KR20170119892A - Natural antiseptic extracted from artemisiae annuae herba and method for producing the same - Google Patents
Natural antiseptic extracted from artemisiae annuae herba and method for producing the same Download PDFInfo
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- KR20170119892A KR20170119892A KR1020160048139A KR20160048139A KR20170119892A KR 20170119892 A KR20170119892 A KR 20170119892A KR 1020160048139 A KR1020160048139 A KR 1020160048139A KR 20160048139 A KR20160048139 A KR 20160048139A KR 20170119892 A KR20170119892 A KR 20170119892A
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- preservative
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Images
Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/12—Asteraceae or Compositae [Aster or Sunflower family], e.g. daisy, pyrethrum, artichoke, lettuce, sunflower, wormwood or tarragon
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3472—Compounds of undetermined constitution obtained from animals or plants
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/282—Artemisia, e.g. wormwood or sagebrush
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/10—Preserving against microbes
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/52—Stabilizers
- A61K2800/524—Preservatives
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- Natural Medicines & Medicinal Plants (AREA)
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- Wood Science & Technology (AREA)
- Gerontology & Geriatric Medicine (AREA)
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- Medicines Containing Plant Substances (AREA)
Abstract
According to one aspect of the present invention, there is provided a natural preservative of Chungho Extract comprising, as an active ingredient, an antimicrobial composition extracted from Artemisiae Annuae Herba.
In a further embodiment of the present invention, the antimicrobial composition comprises an active ingredient obtained through extraction with water ethanol-ethanol and fractionation, and the antimicrobial composition is used as a preservative in cosmetics, pharmaceuticals, foods, , And 0.1 to 30% by weight based on the total weight of the finished product.
According to the present invention, not only does it exhibit a wide spectrum of antimicrobial activity against various fungi, but also has antioxidative and whitening effects. In particular, toxicity and irritation do not occur in the human safety test, There is an effect that the composition can be very usefully used for cosmetics food medicines and the like for the purpose of preservation.
Description
The present invention relates to a natural preservative containing an antibacterial composition extracted from Chung Ho as an effective ingredient. More particularly, the present invention relates to a natural antimicrobial composition which is extracted from Chung Ho and which has excellent antimicrobial activity against Gram-positive and gram-negative bacteria, yeast, And to a method for producing the same.
Preservatives are added to products such as food, cosmetics, household goods, wood, etc., to block or delay the decay of microorganisms, thereby enhancing preservation. Chemically synthesized artificial substances, such as parabens, are exposed to the risk of causing skin allergies and resistant bacteria even if they are ingested in small quantities through human skin like cosmetics.
Currently, preservatives used in cosmetics are composed of synthetic materials which are easy to manufacture due to the nature of the product, low in cost, excellent in formulation stability and preservative oil-repellency.
Representative cosmetic preservatives include Metyl Paraben, Ethyl Paraben, Propyl Paraben, Butyl Paraben, and Benzalkonium Chloride, imidazolidinyl esters, such as paraoxybenzoic acid esters, A chemical such as imidazolidinyl urea, trichlosan, diazolidinyl urea, quaternium-15, phenoxyethanol, and chloroohexidine gluconic acid. Synthetic preservatives.
The problem is that these synthetic preservatives, which make up the vast majority of cosmetic preservatives, are becoming more and more likely to have skin irritants and risk factors, and even the possibility of carcinogenesis through endocrine disruption and hormonal disturbances to be. Particularly, formalin glass-type preservatives including diazolidinyl urea and imidazolidinyl urea have been under review for safety worldwide and their use is on the decline, and hair cleansing products and wash off products And its use is restricted, and in some countries, including Japan, its use is strictly prohibited or its use is strictly restricted. These formalin free-form preservatives are gradually liberated in the formalin group contained therein over time, and the liberated formalin is known to not only cause skin irritation but also cause cancer.
In addition, parabens preservatives such as methylparaben, which are most commonly used in cosmetics and pharmaceuticals, have been reported to be effective against skin allergies (Andrea Counti et al., Contact Dermatitis, 1997, 37, 35-36) Applied Pharmacology, 1998, 153, 12-19.) And the induction of resistant bacteria.
In addition, there is a possibility of new problems such as acute and chronic toxicity and mutagenesis caused by persistent accumulation in the body, which are disallowed within the permitted standards (Shin Dong Hwa, Food Science and Technology, 1990, 23 (4) 68-72)
In recent years, Korea Food and Drug Administration has recognized the serious risks of these preservatives, and in order to ensure safety equivalent to that of Europe, preservatives such as isopropyl paraben, isobutyl paraben, phenyl paraben, benzyl paraben, pentyl Parabens, etc. (2014.08.07, amended by the European Commission to revise the regulations on the use of triclosan and parabens)
Accordingly, there is a growing interest in a natural preservative that is superior to the human body and has excellent antimicrobial activity based on a plant extract having natural preservative ability. Therefore, studies have been conducted to extend the antimicrobial spectrum of synthetic natural preservatives and to reduce side effects and ensure stability.
Korean Patent Registration No. 0782599 (Prior Art 1) and Korean Patent Registration No. 0697183 (Prior Art 2) are known as related art documents.
Prior Art 1 relates to a composite natural preservative prepared by mixing green tea, chamomile, grapefruit, magnolia, propolis, Japanese white bean and willow extract, and exhibits a broad spectrum of antibacterial activity and is excellent in antibacterial activity, It is expected to be useful for cosmetics, pharmaceuticals, food, etc.
Prior Art 2 discloses a process for producing a stearyl ester compound represented by the general formula (1), which comprises the step of extracting an organic solvent with an organic solvent to obtain an organic solvent fraction and then purifying the organic solvent fraction through chromatography or the like, Compounds of formula (1) are proposed. Therefore, it is expected that the novel stearyl ester will be used as a component of a pharmaceutical composition having various activities.
However, according to the above-mentioned prior art documents including various conventional technologies, there is no prior patent technology having antioxidant effect and whitening effect in addition to the antibacterial power while maintaining the basic function of the product. Especially, There is a lot of room for improvement because it is insufficient to apply to various fields.
It is an object of the present invention to overcome the above-mentioned problems of the prior art to provide an antibacterial agent that maximizes the antibacterial function derived from natural products including herbal medicines which can minimize side effects on the human body, The present invention is to provide a natural preservative of Chungho Extract which is effectively extracted from Chungho which has antimicrobial effect, antioxidative effect and whitening effect.
However, the objects of the present invention are not limited to the above-mentioned objects, and other objects not mentioned can be clearly understood by those skilled in the art from the following description.
To achieve the above object, according to one aspect of the present invention, there is provided a natural preservative of Chungho Extract comprising an antibacterial composition extracted from Artemisiae Annuae Herba as an active ingredient.
As a detailed constitution of the present invention, the extraction solvent in the present invention is preferably a solvent selected from the group consisting of water, various alcohols and water, and a mixed solvent of various kinds of alcohols. Examples of the various alcohols include methanol ethanol isopropanol n-propanol n-butanol isobutanol t-butanol and the like, with ethanol being most preferred. More preferably an active ingredient obtained through water, ethanol, ethylacetate extraction and fractionation.
According to another embodiment of the present invention, the antimicrobial composition is used as a natural preservative in cosmetics, pharmaceuticals, food, household goods, etc., and is added in an amount of about 0.1 to 30% by weight based on the total weight of the final product.
According to another aspect of the present invention, there is provided a method of producing a natural preservative according to claim 1, wherein the antibacterial extract is prepared by pulverizing a dried or non-dried crude raw material, adding the solvent to the pulverized product at a weight ratio of 10 to 100 times After which it is filtered or centrifuged to produce the supernatant as a yield.
As a detailed constitution of the present invention, the obtained product is fractionated with water, ethyl acetate, methylene chloride, hexane and butanol after ethanol extraction and used as it is in accordance with the required properties of the finished product (food, Hot air drying, spray drying, and the like.
As described above, according to the present invention, not only does it exhibit a broad spectrum of antimicrobial activity against various fungi, but also has antioxidative and whitening effects. In particular, toxicity and irritation do not occur in the human safety test, There is an effect that the antimicrobial composition of the material can be very usefully used for preserving cosmetics, food, medicine, and the like.
FIG. 1 is a graph showing the results of an identification test for the antioxidative effect of the extract of Cheongho extract according to an embodiment of the present invention,
FIG. 2 is a view showing a test result of the whitening effect of the extract of Cheongho extract according to an embodiment of the present invention,
FIG. 3 is a graph showing the results of a test for confirming the antimicrobial activity against fungi of Cheongho extract according to an embodiment of the present invention,
BRIEF DESCRIPTION OF THE DRAWINGS The accompanying drawings, which are incorporated in and constitute a part of the specification, illustrate exemplary embodiments of the invention and, together with the description, serve to provide a further understanding of the technical idea of the present invention. It should not be construed to be limited to such matters.
Hereinafter, a preferred embodiment of the present invention will be described with reference to the above-mentioned drawings. It is needless to say that the various embodiments described below do not unduly limit the contents of the present invention described in the claims, and it is needless to say that the entire constitution described in each embodiment is not essential as a solution means of the present invention.
Most of the currently used chemical preservative side effects reduce the confidence of users in preservative products, potentially damaging skin safety. For this reason, studies on free radical systems without using chemical preservatives have been actively conducted, and in order to solve the side effects of conventional chemical preservatives and to protect the safety of the products, natural preservatives which have no hypoallergenic, The need for the development of large-scale.
The present invention developed in such a situation relates to a natural preservative containing an antimicrobial composition extracted from Chungho as an active ingredient. Artemisiae Annuae Herba refers to the upper part of Artemisia annua Linne or Artemisia apiacea Hance in the dicotyledonous plants. Artemisin, which is included in Chungho, is known to be 1,200 times stronger than conventional anticancer drugs. Also, it is known that Chungho has an efficacy such as antipyretic analgesic immunity enhancement liver function protection anti-malarial effect.
The antimicrobial composition contained in the natural preservative according to one embodiment of the present invention can be extracted by using water, an organic solvent, or a mixed solvent thereof. In the case of extraction using an organic solvent, an alcohol such as methanol, ethanol, isopropanol, butanol or the like, ethylene acetone ether hexane chloroform ethyl acetate DMF (N, N-dimethylformamide) DMSO (dimethylsulfoxide) It can be extracted with an organic solvent at room temperature or by warming.
From the viewpoint of mass production, the use of water or ethanol as the extraction solvent is preferable, and the high temperature extraction is used in a limited manner because there is a risk of destruction of the active ingredient.
More preferred is ethanol extraction followed by fractionation with ethyl acetate. The extract can be produced 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. The obtained product is used as it is, or it is dried by freeze drying, vacuum drying, hot air drying, spray drying and the like.
The antimicrobial composition (or the natural preservative) of the present invention is preferably contained in an amount of 0.1 to 30% by weight, preferably 0.1 to 20% by weight based on the total weight of the finished product, such as cosmetics, food and medicine.
As such, the natural preservative according to one embodiment of the present invention exhibits excellent antimicrobial activity against various microorganisms, especially bacteria, fungi and yeast because of having a broad spectrum spectrum, and also has antioxidative effect, cytotoxicity and side effects, Skin).
The present invention also provides a composition applicable to cosmetics, food medicines, daily necessities and the like, which comprises an antibacterial composition extracted from Chung Ho. In the cosmetic composition of the present invention, the cosmetic composition is selected from the group consisting of softening oils, longevity convergence lotion, nutritional lotion nutrition cream massage cream essence eye cream eye essence cleansing cream cleansing foam cleansing water various pack powder, body lotion body cream body oil and body essence But it is not limited thereto.
In addition to the antimicrobial composition according to the present invention, it is also possible to add a surfactant, a surfactant, a water-in-oil type or a non-ionic type emulsifier, a sequestering agent, Chelating Agents Preservative Vitamin Blocking Agents Wetting Agents Essential Oil Dye Pigment Hydrophilic or lipophilic Active Agents It may contain adjuvants commonly used in lipid vials or cosmetics.
Further, according to the present invention, there is provided a household article comprising an antibacterial composition extracted from Chungho. There is no particular restriction on the type of household goods. Examples of the application include, but are not limited to, a wet tissue diaper sanitary napkin, a natural pulp polymer (resin) surface nonwoven fabric, and the like.
According to the present invention, there is also provided a food comprising an antimicrobial composition extracted from Chungho. There is no particular restriction on the type of food. Examples of foods to which the composition can be added include meat sausage bread, chocolate candy snacks, confectionery pizza noodles, other noodles, gums, dairy products including ice cream, etc. Various soups Drinks tea Drinks Alcohol drinks and vitamin complexes, All processed foods are included.
The pharmaceutical composition containing the antimicrobial composition extracted from the cheongho according to the present invention may be formulated in the form of an oral preparation external preparation suppository such as an emulsion, a syrup, an aerosol, etc., and a sterilized injection solution, . Examples of the carrier, excipient and diluent which can be contained in the pharmaceutical composition containing the composition include lactose dextrose sucrose sorbitol mannitol xylitol erythritol maltitol starch acacia rubber alginate gelatin calcium phosphate calcium silicate cellulose methylcellulose microcrystalline cellulose polyvinylpyridine Dihydroxybenzoate, talc magnesium stearate, mineral oil, and the like.
In the case of formulation, it is prepared using a diluent or excipient such as filler, extender, wetting agent, disintegrating surfactant usually used. Solid formulations for oral administration include tablets, pills, powders, granules, and the like. Such solid preparations may contain at least one excipient such as starch calcium carbonate sucrose or lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral administration include suspensions, liquid medicines, emulsion, syrups and the like. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, aromatic preservatives and the like may be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous suspensions, emulsion lyophilized formulation suppositories. As non-aqueous solvents and suspensions, injectable esters such as vegetable oil ethyl oleate such as propylene glycol polyethylene glycol olive oil and the like may be used. As a suppository base, witepsol macrogol tween 61 cacao zylarginiglycerol gelatin and the like can be used.
Hereinafter, embodiments of the present invention will be described in detail. However, the following examples are illustrative of the present invention, and the contents of the present invention are not limited to the following examples.
1. Experimental material
1) Reagent
Medium such as NA (Nutrient agar), TSA (Tryptic soy agar), LB (Lactose broth) and PDA (Potato dextrose agar) were purchased from Difco-BBL (USA).
2) Strain
The strains used in the test were four strains classified as bacteria and two strains classified as fungus. That is, Escherichia coli CCARM0237), rugi labor (Pseudomonas Pseudomonas aeruginosa KACC14021), Staphylococcus ( Staphylococcus aureus) aureus KCTC3881) and Bacillus subtilis subtilis KACC14394), fungi Candida albicans KCTC7270, Aspergillus niger KACC40280) was used (CCARM Seoul National University, Korea Antibiotic Resistant Microbial Bank, KACC Rural Development Administration, Agricultural Microbiology Bank, KCTC Korea Biotechnology Research Institute).
2. Experimental Method
1) Extraction and manufacture of Chungho
500 g of well dried chungho was cut into about 1 to 3 mm and extracted with water, 70% ethanol and ethyl acetate, filtered through a 0.45 μm filter, and then concentrated using a vacuum concentrator. Concentrated and then lyophilized and used in the experiment.
2) Disk diffusion antimicrobial experiment
The antimicrobial activity of Cheongho extract was verified by the conventional disk diffusion method. Escherichia coli (E. coli) and Bacillus subtilis (B. subtilis) was the medium used in the culture of LB, rugi labor Pseudomonas (P. aeruginosa) and Staphylococcus aureus (S. aureus) and Candida Albi C. albicans and Aspergillus niger used NA, PDA medium, respectively. Each bacterium was inoculated with 2 to 3 colonies (0.8 × 108 CFU / mL). The extract was dissolved in DMSO at various concentrations, and 10 μl was added to a sterilized paper disk (diameter 8 mm). After the DMSO was completely dried, the microspheres were immersed in a plate medium and kept at 35 캜. After 20 to 24 hours, the clear zone around the paper disk was measured to examine the antimicrobial activity.
3) Measurement of minimum inhibitory concentration (MIC)
MICs were obtained by incubating each bacterium in a solid agar culture medium and then 5 colonies were obtained according to 0.5 McFarland standard (1 × 108 CFU / mL), diluted in MHB medium and inoculated, and maintained at 35 ° C. for 2 to 3 hours. The chrysanthemum extract used in this experiment was titrated to a concentration of 8.0 mg / mL and diluted to 2-fold each. The culture medium containing each bacterium and extract was transferred to a 96-well plate and incubated at 35 ° C for 18-20 hours, and then measured with a Multilabel Counter (1420, Perkin Elmer) at 595 nm to obtain MIC values.
4) Statistical processing
Statistical analysis was performed with ANOVA and Student's t-test. The significance level was set at p <0.05.
< Experimental Example One: Cheongho Antimicrobial Effect of Extracts against Pathogenic Microorganisms (Disk Diffusion)>
In the present invention, the antimicrobial composition was first extracted by using relatively safe water, ethanol, and ethyl acetate solvent, and the medium used for the culture of E. coli and B. subtilis LB, and a total of six microorganisms, Pseudomonas aeruginosa , S. aureus , Candida albicans , and A. niger , The concentration of each extract (antimicrobial composition) was titrated from 5 to 100 mg per mL and tested by disk diffusion method.
As a result, Chungho water extract (an antimicrobial composition extracted from Chungho by using a water solvent) contained E. coli , Bacillus subtilis , Pseudomonas aeruginosa ( P . aeruginosa , S. aureus , and C. albicans showed antimicrobial activity at 10 mg / ml or higher.
[Table 1]
As shown in Table 2, in the case of E. coli , B. subtilis , Staphylococcus aureus ( E. coli ), and B. subtilis S. aureus) are showed an antimicrobial activity at concentration of 5mg / ml, rugi labor (P. aeruginosa Pseudomonas) and Candida albicans (C. albicans) showed an antimicrobial activity at 10mg / ml.
[Table 2]
As shown in Table 3, only S. aureus did not exhibit antimicrobial activity at a concentration of 1 mg / ml in the crude ethyl acetate extract (an antimicrobial composition extracted from Cheongho using an ethyl acetate solvent).
[Table 3]
CCARM is a strain of antibiotics resistant bacteria in Seoul, KACC is a microorganism bank of Rural Development Administration, and KCTC is a strain provided by Korea Research Institute of Bioscience and Biotechnology, Korea, in the "strain number a" of [Table 1] to [Table 3] "Antimicrobial activity (mm) b" is the size (diameter mm) of the zona pellucida.
<Experimental Example 2: Minimum inhibitory concentration (MIC) for the pathogenic microorganisms of Chungho extract>
[Table 4] shows the minimum inhibitory concentrations of the Chungho extract that can kill each bacterium.
[Table 4]
<Experimental Example 3: Effect of DPPH radical scavenging activity on the skin aging inhibition by the extract of Cheongho>
To investigate the antioxidative effect of the mixture of the present invention, 1,1-diphenyl-2-pic rylh-2-ene was prepared according to the method of Kilani et al. (Kilani et al., Environ. Toxicol. Phar. 20: 478-484 ydrazyl (DPPH; Sigma Co., USA). DPPH is widely used to measure the hydrogen donating ability of antioxidants through the measurement of absorbance because it reacts with a substance having an antioxidant activity to decolorize with a dark purple color due to radical scavenging. Arbutin, a well-known antioxidant, was used as a positive control in this experiment.
100 μl of the test substance diluted with methanol (100, 500 and 1,000 μg / ml) was added to each well of a 96-well plate, 100 μl of DPPH solution (0.2 mM DPPH in Methanol) was added, Absorbance was measured at 517 nm using a reader (BioTEK, USA). The radical scavenging ability of each test substance was calculated as the percentage of electron donating ability by the following formula.
Electron donating ability (%) = (1 - absorbance of test substance addition group / absorbance of no addition group) x 100
As shown in the results of Fig. 1, the treatment with purified water (H2O) and ethanol-ethylacetate extract was found to have a better radical scavenging ability than the positive control group, arbutin.
< Experimental Example 4: Check the whitening effect - Tyrosinase Inhibitory activity>
The active assay method described in Ishihara et al., J. Antibiot (Tokyo), 44 (1), p25-32, 1991) was modified to produce a mushroom-derived tyrosinase inhibitory activity involved in melanin biosynthesis and oxidative action Were measured. Concretely, Chungho extracts were added to 50 μl of a 3 mg / ml tyrosine solution (dissolved in 0.1 M phosphate buffer, pH 6.8) and 1 ml of a 50% by volume methanol solution, respectively, to prepare samples according to concentrations. Each of the above concentrations was 10 μg / ml, 100 μg / ml and 250 μg / ml of arbutin of each of the extracts of 10 μg / ml, 100 μg / ml and 250 μg / ml.
50 μl of each of the above samples was placed in a 96-well plate and 50 μl of mushroom extract tyrosinase (Sigma, mushroom, 200 Unit / ml, 0.1 M phosphate buffer, pH 6.8) . After incubation at 37 ° C for 10 minutes, the amount of dopachrome produced was measured at 475 nm using a spectrophotometer (Precision Microplate Reader, Molecular Devices, VersaMax). The experiment was repeated three times. Dopachrome is an intermediate of melanin production and can have a whitening effect when dopachrome production is inhibited. The tyrosinase inhibitory activity was calculated according to the following formula according to Matsuda H et al. Biol. Pharm. Bull. 19 (1), pp 153-156, 1996) using a sample to which no sample was added as a control. The results are shown in Fig.
Tyrosinase inhibitory activity (%) = {(control OD.475 - sample OD.475) / control OD.475} X100
In the above formula 1, OD.475 represents an optical density measurement value at 475 nm.
As shown in FIG. 2, the ethyl acetate extract of Cheongho extract according to the present invention has a superior tyrosinase inhibitory activity as compared to ethanol and water extract, and when compared with arbutin (single substance), a positive control group well known for its whitening effect, The efficacy of Cheongho extract (mixed material) was not much behind, and it was confirmed that it was also excellent in production efficiency.
It can be clearly confirmed that the herbal extract of the present invention exhibits anti-tyrosinase activity in a concentration-dependent manner and exhibits antioxidative and whitening effects.
<Experimental Example 5: Evaluation of cytotoxicity of Chungho extract>
The cytotoxic effects of natural preservatives were studied by MTT using human keratinocyte HaCaT and human normal fibroblast cells. As shown in Fig. 3, in the case of human normal fibroblasts and HaCaT, Chungho extract did not show cytotoxicity at all concentrations. This lack of cytotoxicity suggests the possibility of safe use for food cosmetics, household products, and medicines.
<Experimental Example 6: Preservative effect of cosmetic preparations of Cheongho extract>
To measure the preservative effect of the cosmetic composition containing the antibacterial composition extracted from Chung Ho, a lotion containing 2.5% of the antibacterial composition was prepared as follows. Lotions containing the antimicrobial composition were prepared via conventional methods with the compositions of Table 5 below. [Table 6] shows the test results of the buoyancy test and shows a 70 ~ 82% of buoyancy force compared with commercial products.
[Table 5]
[Table 6]
Here, Control is a conventional commercial preservative or chemical preservative.
<Experimental Example 7: Test for confirming skin stability of Cheongho extract>
As a result of the skin stability test of Chungho extract, it was found that the response to the control was low as shown in Table 8, and thus it was confirmed that the Chungho extract was safe to use on the skin. Table 7 shows patch test criteria.
[Table 7]
Here, the mean skin response (score) = (score × reaction number × 100 × ½) / [3 (maximum score) × total number of subjects (n)]
[Table 8]
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 exemplary embodiments. That is, those skilled in the art will readily understand that many modifications are possible without departing from the gist of the invention. In the following description, well-known functions or constructions relating to the present invention as well as specific combinations of the components of the present invention with respect to the present invention will be described in detail with reference to the accompanying drawings. something to do.
According to the present invention, not only does it exhibit a wide spectrum of antimicrobial activity against various fungi, but also has antioxidative and whitening effects. In particular, toxicity and irritation do not occur in the human safety test, There is an effect that the composition can be very usefully used for cosmetics food medicines and the like for the purpose of preservation.
Claims (5)
Natural Preservative of Chungho Extract containing the antimicrobial composition extracted from Artemisiae Annuae Herba as an active ingredient.
Wherein said antimicrobial composition comprises an active ingredient obtained through water, ethanol, ethyl acetate extraction and fractionation.
The antimicrobial composition is used as a preservative in cosmetics, medicines, food and household products, and is added in an amount of 0.1 to 30% by weight based on the total weight of the finished product.
The obtained product is fractionated with ethanol, extracted with water, ethyl acetate, methylene chloride, hexane and butanol, and used as it is in accordance with the required properties of the finished product (cosmetics, pharmaceuticals, etc.), or by freeze drying, vacuum drying, hot air drying, And then drying the mixture.
The obtained product is fractionated with ethanol, extracted with water, ethyl acetate, methylene chloride, hexane and butanol, and used as it is in accordance with the required properties of the finished product (cosmetics, pharmaceuticals, etc.), or by freeze drying, vacuum drying, hot air drying, And then drying the mixture.
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