KR101602552B1 - Antimicrobial composition comprising the extract of cornus officinalis - Google Patents

Abstract

The present invention relates to an antimicrobial composition comprising a corn oil extract, and more particularly to an antimicrobial and antifungal composition comprising a hydrothermal or ethanol extract of corn oil and an organic solvent fraction thereof. The extract of marine oil of the present invention exhibits an excellent antibacterial activity and is useful as an antibacterial and / or antifungal pharmaceutical composition, a health functional food, a cosmetic composition, an animal feed composition, food, cosmetic or animal food additive, There is an excellent effect that can be used. In addition, the acidic milk extract and fractions of the present invention are excellent in thermal stability and can be easily processed into various forms such as liquid, cream, powder, ring, and tablet without showing loss of activity even in an acidic condition of pH 2 and plasma It can be very useful in pharmaceutical industry, food industry, animal husbandry industry, health industry and cosmetics industry.

Description

ANTIMICROBIAL COMPOSITION COMPRISING THE EXTRACT OF CORNUS OFFICINALIS.

The present invention relates to an antimicrobial composition comprising a corn oil extract, and more particularly to an antimicrobial and antifungal composition comprising a hydrothermal or ethanol extract of corn oil and an organic solvent fraction thereof.

According to the U.S. Center for Disease Control and Prevention, bloodstream infections in hospitals are one of the leading causes of death in the United States. Infections of pathogenic bacteria and fungi through skin and soft tissues are also recognized as universal medical diseases and are mainly caused by trauma or surgery. Many pathogenic bacterial and fungal infections are treated differently depending on the severity of the infection, but successful treatment is possible with oral antimicrobials and topical cleansing. However, the more severe or complex infections that are common in patients with underlying risk factors (e. G., Vascular flow disorders, diabetes) and / or infections caused by difficult or multi-resistant bacilli and fungi, Treatment with antimicrobial agents and removal of invasive surgical tissue may be required. For example, vancomycin, a glycopeptide antibiotic, has been successfully used for severe intra-hospital infections caused by multi-drug-resistant gram-positive bacteria, particularly MRSA, coagulase-negative staphylococcus and enterococcus. Recently, it has been reported that the isolation of strains resistant to vancomycin is increasing, and the current antibiotics are very limited in the treatment of severe infections caused by some of the pathogens. Therefore, there is a growing need for more effective new antibiotics for resistant bacteria.

Meanwhile, Cornus officinalis ) is a medicinal plant belonging to the Comaceae family. Its fruit is in the shape of a central circle around 1.5cm in length. It is also known as a succulent herb, which is prominent in sour taste, and is widely used as a natural food having a nourishing tonic effect. Most of the commercially available products of corn oil, which are currently sold in the market, are dried products with steamed corn oil removed from the seeds, extract liquid beverages with corn oil and corn oil with powder of corn oil, and corn oil seeds and leaves have been used as food It is not registered as a raw material, and there is a restriction to use it as a food.

Therefore, in the case of corn oil, the components and efficacy researches have been conducted mainly on the pulp. It is known that iridoid glycosides such as moronic acid and roganin are known, and the effects known in the private sector include diuretic action, blood pressure strengthening action, antibacterial action, liver protection In one room, the kidneys are seen in the kidneys and the convergent medicine, sweating frequently, and when the urine frequently comes out often. However, up to now, the antidiabetic effects of corn oil seeds (Park, Yeong-gyung, 1995, Chuna-ang J. Pharm (1995) (1995), pp. 219-222). Inhibition of glycation endogenesis in seeds and seeds of marine algae (Lee, G. Y et al., 2008, J. Kor. Soc. 320), but no other biologic activity other than antidiabetic activity is known. In particular, no antimicrobial activity has been reported for the seeds or leaves of corn oil.

As a patent related to corn oil, it is disclosed in Korean Patent Registration No. 10-1358203 (2014-01-27) [composition for preventing or treating thrombosis containing crude oil extract], Registration No. 10-1330687 (2013-11-12 ) As a composition for prevention and treatment of obesity comprising an extract of a medicinal herb as an active ingredient, Registration No. 10-1330688 (2013-11-12) [containing an extract of a medicinal herb as an active ingredient for prevention of diabetes and diabetes As a composition for prevention and treatment], Registration No. 10-0928711 (2009-11-19), [Method for preparing a preserved food based on marine oil], Registration No. 10-0438417 (2004-06-22) [Method for producing fermented acid fermented milk with increased antioxidant activity] and the like have been disclosed as a method for producing health food based on corn oil, and Registration No. 10-1319984 (2013-10-14). However, Leaf-related patents are not known to date.

Accordingly, the present inventors have found strong antimicrobial activity in extracts and fractions of corn oil seeds or leaves while searching for natural materials having antimicrobial activity that can be used for various purposes such as pharmaceutical materials, health functional foods and cosmetics. Accordingly, the present inventors prepared sequential organic solvent fractions after preparing seed oil or leaf extracts with various solvents, and confirmed the strong antibacterial activity of these extracts and fractions, thereby completing the present invention.

KR 10-1358203 B

Accordingly, an object to be solved by the present invention is to provide an antimicrobial composition comprising an antimicrobial active substance extracted from marine oil, in particular, a seed or leaf of marine oil, followed by separation and purification, as an effective ingredient.

In order to solve the above problems, the present invention provides an antimicrobial composition comprising Cornus officinalis extract as an active ingredient.

It is preferable that the above-mentioned extract is a seed or leaf extract of marine oil.

The seed extract of corn oil is preferably an ethanol extract of corn oil seed.

It is preferable that the ethanol extract of the marine oil seeds is at least one of ethyl acetate fraction, butanol fraction and water residue obtained by successively fractionating the ethanol extract with hexene, ethyl acetate and butanol.

Preferably, the seed extract of corn oil is a hydrothermal extract of corn oil seed.

Preferably, the hot-water extract of the marine oil seeds is at least one of hexane fraction, ethyl acetate fraction, butanol fraction and water residue obtained by successively fractionating the hot-water extract with hexene, ethyl acetate and butanol.

The leaf extract of corn oil is preferably an ethanol extract of corn oil.

It is preferable that the ethanol extract of the acidophilus leaves is at least one of hexane fraction, ethyl acetate fraction, butanol fraction and water residue obtained by successively fractionating the ethanol extract with hexene, ethyl acetate and butanol.

It is preferable that the leaf extract of the acidified milk is at least one of a hexane fraction, an ethyl acetate fraction and a butanol fraction obtained by successively fractionating a hot-water extract of a corn oil leaf with hexene, ethyl acetate and butanol.

It is preferable that the antibacterial agent is an antibacterial agent or an antifungal agent.

The antimicrobial composition is preferably selected from the group consisting of a pharmaceutical composition, a health functional food, a cosmetic composition, an animal feed composition, a food additive, a cosmetic additive, an animal feed additive, a detergent and a cleanser.

The extract of mushroom oil of the present invention exhibits excellent antimicrobial activity and is useful as an antimicrobial and / or antifungal pharmaceutical composition, a health functional food, a cosmetic composition, an animal feed composition, food, cosmetic or animal feed additives, various detergents, There is an excellent effect that can be used for applications. In addition, the acidic milk extract and fractions of the present invention are excellent in thermal stability and can be easily processed into various forms such as liquid, cream, powder, ring, and tablet without showing loss of activity even in an acidic condition of pH 2 and plasma It can be very useful in pharmaceutical industry, food industry, animal husbandry industry, health industry and cosmetics industry.

Hereinafter, the present invention will be described in detail.

The present invention relates to a composition exhibiting a strong antimicrobial activity containing a corn oil extract. The inventors of the present invention have found out that the extract of corn oil, especially the marine oil seeds or leaves, is extracted with various solvents such as water, alcohol, ethanol and methanol, and then the organic solvent fraction and the water residue are sequentially prepared with hexene, ethyl acetate and butanol, A strong antibacterial activity was confirmed.

Particularly, hydrothermal and ethanol extracts and their fractions of corn oil seeds showed broad antibacterial activity against Gram positive bacteria, Gram negative bacteria and fungi, and the ethanol extracts and fractions thereof from the leaves of corn oil and the fractions of hydrothermal extract Positive bacterium and gram-negative bacterium, and can be developed as a broad-spectrum antimicrobial agent. Thus, the present invention has been completed.

Accordingly, the present invention provides an antimicrobial composition comprising an extract of corn oil as an active ingredient. The composition may be applied to a pharmaceutical composition, a health functional food, a cosmetic composition, an animal feed composition, a food, a cosmetic or an animal feed additive, a detergent, and various cleaning agents.

In a preferred embodiment, the present invention relates to a method for producing a fermented soybean oil comprising the steps of: Purifying the active ingredient through sequential organic solvent fractions of marine oil seed or leaf extract; And a step of evaluating the hemolysin activity of the substance in the human erythrocyte.

"Corn oil seed extract" contained in the composition of the present invention refers to a seed obtained by extracting seeds of corn oil, and seeds of corn oil include seeds of the entire maturity stage and are obtained after physically removing pulp from the fruit of mature corn oil . The obtained crude oil seeds may be used as is or pulverized powder may be used. Preferably, the powder obtained by pulverizing 100% or less of the vegetable oil seeds is used. In addition, corn oil seeds from which flesh is removed may be heat treated before extraction.

The seed oil containing the antimicrobial active substance of the present invention is not registered as a raw material for food in the domestic foodstuffs industry and contains lectin as a red blood cell coagulation factor and is known to be toxic to the public. At present, corn oil seeds are almost discarded because they have no special purpose, and antimicrobial compositions can be produced economically from corn oil seeds.

In addition, the "acidophylla leaf extract" contained in the composition of the present invention means that the matured leaf of the matured oil is extracted, the leaves of the matured oil contain the leaves of the entire mature stage, and the mature matured fruit oil is recovered before and after the harvest , And does not affect the harvesting of corn oil. As a preferred specific example, the leaves of the above-mentioned leaves may be collected by washing with water, removing the foreign substances, shredding them, or grinding them. When pulverized powder is used, it is preferable to use pulverized powder to 100 mesh or less. In addition, shrubs of safflower leaves may be subjected to a heat treatment such as a fruiting process.

The leaves of corn oil containing the antimicrobial active substance of the present invention are not registered as raw materials for food in the domestic food industry, but there is an example in the private sector where drinking water is concerned about the endless water. At present, the leaves of corn oil are almost not used because they are not used separately, and the antimicrobial composition can be produced economically from the corn oil leaves.

In the present invention, "corn oil seed extract" or "corn oil leaf extract" may be prepared using various extraction solvents. For example, water (cold water, hot water), alcohol, water, or a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, alcohol, propanol, butanol, etc.) can do. Preferably, the extraction is carried out using ethanol or alcohol, and the yield of the active material is the maximum. In the case of economic extraction, it is preferable to use hot water. The corn oil seed or leaf extract of the present invention can be made into powder by a conventional powdering process such as vacuum distillation and freeze drying or spray drying. They maintain their activity even at 100 ° C heat treatment and pH 2 in human body.

The hot-water extract and the ethanol extract and the sequential fraction thereof of the seed oil or seed of the present invention exhibit a strong antimicrobial activity and are useful as a pharmaceutical composition, a health functional food, a cosmetic composition, an animal feed or the like for preventing or treating infection of pathogenic bacteria and fungi Compositions, foods, cosmetics, animal feed additives, and various cleaning agents.

In a preferred embodiment, the antimicrobial composition of the present invention can be applied as a pharmaceutical composition.

The pharmaceutical composition may be formulated into various forms such as powders, granules, tablets, capsules, oral formulations such as suspensions, emulsions, syrups and aerosols, injections of sterilized injection solutions, etc. And can be administered via various routes including oral administration or intravenous, intraperitoneal, subcutaneous, rectal, topical administration, and the like. Such pharmaceutical compositions may further comprise carriers, excipients or diluents, and examples of suitable carriers, excipients or diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, But are not limited to, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, amorphous cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, And the like. In addition, the pharmaceutical composition of the present invention may further include a filler, an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, an antiseptic, and the like.

The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the type of disease, severity, The sensitivity to the drug, the time of administration, the route of administration and the rate of release, the duration of the treatment, factors including co-administered drugs, and other factors well known in the medical arts. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

The effective amount of the component having antimicrobial activity in the pharmaceutical composition of the present invention may vary depending on the age, sex and body weight of the patient, and is generally from 1 to 5,000 mg, preferably from 100 to 3,000 mg per body weight, Or one to three times a day. However, the dosage may not be limited in any way because it may be increased or decreased depending on route of administration, severity of disease, sex, weight, age, and the like. The pharmaceutical composition of the present invention can be administered to a subject through various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections. In the present invention, "administration" means providing a predetermined substance to a patient by any suitable method, and the administration route of the pharmaceutical composition of the present invention is either oral or non-oral May be administered orally. The composition of the present invention may also be administered using any device capable of delivering an effective ingredient to a target cell. In the present invention, the term "object" includes, but is not limited to, human, monkey, cow, horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit or guinea pig , Preferably a mammal, more preferably a human.

In a preferred embodiment, the antimicrobial composition of the present invention can be applied as a health functional food.

The health functional food of the present invention can be used variously for effective foods and beverages which can lower the possibility of infection from a subject having a high risk of infection by a pathogenic microorganism. Examples of food containing the active ingredient having excellent antimicrobial activity of the present invention include various foods, beverages, gums, tea, vitamin complexes, health supplement foods and the like, and may be in the form of powders, granules, tablets, capsules or beverages . The active ingredient of the present invention may generally be added in an amount of 0.01 to 15% by weight of the total food, and the health beverage composition may be added in a proportion of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml. The health functional food of the present invention may contain, as an additional ingredient, a food-acceptable food-aid additive such as natural carbohydrates and various flavors, in addition to containing the above-mentioned compound as an essential ingredient in the indicated ratio. Examples of the natural carbohydrate include sugar sugars such as glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of the flavor agent include tau martin, rebaudioside A, glycyrrhizin, saccharin, and aspartame. The proportion of the above-mentioned flavoring agent is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the health functional food of the present invention. In addition to the above, the health functional food of the present invention may contain various kinds of nutrients, vitamins, minerals, flavors such as synthetic flavors and natural flavors, colorants and heavy stabilizers, pectic acid and its salts, alginic acid and its salts, Thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the health functional food of the present invention may contain flesh for producing natural fruit juice, fruit juice drink, vegetable drink and the like. These components may be used independently or in combination. The ratio of such additives is generally selected in the range of 0.01 to about 20 parts by weight per 100 parts by weight of the active fraction of the present invention.

In a preferred embodiment, the antimicrobial composition of the present invention can be applied as a cosmetic composition.

The cosmetic composition of the present invention can be manufactured in the form of an ingredient having excellent antimicrobial activity of the present invention in the form of an essence, a nutritional cream, a body lotion, a rinse, a shampoo, a body cleanser, a lotion, a skin, . The preparation in this form may be easily carried out according to various production processes known to those skilled in the art.

In a preferred embodiment, the antimicrobial composition of the present invention can be applied as an animal feed composition.

The animal is an organism corresponding to a plant and mainly consumes organic matter as nutrients and refers to the digestion, reproduction and differentiation of the respiratory organs. Specifically, the animal is an animal such as an echinoderm, crustacean, mollusk, fish, amphibian, reptile, Mammals and preferably molluscs such as echinoderms such as echinoderms or sea cucumbers, arthropods including crustaceans such as crabs, shrimps and prawns, cephalopods, gastropods or bivalves such as sea buckets, sea breams, codfish, , Flounder or the like, poultry such as pheasant or chicken, or a mammal such as pig, cattle, or goat.

The composition for animal feed can further comprise cereals, vegetable protein feeds, animal protein feeds, sugar or dairy products in the antimicrobial composition comprising the antimicrobial component of the present invention. The cereal may be specifically wheat, crushed or crushed wheat, oats, barley, corn and rice, and the vegetable protein feed may be one mainly composed of rapeseed, bean and sunflower, and the animal protein feed is specifically May be blood, meat, bone meal, and fish meal, and the sugar or dairy product may specifically be a dry ingredient consisting of various powdered milk and whey powders.

The animal feed composition may further contain ingredients such as nutritional supplements, digestion and absorption enhancers, sex-promoting agents or disease prevention agents.

The antimicrobial composition contained in the composition for animal feed of the present invention may be varied depending on the intended use and the conditions of use of the feed. For example, 0.1 to 100 g of the antimicrobial composition may be contained based on 1 kg of the final feed.

In addition, the feed composition can be made into a light-viscous granulated or granulated material depending on the degree of comminution of the components, which can be fed into a mesh or formed into a desired discrete shape for further processing and packaging, Pelletizing, expanding or extruding processes, and preferably excess water can be dried off for ease of storage.

In a preferred embodiment, the antimicrobial composition of the present invention can be applied as an additive for foods, cosmetics or animal feeds.

The additive is added to impart preservability to food, cosmetics or animal feed used. Since the antimicrobial composition is a natural ingredient and is not harmful to human body, it can be safely applied to foods, cosmetics or animal feeds.

The additive may further include other optional additives selected depending on the purpose and use of the additive, and may be mixed with other active ingredients, such as commonly used additives, or additives such as pigments, surfactants, and preservatives . The additive may be prepared by pulverizing, granulating, tableting or liquefying according to the purpose and use of the additive. For commercialization, a conventional method may be used for packaging.

The amount of the antimicrobial composition to be used may be appropriately used depending on the purpose of use, the application form, the application site, the object to be applied, and the like. For example, the antimicrobial composition may contain 0.01 to 50 parts by weight, The content of the composition is not limited thereto. Since the antimicrobial composition of the present invention is a natural component and is not harmful to human body, it can be used in various amounts as long as the application purpose can be achieved due to the characteristics of the product.

In a preferred embodiment, the present invention can be applied as an application of a cleaning agent.

The detergent may preferably be a detergent for clothes, a dishwasher, a food cleaner, or a household appliance cleaner. The antimicrobial composition is a natural ingredient and is not harmful to human body. Therefore, it is more preferably used for the purpose of washing or antibacterial property of various daily necessities including kitchen utensil or food washing.

The detergent may further contain an adversely selected additive depending on the purpose and use of the detergent. The detergent may be mixed with other active ingredients, such as detergents, and additives such as pigments, surfactants and preservatives. The detergent may be prepared by pulverizing, granulating, tableting or liquefying according to the purpose and use of the detergent, and a conventional method may be used for packaging for commercialization.

The amount of the antimicrobial composition to be used in the cleaning agent may be appropriately used depending on the purpose of use, the application form, the application site, the article to be applied, and the like. For example, the antimicrobial composition may contain 0.01 to 50 parts by weight, The content of the composition is not limited thereto. Since the antimicrobial composition of the present invention is a natural ingredient and is not harmful to human body, it can be used in various amounts as long as its application purpose can be achieved.

As a preferred embodiment, the present invention can be applied as an antibacterial composition for use as a cleansing agent.

The cleaner can be used for hand, oral cavity or ladies' cleanser in a simple manner to prevent infection of a pathogenic microorganism which is easily infected in daily life. Since the antimicrobial composition is a natural ingredient and is not harmful to the human body, it can be safely applied even when it is used as a hand, mouth, and female cleanser.

The cleansing agent may further include other additives optionally selected depending on the purpose and use of the cleansing agent. The cleansing agent may be mixed with other active ingredients, such as commonly used additives, and additives such as coloring matters, surfactants and preservatives . The cleansing agent may be prepared by pulverization, granulation, tableting or liquefying according to the purpose and use of the cleansing agent.

In addition, the amount of the antimicrobial composition to be used in the cleaner may be suitably used according to the purpose of use, application form, application site, and the like. For example, the amount of the antimicrobial composition may be 0.01 to 50 parts by weight, But is not limited thereto. Since the antimicrobial composition of the present invention is a natural component and is not harmful to human body, it can be used in various amounts as long as the application purpose can be achieved due to the characteristics of the product.

Hereinafter, the present invention will be described in more detail with reference to Examples.

< Example >

Example  One

1. Various solvent extraction of corn oil seeds

After separating the seeds from the cornstarch oil harvested in December 2011 in Gyeongbuk province, the pulp was washed with running water to remove the pulp, shredded for 7 days, and crushed into 100 mesh. The extraction solvent was hexane, ethanol (95%), methanol (70%), hot water (70 ℃ and 100 ℃), and 10 times of organic solvent was added to the weight of the crushed seeds. Or 10 times of hot water, and then extracted at 70 ° C or 100 ° C for 30 minutes. The extract was concentrated under reduced pressure to give a powder, which was stored at low temperature until use. Total flavonoid and total polyphenol contents, total sugar and reducing sugar contents of the prepared extracts were measured. As the standard of total flavonoid content, rutin and tannic acid were used as the standard of total polyphenol content. Respectively. Reducing sugar was determined by DNS method and total sugar was quantified by phenol-sulfuric acid method. The extraction efficiency and component analysis results of each extract are shown in Table 1, and the highest yield was obtained in ethanol extraction, but also in hot water extraction. The most significant difference was found in polyphenol contents, and ethanol and methanol extracts showed higher contents of polyphenols than hot water extracts. However, the content of flavonoids was 14.31 ~ 20.06 mg / g except for the extraction with hexene. Considering that the antimicrobial activity of natural products is generally attributed to polyphenols and flavonoids, it was concluded that the high content of polyphenols and flavonoids in corn oil seeds might be associated with strong antimicrobial activity.

Extraction solvent Extraction rate (%) Content (mg / g) Total flavonoid Total polyphenol  Total Reducing sugar Hexen 4.84 1.16 ± 0.11 1.84 + 0.07 40.42 + - 0.83 15.49 + - 0.31 Ethanol (95%) 17.64 20.06 ± 0.22 858.14 + - 3.31 110.01 + - 1.27 92.82 + - 7.47 Methanol (70%) 16.66 14.31 + - 0.72 820.68 + - 5.52 125.34 + - 2.68 108.46 ± 5.19 Hot water (70 ℃) 7.84 15.02 + - 0.14 666.94 ± 0.00 182.90 ± 3.80 163.57 ± 3.20 Hot water (100 ℃) 16.53 14.61 ± 0.58 263.62 + - 2.76 387.75 ± 2.82 274.81 + - 3.05

The ethanol extracts exhibiting the highest extraction efficiency and the hot water (100 ° C) extracts capable of economical extraction were subjected to sequential organic solvent fractionation, and the results are shown in Table 2.

extract Fraction Fraction
(%) Content (mg / g) Total flavonoid Total polyphenol  Total Reducing sugar Ethanol (95%)
extract Hexen 0.1 1.74 ± 0.22 13.60 ± 2.87 27.66 + - 0.84 18.94 ± 2.14 Ethyl acetate 28.2 22.35 + 0.14 673.97 + - 0.10 61.62 + 4.65 54.96 ± 7.63 Butanol 42.1 15.28 ± 0.50 826.15 + - 6.62 113.10 ± 0.56 91.20 ± 5.49 Water residue 26.7 12.73 + - 0.36 528.81 ± 0.00 325.51 + - 0.70 243.53 + - 6.86 Heat number
(100 DEG C)
extract Hexen 10.9 21.84 + 1.44 590.46 + - 7.73 155.12 ± 0.28 87.14 + - 0.76 Ethyl acetate 32.8 17.62 ± 0.79 800.39 + - 3.31 92.39 ± 3.10 74.91 + - 3.81 Butanol 33.7 15.28 ± 0.79 590.46 ± 1.10 112.10 ± 0.28 95.41 + - 1.37 Water residue 22.0 11.05 ± 0.00 175.28 ± 3.31 400.19 + - 1.41 323.34 + - 3.05

The ethyl acetate and butanol fractions were similar to the extracts, but the hexane fractions were 10.9%. The ethyl acetate and butanol fractions were 70.3% and 0.1%, respectively. Respectively. The amount of water residue after fractionation of the organic solvent of the extract was 22.0 ~ 28.2%. The contents of polyphenols and flavonoids in the fractions were as follows: butanol fraction> ethyl acetate fraction> water residue> hexane fraction in the ethanol extract, ethyl acetate fraction, butanol and hexane fraction> Respectively.

2. Corn oil seed extract and Fraction  Antimicrobial activity

Escherichia coli KCTC 1682, Proteus vulgaris KCTC 2433 and Salmonella typhimurium KCTC 1926 were used as Gram-negative bacteria. Staphylococcus aureus KCTC 1916, Listeria monocytogenes KACC 10550, Bacillus spp. subtilis KCTC 1924 was used. Saccharomyces cerevisiae IF0 0233 and Candida albicans KCTC 1940, the causative agent of fungus infections, were used for evaluation of antifungal activity.

First, the case wherein the bacterial activity evaluation, Nutrient broth (Difco Co., USA) and then inoculated to each of the bacteria were cultured at 37 ℃ for 24 hours, by adjusting the respective strain to OD ₆₀₀ 0.1 Nutrient agar (Difco Co. (Diameter 6.5mm, Whatsman No. 2) was added to each well of a sterilized petri dish (9015 mm, Green Cross, Korea) containing the medium . In the case of fungi, Sabouraud dextrose (Difco Co., USA) was cultured in the same manner at 30 ° C for 24 hours, and the size of the growth inhibition rings was measured to evaluate the antimicrobial activity. As control, ampicillin, an antimicrobial agent, and miconazole (Sigma Co., USA), an antifungal agent, were used at a concentration of 1 g / disc, respectively. The size of the growth inhibition ring was determined by the naked eye mm, and is shown in Table 3 as representative results after three or more evaluations.

microbe Growth inhibition ring (mm) Antibiotic
(1 g / disc) Corn oil seed ethanol extract
(500 μg / disc) Corn oil seed hot water extract
(500 μg / disc) MP
Syllable Myconazole Ethanol extract Hexene fraction ethyl
Acetate fraction Butanol fraction water
Residue Hot water extract Hexene fraction ethyl
Acetate fraction Butanol fraction water
Residue Gram negative
Germ EC 10.0 - 10.0 - 12.0 14.0 12.0 9.0 12.0 15.0 13.0 12.0 PV 36.0 - 18.0 - 14.0 20.0 16.0 15.0 16.0 26.0 16.0 18.0 ST 17.0 - 10.0 - 13.0 13.0 10.0 11.0 9.5 10.0 12.0 7.0 Gram positive
Germ SA 28.0 - 14.0 - 18.0 18.0 15.0 12.0 16.0 20.0 18.0 14.0 LM 21.0 - 9.0 - 18.0 14.0 15.0 9.0 17.0 19.0 19.0 12.0 BS 25.5 - 15.0 - 14.0 10.0 10.0 11.0 11.0 12.0 11.0 9.0 Fungus SC - 26.0 10.0 - 16.0 18.0 13.0 10.0 16.0 16.0 17.0 12.0 CA - 28.0 15.0 - 17.0 14.5 18.0 16.0 17.0 16.0 14.0 10.0

EC : Escherichia coli ,

PV : Proteus vulgaris ,

ST : Salmonella typhimurium ,

SA : Staphylococcus aureus ,

LM : Listeria monocytogenes ,

BS : Bacillus subtilis ,

SC : Saccharomyces cerevisiae ,

CA : Candida albicans .

-: No growth inhibition.

As shown in the above Table 3, the ampicillin (1 g / disc) used as an anti-bacterial activity control exhibited a strong inhibitory activity of 10.0-36.0 mm, indicating that the antifungal agent myconezol (1 g / disc ) Showed growth inhibition ring of 28mm against C. albicans . In the case of the extract of corn oil, the ethanol extract (500 g / disc) and its fractions ethyl acetate fraction, butanol fraction, water residue and water extract of the marine oil seeds and all fractions were found to have strong antibacterial and antimicrobial activity And it was found that corn oil seeds contain various antimicrobial active substances. Especially, the hexane fraction of the ethanol extract does not exhibit any antibacterial activity. Therefore, when the extract is prepared after the hexene treatment of the crude oil seeds, or when the active substance in which the hexane-soluble substance is removed after the preparation of the ethanol extract is used, it can be used as an economical broad-spectrum antibiotic . The above results suggest that the extract of corn oil seeds and the active fractions can be effectively used to control foodborne pathogens and pathogenic bacteria.

3. Corn oil seed extract and Fraction  Human red blood cells Hemolytic activity

Table 4 shows the results of evaluating the erythrocyte hemolytic activity of the seed oil samples. First, DMSO and distilled water used as a control had no hemolytic activity, and triton X-100 was found to hemolyze red blood cells at a concentration of 0.1%. At the concentration of 0.5 mg / ml, only 33% or more of hemolytic activity was observed in the ethanol extract and its ethyl acetate fraction in the crude oil seed samples, indicating potential toxicity. Other samples showed no hemolytic activity.

division Sample / Control Human hemolytic hemolytic activity (%) Corn seed oil
Ethanol extract
(0.5 mg / ml) Ethanol extract 33.0 Hexene fraction -2.8 Ethyl acetate fraction 34.0 Butanol fraction 2.8 Water residue 0.9 Corn seed oil
Hot water extract
(0.5 mg / ml) Ethanol extract 3.7 Hexene fraction -4.4 Ethyl acetate fraction 7.6 Butanol fraction 3.6 Water residue -1.7 Control Distilled water 0.0 Control The solvent (DMSO) 1.1 Control Triton X-100 100

When the above results are summarized, the ethanol extract and hot water extract of corn oil seeds showed strong antioxidative activity, and the organic solvent fraction of these extracts showed a very strong antioxidative activity in all fractions except the hexane fraction of the ethanol extract , And various solvent extracts and fractions of marine oil seeds have excellent effects that can be used for prevention and treatment of oxidative stress. The composition containing the same is useful for the prevention and treatment of oxidative stress related diseases in the cosmetics industry, food industry and pharmaceutical industry Can be used as material.

4. Stability of seed oil seed active material

The crude extracts of the crude oil seed oil and hot-water extract of the above-mentioned Example were treated at 100 ° C for 1 hour, 2 hours at pH 2 (0.01M HCl), and 2 hours at the plasma, respectively. , It was confirmed that the antioxidant active ingredients of the extract showed high stability in acid, heat, and plasma treatment (no result).

Example  2

1. Various solvent extraction of corn oil leaves

In October ~ November 2011, leaves of corn oil were collected from Gyeongsangbuk-do, Gyeongsangbuk-do Province, and the water was washed with water to remove foreign matter. After shaking for 7 days, it was crushed into 100 mesh and used for extraction. The extraction solvent used was hot water and ethanol (95%). After 10 times as much organic solvent as the weight of the crushed leaf, it was extracted 3 times at room temperature for 24 hours or 10 times of hot water, Respectively. The extract was concentrated under reduced pressure to give a powder, which was stored at low temperature until use. Total flavonoid, total polyphenol, total sugar, and reducing sugar content of the prepared extracts were measured. As the standard of total flavonoid content, rutin and tannic acid were used as the standard of total polyphenol content. Respectively. Reducing sugar was determined by DNS method and total sugar was quantified by phenol-sulfuric acid method. The extraction efficiency and component analysis results of each extract are shown in Table 5. The extraction efficiency of hot water extraction was about twice as high as that of ethanol extraction. In the case of the fraction of the hot water extract, the water residue after the fractionation of organic solvent accounted for 47.9% of the extract, whereas the fraction of the ethanol extract accounted for 13.0% of the water residue after the fractionation of the organic solvent, I could see the difference. The major difference in the extracts was found in the content of polyphenol, and the content of hot - water extract was 2.2 times higher than that of ethanol extract. In the case of the ethyl acetate fraction of the hot water extract, fractional yields were as low as 7.3%, while very high polyphenol and flavonoid contents of 489 mg / g and 60.4 mg / g were obtained. Ethyl acetate> butanol> water residues> Respectively. In the case of polyphenol, the water residue (178.95 mg / g) after the solvent fractionation of the ethanol extract was higher than that of ethyl acetate (129.78 mg / g) in the ethyl acetate fraction, Respectively. Therefore, in the case of polyphenol content, ethyl acetate> butanol> hexene fraction in order of water residues, ethyl acetate> butanol> water residue> hexene fraction in order of flavonoid content. These results indicate that the acidophilus leaves contain a high concentration of water-soluble polyphenols and a large amount of fat-soluble polyphenols. Considering that the antimicrobial activity of natural products is generally attributed to polyphenols and flavonoids, very high levels of polyphenols and flavonoids in the leaves of corn oil were thought to be associated with strong antimicrobial activity.

extraction
menstruum Extract / fraction Extraction rate /
Fraction (%) Content (mg / g) Total flavonoid Total polyphenol  Total Reducing sugar Ethanol (95%) Ethanol extract 13.7 32.87 ± 1.19 215.63 ± 10.98 218.34 ± 12.02 208.40 + - 0.55 Hexen 14.0 4.23 + - 0.43 11.81 ± 1.05 104.04 + 0.14 20.77 ± 1.68 Ethyl acetate 35.0 60.41 + - 2.30 489.94 + - 5.52 178.32 + - 1.27 78.80 +/- 10.83 Butanol 37.0 33.34 + - 0.14 199.24 + - 4.97 501.26 ± 2.82 491.59 ± 2.29 Water residue 13.0 17.67 ± 0.29 37.44 ± 0.28 154.72 ± 0.28 118.49 + - 6.71 Heat number
(100 DEG C) Hot water extract 26.8 33.54 1.51 98.22 + - 2.55 317.32 + - 5.31 275.93 + - 1.49 Hexen 1.0 2.96 ± 0.22 25.15 + - 2.70 141.37 + - 8.17 42.88 + - 4.27 Ethyl acetate 7.3 36.59 ± 1.01 129.78 + - 4.97 418.61 + - 4.20 64.02 + - 5.19 Butanol 42.0 15.28 ± 0.65 93.10 ± 4.97 693.93 + - 3.52 583.26 ± 7.63 Water residue 47.9 13.39 ± 0.86 178.95 ± 3.86 585.89 + - 4.22 499.14 + - 3.81

2. Corn oil leaf extract and Fraction  Antimicrobial activity

Escherichia coli KCTC 1682, Proteus vulgaris KCTC 2433 and Salmonella typhimurium KCTC 1926 were used as Gram negative bacteria. Staphylococcus aureus KCTC 1916 and Listeria monocytogenes KACC 10550 were used as Gram positive bacteria. , Bacillus subtilis KCTC 1924 were used. Saccharomyces cerevisiae IF0 0233 and Candida albicans KCTC 1940, the causative agent of fungus infections, were used for evaluation of antifungal activity.

First, the case wherein the bacterial activity evaluation, Nutrient broth (Difco Co., USA) and then inoculated to each of the bacteria were cultured at 37 ℃ for 24 hours, by adjusting the respective strain to OD ₆₀₀ 0.1 Nutrient agar (Difco Co. (Diameter 6.5mm, Whatsman No. 2) was added to each well of a sterilized petri dish (9015 mm, Green Cross, Korea) containing the medium . In the case of fungi, Sabouraud dextrose (Difco Co., USA) was cultured in the same manner at 30 ° C for 24 hours, and the size of the growth inhibition rings was measured to evaluate the antimicrobial activity. As control, ampicillin, an antimicrobial agent, and miconazole (Sigma Co., USA), an antifungal agent, were used at a concentration of 1 g / disc, respectively. The size of the growth inhibition ring was determined by the naked eye mm, and is shown in Table 6 as representative results after three or more evaluations.

microbe Growth inhibition ring (mm) Antibiotic
(1 g / disc) Corn oil leaves ethanol extract
(500 μg / disc) Hot water extract of corn oil leaves
(500 μg / disc) MP
Syllable Myconazole Ethanol extract Hexene fraction ethyl
Acetate fraction Butanol fraction water
Residue Hot water extract Hexene fraction ethyl
Acetate fraction Butanol fraction water
Residue Gram negative
Germ EC 10.0 - - 8.0 10.0 9.0 - - 8.5 11.0 - - PV 36.0 - - 14.0 12.0 10.0 13.0 - 8.0 13.0 - - ST 17.0 - - 10.0 8.0 12.0 - - 10.0 8.0 - - Gram positive
Germ SA 28.0 - 12.0 23.0 17.0 10.0 15.0 - 18.0 18.0 14.0 - LM 21.0 - 8.5 14.0 16.0 10.0 16.0 - 11.0 19.0 9.0 - BS 25.5 - 9.0 9.0 10.0 9.0 9.0 - 8.0 11.0 - - Fungus SC - 26.0 - - - - - - 8.0 - - - CA - 28.0 - - - - - - 8.0 - - -

EC : Escherichia coli ,

PV : Proteus vulgaris ,

ST : Salmonella typhimurium ,

SA : Staphylococcus aureus ,

LM : Listeria monocytogenes ,

BS : Bacillus subtilis ,

SC : Saccharomyces cerevisiae ,

CA : Candida albicans .

-: No growth inhibition.

As shown in Table 6, the strong antifungal activity of amphylline (1 g / disc) used as an anti-bacterial activity control was confirmed to be 10.0 ~ 36.0 mm, The growth inhibition ring was 28 mm against C. albicans . The ethanol extract (500 g / disc) showed antimicrobial activity only against Gram - positive bacteria, whereas the hydrothermal extract (500 g / disc) showed no antimicrobial activity against Gram - positive, negative bacteria and fungi. However, the fraction of ethanol extract showed broad antimicrobial activity against gram negative bacteria and gram positive bacteria in the hexene, ethyl acetate and butanol fractions except the water residue, and especially the hexene and ethyl acetate fractions showed excellent antimicrobial activity . On the other hand, hydrothermal extract did not show antimicrobial activity in the extract, but strong broad antimicrobial activity was confirmed in the hexene and ethyl acetate fractions. Therefore, it was confirmed that the hexane and ethyl acetate fractions of the ethanol extract and hot water extract of the corn oil leaves exhibit a strong antimicrobial activity, and that it can be developed as a composition for prevention and treatment of bacterial infection including these fractions.

3. Corn oil leaf extract and Fraction  Human red blood cells Hemolytic activity

Table 7 shows the results of evaluating the erythrocyte hemolytic activity of the extracts and fractions of corn oil. First, DMSO and distilled water used as a control had no hemolytic activity, and triton X-100 was found to hemolyze red blood cells at a concentration of 0.1%. At 0.5 mg / ml concentration, ethanol extract alone showed 17.2% hemolytic activity in leaves of corn oil, and other samples showed little hemolytic activity.

division Sample / Control Human hemolytic hemolytic activity (%) Leaves of corn oil
Ethanol extract
(0.5 mg / ml) Ethanol extract 17.2 Hexene fraction -3.6 Ethyl acetate fraction 6.8 Butanol fraction -2.1 Water residue -4.4 Leaves of corn oil
Hot water extract
(0.5 mg / ml) Ethanol extract -5.2 Hexene fraction -1.2 Ethyl acetate fraction -2.9 Butanol fraction -5.1 Water residue -5.4 Control Distilled water 0.0 Control The solvent (DMSO) 1.1 Control Triton X-100 100

Therefore, when the above results are summarized, the ethanol extract and hot water extract of corn oil leaves contain various antioxidant active substances. Particularly, ethanol and hot water extract of the corn oil leaves have side effects such as erythrocyte hemolytic activity And exhibits a strong antioxidative activity at the same time, thus showing an excellent effect that can be used for prevention and treatment of diseases related to oxidative stress.

4. Stability of Active Ingredients of Acanthopanax oleic Acid

The antioxidant activity of the extracts of the leaves of the leaves of corn oil and the ethylacetate fractions of the hot-water extract were measured at 100 ° C for 1 hour, 2 hours at pH 2 (0.01M HCl) and 2 hours at the plasma, It was confirmed that the antioxidant active ingredients of the extract showed high stability in acid, heat, and plasma treatment (no result).

Claims (11)

An antimicrobial composition comprising a hexene or ethyl acetate fraction of a hydrothermal extract of Cornus officinalis leaves. A pharmaceutical composition for preventing or treating a bacterial disease comprising a hexene or ethyl acetate fraction of a hydrothermal extract of Cornus officinalis leaf. A health functional food for preventing or improving a bacterial disease comprising a hexene or ethyl acetate fraction of a hydrothermal extract of Cornus officinalis leaf. Food additive comprising hexane or ethyl acetate fraction of hydrothermal extract of Cornus officinalis leaves. A cosmetic additive comprising a hexene or ethyl acetate fraction of a hydrothermal extract of Cornus officinalis leaf. Feed additive comprising hexane or ethyl acetate fraction of hydrothermal extract of Cornus officinalis leaf. A detergent comprising a hexene or ethyl acetate fraction of a hydrothermal extract of Cornus officinalis leaves. A cleansing agent comprising a hexene or ethyl acetate fraction of a hydrothermal extract of Cornus officinalis leaf. delete delete delete