KR102086166B1 - Antimicrobial composition comprising the extracts of rambutan peel - Google Patents

Abstract

The present invention relates to an antimicrobial composition comprising a bark extract of rambutan ( Nemphelium lappaceum ) fruit, and more particularly, to an antimicrobial composition having excellent antibacterial activity, including a hexene fraction or an ethyl acetate fraction of rambutan bark extract. will be. Rambutan peel extract of the present invention exhibits excellent antimicrobial activity, and can be used for antimicrobial pharmaceutical compositions, health functional foods, cosmetic compositions, animal feed compositions, food, cosmetic or animal feed additives, various cleaning agents, and various cleaning agents. It works. In addition, the rambutan shell extract of the present invention is excellent in thermal stability, do not show the loss of activity in the acidic conditions and plasma of pH 2, can be easily processed into various forms, such as liquid, cream, powder, pills, tablets It can be very useful in the pharmaceutical, food, livestock, health and cosmetic industries.

Description

Antibacterial composition containing rambutan peel extract as an active ingredient {ANTIMICROBIAL COMPOSITION COMPRISING THE EXTRACTS OF RAMBUTAN PEEL}

Rambutan ( Nephelium) lappaceum ) relates to an antimicrobial composition comprising a bark extract of a fruit, and more particularly, to an antimicrobial composition having excellent antibacterial activity, including a hexene fraction or an ethyl acetate fraction of rambutan peel extract.

Humans have used various antimicrobials and preservatives to defend against the invasion of pathogenic microorganisms and to control food spoilage microorganisms. However, chemical antibacterial and preservatives show excellent antimicrobial activity against various bacteria and fungi, while long-term use causes many side effects such as allergy redness, vascular edema, bone marrow suppression, anemia, gastritis, and degenerative brain diseases. do. In fact, the US has banned the use of antibiotics with severe side effects such as chloramphenicol. Therefore, various studies are being conducted on the development of natural antibacterial agents using natural products having long-term use while ensuring safety.

Meanwhile, according to the U.S. Center for Disease Control and Prevention, hospital blood flow is one of the leading causes of death in the United States. In addition, infection of pathogenic bacteria and fungi through skin and soft tissues is recognized as a universal medical disease, and is understood to be mainly caused by trauma or surgery. Many pathogenic bacteria and fungi infections differ in their treatment depending on the severity of the infection, but can be successfully treated with oral antimicrobial agents and topical cleansing. However, the more severe or complex infections that commonly occur in patients with underlying risk factors (eg vascular blood flow abnormalities, diabetes) and / or infections caused by bacteria and fungi that are difficult or multi-resistant to treat are venous Antimicrobial treatment and invasive surgical tissue removal may be necessary. For example, glycopeptide antibiotic vancomycin has been used successfully for severe in-hospital infections caused by multi-drug-resistant Gram-positive bacteria, in particular MRSA, coagulase-negative Staphylococcus and Enterococcus. However, it has recently been reported that the isolation of strains resistant to vancomycin is increasing, and the current antibiotics are very limited in the treatment of severe infection caused by some of the pathogens. Thus, there is an increasing need for the development of new antibiotics that are more effective against increasingly resistant bacteria.

On the other hand, the consumption of fruit is rapidly increasing due to the increase of population, and a large amount of fruit peels and seeds are generated as waste. Therefore, efficient reuse studies of shells and the like that are being discarded are urgently needed (Cheok CY et al., 2016, Critical Reviews in Food Science and Nutrition, http://dx.doi.org/10.1080/ 10408398.2016.1176009). Rambutan ( Nephelium lappaceum ) is one of the most bark-bearing tropical fruits, belongs to the Sapindaceae, with a tree height of 10-15 m, leaves alternate, 5-7 pairs of upper right lobes, thick and It is shiny and has hair on its back when young. Flowers bloom in April-June, fruits are gathered in 10-12 pieces. They are the size of small eggs, ripen in July-August, and have fleshy processes. The origin of Rambutan originated from the Malay word "rambut" and means "headed". In fact, Rambutan is one of the most well-known fruits of tropical fruit in Southeast Asia, and its largest producer is Thailand. Cultivated fruits are used to remove the skin and seeds and are usually used for raw food, as well as canned syrup, as well as in jam or cooked form.

In addition, rambutan is a low-calorie fruit, containing 80% water, 17.5% carbohydrates, 1.2% fat, and 1% protein, representing 79Kcal / 100g, and containing large amounts of vitamin C, phosphorus, potassium, calcium, and magnesium. It is known. Seeds contain 38% of the lipids, oleic acid (42.0%) and arachidic acid (34.3%) are known as the major constituents (ManaM et al., 2013. J. Oliec Sci. 62, 335-343). The skin of rambutan contains tannin and saponin and is used as a herbal medicine. Recently, the main ingredient of the skin was geraniin (Elendran S et al., 2015, Pharm Biol. 53, 1719-1726). The geraniin accounts for 30% of the ethanol extract of the rambutan shell, has anticancer and antidiabetic activity and is known to be effective in preventing hypertension (Elendran S et al., 2015, Pharm Biol. 53, 1719-1726). Corilagin and ellagic acid have also been reported from the bark (Thitilertdecha N et al., 2010. Molecules. 15, 1453-1465). In addition, the roots of rambutan are used for the treatment of colds, the leaves as poultices, and the bark is used as a remedy to relieve muscles when the tongue develops, but the antimicrobial activity of rambutan bark extract is not known at all.

The physiological activities of rambutan and rambutan shells known to date include antioxidant activity (Zhuang Y et al., 2017, Food Chem Toxicol. S0278-6915, 30030-30033), fatty acid synthesis inhibitory effect (Zhao YX et al., 2011, Carbohydr Res. 346, 1302-1306) and cancer cell growth inhibitory activity (Fang EF et al., 2015, Appl Biochem Biotechnol. 175, 3828-3839).

In addition, patents related to rambutan are known in the Republic of Korea Patent No. 10-1393007 [cosmetic composition for preventing skin aging and wrinkle improvement containing rambutan and rich extract as an active ingredient], registered patent No. 10-1411947 Japanese Patent Laid-Open Publication No. 10-2006-0007083 discloses [Whitening Cosmetic Composition and Rambutan Extract Method Containing Rambutan Extract] in Japanese Patent Laid-Open No. 4-244004. No. [Cosmetic compositions containing a skin extract of rambutane and having a whitening effect] are disclosed. However, the antimicrobial activity of rambutan peel extract is not known at all.

KR 10-1393007 B KR 10-1411947 B KR 10-2006-0007083 A JP 1992-244004 A

 Antioxidant and Antibacteiral activities of Nephelium lappaceum L. extracts. LWT-Food Sci. Technol. 2008. 41, 2029-2035.

The present invention has been made to solve the problems of the prior art as described above, the problem to be solved in the present invention is to prepare a rambutan shell extract, to provide an antimicrobial composition as an active ingredient.

In order to solve the above problems, the present invention provides an antimicrobial composition containing rambutan ( Nephelium lappaceum ) bark extract as an active ingredient.

The rambutan shell extract is preferably an ethanol extract of the rambutan shell.

The rambutan shell extract is preferably a hexene fraction or an ethyl acetate fraction of the rambutan shell ethanol extract.

The antimicrobial composition preferably has antimicrobial activity against Gram-positive bacteria.

The antimicrobial composition is preferably selected from the group consisting of pharmaceutical compositions, health functional foods, cosmetic compositions, animal feed compositions, food additives, cosmetic additives, animal feed additives, cleaning agents and cleaning agents.

Rambutan peel extract of the present invention exhibits excellent antimicrobial activity, and can be used for antimicrobial pharmaceutical compositions, health functional foods, cosmetic compositions, animal feed compositions, food, cosmetic or animal feed additives, various cleaning agents and various cleaning agents. It works. In addition, the rambutan shell extract of the present invention is excellent in thermal stability, do not show the loss of activity in the acidic conditions and plasma of pH 2, can be easily processed into various forms, such as liquid, cream, powder, pills, tablets It can be very useful in the pharmaceutical, food, livestock, health and cosmetic industries.

1 shows the entire rambutan fruit,
2 shows the pulp portion,
3 shows the shell area,
4 is a photograph showing a seed site.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail.

The present invention relates to a composition exhibiting potent antimicrobial activity containing rambutan shell extract. The inventors of the present invention prepared each fraction and water residue obtained by sequentially fractionating the ethanol extract of rambutan shell and its organic solvent of hexene, ethyl acetate, butanol, and confirmed the strong antibacterial activity in the double hexene fraction and ethyl acetate fraction It was.

In particular, the active fractions exhibited strong antimicrobial activity against Gram-positive bacteria, confirming that it can be developed as a wide range of antimicrobial agents against Gram-positive bacterial infections, and completed the present invention.

Therefore, the present invention is rambutan ( Nephelium) lappaceum ) provides an antimicrobial composition containing the bark extract as an active ingredient.

The rambutan shell extract is preferably an ethanol extract of the rambutan shell.

The rambutan shell extract is preferably a hexene fraction or an ethyl acetate fraction of the rambutan shell ethanol extract.

The antimicrobial composition preferably has antimicrobial activity against Gram-positive bacteria.

The antimicrobial composition is preferably selected from the group consisting of pharmaceutical compositions, health functional foods, cosmetic compositions, animal feed compositions, food additives, cosmetic additives, animal feed additives, cleaning agents and cleaning agents.

In a preferred embodiment, the present invention comprises the steps of separating the skin, pulp, seeds from selected, washed rambutan fruit; Preparing an extract from the separated shell; Purifying the active fraction through the sequential organic solvent fraction of rambutan shell extract; It may include a step of evaluating the stability of the active fraction.

"Rambutan shell extract" included in the composition of the present invention is a step of recovering the shell from the imported rambutan fruit, extracting the shell with a solvent, and filtering the extract using a filter network of 0.06mm or less, and concentrating it under reduced pressure It can be obtained by.

In the present invention, "rambutan shell extract" can be prepared using various extraction solvents. For example, it is prepared using water (cold water, hot water), alcohol, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, alcohol, propanol, butanol, etc.), a mixed solvent of the lower alcohol and water, and the like. can do. Preferably it shows the maximum yield of active material when extracted with ethanol or alcohol.

In addition, the "rambutan shell extract" of the present invention may be present as an active fraction by an additional organic solvent fraction after the extraction process using the extraction solvent. In a preferred embodiment, the rambutan shell ethanol extract of the present invention can be obtained as a water residue after a hexene fraction, an ethyl acetate fraction, a butanol fraction and an organic solvent fraction by performing sequential organic solvent fractions of hexene, ethyl acetate and butanol.

Rambutan shell extract of the present invention and organic solvent fractions thereof may be prepared into a powder through a conventional powdering process such as distillation under reduced pressure and lyophilization or spray drying. They remain active even at a heat treatment of 100 ° C. and pH within the human stomach at pH 2.

Rambutan bark extract and active fractions thereof of the present invention exhibit potent antimicrobial activity such as pharmaceutical compositions, health functional foods, cosmetic compositions, animal feed compositions, food, cosmetics, animal feed additives and the like, which are involved in the prevention or treatment of infections of pathogenic bacteria and It can be used as a material for various cleaning agents.

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

The pharmaceutical composition may be formulated in various forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, injections of sterile injectable solutions, etc. It may be used orally or may be administered through various routes including intravenous, intraperitoneal, subcutaneous, rectal, topical, and the like. Such pharmaceutical compositions may further include carriers, excipients or diluents, and examples of suitable carriers, excipients or diluents that may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, Starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, amorphous cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil Etc. can be mentioned. In addition, the pharmaceutical composition of the present invention may further include a filler, an anticoagulant, a lubricant, a humectant, a perfume, an emulsifier, a preservative, 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 an effective dose level means the type, severity, activity of the drug, Sensitivity to drug, time of administration, route of administration and rate of release, duration of treatment, factors including concurrent use of drugs, and other factors well known in the medical arts. The pharmaceutical compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered as single or multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can achieve the maximum effect with a minimum amount without side effects, which can be readily determined by one 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 weight of the patient, and generally 1 to 5,000 mg per body weight, preferably 100 to 3,000 mg daily or every other day Or divided into 1 to 3 times a day. However, since the dose may be increased or decreased depending on the route of administration, the severity of the disease, sex, weight, age, etc., the above dosage does not limit the scope of the present invention by any method. The pharmaceutical composition of the present invention can be administered to a subject through various routes. All modes of administration can be expected, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intra-cerebroventricular injection. As used herein, "administration" means providing a patient with any substance by any suitable method, wherein the route of administration of the pharmaceutical composition of the present invention is oral or parenteral via all common routes as long as the target tissue can be reached. Oral administration. In addition, the composition of the present invention may be administered using any device capable of delivering an active ingredient to a target cell. "Subject" in the present invention is not particularly limited, but includes, for example, humans, monkeys, cattle, horses, sheep, pigs, chickens, turkeys, quails, cats, dogs, mice, rats, rabbits or guinea pigs. And preferably mammals, and more preferably humans.

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

The health functional food of the present invention can be used in various ways such as effective foods and beverages that can reduce the possibility of infection from a subject of high concern of pathogenic microorganisms. Examples of the food containing the active ingredient having excellent antimicrobial activity of the present invention include various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and are powders, granules, tablets, capsules, or beverages. Can be used as The active ingredient of the present invention may generally be added at 0.01 to 15% by weight of the total food weight, the health beverage composition may be added at a ratio of 0.02 to 10g, preferably 0.3 to 1g based on 100 ml. In addition to containing the compound as an essential ingredient in the indicated proportions, the health functional food of the present invention may contain as food additives food additives such as natural carbohydrates and various flavoring agents. Examples of the natural carbohydrates include conventional sugars such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavourant, tau martin, rebaudioside A, glycyrrhizin, saccharin, aspartame and the like can be used. The proportion of the flavoring agent is generally used in the range of 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 includes various nutrients, vitamins, minerals, synthetic flavors and natural flavoring agents, colorants and neutralizing agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloids Thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks, and the like. In addition, the health functional food of the present invention may contain flesh for preparing natural fruit juice, fruit juice beverage, vegetable beverage and the like. These components can be used independently or in combination. The proportion 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 use of the cosmetic composition.

The cosmetic composition of the present invention may be prepared in the form of components having excellent antimicrobial activity of the present invention, for example, essence, nutrition cream, body lotion, rinse, shampoo, body cleanser, lotion, skin, mask pack, It is not limited to this. Preparation in this form may be readily performed according to various manufacturing processes known to those skilled in the art.

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

The animal is a biological group corresponding to plants, mainly ingesting organic matter as nutrients, and means that digestion, embryogenicity, and respiratory organs are differentiated, specifically, echinoderm, crustaceans, molluscs, fish, amphibians, reptiles, birds, It may be a mammal, preferably mollusks such as arthropods, cephalopods, gastropods or bivalve shells, including shellfish such as sea urchins or sea cucumbers, crustaceans such as crabs, shrimps and lobsters, red snapper, sea bream, cod, and flounder , Birds such as flounder, birds including poultry such as pheasants or chickens, or mammals such as pigs, cattle, goats.

The animal feed composition may further include grain, vegetable protein feed, animal protein feed, sugar or dairy product in the antimicrobial composition comprising the antimicrobial component of the present invention. The grains may be specifically crushed or crushed wheat, oats, barley, corn and rice, and the vegetable protein feed may be specifically based on rapeseed, soybean and sunflower, the animal protein feed specifically May be blood meal, meat meal, bone meal and fish meal, and the sugar or dairy product may specifically be a dry ingredient consisting of various powdered milk and whey powder.

The animal feed composition may further be used with ingredients such as nutritional supplements, digestion and absorption enhancers, sex stimulants or disease prevention agents.

The antimicrobial composition included in the composition for animal feed of the present invention may vary depending on the purpose and conditions of use of the feed. For example, the antimicrobial composition may be included in an amount of 0.1 g to 100 g based on 1 kg of the final produced feed.

In addition, the feed composition may be made of a viscous granulated or granular material depending on the degree of grinding of the components, and the composition may be fed into a mesh or formed into a desired discrete shape for further processing and packaging, It may be subjected to pelletization, expansion or extrusion processes, and preferably excess water may be dried off for ease of storage.

In a preferred embodiment, the antimicrobial composition of the invention can be applied as a use of additives for food, cosmetics or animal feed.

The additive is added to impart preservation to the food, cosmetics or animal feed used, the antimicrobial composition is a natural ingredient, so it can be safely applied to food, cosmetics or animal feed.

The additive may further include an additional additional additive selected appropriately according to the purpose of use and use thereof, 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 powdering, granulating, tableting or liquefying according to the purpose and use thereof, and a conventional method may be used for packaging for commercialization.

In addition, the amount of the antimicrobial composition used in the additive may be appropriately used according to the purpose of use, application form, application site, target object, and the like, and may include, for example, 0.01 part by weight to 50 parts by weight based on the total composition. The content of the composition is not limited thereto. Since the antimicrobial composition of the present invention is a natural ingredient, which is not harmful to the human body, it can be used in various amounts as long as the purpose of application can be achieved due to the characteristics of the product.

In a preferred embodiment, the invention antimicrobial composition can be applied as a use of the cleaning agent.

The detergent may be preferably a detergent for clothes, a dish cleaner, a food cleaner, a household appliance cleaner. Since the antimicrobial composition is not harmful to the human body as a natural component, more preferably, it may be used for the purpose of washing or providing antimicrobial properties of various household products including kitchenware or food washing.

The cleaning agent may further include an appropriately selected additive according to the purpose of use and use thereof, and may be mixed with other active ingredients such as commonly used cleaning agents or additives such as colorants, surfactants, and preservatives. The detergent may be prepared by powdering, granulating, tableting or liquefying according to the purpose and use thereof, and a conventional method may be used for packaging for commercialization.

In addition, the amount of the antimicrobial composition used in the cleaning agent may be suitably used according to the purpose of use, application form, application site, target object, and the like, and may include, for example, 0.01 part by weight to 50 parts by weight based on the total composition. The content of the composition is not limited thereto. Since the antimicrobial composition of the present invention is a natural ingredient, which is not harmful to the human body, it can be used in various amounts as long as the purpose of application can be achieved due to the characteristics of the product.

In a preferred embodiment, the invention antimicrobial compositions can be applied as a cleaner.

The cleaning agent may be used in a simple method such as hand, mouth or feminine cleaner in a simple way to prevent the infection of the pathogenic microorganisms that are commonly susceptible to daily life. Since the antimicrobial composition is not harmful to the human body as a natural ingredient, it can be safely applied even when used as a hand, mouth, or feminine cleaner.

The cleaner may further include an additional additional additive selected according to the purpose and use thereof, and may be mixed with other active ingredients such as commonly used additives or additives such as colorants, surfactants, and preservatives. . The cleaning agent may be prepared by powdering, granulating, tableting or liquefying depending on the purpose and purpose of use, but in the case of a liquid type, it may be conveniently used.

In addition, the amount of the antimicrobial composition used in the cleaning agent may be suitably used according to the purpose of use, application form, and application site, and may include, for example, 0.01 part by weight to 50 parts by weight based on the total composition. It is not limited to this. Since the antimicrobial composition of the present invention is a natural ingredient, which is not harmful to the human body, it can be used in various amounts as long as it can achieve an application purpose due to the characteristics of the product.

Hereinafter, the present invention will be described in more detail with reference to Examples. The following examples are only preferred embodiments of the present invention, and the scope of the present invention is not limited to the following examples.

EXAMPLE

Example  One: Rambutan  Preparation of extract for each part

After purchasing Thai rambutan fruit (moisture content 80%), foreign matters were removed, and each was divided into peel, pulp, and seed (FIG. 1), and this was used to prepare an ethanol extract. In preparing the ethanol extract, 10 times ethanol (95%, Deoksan, Korea) was added to each sample, extracted once at room temperature, the extracts were collected and filtered, and concentrated under reduced pressure to prepare a powder. Component analysis of the prepared extracts determined the total polyphenols, total flavonoids, total sugars and reducing sugars. The total polyphenol content was added to 400μl of the extracted sample solution, 50μl of Folin-ciocalteau, 100μl of Na ₂ CO ₃ saturated solution, which was left at room temperature for 1 hour and absorbance at 725nm. Tannic acid was used as the standard reagent. For the total flavonoid content, each sample was extracted by stirring for 18 hours with methanol, 400 ml of the filtered extract sample was added 4 ml of 90% diethylene glycol, 40 µl of 1N NaOH was added thereto, and the absorbance was measured at 420 nm after 1 hour of reaction at 37 ° C. Rutin was used as a standard reagent. Reducing sugars were quantified by DNS method and total sugars by phenol-sulfuric acid method.

TABLE 1 Rambutan  Yield and Composition of Extracts by Parts

As a result, as shown in Table 1, the extraction yield of each part of the rambutan fruit was found in the order of peel> pulp> seed, and showed a high yield of 7.33% in the peel. As a result of the analysis of the total polyphenol content, the bark extract showed a very high total polyphenol content (284.6 mg / g) and a relatively low total flavonoid content. On the other hand, the seeds showed low total sugar and reducing sugar content, but showed 12.6 mg / g total polyphenol and 4.6 mg / g total flavonoid.

Example  2: Rambutan  Evaluation of Antimicrobial Activity of Extracts by Parts

The antibacterial activity of the ethanol extracts of various parts of rambutan obtained in Example 1 was evaluated, and the results are shown in Table 2. As a strain used for the evaluation of antimicrobial activity, Escherichia as a Gram-negative bacterium coli KCTC 1682, Proteus vulgaris KCTC 2433, Pseudomonas aeruginosa KACC 10186, Salmonella typhimurium KCTC 1926, and Gram-positive bacteria were Staphylococcus aureus KCTC 1916, Staphylococcus epidermidis ATCC 12228, Listeria monocytogenes KACC 10550 and Bacillus subtilis KCTC 1924 were used. Meanwhile, for the evaluation of antifungal activity, Saccharomyces cerevisiae IF0 0233 and Candida causative agent Candida albicans KCTC 1940 was used.

First, in the case of antibacterial activity evaluation, each bacteria were inoculated to Nutrient broth (Difco Co., USA) and incubated at 37 ° C. for 24 hours, and then each strain was adjusted to OD ₆₀₀ 0.1 to adjust Nutrient agar (Difco Co. , USA) 100 μl each in a sterile petri dish (90 mm, green cross, Korea) containing medium, 5 μl of each sample was added to a sterile disc-paper (6.5 mm diameter, Whatsman No. 2), at 37 ° C. for 24 hours. In the case of fungi, after culturing for 24 hours at 30 ℃ in the same manner using Sabouraud dextrose (Difco Co. USA), the growth inhibitory ring was measured to evaluate the antimicrobial activity. As a control, antimicrobial ampicillin and antifungal myconazole (miconazole, Sigma Co., USA) were used at concentrations of 1 μg / disc, respectively. Measured in mm, and represented as a representative result after three or more evaluations.

TABLE 2 Rambutan  Antimicrobial Activity of Extracts by Parts

As a result, as shown in Table 2, in the case of empicillin (1 μg / disc) used as an antibacterial activity control, growth of 20.0 ~ 30.0mm in all bacteria except E. coli and P. aeruginosa Inhibitory ring showed potent antibacterial activity. Mycoazole (1μg / disc), an antifungal agent, showed growth inhibition of 15.5mm and 18.0mm for C. albicans and S. cerevisiae , respectively. On the other hand, in the case of rambutan skin extract exhibited a growth inhibition ring of 14.0mm for Staphylococcus epidermidis and Staphylococcus aureus at 500μg / disc density, Proteus The growth inhibitory ring of 13.5mm was shown for vulgaris . On the other hand, rambutan pulp and seed extract showed no antimicrobial activity. In addition, all rambutan samples showed no antifungal activity against the fungi used. These results indicate that rambutan bark extract can be efficiently applied to the control of some Gram-positive bacteria.

Example 3 Preparation of Sequential Organic Solvent Fractions from Rambutan Shell Extracts and Their Useful Component Analysis

Based on the above results, a large amount of rambutan shell ethanol extract was prepared, and then organic solvent fractionation was performed sequentially with hexene, ethyl acetate and butanol, and finally water residue was recovered. Organic solvent fraction efficiency and component analysis results are shown in Table 3. Component analysis measured the total polyphenol, total flavonoid, total sugar and reducing sugar content in the same manner as in Example 1.

TABLE 3 Rambutan  Bark Ethanol Extract and Its Sequential Fraction  Yield and ingredient analysis

As shown in Table 3, among the fractions of the rambutan shell extract, butanol fraction was the highest fractionation efficiency of 63.8%, ethyl acetate fraction and water residue accounted for 28.5% and 5.3% of the extract, respectively. On the other hand, the hexene fraction accounted for 1% of the extract, it was found that the rambutan shell extract has a very low fat-soluble oil component. Thus, 0.073 g of hexene fraction and 2.08 g of ethyl acetate fraction could be recovered from 100 g of rambutan shell.

As a result of component analysis of the fractions, the total polyphenol content was found in the order of ethyl acetate fraction, butanol fraction, water residue, hexene fraction. In addition, the total flavonoid content was also highest in the ethyl acetate fraction, but the butanol fraction and water residue showed a similar 8.5 ~ 9.9 mg / g. On the other hand, the total sugar and reducing sugar contents were highest in the butanol fraction, respectively, followed by water residues. Therefore, the hexene fraction and ethyl acetate fraction of rambutan shell extract were expected to be the main component of the phenolic glycosides, it was determined to exhibit a variety of physiological activity, it was confirmed that the actual strong antioxidant activity.

Example  4: Rambutan  Sequential Extract of Bark Extract Organic solvent Fraction  Antimicrobial Activity Assessment

The antimicrobial activity of the sequential organic solvent fractions of the rambutan shell ethanol extract prepared in Example 3 was evaluated, and the results are shown in Table 4.

TABLE 4 Rambutan  Sequential of Bark Ethanol Extract Organic solvent Fraction  Antimicrobial activity

As shown in Table 4, empicillin (1 μg / disc) used as an antibacterial activity control showed strong antibacterial activity against bacteria except E. coli and P. aeruginosa . In addition, the antifungal myconazole (1 μg / disc) also showed excellent antifungal activity against the fungi used. Among the fractions of the rambutan shell ethanol extract, only hexene fraction and ethyl acetate fraction showed excellent antibacterial activity.In particular, unlike the ethanol extract of the rambutan shell, Listeria Monocytogens and Bacillus subtilis also showed potent antibacterial activity. The results suggest that rambutan shell extract, its hexene fraction and ethyl acetate fraction can be effectively used for the control of food poisoning and pathogenic bacteria.

Example  5: Rambutan  Bark Extract and Its Hexene Fractions and  Ethyl acetate Fraction  Acid and Thermal Stability Assessment

The rambutan shell extracts obtained in Examples 1 and 3, hexene fractions and ethyl acetate fractions thereof were confirmed for thermal and acid stability against antimicrobial activity. The samples did not show a significant decrease in antimicrobial activity after 1 hour heat treatment at 100 ° C. and 1 hour treatment at pH 2 (0.01 M HCl), indicating high thermal and acid stability. Therefore, the rambutan shell extract and its active fractions are expected to maintain excellent antimicrobial activity even under various processing processes.

Claims (5)

A pharmaceutical composition for the prevention or treatment of Listeriosis containing hexene fraction or ethyl acetate fraction of Rambutan ( Nephelium lappaceum ) bark ethanol extract as an active ingredient. A dietary supplement for the prevention or improvement of Listeriosis containing hexene fraction or ethyl acetate fraction of Rambutan ( Nephelium lappaceum ) bark ethanol extract as an active ingredient. Animal feed additive for the prevention or amelioration of Listeriosis containing hexene fraction or ethyl acetate fraction of the ethanol extract of the rambutan ( Nephelium lappaceum ) bark. delete delete

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