the content of the preservative in the cosmetic composition as much as possible and replace the chemical preservative such as not using a synthetic preservative.

The requirements of these natural preservatives should be able to prevent hypoallergenic, low toxicity and resistance to the skin, as well as antimicrobial activity against microorganisms such as bacteria and fungi. However, some raw materials of natural preservatives known to date are too dark and have a strong raw material pick-up. Therefore, they are limited or unsuitable for use in cosmetics requiring a good quality product, or they lower the viscosity of cosmetics and affect the emulsifying system. It was difficult to use. In addition, it has a narrow antimicrobial spectrum as compared with a chemical preservative, and its physicochemical stability against heat, light, and oxygen is very low, so that the antimicrobial active ingredient is decomposed during the distribution period, The problem is that most of them are not commercialized due to limitations as natural preservatives.

The inventors of the present invention have made intensive studies on natural plant extracts having antimicrobial activity and found that when carbacol and / or oregano extract and Ghazu extract are mixed as an active ingredient, synergy The present inventors have completed the present invention in order to provide an antimicrobial composition containing the same as an active ingredient and a cosmetic composition containing the antimicrobial composition.

An object of the present invention is to provide an antimicrobial composition comprising a natural plant extract excellent in antimicrobial activity against microorganisms.

It is an object of the present invention to provide a natural preservative containing said antimicrobial composition.

It is also an object of the present invention to provide a cosmetic composition containing the antimicrobial composition as a preservative component.

The present invention provides an antibacterial composition comprising carbacol and GAZA extract as an active ingredient.

The antimicrobial composition according to an embodiment of the present invention may contain an oregano extract and a Ghazu extract containing carbacol as an active ingredient.

The antimicrobial composition according to an embodiment of the present invention may further include one or more extracts selected from the group consisting of Lilium extract, green tea extract, orchid extract, strawberry herb extract, Parsley extract, and the like, but is not limited thereto.

The antimicrobial composition according to an embodiment of the present invention has an antibacterial activity against a microorganism selected from bacteria and fungi. When a mixture of carbacol and / or oregano extract and Gadja extract is used, Synergy effects on the antimicrobial activity against fungi such as Candida albicans and Aspergillus niger.

The present invention also provides a cosmetic composition comprising the antimicrobial composition as a preservative component and a natural preservative.

The antimicrobial composition according to the present invention has no toxicity to the human body and exhibits an excellent antibacterial effect and is expected to be useful as a natural preservative ingredient in cosmetics, foods, and quasi-drugs by replacing conventional synthetic preservatives.

Fig. 1 shows the result of measuring the preservative activity of the antimicrobial composition prepared in Example 1. Fig.
FIG. 2 shows the minimum inhibitory concentration (MIC) for each microorganism of the antimicrobial composition prepared in Example 1. FIG.
Fig. 3 shows the skin irritation test results of the antimicrobial composition prepared in Example 1. Fig.

The antimicrobial composition comprising the natural plant extract according to the present invention will be described in detail below. Unless otherwise defined in the technical terms and scientific terms used herein, the person skilled in the art will understand In the following description, well-known functions and constructions that may unnecessarily obscure the gist of the present invention will not be described.

It is needless to say that "carbacol" in the present invention can be isolated and purified from natural sources, used commercially or used by chemical synthesis methods known in the art. The separation tablets from nature are Carpenter's Weed, Achillea millefolium, Common Couch, Agropyronrepens, Dill, Anethum graveolens, Chinese angelica, Angelicasinensis, Artemisia annua, Artemisia cina, Boldo, Boldeafragrans, Black Tea, Camelliasinensis, Caraway, Carumcarvi, Bachelor's buttons, Chrysanthemum parthenium, Camphor, Cinnamomum camphora, American Dittany, Cunilaoriganoides, Akgiritotu, Dictamnusalbus, Bitter root, Gentianalutea, Licorice, Glycyrrhizaglabra, Dokudami, Houttuyniacordata, , Azop (Hyssopusofficinalis), Black Walnut (Juglansnigra), Sabine (Juniperussabina), Bay, Laurus nobilis, Lovage (Levisticumofficinale) , Gold-and-silver (Lonicera japonica), Barbados lilac, Meliaazedarach, European pennyroyal, Menthapulegium, Fish mint, Mentha spicata, Lemon bergamot, Monardacitriodora, There are two species of bee bale, Monarda didyma, Horse mint, Monardapunctata, Common myrtle, Myrtuscommunis, Cat-mint, Nepetacataria, Basil, Ocimum basilicum, Chinese-celery , Oenanthejavanica, Oregano, Origanum onites, Bible hyssop, Origanumsyriacum, Greek oregano, Origanumvulgarehirtum, Black Pepper, Pipernigrum, Asian plantain, Plantagoasiatica, Rosemary Plant, Rosmarinus officinalis, Garden Savory, Saturejahortensis, Savory, Saturejamontana, peppertree, Schinusmolle California, Honey Leaf, Steviarebaudiana, Thymus vulgaris, Thymus vulgaris, Funk's thyme, Thymus x citriodorus, Spanish thyme, Thymuszygis, Azoan (Indian date, Tamarindus indica), Breckland Thyme plant, Thymusserpyllum, aJwain, Trachyspermumammi), Common Valerian (Valeriana officinalis) or corn (Corn, Zea Mays) by a solvent extraction method and chromatographic separation method known in the art. For example, the extraction of carbacol from the plant may be carried out in a solvent such as water, ethanol, methanol, propanol, isopropanol, butanol, etc .; alcohols having 1 to 6 carbon atoms, acetone, ether, chloroform, ethyl acetate, methylene chloride, hexane, An organic solvent such as methanol, ethanol, isopropanol, petroleum ether, diethyl ether, benzene and the like or a mixed solvent thereof. In addition, a fraction according to polarity is obtained from the extract by a separation method using chromatography known in the art, for example, silica gel column chromatography, and the separated specific fractions are further purified by reversed phase column chromatography and high performance liquid chromatography (HPLC) method or the like.

The official name of "oregano" in the present invention is Origanum vulgare L., the most common species of Lamiaceae flowering Origanum. Oregano is a perennial herb that is native to the western and southwestern parts of Eurasia and the Mediterranean region. It is a plant that grows up to 20-80 cm and has a 1-4 cm long camellia. Thus, the oregano extract in the present invention may be origanum vulgare extract using oregano leaves and stem, but is not limited thereto. In addition, the oregano extract according to the present invention contains carvacrol as a main indicator substance (containing 90 wt% or more), and the other ingredients include sabinene, 1,8-cineole terpineol,? -terpinene,? -terpinene, caryophyllene, geracrene D, thymol (? -terpinene),? -terpinene, , p-cimene, myrcene, spathulenol, and the like, but the present invention is not limited thereto.

In the present invention, "Gaza" is a fruit of Gaza (Terminalia chebula Retz.) Or Terminalia chebula Retz. Var. Tomentella Kurt. The flowering period is from June to August and the fertilization period is 8 It is from October to October, and in the fall, mature fruit is picked and dried in the sun. The fruit-like phase of dried Gaza is long egg-shaped or egg-shaped and has a length of 2.5 cm to 3.0 cm and a diameter of 1.5 cm to 2.5 cm. The outer surface is yellowish brown to brownish in color, with five prominent wrinkles vertically, with irregular wrinkles therebetween, and there is a scar on the base. The vagina is hard, the side cut off is dark brown, and the thickness of the flesh is 2mm to 5mm. The periplasm is about 5 mm thick, yellowish brown, the quality is very hard, there is a brown suture line, and there is one seed of about 5 mm in diameter at the center. Let go have a slight peculiar smell, taste a little bitterly to wear. Thus, the Ghazi extract in the present invention may be a terminalia chebula fruit extract, but the present invention is not limited thereto. The compounds of the present invention are preferably selected from the group consisting of Chebulic acid, Chebulagic acid, Chebulinic acid, Corilagn, Punicalin, Punicalagin and Methyl neochebulagate, and the like.

In the present invention, the term " glutinous "is a perennial herb belonging to Magnoliaceae, including Magnolia obovata Thunberg, Magnolia officinale Rehder et Wils and Machilus thunbergii Sieb. Et Zucc, . Thus, the extract of the present invention may be Magnolia obovata bark extract, but not limited thereto.

The term "green tea" in the present invention has not only been consumed as a symbolic tea since BC, but recently, the pharmacological mechanism of the various components contained in green tea has gradually become known, and thus the value of the plant has been recognized. Camellia sinensis It is extracted from the leaves and leaves of Camellia Sininsis by treatment with oxidative enzymes present in the leaves of the tea leaves with pyrogen or steam. Accordingly, the green tea extract of the present invention may be Camellia sinensis leaf extract, but is not limited thereto.

The term "orchid" in the present invention is a perennial herbaceous plant belonging to the genus Orchidaceae. Specifically, the term " orchidaceae " refers to a plant belonging to the genus Acanthopipium Blume, Acianthus, Aerides, But are not limited to, Angraecum, Amitostigma, Anoectochilus, Ansellia, Aplectrum, Arethusa, Arundina, Arundina, Blume, Ascocentrum, Brassia, Beltilla, Bulbophyllum, Calanthe, Calypso, Cattleya, Cattleya, And are classified into three genera: Cephalanthera, Coelogyne, Coeloglossum, Corallorhiza, Cremastra, Cymbidium, (Cypripedium), Crytopodium, Dendrobium, Elle kies (El eorchis, Epidendrum, Epipogium, Eria, Eulophia, Galeola, Goodyera, and Geodolum. Geodorum, Gymnadenia, Habenaria, Hancockia, Herminium, Kitigorchis, Linaria, Lepariaceae, Listeria, Lycaste, Macodes, Malaxis Soland., Microstylis, Neofinetia, and the like. , Neottia, Oncidium, Oreorchis, Phalaenopsis, Phagmopedium, Pholidota, Plataneta, and the like. Platanthera, Pogonia, Rhynchostylis, Sarcanthus, Spiranthes, Spa sproutis Bloom (sp.), Refers to an orchid plant belonging to the genus such as Spathoglottis Blume, Tipularia, Tainia, Vanda, Vexillabium, Zygopetalum and the like . Accordingly, the orchid extract of the present invention may be Ndrobium Nobile extract extracted from leaves of Ndrobium Nobile, but is not limited thereto.

The term "straw sandalwood" in the present invention belongs to Rosaceae as a perennial perennial herbaceous plant, and it is known that 10 species are distributed in the temperate zone of the Northern Hemisphere and Brazil and South America. In Korea, it grows frequently in the roadside of the mountains and fields, and it is also called Seonghakcho, Yonga, Yongaeko, Hwanghaeko, Hwangyongjang, Gijunseok. Its height grows about 15 ~ 60 cm. It has white hairs all over, with a leaflet shape or a long elliptical leaflet staggered. Yellow flowers bloom in June ~ August. Thus, the extract of the present invention may be Agrimonia Pilosa Extract using leaves of straws, but the present invention is not limited thereto.

In addition, in the present invention, "parsley" is a ladybug and an evergreen herbaceous shrub, and a stem of 30-80 cm high from the tropics is branched from the base. Leaves are alternate and wide elliptical with wavy sawtooth on edge. Accordingly, the Pachiri extract in the present invention may be Pachostemon Cablin leaf extract, but is not limited thereto.

In addition, the extract according to the present invention is extracted according to a conventional method known in the art by adding an extraction solvent to at least one selected from plant leaves, flowers, stems, branches, roots, outposts, bark and fruits. That is, it may be extracted by methods such as cold extraction, ultrasonic extraction, reflux cooling extraction, hot water extraction, room temperature extraction, warm-up extraction, supercritical extraction, etc. using a conventional solvent under ordinary temperature and pressure conditions, It is not. The extraction solvent may be, but is not limited to, water, anhydrous or lower alcohol having 1 to 7 carbon atoms, hexane, chloroform, ethyl ether, acetone, ethyl acetate, butyl acetate, 1,3-butylene glycol, Propylene glycol, and 1,3-propylene glycol. Of these, lower alcohols and water selected from methanol, ethanol, propanol and butanol are preferable. In addition, the extract extracted with the above-mentioned extraction solvent includes all the extracts, fractions and purified products obtained in each step of fractionation or purification, their diluted solution, concentrated solution or dried product, and if necessary, they are subjected to freeze-drying, vacuum drying , Dried by hot air drying, spray drying, vacuum drying, foam drying, high frequency drying, infrared drying, and the like, and further pulverizing the dried extract. Accordingly, the extract is characterized by having a liquid phase, a concentrated solid phase, and a lyophilized powder phase, as the extract has additional steps such as drying and grinding according to the case after filtration.

The term "antibacterial" in the present invention means an ability to resist bacteria and means all the mechanisms for defending against the action of microorganisms such as bacteria or fungi. Herein, the term "bacterium " refers to an organism belonging to bacteria, which is the most prolific species among living organisms, and may be a pathogenic bacterium. The term" fungal "refers to at least 72,000 species including fungi, yeast, And microorganisms belonging to the same genus.

The present invention provides an antibacterial composition comprising carbacol and GAZA extract as an active ingredient. When the above-mentioned carbacol and Gadia extract were used as a single component, the antibacterial activity against bacteria and fungi was generally low, and the antibacterial activity against Pseudomonas aeruginosa and yeast (Candida albicans) was remarkably low, It was possible to maximize the antimicrobial activity against fungi when used in combination with an extract.

As an example of a preferred embodiment of the present invention, it may be an antimicrobial composition containing an oregano extract and a Ghazu extract containing the carbacol as an active ingredient. However, if the extract contains carbacol as a main component in the antimicrobial composition, it is not limited to oregano extract Do not.

That is, the antimicrobial composition according to the present invention is capable of exhibiting excellent antibacterial activity against bacteria and fungi. In particular, the antimicrobial composition according to the present invention can be applied to bacteria such as Pseudomonas aeruginosa and Candida albicans, Aspergillus niger, Etc., exhibit an excellent synergistic effect against the antimicrobial activity used as a single component.

The carbacol and / or oregano extract according to an embodiment of the present invention has a liquid oily form and exhibits a low solubility in water. However, the solubility in water can be drastically increased by the Ghazu extract used together. This effect is expected to result from changes in the physical properties of the GABA extract, which are generated by the action of carbacol and / or oregano extract as solubilizers in water.

The antimicrobial composition according to the present invention may further comprise various extracts which can give synergy to the main effect within a range not impairing the intended main effect of the present invention. At this time, a preferred example of the extract may be at least one extract selected from the group consisting of the extract of Leek extract, green tea extract, orchid extract, strawberry seed extract, and Parsley extract, but is not limited thereto.

In addition, the antimicrobial composition according to an embodiment of the present invention may be prepared by diluting the above-mentioned carbacol and / or oregano extract and GAZA extract with one or more solvents selected from glycerin, propanediol, butanediol, hexanediol and octanediol And preferred examples thereof include glycerin, 1,2-propanediol, 1,3-propanediol, 1,3-butanediol, 1,4-butanediol, 1,6-hexanediol, octanediol and methylpropanediol But may be more than one.

The antimicrobial composition according to the present invention has no adverse effects on the skin, can provide excellent preservation power for a long period of time, and exhibits excellent antimicrobial activity against bacteria and fungi. The bacteria and fungi include, but are not limited to, Escherichia coli, Pseudomonas aeruginosa, Staphlyococcus aureus, Candida albicans, Aspergillus niger, niger, Bacillus megaterium, and Exophiala moniliae. Examples of the microorganism include bacteria such as Pseudomonas aeruginosa, Candida albicans, Aspergillus oryzae, And exhibits remarkable antimicrobial activity against fungi such as Aspergillus niger.

The minimum inhibitory concentration (MIC, v / v) according to the present invention means the minimum concentration that inhibits the growth of microorganisms by an antimicrobial substance or the like. The antimicrobial composition according to the present invention has a minimum development inhibitory concentration (MIC) of 0.1 to 100 μl / ml (0.01 to 10.0% v / v) for the microorganism. In particular, the antimicrobial composition according to the present invention exhibits a minimum development inhibition concentration of Candida albicans at a concentration of 2 μl / ml (MIC value, 0.2% v / v) or less, and Aspergillus niger, (MIC value, 0.1% v / v) to a minimum development inhibitory concentration of 1 μl / ml. In the minimum development inhibition concentration, 1.0 占 퐇 / ml may be equivalent to 0.1% v / v.

The antimicrobial composition according to an embodiment of the present invention may preferably be mixed with the oregano extract and the Ghazu extract at an effective weight ratio of 1: 0.1 to 1: 1 by weight as an effective ingredient in the solvent, And they can be mixed and used at the same weight ratio. The antimicrobial composition may be a mixture of 0.1 to 10% by weight of the active ingredient with respect to the total weight of the solvent, and this is prior to the present invention.

The antimicrobial composition according to the present invention may further comprise one or more extracts selected from the group consisting of an extract of Zygomycetum extract, green tea extract, orchid extract, Zygomycethea herb extract, Parsley extract and the like for further antibacterial activity and excellent stability according to the use conditions Of course. At this time, each of the extracts may be used in an amount of 0.1 to 15% by weight based on the total weight of the antimicrobial composition. In addition, the above stability may be not only stability for long-term storage but also stability not causing changes in product properties due to destruction of emulsion or emulsion and change in product pH when used in water or formulations.

According to another aspect of the present invention, there is provided a cosmetic composition comprising the antimicrobial composition as a preservative component.

The cosmetic composition according to one embodiment of the present invention can be used without limitation in the content range that can exhibit the antimicrobial activity of the antimicrobial composition but can be used in a range of 0.5 to 5 wt% But it is not limited thereto.

In addition, the cosmetic composition according to one embodiment of the present invention may be prepared in the form of a general emulsified formulation, a solubilized formulation, or the like, using a conventionally known manufacturing method, in addition to the manufacturing method specifically disclosed in the present invention. The cosmetic composition may be appropriately selected according to the purpose. For example, the cosmetic composition may be selected from the group consisting of softening agents, convergent lotion, nutritional lotion, eye cream, nutritional cream, massage cream, cleansing cream, cleansing foam, cleansing water, But the present invention is not limited thereto. The cosmetic composition of the present invention may be formulated into a formulation selected from the group consisting of cosmetics, essences, packs, and the like, and it does not cause creaming phenomenon or flocculation phenomenon due to high- It is not possible to find a phenomenon such as discoloration or swelling caused by the above-mentioned problems.

According to another aspect of the present invention, the present invention provides a natural preservative comprising the antimicrobial composition. That is, the antimicrobial composition according to the present invention can be substituted for a synthetic preservative used in foods, quasi-drugs, etc. in addition to a cosmetic composition.

The natural preservative according to an embodiment of the present invention not only shows antimicrobial spectrum equal to or more than comparable to conventionally used parabens such as chemical preservatives, but also can be used by deep color and raw material intake which has been pointed out as a problem of conventional natural preservatives And limitations of the range.

Hereinafter, the antimicrobial composition including the natural plant extract according to one example of the present invention will be described in more detail by way of examples. It should be understood, however, that the invention is not limited to the disclosed embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the invention. In addition, the unit of the additives not specifically described in the specification may be% by weight.

(Example 1)

Step A . Manufacture of Gadja Extract

The pulverized gauze having a dry weight of 100 g was heated and refluxed in an aqueous 70% (v / v) ethanol solution for 12 hours, cooled, and then filtered through a filter paper having a permeation size of 1.2 탆. The filtered extract was concentrated under reduced pressure at 50 DEG C or lower, and then completely dried using a vacuum dryer to obtain 50 g of a GAZA extract.

Step B . Preparation of antimicrobial composition

The antibacterial composition was prepared with the composition shown in the following Table 1, using the Ghassae extract obtained by the above method.

(Example 2)

a step . Manufacture of strawberry herb extract

100 g of the dry weight of the crushed strawberry greens was heated and refluxed with a 70% (v / v) aqueous ethanol solution for 12 hours, cooled, and then filtered through a filter paper having a size of 1.2 탆 penetration. The filtrate was concentrated under reduced pressure at 50 DEG C or lower, and then completely dried using a vacuum dryer to obtain 12 g of a strawberry herb extract.

Step b . Preparation of antimicrobial composition

An antimicrobial composition was prepared in the same manner as in Example 1, except that the extract further contained a strawberry herb extract. The composition thereof is shown in Table 1 below.

(Example 3)

a step . Manufacture of orchid extract

The pulverized orchids having a dry weight of 100 g were heated and refluxed for 12 hours in an aqueous 70% (v / v) ethanol solution, followed by cooling and filtration with a filter paper having a permeation size of 1.2 mu m. The filtered extract was concentrated under reduced pressure at 50 DEG C or lower and completely dried using a vacuum dryer to obtain 19 g of an orchid extract.

Step b . Preparation of antimicrobial composition

An antimicrobial composition was prepared in the same manner as in Example 1 except that the composition further contained an orchid extract. The composition thereof is shown in Table 1 below.

(Example 4)

a step . Manufacture of green tea extract

The ground green tea having a dry weight of 100 g was heated and refluxed with a 70% (v / v) aqueous ethanol solution for 12 hours, cooled, and then filtered with a filter paper having a size of 1.2 탆 penetration. The filtered extract was concentrated under reduced pressure at 50 DEG C or lower and completely dried using a vacuum dryer to obtain 21 g of green tea extract.

Step b . Preparation of antimicrobial composition

An antimicrobial composition was prepared in the same manner as in Example 1, except that the green tea extract was further contained. The composition thereof is shown in Table 1 below.

(Example 5)

a step . Preparation of extract

The pulverized latex having a dry weight of 100 g was heated and refluxed with an aqueous 70% (v / v) ethanol solution for 12 hours, cooled, and then filtered with a filter paper having a size of 1.2 탆 penetration. The filtered extract was concentrated under reduced pressure at 50 DEG C or lower, and then completely dried using a vacuum dryer to obtain 4 g of anextra extract.

Step b . Preparation of antimicrobial composition

An antimicrobial composition was prepared in the same manner as described in Example 1, except that it further contained an extract of Lilac. The composition thereof is shown in Table 1 below.

(Example 6)

a step . Parsley  Preparation of extract

The pulverized pulpy having a dry weight of 100 g was heated and refluxed with a 70% (v / v) aqueous ethanol solution for 12 hours, cooled, and then filtered with a filter paper having a size of 1.2 μm. The filtered extract was concentrated under reduced pressure at 50 DEG C or lower, and then completely dried using a vacuum dryer to obtain 10 g of Parsley extract.

Step b . Preparation of antimicrobial composition

An antimicrobial composition was prepared in the same manner as described in Example 1, except that the extract further contained Parsley extract. The composition thereof is shown in Table 1 below.

(Comparative Example 1)

a step . Preparation of antimicrobial composition

An antimicrobial composition was prepared in the same manner as in Example 1, except that the extract was not used. The composition thereof is shown in Table 1 below.

(Comparative Example 2)

a step . Preparation of antimicrobial composition

An antimicrobial composition was prepared in the same manner as in Example 1, except that carbacol was not used. The composition thereof is shown in Table 1 below.

(Comparative Example 3)

a step . Preparation of antimicrobial composition

An antimicrobial composition was prepared in the same manner as in Example 1, except that the extract of Lycoris sp. The composition thereof is shown in Table 1 below.

In order to measure the antimicrobial effect of the antimicrobial composition prepared in the above Examples and Comparative Examples, the minimum inhibitory concentration (MIC) for each microorganism was measured by the following method, and the results are shown in Tables 2 and 2 below And its antibacterial activity was measured using the following method, and the results are shown in Table 3 and FIG. 1 below. The cytotoxicity of the antimicrobial composition prepared in Example 1 was evaluated by the following method. The results are shown in Table 4 below.

In addition, the skin irritation test for the antimicrobial composition prepared in the above examples was carried out by the following method, and the results are shown in FIG.

1. Minimum development inhibition concentration ( MIC )

Staphylococcus aureus ATCC 6538P as gram-positive bacteria, Escherichia coli ATCC 8739 as gram-negative bacteria, Pseudomonas aeruginosa as a gram-positive bacterium were used to measure the antibacterial effect of the antimicrobial composition prepared from the above Examples and Comparative Examples. Candida albicans ATCC 10231 and Aspergillus niger ATCC 22343 were purchased from Korea Biotechnology Research Center (KCTC, Korea) as yeast. Nutrient broth solutions were prepared and sterilized at 121 ° C for 15 min. Each of the cultured strains was inoculated into nutrient broth (10 ml) and cultured in a shaking incubator at 24 And cultured for a period of time to prepare a strain for an antibacterial experiment.

In order to measure the minimum inhibitory concentration of the antimicrobial composition inhibiting the growth of the microbial strain, the strain cultured in the sterilized medium was cultured at a concentration of 1 × 10 6 cfu / ml for bacteria (bacteria) and 1 × 10 5 cfu / ml for fungi Were cultured at 35 ° C for 2 days, and fungi were cultured at 25 ° C for 5 days and observed as a control. The antimicrobial composition was added to the medium so as to be 1% by weight, and then sterilized. The bacteria were inoculated at 1 × 10 6 cfu / ml for fungi and 1 × 10 5 cfu / ml for fungi, For 2 days, and fungi were cultured at 25 ° C for 5 days and observed as an experimental group.

To confirm the presence or absence of the growth of each strain in the culture medium of the cultured control and experimental groups, approximately 300 μl of the sample was taken from the aseptic space and dispensed into a sterilized 96-well microplate. Using a spectrophotometer, The minimum inhibitory concentration (MIC) was determined by measuring the minimum concentration at which low proliferation was observed for each strain compared with the control group. The results are shown in the following table 2.

As shown in Table 2 and FIG. 2, the antimicrobial composition according to the present invention exhibited excellent antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus and Escherichia coli at a concentration of 0.4 wt% or less, % Or less of the antimicrobial activity. In particular, it was confirmed that the antibacterial activity of carbacol and gadia extract alone was not significantly improved against P. aeruginosa but was remarkably improved when mixed with GAZA extract. In addition, Candida albicans, Aspergillus oryzae, and the like. In addition, in the case of an antimicrobial composition comprising oregano extract and Gadja extract having carbacol as a main indicator substance, it was confirmed that the synergistic effect on the antimicrobial activity was remarkable in combination with other ingredients. In the case of Comparative Example 1, which is used alone as carbacol, the solubility in water was so low that it had to be shaken for re-mixing at the time of use.

2. Measurement of antimicrobial activity (preservative activity)

In order to measure the antimicrobial activity of the antimicrobial composition prepared in Example 1, Staphylococcus aureus ATCC 6538P was used as a gram positive bacterium, Escherichia coli ATCC 8739 was used as a gram negative bacterium, Pseudomonas aeruginosa ATCC 9027 Candida albicans ATCC 10231 and Aspergillus niger ATCC 22343 were used as yeast.

Approximately 50 μl of each of the cultivated strains was added to a solidified plate medium, and the mixture was dispensed into a plate culture medium and uniformly smoothed using a spreader. About 40 μl of the antimicrobial composition was absorbed into a paper disk having a diameter of 10 mm, the solvent was dried, the microorganism was closely adhered to the surface of the medium on which the strain had been coated, and cultured in an incubator at 30 ° C. for 48 hours or more. And the size of the clear zone formed in the test piece was measured to confirm the antibacterial effect. Since microorganisms can not grow near the paper disk, the larger the average diameter value, the better the antimicrobial activity. Table 3 and FIG. 1 show the results of analysis by using DMSO (dimethyl sulfoxide), propanediol (PD) and methyl paraben (MP) as control groups in the same manner.

As shown in Table 3 and FIG. 1, the antimicrobial composition according to the present invention showed excellent antibacterial activity in various combinations including carbacol and GAZA extract. In particular, antimicrobial activity against Candida albicans and Aspergillus niger was remarkable.

3. Cytotoxicity Assessment

The cell toxicity in HaCaT cells and HDF-N cells was measured using Example 1, which is the basis of the antimicrobial composition according to the present invention. HaCaT cells were seeded at a density of 5 × 10 4 cells per well in a 96-well plate and at a density of 6 × 10 3 cells per well in a 96-well plate for HDF-N cells. The medium was removed and washed with DPBS. After 12 hours of starvation, the test solution and fresh medium (except media supplement) were added and cultured for 24 hours. After washing with PBS, the WST-1 reaction solution was diluted 1/10 in the supplemented medium and treated with 100 μl of each solution. , And the absorbance was measured at 450 nm. As a result, survival rates of HaCaT cells and HDF-N cells were 93.1% and 98.3%, respectively, at the maximum concentration of 5%. Accordingly, the antimicrobial composition according to the present invention is expected to be able to exhibit antimicrobial activity effectively by substituting a chemical preservative for cosmetic composition, food, and pharmaceutical formulation (see Table 4).

4. Skin irritation test of antimicrobial composition

Using Example 1, which is the basis of the antimicrobial composition according to the present invention, the antimicrobial composition was diluted to 5 wt% and 10 wt% in purified water to conduct a skin irritation test. Toxicologically, 11 healthy adults were conducted according to the CTFA guidelines (The Cosmetic, Toxic and Fragrance Association, Inc. Washington, DC, 20036, 1991). After dropping 20 μl of the sample and solution into a finn chamber, the sample and the solution were placed on the arm of the human body to be tested and fixed with a tape. After 24 hours, 48 hours, and 72 hours of application, the patches were removed, and after 2 hours, the skin response of the test site was evaluated according to the following criteria (no 0-stimulation; 0.5- 1 - mild irritation such as weak erythema, 2 - pronounced erythema accompanied by papule and edema, and 3: severe irritation such as severe erythema accompanied by blisters). At this time, purified water was used as a control group of the skin irritation experiment.

As a result, all of the antimicrobial compositions according to the present invention showed no irritation, indicating that the irritation index was 0 (see FIG. 3).

(Examples 7-9)

A cosmetic composition (serum) was prepared with the composition shown in Table 5 below. The raw materials 1 to 4 were added to purified water, stirred at room temperature (23 ° C) for 10 minutes, and the raw material 5 was added to prepare a transparent serum preparation.

(Comparative Example 4)

A transparent serum formulation was prepared in the same manner as in Example 7 except that methylparaben was used in the same amount instead of the antimicrobial composition prepared in Example 1.

(Examples 10-12)

A cosmetic composition (cream-in-water type formulation) was prepared with the composition shown in Table 6 below. The raw materials 1 to 3 were heated and adjusted to 70 ° C to prepare a water component (phase A), and the raw materials 4 to 12 were heated and adjusted to 70 ° C to prepare a milk ingredient (phase B). The oil component was added to the water component, homogenized for 3 minutes with a homomixer (3000 rpm), defoamed, filtered and cooled to prepare an underwater type cream formulation.

(Comparative Example 5)

An underwater type cream formulation was prepared in the same manner as in Example 10, except that the same amount of methylparaben was used instead of the antimicrobial composition prepared in Example 1.

 The antibacterial effect of the cosmetic composition containing the antimicrobial composition prepared in the above Examples was measured by the following method, and the results are shown in Table 7 below.

5. Cosmetics  Confirming the antibacterial effect of the composition

To confirm the antimicrobial effect of the cosmetic composition containing the antimicrobial composition, the test method of the microbiology guidelines of the Cosmetic, Toility and Fragrance Association, Inc. (CTFA) was used. Escherichia coli ATCC 8739, Pseudomonas aeruginosa ATCC 9027, and Candida albicans were used as gram-positive bacteria, Staphylococcus aureus ATCC 6538P, Gram-negative bacteria, (Candida albicans ATCC 10231), and Aspergillus niger ATCC 22343 were used.

A suspension in which three kinds of bacteria (E. coli, P. aeruginosa, S. aureus) and two kinds of fungi (C. albicans, A. niger) were mixed was injected into a vial containing 10 g of each cosmetic composition, / g and 1 × 10 5 cfu / g, and then 1 g of the sample was taken under sterile conditions at 4 hours, 1 day, 2 days, 3 days, and 7 days after storage at 25 ° C. and appropriately diluted with a diluent Bacteria were cultured on Nutrient agar (Difco) and fungi were cultured on potato dextrose agar (Difoc) medium at 35 ℃ and 25 ℃ for 24 ~ 72 hours, respectively. Respectively.

As shown in Table 7, it was confirmed that the antiseptic effect of the cosmetic composition containing the antimicrobial composition according to the present invention is equivalent to or better than the comparative example using the conventional chemical preservative (methylparaben). That is, when the antimicrobial composition according to the present invention is used in a cosmetic composition, it is possible to realize an excellent antibacterial effect.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, Those skilled in the art will recognize that many modifications and variations are possible in light of the above teachings.

Accordingly, the spirit of the present invention should not be construed as being limited to the embodiments described, and all of the equivalents or equivalents of the claims, as well as the following claims, belong to the scope of the present invention .

Claims (11)

Wherein the composition contains carbacol and GAZA extract as an active ingredient. The method according to claim 1,
An antimicrobial composition comprising the oregano extract and the Ghazu extract as an active ingredient. The method according to claim 1,
Wherein the antimicrobial composition further comprises one or more extracts selected from the group consisting of Lilac extract, green tea extract, orchid extract, strawberry herb extract and Parsley extract. The method according to claim 1,
Wherein the antimicrobial composition further comprises at least one selected from glycerin, propanediol, butanediol, hexanediol, octanediol, and methylpropanediol. The method according to claim 1,
Wherein the antimicrobial composition has an antimicrobial activity against a microorganism selected from bacteria and fungi. 6. The method of claim 5,
Wherein the microorganism is at least one selected from the group consisting of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, Aspergillus niger, Bacillus megaterium, and Exopiala monileri. 6. The method of claim 5,
Wherein the antimicrobial composition has a minimum development inhibitory concentration (MIC) of 0.1 to 100 占 퐇 / ml for the microorganism. A cosmetic composition comprising a solubilizable antimicrobial composition according to any one of claims 1 to 7 as a preservative. 9. The method of claim 8,
Wherein the cosmetic composition comprises 0.1 to 10% by weight of the antimicrobial composition. 9. The method of claim 8,
Wherein the cosmetic composition is formulated with a softening agent, a convergent lotion, a nutritional lotion, an eye cream, a nutritional cream, a massage cream, a cleansing cream, a cleansing foam, a cleansing water, a powder, an essence or a pack. A natural preservative comprising the solubilizable antimicrobial composition of any one of claims 1 to 7.

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