KR101806800B1 - Complex natural preservative comprising scutellaria baicalensis extract and method manufacture the same - Google Patents

Abstract

The present invention relates to a composite natural preservative comprising a composition containing at least any one of gold, chrysolophthy, lantern leaf, lavender, and rosemary extract, and a method for producing the same. The natural natural preservative comprises a natural extracting antimicrobial composition derived from a natural extract having low side- As an antiseptic, an antimicrobial composition extracted from a mixture of Skullcap, Scutellaria baicalensis and Artemisia annua is provided as an active ingredient, so that no toxicity or irritation occurs in the human safety test, The antimicrobial composition containing the extract from gold can be usefully used in cosmetics, foods and medicines for preservation purposes.

Description

[0001] The present invention relates to a natural natural preservative comprising a gold extract and a method for producing the natural natural preservative comprising scutellaria baicalensis extract,

The present invention relates to a complex natural preservative having a composition containing a natural extract and a method of preparing the same. More particularly, the present invention relates to a complex natural preservative comprising a composition containing at least one of gold, chrysolophy, Preservatives and methods for their production.

Due to socio-economic development, the importance of cosmetics and related industries has increased as one of personal management, and consumers' interest in the quality of cosmetics and personal care products has also been growing. That is, as the income increases, the cleanliness becomes more important, and the interest in the appearance is heightened, the amount of various cosmetics such as skins, lotions, creams, essences, shampoos, soaps, toothpastes, and detergents is increasing.

Such cosmetics are frequently opened, frequently used, and have a characteristic of being easily contaminated, and are products that require the use of preservatives.

Preservatives used in cosmetics mainly consist of preservatives using synthetic raw materials. Synthetic preservatives are problematic in that they do not completely eliminate side effects on skin even though they are excellent in preservative ability, thus affecting the health of users. In recent years, there has been a growing demand for cosmetics using natural or organic natural materials as a raw material.

In other words, preservatives that prevent decay by microorganisms are essential to maintain the quality of many products such as foods, medicines, and cosmetics for a long time. Parabens preservatives, which are safely used as cosmetics and medicines, are safely used as conventional preservatives because of their potential for skin allergies (Andrea Counti et al., Contact Dermatitis, 1997, 37; 35-36) and environmental hormones (Edwin et al , Toxicology and applied pharmacology, 1998, 153, 12-19) and resistance to inducing resistant bacteria.

In the case of cosmetics, it is essential to use the cosmetic directly, since it is used for a relatively long period of time after the package is opened and the characteristics of the product deteriorate easily. Accordingly, conventionally, a certain amount of a synthetic preservative prepared by synthesizing a chemical substance is added to cosmetics to prevent the deterioration and corruption of cosmetics. Although these preservatives are excellent in preservative effect on cosmetics, they are chemical products made by synthesizing chemical substances. Therefore, they affect the health of users of cosmetics. Naturally, they are placed with the needs and social trends of cosmetics users who value health and wellbeing .

In addition, most of the natural active materials have not been commercialized because of color, stability, narrow antibacterial spectrum, problems of form, and the like. In addition, hinokitiol, Magnolol, Magnolol extract, DF-100 and so on. It is known that the antimicrobial activity of DF-100 prior to product development is due to the organic acid contained in DF-100 and benzethonium chloride, which is a synthetic preservative, and other natural antimicrobial agents are known to have low or narrow antimicrobial spectrum Or the limit of the commercial range due to physical properties, and so there is an urgent need for a natural antimicrobial agent which is more advanced and can be developed into a product. Therefore, studies on natural preservatives having a wide spectrum of antimicrobial spectrum and stability of form have been actively carried out while reducing side effects of conventional synthetic preservatives.

Examples of such techniques are described in documents 1 and 2 below.

For example, Patent Document 1 discloses a natural preservative composition comprising 10 to 40 parts by weight of a golden extract, 10 to 30 parts by weight of a mulberry extract, and 15 to 40 parts by weight of a grapefruit seed extract.

Also, Patent Document 2 discloses a natural antimicrobial preservative composition containing a mixture of a yulgok extract, a yellowtail extract, and a tea extract as an active ingredient, wherein the weight ratio of the Yulgok extract: the Yulchil extract: the dragon tea extract is 2: 1: < RTI ID = 0.0 > 2. ≪ / RTI >

Patent Document 3 discloses a method of producing a fermented citron extract by mixing citron seeds in a culture medium for lactic acid bacteria and fermenting the fermented citron seed extract to obtain a citron extract from the citron; And 1 to 60% by weight of the fermented Yuzu seed extract, 1 to 30% by weight of Yuza seed extract, 1 to 80% by weight of the golden extract and 1 to 60% by weight of the octopus fennel extract ≪ / RTI >

Korean Registered Patent No. 10-0762303 (registered on September 20, 2007) Korean Registered Patent No. 10-1525976 (Registered on May 29, 2015) Korean Registered Patent No. 10-1532005 (Registered on June 22, 2015)

Benzoic acid, which is mainly used in cosmetics, and parabens, imidazole urea, and chlorphenesin, which are used in external preparations including cosmetics, are excellent in formulation stability and preservative efficacy but can cause toxicity, skin irritation and allergy.

In addition, although the preservative of a natural product has been proposed in the above-mentioned conventional art, there has been a problem that the extraction process is troublesome and the repelling force is lowered.

The object of the present invention is to solve the above-mentioned problems, and it is an object of the present invention to provide a gold extract suitable for use in organic cosmetics, since it is made of a material derived from natural materials and has no side effects on human body, And a method for producing the same.

Another object of the present invention is to provide a complex natural preservative comprising a gold extract having excellent antimicrobial activity against Gram-positive and Gram-negative bacteria, yeast and mold, and a process for producing the same.

Another object of the present invention is to provide a complex natural preservative which is safe for human body having a wide range of antimicrobial spectrum and excellent antimicrobial activity by blending with plant extracts at an optimal mixing ratio and a method for producing the same.

In order to attain the above object, the natural preservative according to the present invention is a natural preservative containing an antimicrobial composition derived from a natural extract derived from a natural extract having low side effects and toxicity. The antimicrobial composition comprises a extract of Skullcap (Scutellaria baicalensis) Artemisia annua extract as an active ingredient and the extraction is carried out with 70% ethanol, followed by sequential fractional extraction with EtOAc, n-BuOH and CH ₂ Cl ₂ , and the antimicrobial composition is applied to 100 parts by weight of the composition 20 to 50 parts by weight of gold extracted with EtOAc, 20 to 50 parts by weight of gold extracted with CH ₂ Cl ₂ , and 20 to 50 parts by weight of chrysol extracted with CH ₂ Cl ₂ .

Further, the natural preservative according to the present invention is characterized by further comprising an antimicrobial composition extracted from at least one of Neem leaves, Azadirachta indica, Lavender, Lavandula angustifolia and Rosemary (Rosmarinus officinalis).

In the natural preservative according to the present invention, the antimicrobial composition is contained in any one of cosmetics, foods or medicines.

In the natural preservative according to the present invention, the antimicrobial composition contains 0.1 to 30 parts by weight per 100 parts by weight of the finished product of cosmetics, food or pharmaceuticals.

The natural preservative according to the present invention is characterized in that the extraction is carried out with purified water, alcohol, 70% ethanol, 100% ethanol, ethyl acetate or dimethylchloromethane.

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In order to achieve the above object, the present invention provides a method for producing a natural preservative containing an antimicrobial composition derived from a natural extract having low side effects and toxicity, comprising the steps of: (a) (B) drying the herb prepared in step (a) with purified water, alcohol, 70% ethanol, 100% ethanol, ethyl acetate or dimethylchloromethane extracting a component, (c) in CH ₂ Cl extraction in a comprising the step of filtration and concentrated to the active ingredient, wherein the step (b) extraction in step (b) is a solvent having different polarities from fraction funnel _2, EtOAc, and BuOH fraction by running successively with purified water, and the gold is extracted in the Chung Ho is after the run with 70% ethanol, sequentially extracted fraction by EtOAc, n-BuOH and CH ₂ Cl ₂ , The antimicrobial composition comprises a the Chung Ho 20-50 wt extracted with golden 20 to 50 parts by weight, CH ₂ Cl golden 20 to 50 parts by weight and extracted with _2, and CH ₂ Cl ₂ and extracted with EtOAc for 100 parts by weight of the composition .

As described above, the natural preservative composition according to the present invention and the preparation method thereof show wide spectrum of antibacterial activity against various fungi and do not cause toxicity or irritation in the human safety test, Can be advantageously used for cosmetics, foods and medicines for the purpose of preserving the antimicrobial composition.

According to the composite natural preservative containing the gold extract according to the present invention and the method for producing the same, since the antimicrobial composition containing a golden extract having excellent safety as an active ingredient as a natural product and having an excellent antibacterial effect is used, It is possible to provide a preservative effect to medicines, detergents and the like.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a process diagram for explaining a herb extract for producing a natural preservative applied to the present invention;
FIG. 2 is a view showing a process for producing a fraction extract,
3 is a photograph showing the results of antibacterial effect on Bacillus subtilis,
4 is a photograph showing the results of the antibacterial effect on Escherichia coli,
FIG. 5 is a photograph showing the results of the antibacterial effect against Staphylococcus aureus,
6 is a photograph showing the results of antibacterial effect on Pseudomonas aeruginosa,
FIG. 7 is a photograph showing the results of antibacterial effect against Candida albicans, a fungus,
8 is a photograph showing the results of antibacterial effect against Aspergillus niger,
9 is a photograph showing the antimicrobial effect of the 70% ethanol extract by fractionation,
FIG. 10 is a photograph showing a state in which the cells were cultured for 1 to 6 days.

These and other objects and novel features of the present invention will become more apparent from the description of the present specification and the accompanying drawings.

Medicinal herbs used in natural preservatives containing the antimicrobial composition of the present invention as an active ingredient include herbs such as Skullcap, Scutellaria baicalensis, Artemisia annua, Neem leaves, Azadirachta indica, (Lavender, Lavandula angustifolia), and rosemary (Rosemary, Rosmarinus officinalis). Hereinafter, the term " herb " means any of gold, chungho, linden, lavender, rosemary or a mixture thereof unless the type of the herb is specified.

The gold (Skullcap, Scutellaria baicalensis) applied to the present invention is a root of a perennial herbaceous plant such as spiny lentil, which is removed as it is. Gold contains more than 35% of flavonoids such as Baikal, Baikal and Wigonin. This component has anticarcinogenic, antiviral and antioxidant properties as well as anti-inflammatory effects (Kim, EN et al., 2008), and gold as a herbal medicine exhibits various pharmacological effects such as chewing and detoxification in oriental medicine (Yang, JH et al. , 2006).

The Artemisiae Annuae Herba applied to the present invention means the upper part of the Artemisia annua Linne or Artemisia apiacea Hance of the Cercopithecidae. Artemisin, which can kill and kill only the cancer cells contained in Chung Ho, is known to be 1,200 times more sedative than conventional anticancer drugs. In addition, Chung Ho is known to have the effects of antipyretic analgesic, immune enhancement, protection of liver function, and anti-malarial action.

Neem leaf (Azadirachta indica) refers to the leaf part of Azadirachta indica belonging to the Merlot. Nimyeo is an evergreen broad-leaved arboreous tree, originated from India, Pakistan and Sri Lanka, and has been widely used for many years as a private medical center. Malaria, encephalitis and other diseases, especially in the treatment of omia, smallpox, and external parasites. The mulberry leaf extract contains azadirachtin and is a safe and safe ingredient for infants.

Lavender (Lavender, Lavandula angustifolia) is the origin of the Mediterranean coast, its stems are dull square and united, the leaves are turned or face to face, and the perfume taken from flowers and plants is used as a raw material for perfumes and cosmetics, Not used to treat headaches or nervous stability, was used for sterilization and insect repellent.

Rosemary (Rosemary, Rosmarinus officinalis) is a kind of herb that is rich in vitamins and minerals, contains various pharmacological ingredients, digesting, diuretic, sterilizing and antibacterial.

The herb (herb) contained in the natural preservative according to an embodiment of the present invention is at least one or more of gold, chrysolophyta, linden, rosemary, and rosemary. The antimicrobial composition extracted from the herb contains water, an organic solvent, Can be extracted using a solvent.

In the case of extraction using an organic solvent, an alcohol such as methanol, ethanol, isopropanol or butanol, an alcohol such as ethylene, acetone, ether, hexane, chloroform, ethyl acetate, DMF (N, N-dimethylformamide), DMSO Side), a mixture thereof, or the like, at room temperature or with heating, and then may be extracted with an organic solvent. Preferably, water (purified water) or ethanol is used as the extraction solvent. More preferred is ethanol extraction followed by fractionation with water (purified water), ethyl acetate, methylene chloride, hexane and butanol. The extract can be prepared by pulverizing a dry or non-dried raw material, adding the solvent to the pulverized product at a weight ratio of 10 to 100 times and immersing it in the pulverized product, followed by filtration or centrifugation to obtain a supernatant.

In the case of high-temperature extraction, the raw material is added to a water / organic solvent and heated at a temperature of about 70 ° C or higher, preferably about 80 ° C or higher, more preferably 90 ° C or higher for 1 hour or more, preferably 2 hours or more, Heating for 4 hours or longer, and filtering the heated extract. When heating at a high temperature, the time is reduced, but the active ingredient may be destroyed and lost. When heated at a temperature lower than the above temperature, the effective ingredient of the mixture is not extracted completely, so it is important to adjust the temperature and time appropriately.

The extract obtained by heat extraction or cold extraction is filtered to remove suspended solid particles by filtration, for example, nylon or the like, or filtered using a freeze filtration method or the like, or the extract may be used as it is, Drying, spray drying and the like.

The antimicrobial composition (or natural preservative) extracted from gold may contain 0.1 to 30 parts by weight, preferably 0.1 to 20 parts by weight, based on 100 parts by weight of the finished product such as cosmetics, foods, medicines and the like.

The natural preservative according to one embodiment of the present invention exhibits excellent antimicrobial activity against bacteria, yeast, especially fungi, and is safe for human skin. As described above, the natural preservative according to the present invention having a wide spectrum of preservation is used in cosmetics, foods, and medicines, so that the existing chemical synthetic natural preservative does not exhibit any adverse effects on the human body and has excellent repellency.

In the cosmetic composition of the present invention, the cosmetic composition may be selected from the group consisting of softening agents, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, Creams, body oils, and body essences, but are not limited thereto.

In addition to the antimicrobial composition of the present invention, it is also possible to add to the antimicrobial composition a composition containing at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softening agents, antioxidants, suspending agents, stabilizers, foaming agents, Or any other conventionally used in cosmetics such as nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, barrier agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, ≪ RTI ID = 0.0 > ingredients. ≪ / RTI >

The present invention also provides dairy products comprising antimicrobial compositions extracted from gold. There is no particular restriction on the type of household goods. Examples of applications include, but are not limited to, wet tissues, diapers, sanitary napkins and natural pulp, polymers (resins), cotton, nonwoven fabrics and the like.

The present invention also provides a food comprising an antimicrobial composition extracted from gold. There is no particular restriction on the type of food. Examples of foods to which the composition can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, soups, Drinks, alcoholic beverages, and vitamin complexes, all of which are processed foods in a conventional sense.

Hereinafter, an embodiment of a process for producing a complex natural preservative containing a gold extract according to the present invention will be described with reference to FIG. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

FIG. 1 is a process diagram for explaining a herb extract for producing a natural preservative applied to the present invention, and FIG. 2 is a diagram showing a process for producing a fraction extract.

First, as shown in FIG. 1, golden, chungho, lantern, lavender, and rosemary were prepared as a hub and dried (S10).

In step S 20, 500 g of the sample, in which at least one of flesh leaves, lavender, and rosemary are cut and mixed in the dried gold, chungho, golden, and chungho are extracted with purified water, 70% ethanol, 100% ethanol or ethyl acetate, respectively (S20) .

The extraction in the step S20 can be carried out by refluxing three times at 80 DEG C for 2 hours after adding, for example, 10 times of purified water (w / v) of the sample to a flask.

Thereafter, the mixture was filtered through a 0.45 mu m filter, and then concentrated using a vacuum concentrator (S30). The step S30 is, for example, twice filtered with Whatman NO.2 filter paper and then concentrated under reduced pressure using a rotary vacuum evaporator.

As shown in FIG. 2, the concentrate obtained through extraction of hot water, 70% EtOH, 100% EtOH and EtOAc was dissolved in sterilized DMSO (dimethylsulfoxide) solution, / ml and used for the experiment.

Yeast, fungi, and the like. Thus, a 70% EtOH extract was obtained for the fraction, and then fractionated with a solvent as shown in FIG.

That is, as shown in FIG. 2, the respective extracts were sequentially fractionated with a solvent (CH ₂ Cl ₂ , EtOAc, BuOH, water) having different polarities from the fractional oocyte, The solvent was completely removed by concentration on an evaporator. That is, in the present invention, ethanol (70% EtOH) extract was fractionated sequentially using ethyl acetate, dichloromethane and butanol using an open column. The thus-obtained antimicrobial active substance was concentrated and spray-dried.

The concentrations of each of the concentrates were adjusted by using a sterilized DMSO solution as a solvent, and the antimicrobial activity of each strain was measured.

The final concentration of the extract is shown in Table 1.

The antimicrobial activity of the complex natural preservative according to the present invention was measured by concentration and then freeze-dried to be used in the experiment.

As a reagent for the assays of the present invention, NA (Nutrient agar), TSA (Tryptic soy agar), LB (Lactose broth) and PDA (Potato dextrose agar) were used. These media were purchased from Difco-BBL Were purchased and used. For example, the medium used for the cultivation of E. coli and B. subtilis was LB, and Pseudomonas aeruginosa and S. aureus, Candida albicans and Aspergillus niger used NA and PDA media, respectively.

That is, an agar plate is prepared for the antibacterial activity test of the complex natural preservative according to the present invention.

To check for contamination of the medium, remove the water from the plate and place it in an incubator at 37 ° C for 2 to 3 hours before the experiment. To determine the proper concentration of the strain, dilute it to about 10 ^-1 , 10 ^-2 , 10 ^-3 , and cultivate before this experiment to confirm that it grows to the proper concentration on the plate.

Next, the upper culture medium (0.7% agar) is prepared, and the diluted strains are quickly added and mixed thoroughly. The upper layer medium containing the strain is poured evenly over the lower layer medium. For preliminary culture, pre-culture for 2 to 3 hours to harden the upper layer medium.

The herb samples diluted to various concentrations are spotted on a pre-incubated plate (spot lawn method). The samples showing activity as spots were selected and reaffirmed by the paper disc diffusion method.

In the agar diffusion method, the diluted strains are plated on an agar plate at 200 each. Place the silver foil on the 8mm disc which is sterilized by using tweezers. After that, 20 ~ 40 ㎕ of 5 kinds of herbs such as golden, blue, lemon, lavender, and rosemary are prepared by drying.

Pick up the loaded disk with sterilized tweezers and place on a strained plate.

The strains used in the test according to the present invention were four strains classified as bacteria, one strain classified as yeast, and one strain classified as fungus. Escherichia coli CCARM0237, Pseudomonas aeruginosa KACC14021, Staphylococcus aureus KCTC3881 and Bacillus subtilis KACC 14394, fungi Candida albicans KCTC7270, , And Aspergillus niger KACC40280 (CCARM Seoul National University), KACC Agricultural Microbiology Bank, KCTC Korea Research Institute of Bioscience and Biotechnology). Each bacterium was inoculated with 1 × 10 ⁸ CFU / mL. The extracts were dissolved in DMSO at various concentrations, and 10 μl was added to a sterilized paper disk (diameter 8 mm). DMSO was completely dried and then plated on a plate medium. The plate was kept at 28 to 37 ° C, and after 1, 2, and 3 days, the clear zone around the paper disk was measured to examine the antibacterial activity.

Five kinds of the above-mentioned test strains are cultured and the cultured strains are diluted to 10 < ^{-2 &} gt ^{; to- 3} . Then, the cells were transferred to a 96-well plate and cultured at 37 ° C for 24 hours at intervals of 3 hours (0, 3, 6, 9, 12, 15, 18, 21, 24 hours) The growth inhibition curve and MIC (Minimum Inhibitory Concentration) were determined by measuring with a labeling meter (Multilabel Counter: 1420, Perkin Elmer).

The culture conditions of the strain are incubated (18-24 hr) until a clear zone appears in an incubator at 28-37 ° C. In case of mold, cultivate for 3-7 days.

Table 2 below shows the relationship between the strains used and the culture medium.

The antimicrobial effect of the herbal hot-water extract according to the present invention was tested as follows.

50 g of each of gold, Chungho, femyeop, lavender and rosemary were immersed in 10 times (500 ml) of the third distilled water. After heating at 100 ° C for 2 hours, the mixture was slowly cooled and extracted. The extract was filtered again under reduced pressure, and the extract was concentrated to 40 rpm at 80 rpm using a rotary pressure reducer.

The extracts were dissolved in DMSO at concentrations of 5,000 ppm, 10,000 ppm, and 100,000 ppm. As a control, the extract was used as dissolved DMSO.

[Experimental Example 1]

In Experimental Example 1, antimicrobial activity of gold, chrysolophus, femme, lavender and rosemary extract was measured by disk diffusion method.

In the present invention, the antimicrobial composition was extracted using relatively safe water, ethanol, and ethyl acetate as a solvent for gold, Chungho, linden, lavender, and rosemary, and the culture of E. coli and Bacillus subtilis The medium used was LB, and the culture medium of P. aeruginosa, S. aureus, L. monocytogenes and Aspergillus niger Six antimicrobials were tested by disk diffusion method, titrating the concentration of each extract (antimicrobial composition) at 5 ~ 100mg / ml.

Here, the control group is a conventional commercial preservative or a chemical preservative.

As a result, the water, ethanol, and ethyl acetate extracts of Chungho, Mysyne, and Lavender were identified as E. coli, Pseudomonas aeruginosa, Bacillus subtilis (B . subtilis) showed antimicrobial activity.

Table 3 shows the antimicrobial activity in the hot-water extract.

As shown in Table 3, the highest concentration of 100,000 ppm of antimicrobial activity against E. coli strains was observed in the hot water extracts of Chungho, Myon leaf and Lavender.

Next, extraction and antimicrobial activity of the herb with 70% ethanol was tested.

First, 100 g of each of the herbs were immersed in 10-fold (1.0 L) volume of 70% ethanol, then rotated and stirred at 160 rpm and extracted for 24 hours.

This extract was sieved again, and the supernatant was filtered under reduced pressure. The extract was concentrated using a rotary vacuum at 40 rpm at 80 rpm. The concentrate was dissolved in DMSO at a concentration of 5, 10, 100 mg / ml (5000, 10000, 100000 ppm) and then used.

The control group was DMSO without extract, and the MIC value of each extract was measured.

Table 4 shows the antimicrobial activity in the 70% ethanol extract.

As can be seen in Table 4, among the five samples extracted with 70% ethanol, gold showed high antimicrobial activity against all strains at 100,000 ppm concentration. Lavender and rosemary showed high antimicrobial activity at 100,000 ppm concentration in strains except A. niger. Cheongho showed antimicrobial activity in strains except A. niger and P. aeruginosa at 100,000 ppm concentration. Mycobacteria exhibited antimicrobial activity at a concentration of 100,000 ppm in strains other than E. coli and A. niger.

Table 5 shows the MIC values of the 70% ethanol extract.

As a result of MIC measurement, A. niger did not grow the strain including the control group. The gold showed lower inhibitory concentration than the other herbs.

Next, to enhance the reliability of the experiment, the antimicrobial activity against the 70% ethanol extract was performed again, and the antibacterial activity of the extract using 100% ethanol and ethyl acetate was tested.

The extraction and antimicrobial activity tests with 70% / 100% ethanol and ethyl acetate were as follows.

First, 100 g of each of the herbs was immersed in 10-fold (1.0 L) volume of 70% / 100% ethanol and ethyl acetate, respectively. Then, each of the extracts was subjected to rotary stirring at 160 rpm for 24 hours, and each of the extracts was sieved again. The supernatant was filtered under reduced pressure, and each of the extracts was concentrated at 40 rpm at 80 rpm using a rotary pressure reducer. Water was dissolved in DMSO at concentrations of 1, 5, 10, and 50 mg / ml.

In the control group, DMSO without the extract, 0.4% Ethylparaben, europa, and phenoxyethanol were applied.

The antimicrobial effects of 70% ethanol, 100% ethanol and ethyl acetate extract are shown in Tables 6-11 and 3-8.

Table 6 and FIG. 3 show the results of the antimicrobial effect against Bacillus subtilis. Table 7 and FIG. 4 show the results of the antimicrobial effect against Escherichia coli, and Table 8 and FIG. 5 show the results of the antibacterial activity against Bacillus subtilis Table 9 and FIG. 6 show the results of the antimicrobial effect against Pseudomonas aeruginosa, and Table 10 and FIG. 7 show the results of the antimicrobial effect against Staphylococcus aureus (Candida albicans Candida albicans), and Table 11 and Fig. 8 show the results of antibacterial effects against Aspergillus niger.

As shown in Tables 3 to 8, the extracts of Bacillus subtilis (B. subtilis) showed antimicrobial activity, and the extracts of golden, rosemary, rosemary, The antimicrobial activity against S. aureus was observed at a concentration of 5 mg / ml or more. That is, the antimicrobial activity against the strains other than Bacillus subtilis (B. subtilis) was weak or scarcely exhibited, and the fraction was extracted.

[Experimental Example 2]

Experimental Example 2 measured the antimicrobial activity of extracts of 70% ethanol extract (EtOAc, n-BuOH, CH ₂ Cl ₂ ) against gold, Chungho, linden leaves, lavender and rosemary.

First, to extract the fraction as shown in FIG. 2, five kinds of herbs of golden, blueberry, linden, lavender and rosemary were extracted with 70% ethanol.

In the 70% ethanol extract using a fraction funnel water: CH ₂ 1 ratio of Cl _2: by the addition of 1 to fraction extract layer (layer) and water layer (layer) by a CH ₂ Cl ₂ CH ₂ Cl ₂ To obtain a fractional extract. The same amount of EtOAc of the obtained water layer was added, and the mixture was divided into an extract layer and a water layer by EtOAc to obtain a fraction extract with EtOAc.

The same amount of BuOH as the obtained water layer was added, and the mixture was divided into an extract layer and a water layer by BuOH to obtain a fractional extract with BuOH.

Each of the obtained EtOAc, n-BuOH, and CH ₂ Cl ₂ extracts was dissolved in DMSO.

The dissolved samples were first subjected to the spot lawn method and the samples showing antibacterial activity in the spot lawn method were selected. The selected samples were subjected to the disk diffusion method as in Experimental Example 1.

The antimicrobial effect of the fraction extraction (EtOAc, n-BuOH, CH ₂ Cl ₂ ) of the 70% ethanol extract as described above was as shown in Table 12 and FIG.

Table 12 and FIG. 9 show the antimicrobial effect by fractionation of the 70% ethanol extract.

As shown in Table 12, it can be seen that the golden ethyl acetate fraction extract showed antimicrobial activity in pathogenic microorganisms except Bacillus subtilis and A. niger strains. According to the invention, the fraction of the extract ChungHo CH ₂ Cl _2, EtOAc Golden, Golden CH ₂ Cl ₂ of these 3 kinds of extract fraction of Chung Ho and gold was confirmed to be a high antimicrobial activity. That is, the extraction from the mixture of gold and charcoal was carried out with 70% ethanol, followed by fraction extraction with EtOAc, n-BuOH, CH ₂ Cl ₂ , and the antimicrobial composition was extracted with EtOAc for 100 parts by weight of the composition to include gold from 20 to 50 parts by weight, CH ₂ golden 20 to 50 parts by weight and extracted with ₂ Cl, CH ₂ Cl ₂ and extracted with a ChungHo 20 to 50 parts by weight was higher for the antibacterial activity.

[Experimental Example 3]

For the exact experiment of Experimental Example 2, the antimicrobial activity measurement for the preparation of the optimum mixing ratio of the composite natural preservative was conducted by the disk diffusion method. That is, in Experimental Example 2 The gold 20 composition and extracted with EtOAc for 100 parts by weight of conducted by to 50 parts by weight, CH ₂ Cl golden 20 to 50 parts by weight and extracted with _2, CH _2, the Chung Ho 20-50 wt extracted with Cl ₂ The antimicrobial activity was measured at various mixing ratios to produce an optimum mixing ratio among the extracts containing the fractions.

20 to 50 parts by weight, 20 to 50 parts by weight, and 20 to 50 parts by weight, respectively, of Chungho CH ₂ Cl ₂ , gold EtOAc and gold CH ₂ Cl _2, which are the three samples obtained in Experimental Example 2, Respectively.

Table 13 shows the results of the antimicrobial effect of the selected fractionated extract mixture of purified CH ₂ Cl ₂ , golden EtOAc, and golden CH ₂ Cl ₂ .

These three samples were mixed at various ratios as shown in Table 13, and as a result, no. 1, 2, 3, 4, 5, 8, and 9 showed high antimicrobial activity. C. albicans showed high antimicrobial activity in all mixtures. In the case of B. subtilis, S. aureus, and P. auroginosa, the specimen was spotted and redisplayed by disc diffusion method Respectively.

[Experimental Example 4]

Based on the results obtained in Experimental Example 3, the antimicrobial activity for the preparation of the optimal mixing ratio of the composite natural preservative was measured by the disk diffusion method. Namely, no. 2, 3, 4, 5, 8, and 9 were selected, and the mixture of the selected six species was confirmed by the disk diffusion method. The results are shown in Table 14.

In Table 14, A represents CH ₂ Cl ₂ (single), B represents golden EtOAc (single), C represents golden CH ₂ Cl ₂ (single), and C + represents control (DMSO).

As shown in Table 14, the antibacterial activity against B. subtilis, C. albicans, and S. aureus strains was high. In addition, the activity of A (CH ₂ Cl ₂ ) for B. subtilis, C. albicans, S. aureus, E. coli, and P. aeruginosa strains was lower than that of the gold single mixture. The antimicrobial activities of gold single (EtOAc, CH ₂ Cl ₂ ) on B. subtilis, C. albicans, S. aureus, E. coli and P. aeruginosa strains were similar to those of mixtures. The antimicrobial activity against A. niger was confirmed, although the antimicrobial activity against A. niger was confirmed.

That is, when cultured for 1 to 6 days and observed, it was confirmed that the antimicrobial activity against A. niger of Mixtures 1 to 4 was excellent, as shown in FIG. Fig. 10 is a photograph showing a state of observation after 1 to 6 days of culturing.

Through the above results, no. 2, 3, and 4 were selected. The antimicrobial activities of gold were significantly different from those of bluegrass. In order to increase the reliability, the antimicrobial activity effects of the three kinds of fraction extracts according to their concentrations and the antimicrobial activity effects of these mixtures are shown in Tables 15 to 17 and the results are shown for MIC values.

Table 15 shows the antimicrobial activity effect by concentration of the fractions, Table 16 shows the antimicrobial activity effect of the mixture, and Table 17 shows the MIC value (%) of the mixture.

As a result of the above-described experiment, 3 and 4 showed high antimicrobial activity. That is, the antimicrobial composition extracted from gold and chrysanthemum showed antimicrobial activity.

[Experimental Example 5]

Next, the primary skin irritation of the complex natural preservative containing the gold extract according to the present invention was tested.

As shown in Table 18, the composite natural preservative containing the gold extract according to the present invention was found to have low responsiveness to the general control. Therefore, it was confirmed that the complex natural preservative containing gold extract was safe to use on skin. Table 19 shows the patch test criteria.

In Table 18, Response = {? (Grade 占 No. of Responders)} / {4 (Maximum grade) 占 n (Total subjects) 占 100 占 1/2

That is, the composite natural preservative containing the gold extract according to the present invention was dissolved in petrolatum to a final concentration of 5%, and then absorbed into the patch, and the patch was applied to 30 subjects and then removed in 48 hours. The reaction of the skin for 24 hours was observed and evaluated. The composite natural preservative according to the present invention was found to be a reducing substance in terms of human skin sensitivity, in which no skin reaction could be observed in all subjects.

Although the present invention has been described in detail with reference to the above embodiments, it is needless to say that the present invention is not limited to the above-described embodiments, and various modifications may be made without departing from the spirit of the present invention.

By using the complex natural preservative containing the gold extract according to the present invention and the preparation method thereof, it can be used for cosmetics, foods and medicines for preservative purposes.

Claims (7)

A natural preservative containing an antimicrobial composition of natural extract derived from a natural extract having low side effects and low toxicity,
The antimicrobial composition comprises an extract of gold (Scutellaria baicalensis) and an extract of Artemisia annua as an active ingredient,
The extraction is carried out with 70% ethanol, followed by sequential fractional extraction with EtOAc, n-BuOH and CH ₂ Cl ₂ ,
The antimicrobial composition which comprises an a ChungHo 20 to 50 parts by weight and extracted with the gold 20 to 50 parts by weight, CH ₂ Cl golden 20 to 50 parts by weight and extracted with _2, and CH ₂ Cl ₂ and extracted with EtOAc for 100 parts by weight of the composition Lt; / RTI > The method of claim 1,
Characterized in that it further comprises an antimicrobial composition extracted from at least one of Neem leaves, Azadirachta indica, Lavender, Lavandula angustifolia and Rosemary (Rosmarinus officinalis). The method of claim 1,
Wherein the antimicrobial composition is contained in any one of cosmetics, foods or medicines. The method of claim 1,
Wherein the antimicrobial composition comprises 0.1 to 30 parts by weight per 100 parts by weight of the finished product of cosmetics, food or pharmaceuticals. delete delete A method for producing a natural preservative containing an antimicrobial composition of natural extract derived from a natural extract having low side effects and low toxicity,
(a) a step of providing and drying golden, chungho, lantern, lavender and rosemary as herbs, respectively,
(b) extracting an active ingredient from the herb prepared in step (a) using purified water, alcohol, 70% ethanol, 100% ethanol, ethyl acetate or dimethylchloromethane,
(c) filtering and concentrating the active ingredient extracted in step (b)
The extraction in step (b) is carried out in sequential fractions with CH ₂ Cl ₂ , EtOAc, BuOH and purified water, which are different polar solvents in the fractional column,
The extracts of gold and blue are run with 70% ethanol, then sequentially extracted with EtOAc, n-BuOH and CH ₂ Cl ₂ ,
The antimicrobial composition which comprises an a ChungHo 20 to 50 parts by weight and extracted with the gold 20 to 50 parts by weight, CH ₂ Cl golden 20 to 50 parts by weight and extracted with _2, and CH ₂ Cl ₂ and extracted with EtOAc for 100 parts by weight of the composition ≪ / RTI >

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