KR101483749B1 - Whitening cosmetic composition and Antibacterial composition containing the extract of Hippophae rhamnoides L. - Google Patents

Abstract

The present invention relates to a whitening cosmetic composition and an antimicrobial composition comprising a vitamin Hippophae rhamnoides L. extract. In particular, a fruit extract of a vitamin tree, preferably an ethanol extract of a vitamin Nut fruit, The present invention also relates to an antimicrobial composition which can further enhance the antimicrobial effect by including the extract of the rhizomes with the vitamin-tree extract.

Description

(Whitening cosmetic composition and Antibacterial composition containing extract of Hippophae rhamnoides L.)

The present invention relates to a whitening cosmetic composition and an antimicrobial composition comprising a vitamin Hippophae rhamnoides L. extract, and more particularly to an extract having excellent antimicrobial activity from a vitamin tree.

Conventionally, synthetic organic antibacterial agents or inorganic antibacterial agents have been used to inhibit the activity of harmful microorganisms. However, existing synthetic organic antibacterial agents are highly irritating to eyes, skin and olfactory sense of humans, and have a weak antibacterial activity against Gram-negative bacteria (presence of cell membrane). The inorganic antimicrobial agent generally has a weak antibacterial effect, and particularly has insufficient antibacterial activity against fungi, and has a disadvantage in that the antibacterial activity is rapidly reduced upon contact with moisture.

At present, representative natural antimicrobial agents that can be used include niacin and polylysine, which are bacteriocins of lactic acid bacteria. However, they are required to have a large amount for antibacterial activity, and they are expensive. In addition, chitosan is more effective against Gram-positive bacteria than Gram-negative bacteria, but exhibits relatively weak antibacterial activity against fungi and has little effect on genus containing chitosan in the cell wall.

In this regard, Korean Patent Publication No. 90-17490 discloses a pharmaceutical composition suitable for the treatment of fungal skin diseases by topical administration in order to prevent and cure bacterial or fungal infections. Examples thereof include powders, solutions, suspensions, A gel, a paste, an ointment or a tincture, and a cosmetic composition. However, when the composition of the above-mentioned formulation is applied to the affected part, it is required to apply a large amount of the drug when the application site is wide, and it is difficult to control the application amount uniformly. In the case of semi- ) There is a problem of sticking to the coating after application to the skin.

Further, in the case of an oral antibiotic or antifungal agent, since the drug is circulated throughout the body, it is necessary not only to administer the drug more than necessary but also to cause side effects due to long-term use in order to exhibit its effect. Particularly, toxicities in the body such as causing side effects at the time of long-term use of triazole-based drugs have been questioned.

Hippophae rhamnoides L. is a shrub belonging to the family Borneo, and contains more than 100 kinds of ingredients. Especially, it contains a lot of effective ingredients such as vitamins (A, B, C, E, F and K) have. It grows well in the cold of winter and the high temperatures of summer, and has a strong adaptability that grows well in the barren zone. It has already been reported that antioxidant activity of Sanjia wood is investigated, and fatty acid content, cancer prevention, and immune system of Sanjay tree are being studied. Beta carotene is contained in the fruit tree. In addition, the active ingredient is measured by instrumental analysis of Sanjia leaves, and various foods are made using Sanjia wood, but the research is not active in Korea yet.

In this regard, Korean Patent Publication No. 2010-0054772 (entitled Vitamin tree extract and fractions thereof) discloses an antioxidant comprising an extract of Sanjay tree, a root or stem extract or an organic solvent fraction thereof as an active ingredient. However, when the leaves, roots or stem extracts of vitamin trees are used, not only the cell survival rate is low but also the tyrosinase inhibitory effect, melanin formation inhibitory effect and antibacterial activity are insufficient, so that cosmetic composition or antibacterial composition It is not suitable for commercialization.

Accordingly, there is always a demand for natural extracts and compositions which not only have a high cell survival rate, but also have a tyrosinase inhibitory effect, a melanin production inhibitory effect and an antibacterial activity.

The object of the present invention is to obtain a vitamin A tree extract having excellent cell survival rate, tyrosinase activity inhibiting effect and melanin formation inhibiting effect.

The present invention also provides a vitamin A tree extract having excellent antimicrobial activity.

In order to accomplish the above object, the present invention is a whitening cosmetic composition comprising a fruit extract of vitamin wood or a fraction of the extract as an active ingredient.

Another embodiment of the present invention is an antimicrobial composition comprising a fruit extract of vitamin wood or a fraction of the extract as an active ingredient.

Here, the fruit extract of the vitamin tree is preferably an ethanol extract of vitamin Nut fruit.

Yet another embodiment of the present invention is an antimicrobial composition comprising an extract of vitamins and ginseng extract or a fraction of the extract as an active ingredient.

Here, the vitamin tree extract is preferably an ethanol extract of vitamin Nut fruit.

It is more preferable that the vitamin tree and the herringbone extract include 0.5 to 1.5 parts by weight of the herbal extract in relation to 1 part by weight of the vitamin tree extract.

The details of other embodiments are included in the detailed description and drawings.

The present invention is more suitable as a cosmetic composition for whitening because it contains a fruit extract of vitamin tree and thus has superior cell survival rate, tyrosinase activity inhibitory effect and melanin formation inhibitory effect than leaf extract.

Further, the present invention is more suitable as an antimicrobial composition because it has better antimicrobial activity than hot-water extract by including ethanol extract of vitamin Nut.

In addition, the present invention has an antimicrobial effect that is remarkably improved by including a herbal extract with a vitamin-tree extract.

1 is a graph showing an example of experimental results on cell viability of vitamin A nut and leaf extract according to the present invention,
FIG. 2 is a graph showing an example of experimental results on the antioxidant effect of vitamin A fruit extract according to the present invention,
FIG. 3 is a graph showing an example of experimental results on the antioxidant efficacy of the vitamin-tree leaf extract according to the present invention,
FIG. 4 is a graph showing an example of experimental results on inhibition of tyrosinase activity of vitamin Nut and leaf extract according to the present invention,
FIG. 5 is a graph showing an example of experimental results on inhibition of melanin formation by the vitamin tree leaf extract according to the present invention,
FIG. 6 is a graph showing an example of experimental results on the melanin production inhibitory effect of the vitamin Nut extract of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS The present invention is capable of various modifications and various embodiments, and specific embodiments are illustrated in the drawings and will be described in detail in the detailed description. It is to be understood, however, that the invention is not to be limited to the specific embodiments, but includes all modifications, equivalents, and alternatives falling within the spirit and scope of the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

The terminology used in this application is used only to describe a specific embodiment and is not intended to limit the invention. The singular expressions include plural expressions unless the context clearly dictates otherwise. In the present application, the terms "comprises" or "having" and the like are used to specify that there is a feature, a number, a step, an operation, an element, a component or a combination thereof described in the specification, But do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, or combinations thereof.

The terms first, second, etc. may be used to describe various components, but the components should not be limited by the terms. The terms are used only for the purpose of distinguishing one component from another.

The present invention provides a vitamin A tree extract, more preferably a fruit extract of a vitamin A tree.

The inventors of the present invention continued research to obtain an extract having a high cell survival rate from a natural substance, an inhibitory effect on tyrosinase activity, an inhibitory effect on melanin production, and an antibacterial activity, and a vitamin extract, especially a vitamin extract The inventors of the present invention have completed the present invention after confirming that they have superior cell survival rate, tyrosinase activity inhibitory effect, melanin production inhibitory effect and antibacterial effect.

The vitamin tree can be used without limitation, such as cultivated or commercially available. In addition, the extract of Liliaceae according to the present invention can be prepared as follows. The fruit is washed cleanly with water, dried in the shade and room temperature for 7 days, and the dried fruit is pulverized, put into an extraction vessel, and then extracted with an extraction solvent at a suitable temperature for a certain period of time. The extraction solvent is preferably a solvent selected from water, an alcohol or a mixture thereof, preferably a C1 to C4 lower alcohol or a mixed solvent thereof, more preferably methanol, but is not limited thereto. The amount of the extraction solvent is 2 to 10 times, preferably 4 times, the dry weight of the corn cob. The extraction method may be an extraction method such as an immersion extraction method, a heat extraction method, a reflux cooling extraction method, and an ultrasonic extraction method. Preferably, the extraction method may be performed once to five times by an immersion extraction method. The temperature at the time of extraction is preferably 20 ° C to 30 ° C, more preferably 20 ° C to 25 ° C at room temperature. The extraction time is 5 to 10 days, preferably 7 days. After obtaining the above-mentioned extract, the solid content is removed using a filter paper or the like, the suspension is centrifuged, and the supernatant can be filtrated under reduced pressure.

In addition, the present invention provides fractions obtained in each step by extracting the above-mentioned extract from the above-mentioned extracts in the order of n-hexane, ethyl acetate and n-butanol. The fraction solvent for obtaining fractions of the above extract is selected from the group consisting of n-hexane, ethyl acetate and n-butanol, and is usually fractionated using a separatory funnel. The fraction may be obtained from the extract by repeating the fractionation process one to five times, preferably three times, and may be fractionated and then concentrated under reduced pressure. In the example of the present invention, the above-described extract of the Japanese apricot tree was suspended in distilled water, followed by fractionation in the order of n-hexane, ethyl acetate and n-butanol, followed by repeating this three times and each solvent fraction was concentrated under reduced pressure

 Accordingly, the present invention is a whitening cosmetic composition comprising a fruit extract of the vitamin tree or a fraction of the extract as an active ingredient.

The present invention has excellent cell survival rate (see FIG. 1), tyrosinase activity inhibitory effect (see FIG. 4) and melanin formation inhibitory effect (see FIG. 5 and FIG. 6) Therefore, it is more suitable as a cosmetic composition for whitening.

The whitening cosmetic composition may include, but is not limited to, lotions, ointments, gels, creams, patches or sprays. In preparing the cosmetic for skin whitening containing the vitamin nuts extract or the fraction thereof of the present invention, 1 to 15 parts by weight of the vitamin nuts extract or the fraction thereof of the present invention is added to the composition for external application for skin, May be added in an amount of 2 or 10 parts by weight. The external composition for skin of the present invention may further contain, in addition to the vitamin Nutrient extract of the present invention or its fractions, a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, a softener, an antioxidant, a suspending agent, a stabilizer, A preservative, a vitamin, a barrier, a wetting agent, a essential oil, a dye, a pigment, a hydrophilic or lipophilic active agent, a lipid vesicle or In the field of dermatology such as any other ingredient commonly used in external preparations for skin

And may contain commonly used adjuvants. The components can also be introduced in amounts commonly used in the field of dermatology.

Another embodiment of the present invention is an antimicrobial composition comprising a fruit extract of vitamin wood or a fraction of the extract as an active ingredient.

The antimicrobial composition is characterized by exhibiting excellent antimicrobial activity against pathogenic bacteria and the like. The pathogenic bacteria include Gram-positive bacteria and Gram-negative bacteria, for example, Escherichia coli, Staphylococcus aureus, which is Staphylococcus aureus, and Pseudomonas aeruginosa, which is Pseudomonas aeruginosa. It is one or more selected from the group. The Escherichia coli is an indicator bacterium for food contamination, Staphylococcus aureus is a causative agent of food poisoning, and Pseudomonas aeruginosa is the only species having pathogenicity to humans, and acts as a pathogen when deficient in defense or mixed infections, Of meningitis, meningitis, necrotizing pneumonia, ocular disruption, urinary tract infection and the like.

In the examples of the present invention, E. coli, Staphylococcus aureus, and Pseudomonas aeruginosa, E. coli, E. coli, and C. fungi, C. albicans and A. niger were tested for their antibacterial effect, Were found to have excellent antimicrobial activity against the three bacterial species. In particular, the ethanol extracts of the vitamin-wood extracts were more effective than the hot-water extracts of the vitamins (see Table 2 and Table 3), and thus the fruit extract of the vitamin tree is preferably an ethanol extract of vitamin- Among them, 70% ethanol extract of vitamin Nut fruit is more preferable. The present invention is more suitable as an antimicrobial composition because it has better antibacterial activity than hot-water extract by including ethanol extract of vitamin Nut fruit.

Accordingly, the present invention provides a natural preservative or a preservative containing the above vitamin A fruit extract or a fraction thereof as an active ingredient.

The natural preservatives or preservatives of the present invention exhibit excellent antimicrobial activity over a wide range of microorganisms, particularly pathogenic bacteria. The natural preservatives or preservatives of the present invention may contain, in addition to the above-mentioned vitamin Nut fruit extract or its fractions, additional ingredients as long as they do not inhibit their preservative activity. As described above, the natural preservative according to the present invention having a broad spectrum of preservation can be widely used for the purpose of preservation in medicines, cosmetics, foods, fibers, household goods, etc., Increase stability. When the natural preservative of the present invention is contained in the above-mentioned products, the amount of the natural preservative is 0.001 to 30% by weight, preferably 0.001 to 20% by weight, more preferably 0.001 to 5% by weight Range. To prepare a product comprising the natural preservative of the present invention, one may make suitable formulations and additives as is well known to those skilled in the art.

In addition, the present invention may be an antioxidant containing vitamin A fruit extract or a fraction of the extract as an active ingredient. The fruit extract of the vitamin tree has an excellent antioxidative effect (see FIG. 2 and FIG. 3) and is suitable for use as an antioxidant.

The present invention also provides a pharmaceutical composition for the prevention and treatment of diseases in which excessive amounts of active oxygen, which is an active ingredient, are accumulated by the above-mentioned vitamin nuts extract or its fractions. The present invention also provides a pharmaceutical composition for preventing or treating aging, which contains the above vitamin A fruit extract or a fraction thereof as an active ingredient. The vitamin A nut extract or the fraction thereof of the present invention exhibits antioxidative activity and thus can be effectively used as a pharmaceutical composition for preventing or treating diseases in which effective oxygen is accumulated in an excessive amount, or a pharmaceutical composition for preventing aging. At this time, the disease in which the active oxygen accumulates and is caused by excessive accumulation is selected from the group consisting of liver disease, stroke, myocardial infarction, diabetic vascular disorder, hyperlipidemia, acute inflammation, rheumatism and cancer. The composition may be used alone or in combination with methods using surgery, hormone therapy, drug therapy, and biological response modifiers, as well as prevention and treatment of diseases caused by accumulation of excessive amounts of active oxygen, for prevention of aging.

In addition, the present invention provides a cosmetic for preventing skin aging comprising the vitamin Nut fruit extract or a fraction thereof.

In addition, the present invention provides an antibiotic comprising the above vitamin A fruit extract or a fraction thereof as an active ingredient.

In addition to the above, vitamin A fruit extract or its fractions of the present invention may be used as a nutritional supplement, a vitamin, an electrolyte, a flavoring agent, a colorant, a pectic acid and its salt, an alginic acid and its salt, an organic acid, a protective colloid thickener, Preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the vitamin nuts extract or its fractions or derivatives thereof of the present invention may contain flesh for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Yet another embodiment of the present invention is an antimicrobial composition comprising a vitamin A extract and a mulberry extract or a fraction of the extract as an active ingredient.

The term " Moutan Root Bark "in the present invention is a root shell of a peony (name: Paeonia Suffruticosa Andrews), which is harmless to the human body used as a medicinal material in Oriental medicine. Ingredients include paeonoside (paeonol glucoside), paeonolide (paeonol-rhamnoglucoside), and paonol, which degrades during storage to produce sugars and patois. It is also possible to use paeoniflorin, oxypaeoniflorin, benzoylpaeoniflorin, paeonolide tannin, procyanidin B1, benzoyloxypaeoniflorin, It contains ingredients such as paeonin, astragalin, pelargonin and so on. These components were not only found in the muscle but also in the neck area. It is presumed that the anti-inflammatory action as a former hemolytic agent is related to the monoterpene glycosides component. It has been found that phenol has antimicrobial action, and that it inhibits the proliferation of Escherichia coli, Staphylococcus, Streptococcus, Bacillus, etc. at a dilution concentration of 1,500 to 2,000 times in vitro. Benzoylphenofuran, benzoyloxyphenofuran and the like have an inhibitory action on histamine release in mast cells, and phenolide tannin has an antiviral action. Ferniflorin inhibits platelet aggregation and reduces fibrinogen. The herbal medicine is used as an antihemorrhagic medicine in the herbal medicine, and its efficacy is aimed at the inflammation of the vasculature of the lower abdomen, pain due to congestion, fever, pyuria and hemorrhage. Especially in the gynecological area, Inflammation of the adnexa, congestion, traction, anti-inflammation, analgesia, and has the effect of spine, hemorrhoids, appendicitis are also applied.

The inventors of the present invention completed the present invention after confirming that antimicrobial activity was further increased when the extract of Mulberry extract was included together with other natural products while studying a method of further enhancing the antimicrobial activity of vitamin A tree extracts.

Here, the vitamin tree extract is preferably an ethanol extract of vitamin A nutrients as described above.

The present invention is characterized in that the vitamin tree extract and the herbal extract are used as an active ingredient. Specifically, the extract may be a mixed extract prepared by preparing each of the extracts of the above-mentioned vitamins and ginseng and then mixing them at an appropriate ratio. The above-mentioned mixed extract has an antimicrobial effect which is remarkably improved as compared with that obtained by extracting each of the vitamins or wood vinegar.

It is more preferable that the vitamins and mulberry extracts contain 0.5 to 1.5 parts by weight of the Mulberry Extract with respect to 1 part by weight of the vitamin tree extract. If the amount is less than the above range, the synergistic effect due to the extract of Myrrh can be insufficient and the effect of increasing the antimicrobial activity is not high even if it exceeds the above range.

The vitamins and mulberry extracts of the present invention may be, but are not limited to, ethanol dry extracts or soft extracts. The term "dry extract" in the present invention means a preparation prepared by adding an appropriate precipitant to a crude drug raw material, leaching the mixture for a predetermined period of time by a cold, warming or percolation method, filtering the resulting liquid, . Dry extracts can be made into crumbable lumps, granules or powder. The term "soft-drink extract" in the present invention means that the extract obtained by adding a suitable solvent (purified water, ethanol, etc.) to the raw material of herbal medicine for a certain period of time and concentrating the extract to obtain the same degree of lightness as syrup.

The present invention may be better understood by the following examples, which are for the purpose of illustrating the invention and are not intended to limit the scope of protection defined by the appended claims.

Example  One: Vitamin tree  Preparation of extract

Example  1-1: Vitamin tree  Preparation of fruit and leaves

The fruit and leaves of the vitamin tree were washed, dried for 5 days at room temperature, and then pulverized.

Example  1-2: Vitamin tree Heat number  Preparation of extract

The fruit of vitamin tree prepared in Example 1-1 and 200 g of leaf powder were each added to 2 L of distilled water for 3 hours and stirred at a magnetic bar for 24 hours at 60 ° C. and then only the supernatant was recovered by high speed centrifugation. After filtration through paper, freeze-drying was carried out to solidify it.

Example  1-3: Vitamin tree  Preparation of ethanol extract

Each of the vitamins and 200 g of leaf powder prepared in Example 1-1 was added to 2 L of a 70% ethanol solution, stirred at room temperature for 24 hours with a magnetic bar, sonicated at 30 ° C for 15 minutes, filtered After filtration through paper, freeze-drying was carried out.

Experimental Example  One : Vitamin tree  Efficacy test for extract

Experimental Example  1-1: Cell viability test

The toxicity test was carried out through the cell survival rate of melanocyte (B16F10 cell line) of the extract prepared according to Example 1-2. First, the extract was diluted with various concentrations (0, 5, 10, 20, 50, 100, 500, 1000, 1500, 2000 / / ml) and incubated in a CO ₂ incubator at 37 ° C in a B16F10 cell line Time. Then, 50 μl of 1/10 volume of CCK-8 assay solution was added to each well and incubated in a CO ₂ incubator at 37 ° C for 1 hour. Then, the raw data were determined at 480 nm with an ELISA leader, and the viability measurement was calculated. In the case of living cells, NAD-NADH is synthesized to react with CCK-8 solution, resulting in red discoloration and reading at OD 450 nm. In the case of dead cells, the amount of NAD-NADH is low and the absorbance value is low. The absorbance values of the cells and the CCK-8 solution as negative controls and the absorbance values of the material and CCK-8 solution (by concentration) were used as blank values.

As a result, as shown in Fig. 1, the survival rate of vitamin A fruit extract was 50% or more in the concentration range of 0 ~ 2000 ㎍ / ㎖, and the survival rate of leaf extract was abruptly increased in the concentration range of 500 ~ 2000 ㎍ / .

Experimental Example  1-2: Antioxidant efficacy test

The antioxidant activity of the extract prepared according to Example 1-2 was measured at various concentrations (100, 300, 500, 700, 1000 μg), and vitamin was used as a positive control in the same amount as the extract. The extract was diluted with methanol by concentration and mixed with DPPH solution (0.2 mM DPPH in Methanol) at a ratio of 1: 1 (100 μl), followed by reaction at room temperature for 10 minutes, and then the absorbance was measured at OD 517 nm. The absorbance (OD 517 nm) value of each of "S" mixed with DPPH solution and "B1" mixed with only methanol and DPPH solution, "C" mixed with only methanol and DPPH solution and "B2" And the like.

DPPH scavenging activity (%) = [1- (S-Bl) / (C-B2)] X 100 (%)

As a result, as shown in FIG. 2 and FIG. 3, the antioxidant efficacy was similar to that of the positive control vitamin and the extract of the present invention (leaf, fruit).

Experimental Example  1-3: Tyrosinase  Active inhibition efficacy test

The inhibitory effect of the extract prepared according to Example 1-2 on the tyrosinase activity was measured at various concentrations (5, 10, 20, 50, 100, 500 / / ml) and as a positive control, Respectively. The extract was dissolved in a suitable solvent such as ethanol or DMSO to make a 1% solution (sample concentration: at least 5 concentration ranges). In the test tube, 220ul of 0.1M phosphate buffer (pH 6.5), 20ul of the sample solution and 20ul of the mushroom tyrosinase (2000U / ml) were put in order. To this solution was added 40 ul of 1.5 mM tyrosine solution, and the mixture was reacted at 37 DEG C for 10 to 15 minutes. The absorbance was measured at 490 nm using an ELISA reader. A 0.1M phosphate buffer solution (pH 6.5) was added to the sample solution instead of the sample solution. The concentration of the sample (IC 50) when the inhibition rate was 50% was calculated using an appropriate program.

(%) = 100 - ((b-b ') / (a-a')) * 100. a (absorbance after reaction of the blank), b (absorbance after reaction of sample solution), a ', b' (absorbance measured by substituting buffer instead of tyrosinase)

As a result, as shown in FIG. 4, the activity of tyrosinase of the extract according to the present invention was lower than that of the positive control group, and the vitamin extracts of the leaves of vitamins showed a concentration-dependent inhibitory effect of tyrosinase .

Experimental Example  1-4: Test for inhibiting melanin formation

The treatment concentration was determined in a concentration range without cytotoxicity. The treatment concentration of the extract prepared according to Example 1-2 was used in an amount of 5, 10, 20, 50, and 100 μg / ml. Arbutin was used as a positive control in the same amount as the extract Respectively. The B16F10 cell line is a cell line that synthesizes melanin. The melanin cell line was seeded after 3 x 10 ^ 5 cell counts. After confirming that at least 80% of the melanocytes were attached (16 hours), the extract was treated for 48 hours by concentration. After the culture was completed, the medium was removed, washed with PBS (Phosphated Buffer Saline), and treated with trypsin to recover the cells. Then, the recovered cells were counted using a hematocytometer, centrifuged at 5,000 to 10,000 rpm for 10 minutes, and the supernatant was removed to obtain pellets. After addition of 200 μl of 1N NaOH (containing 10% DMSO), the melanin was completely dissolved by standing at 80 ° C for 1 hour, and the absorbance at 405 nm was determined.

As a result, as shown in Fig. 5 and Fig. 6, the inhibitory effect of vitamin-tree leaf extract on melanin formation was lower than that of arbutin, which is a positive control group. Compared with the positive control group arbutin, Melanin inhibitory activity was confirmed to be effective.

Experimental Example  1-5: Antimicrobial efficacy test

The antimicrobial activity of the hot-water extract of the vitamin Nutrient prepared according to Example 1-2 and the ethanol extract of the vitamin Nutrient prepared according to Example 1-3 was tested. The test strains were Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 8739, Pseudomonas aerugnosa ATCC 9027, Candida albicans ATCC 10231, Aspergillus niger ATCC 16404 were used. For the test method , E. coli , S. aureus and P. aerugnosa were inoculated into soybean casein digestion broth (Trypticase Soy Broth) and cultured at 30 ~ 35 ℃ for 24 hours. Of the fungi, C. albicans was inoculated into Sabouraud Dextrose broth and incubated at 20-25 ° C for 48 hours. The culture broth was centrifuged, the medium was removed, washed with sterile physiological saline, and adjusted to a concentration of 1 to 9x108 cells / ml, and used as the inoculum. A. niger was inoculated on a Sabouraud Dextrose Agar and incubated at 20-25 ° C for 7 days. The cultured A. niger was used as inoculum with 1 ~ 9 × 108 cells / ml spore suspension using 0.05% polysorbate 80-added sterile physiological saline solution. Each test solution was inoculated in 20.0 mL of each test solution to be inoculated with 105 ~ 106 cells / mL, and then stored at room temperature in a dark place. The number of viable cells was measured on the day 0, 14, and 28 after inoculation of each sample in storage, and the change in color and smell was observed and the results were recorded. However, in the initial dilution stage of all experiments, neutralization was performed using D / E Neutralizing Broth (DIFCO), and sterilized physiological saline was used as a diluent.

The cultures were incubated at 30 ~ 35 ℃ for 24 hours, and C. albicans were incubated at 20 ~ 25 ℃ for 48 hours and A. niger at 20 ~ 25 ℃ for 7 days. When bacteria were grown in the medium after cultivation, the number of bacteria on the medium was multiplied by the dilution factor. When the bacteria did not proliferate in the medium, the result was multiplied by the dilution factor in the neutralization stage and expressed as "less than 10 (<10)".

The number of viable cells was calculated as "N (live cells) = C (number of colonies) x D (dilution factor) x V (dilution ratio of D / E Neutralizing Broth used in the extraction step (10 times)").

Table 1 below shows the criteria for preservation.

The results are shown in Tables 2 and 3 below.

division E. coli S. aureus P. aeruginosa C. albicans A. niger Initial inoculation (0 day) 4.8 x 10 ⁶ 4.1 x 10 ⁶ 1.3 x 10 ⁶ 2.0 x 10 ⁵ 2.3 x 10 ⁵ After 7 days, the number of bacteria 3.5 x 10 ³ <10 <10 5.2 x 10 ⁵ 3.2 x 10 ⁵ After 14 days, the number of bacteria <10 <10 <10 5.1 x 10 ⁵ 2.1 x 10 ⁵ After 28 days, the number of bacteria <10 <10 <10 3.6 x 10 ⁵ 1.0 x 10 ⁵

(Unit: CFU / ml)

As shown in Table 2 above, the 70% ethanol extract of vitamin Nut was excellent in antibacterial effect against three kinds of E. coli , S. aureus and P. aerinosa , , it was confirmed that the preservative efficacy low for the yeast fungi (C. albicans) and filamentous fungi (A. niger).

division E. coli S. aureus P. aeruginosa C. albicans A. niger Initial inoculation (0 day) 4.8 x 10 ⁶ 4.1 x 10 ⁶ 1.3 x 10 ⁶ 2.0 x 10 ⁵ 2.3 x 10 ⁵ After 7 days, the number of bacteria 2.2 x 10 ⁶ 3.8 x 10 ⁶ 4.1 x 10 ⁶ 3.2 x 10 ⁵ 1.6 x 10 ⁵ After 14 days, the number of bacteria 1.6 x 10 ⁶ 2.0 x 10 ⁶ 2.1 x 10 ⁶ 4.7 x 10 ⁵ 3.5 x 10 ⁵ After 28 days, the number of bacteria 3.4 x 10 ⁶ 3.7 x 10 ⁶ 5.1 x 10 ⁶ 1.6 x 10 ⁵ 2.8 x 10 ⁵

(Unit: CFU / ml)

In addition, as shown in Table 3, in the case of the hot-water extract of vitamin Nutrition, E. coli , S. aureus , P. aeruginosa and fungal yeast ( C. albicans ) And A. niger were found to be low in antibacterial and antiseptic properties.

Example  2: Vitamin tree  &Lt; / RTI &gt;

Vitamin tree extracts were prepared from the ethanol extracts of vitamin A nutrients prepared according to Examples 1-3.

And, it was purchased as medicinally sold as root shell with peony (Paeoniaceae, Paeonia suffruticosa). The sheath was completely dried, pulverized and passed through a 40 mesh net (1.18 mm mesh). 350 ml of distilled water was added to 30 g of wood pulp powder, followed by shaking incubation at 60 DEG C for 24 hours, followed by centrifugation at 6,000 rpm for 1 hour. The resulting extract was filtered through 0.45 占 퐉 and lyophilized to obtain a herbal extract powder.

Each of the extracts thus prepared was mixed at different ratios (1: 0.1, 1: 1.0, 1: 2.0) to obtain a mixture of vitamins and mulberry leaves Mixed extracts were prepared.

Comparative Example  One: Vitamin tree  And licorice mixed extract

A mixed extract of vitamin tree and licorice was prepared in the same manner as in Example 2, except that Licorice was used instead of Mulberry in Example 2.

Comparative Example  2: Vitamin tree  And Black garlic  Preparation of mixed extracts

A mixed extract of vitamin wood and black garlic was prepared in the same manner as in Example 2, except that black garlic was used instead of mulberry in Example 2.

Experimental Example  2 : Vitamin tree  Antibacterial test for mixed extracts

The mixed extracts prepared according to Example 2 and Comparative Examples 1 and 2 were subjected to an antibacterial test. The antibacterial activity test was carried out in the same manner as in Experimental Example 1-5, and the test strains used were the same.

The results are shown in Table 4 below. Table 4 shows the number of bacteria at the initial inoculation and the number of bacteria after 14 days.

division E. coli S. aureus P. aeruginosa C. albicans A. niger Initial inoculation (0 day) 3.6 x 10 ⁶ 4.7 x 10 ⁶ 2.9 x 10 ⁶ 1.8 x 10 ⁵ 4.5 x 10 ⁵ Example 2-1 (1: 0.1) <10 <1 <1 <10 1.2 x 10 ³ Example 2-2 (1: 1.0) <1 <1 <1 <10 <10 Example 2-3 (1: 2.0) <1 <1 <1 <10 <10 Comparative Example 1 <10 <10 <10 1.1 x 10 ³ 2.7 x 10 ⁵ Comparative Example 2 <10 <10 <10 2.5 x 10 ³ 1.3 x 10 ⁵

(Unit: CFU / ml)

As shown in Table 4, it was confirmed that the number of bacilli was significantly reduced after 14 days compared with Comparative Examples 1 and 2 in the vitamin tree and Mulberry extract according to the present example.

Particularly, in Examples 2-2 and 2-3, it was found that they have high antimicrobial activity against fungal yeast ( C. albicans ) and A. niger , which is considered to be an influence of the herbal extract.

While the present invention has been particularly shown and described with reference to preferred embodiments thereof, it is to be understood that the invention is not limited to the disclosed embodiments, but, on the contrary, It will be apparent to those skilled in the art that changes may be made.

Claims (6)

delete delete delete 70% ethanol extract of vitamin Nuts; And a herbal extract,
Wherein the herbal extract comprises 0.5 to 1.5 parts by weight of the extract of vitamin A nutrient with respect to 1 part by weight of the ethanol extract. delete delete

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