KR20230058291A - Novel Method for Preparing Immature Sword Bean Pod Extracts Having Increased Rutin Content and Functionality, and Immature Sword Bean Pod Extracts Prepared Thereby - Google Patents

Abstract

The present invention relates to a method for preparing a Dodu pod extract with increased rutin content and functionality and an extract prepared thereby, wherein the Dodu pod extract with increased rutin content and functionality prepared according to the method of the present invention has excellent ABTS scavenging and DPPH scavenging And since it shows an effect of increasing the total polyphenol content and an excellent nitric oxide production inhibitory effect, it can be usefully used as an antioxidant and anti-inflammatory composition.

Description

Manufacturing method of pod pod extract with increased rutin content and functionality and extract prepared thereby

The present invention relates to a method for preparing a Dodu pod extract with increased rutin content and functionality, and an extract prepared thereby.

As we enter the well-being era, interest in functional foods that are good for health is growing, and accordingly, efforts are being made in the agri-food industry everywhere to create foods that have useful high-quality ingredients as well as effects that affect human health. Convenience processed foods are consumed frequently due to busy daily life, and when choosing convenience processed foods, people want to maintain health by consuming foods made of functional materials or containing functional ingredients that can help health for well-aging. Therefore, consumer needs are expanding to include foods made from healthy raw materials containing functional and nutritional components, food raw materials and country of origin.

Reflecting these consumer trends, the food industry is concentrating on discovering raw materials with excellent functionality that can be used for foods with excellent physiological activity and developing products using them, developing various products that include functionality and replacing domestic products with functionality. Efforts are being made to discover raw materials.

Dodu (sword bean, Canavalia gladiata , Canavalia ensiformis ) is a dicotyledonous plant, a climbing annual plant belonging to the rosemary legume family, and is called sword bean or sword bean because the bean pod looks like a sword bean, and has been consumed as tea or cooked vegetables at home and abroad. It belongs to the leguminous family and is a climbing annual herb. Its main cultivation area is in Africa, Southeast Asia, etc., but due to climate warming, it is introduced and cultivated in Korea.

Soybeans have been mainly eaten in the form of soybeans, but due to the nature of their growth, the maturity rate of soybeans rapidly decreases when flowering after August. there is a problem.

As a result, it was recently registered as a new food raw material by the Ministry of Food and Drug Safety in order to utilize the unripe pods, laying the groundwork for developing general food and functional food raw materials.

However, as it has recently been registered as a new food ingredient, as a commercialized product, it has not been eaten for a short period of time, so research for industrialization and functionality of Dodu pods has not been conducted. Research to improve the utilization and practicality of materials is needed.

In addition, as the functional labeling system for general foods and agricultural products has recently been implemented, it is necessary to make efforts to industrialize materials for the globalization of domestic agricultural products, such as entering the global market and securing competitiveness through the preparation of scientific basis data.

Newly registered food raw materials have excellent food value, but there are limits to commercialization and development of the material as food due to insufficient industrialization research and functional research and lack of scientific basis. It is urgently needed to expand research to discover functionality and improve the usability of materials.

The pods of these soybeans (see FIG. 1) are defined as limited to young (soft, before swelling) pods having soft tissue as food raw materials in the food ingredient DB provided by the Ministry of Food and Drug Safety.

Research on dodu pods, which have been reported to contain a large amount of nutrients and effective ingredients, and have recently been newly registered as food raw materials, is still extremely limited. However, it is limited in its application to various products such as baking, preference food, functional food, and cosmetics.

Therefore, by developing a method for efficiently increasing the content and efficacy of a specific active ingredient in soybean pods that can be used as a food material with high added value, the range of use as a food material such as health functional food can be expanded.

Republic of Korea Patent No. 10-2248056 Republic of Korea Patent No. 10-1476218

Under these circumstances, the present inventors intensively researched to develop an active ingredient and a method for increasing the efficacy of canavalia gladiata (Jacq.) DC.) pod extract, which is a useful material as a food raw material. As a result, the present inventors completed the present invention by identifying that it is possible to increase the content of rutin, an active ingredient of soybean pods, and prepare an extract with increased functional efficacy such as antioxidant and anti-inflammatory properties through optimal treatment conditions. .

Accordingly, one object of the present invention is to provide a method for preparing a Dodu pod extract with enhanced rutin content and functionality.

In addition, another object of the present invention is to provide a Dodu pod extract with improved rutin content and functionality, prepared by the above method.

In addition, another object of the present invention is to provide an antioxidant or anti-inflammatory health functional food containing the rutin content and functionality of a Dodu pod extract as an active ingredient.

In addition, another object of the present invention is to provide a cosmetic composition for anti-oxidation or anti-inflammatory use, which contains, as an active ingredient, a Dodu pod extract with enhanced rutin content and functionality.

In addition, another object of the present invention is to provide a pharmaceutical composition for antioxidant and anti-inflammatory containing the rutin content and functionality of Dodu pod extract as an active ingredient.

In addition, another object of the present invention is to provide an antioxidant and anti-inflammatory feed additive containing the rutin content and functionality of a Dodu pod extract as an active ingredient.

Other objects and advantages of the present invention will become more apparent from the following detailed description and claims.

The terms used herein are used for descriptive purposes only and should not be construed as limiting. Singular expressions include plural expressions unless the context clearly dictates otherwise. In this specification, terms such as "include" or "have" are intended to designate that there is a feature, number, step, operation, component, part, or combination thereof described in the specification, but one or more other features It should be understood that the presence or addition of numbers, steps, operations, components, parts, or combinations thereof is not precluded.

In addition, unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which the embodiment belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the related art, and unless explicitly defined in the present application, they should not be interpreted in an ideal or excessively formal meaning. don't

Hereinafter, the present invention will be described in detail.

According to one aspect of the present invention, the present invention provides a method for producing an extract of Dodu pod with enhanced rutin content and functionality of Formula 1, comprising the step of extracting by adding a solvent to dry powder of Dodu pod :

[Formula 1]

.

.

The functionalities are antioxidant activity and anti-inflammatory activity.

According to one embodiment of the present invention, the manufacturing method of the present invention may include the steps of sorting, washing, and drying the soybean pods by any method known in the art before performing the extraction step.

In addition, according to one embodiment of the present invention, the present invention, the step of filtering the extracted extract after the extraction step; And drying the filtrate to powder; may additionally include.

More specifically, the method for preparing the extract of the present invention for achieving the above object includes removing moisture from the selected and washed soybean pods; Cutting the water-removed soybean pods and then drying and powdering them; Extracting after adding a solvent to the dried bean pod powder; Filtering the extracted extract after centrifugation; and drying and pulverizing the filtrate.

The soybean pods can be cut into 0.3 to 5 cm, and dried at 50 to 120 ° C for 1 to 96 hours.

In the extraction step, the solvent may be added in an amount of 500 to 5,000 parts by weight based on 100 parts by weight of the dried flax pods.

The extraction time may be performed for 1 to 100 hours, and the extraction temperature may be 10 to 120 °C.

The centrifugation may be performed for 1 to 30 minutes at 1,000 to 10,000 g.

The pore size of the filtration membrane may be 0.01 to 10 μm.

The powdering may be spray-dried, freeze-dried, vacuum-dried, or hot-air-dried.

The dodu pod may be a tool angle or a dodu.

The present invention relates to a method for preparing an extract with enhanced rutin content and functionality from soybean pods, wherein the extract secures safety as a food ingredient and has enhanced rutin content and functionality.

According to the method of the present invention, not only can the rutin content be significantly increased from the same amount of raw material (bean pea pod), but even if the same amount of bean pod extract is used, soybean extracted without treatment by the method of the present invention Excellent antioxidant and anti-inflammatory effects were confirmed compared to the pod extract.

In this specification, the term 'extract' used while referring to the Dodu pod extract refers to formulations (e.g., , Powdered) has the meaning of including processed bean pods.

In the case where the Dodu pod extract used in the present invention is obtained by treating the Dodu pod treated with the method of the present invention with an extraction solvent, various extraction solvents may be used. Preferably, a polar solvent or a non-polar solvent may be used. Suitable as polar solvents are (i) water, (ii) alcohols (preferably methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol), (iii) acetic acid, (iv) dimethyl-formamide (DMFO) and (v) dimethyl sulfoxide (DMSO). Suitable as non-polar solvents are acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkanes, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1- Penten, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane, chlorobenzene, benzene, diethyl ether, diethyl sulfide, chloroform, dichloro methane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride and THF.

Preferably, the extraction solvent used in the present invention is (a) water, (b) anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, (c) a mixed solvent of the lower alcohol and water, (d) acetone, ( e) ethyl acetate, (f) chloroform, (g) butyl acetate, (h) 1,3-butylene glycol, (i) hexane and (j) diethyl ether, more preferably methanol, ethanol, It is at least one lower alcohol having 1 to 4 carbon atoms selected from the group consisting of propanol, isopropanol, butanol and isobutanol, most preferably ethanol.

The ethanol is 10% to 90% (v / v) fermented alcohol, preferably 30% fermented alcohol.

As used herein, the term extract has a common meaning as a crude extract in the art as described above, but also includes a fraction obtained by further fractionating the extract in a broad sense. That is, the Dodu pod extract includes not only those obtained using the above-described extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, separation by various chromatography (made for separation according to size, charge, hydrophobicity or affinity), etc. The fraction obtained through the purification method is also included in the Dodu pod extract of the present invention.

The Dodu pod extract used in the present invention may be prepared in a powder state by an additional process such as distillation under reduced pressure and freeze drying or spray drying.

In the present invention, the method for preparing the extract of the Dodu pods of the present invention includes the steps of removing moisture from the selected and washed Dodu pods; Cutting the water-removed soybean pods and then drying them into powder; Extracting after adding a solvent to the dried bean pod powder; Filtering the extracted extract after centrifugation; and drying and pulverizing the filtrate.

First, the soybean pods are washed with an aeration type or swirl type washer for 1 to 2 minutes, and then the water in the washed soybean pods is removed with an air brower.

Pesticide components, heavy metals, fungi, etc. remaining on the surface of the pods are removed through an aeration type or swirl type washer.

Water must be removed so that the water content on the surface of the washed soybean pods is 2 to 10%. It is added and dried for 1 to 2 hours to obtain a bean pod having the above moisture content.

If the moisture content of the pods is less than the lower limit above, some of the skin may be removed when removing moisture with an air blower, and if it exceeds the upper limit above, the subsequent drying time becomes longer, resulting in reduced workability and hardening of the tissue. there is.

As the soybean pods of the present invention, soybean pods or soybean pods can be used.

The soybean pod refers to the entire bean shell in which soybean is contained, and in the present invention, it is preferably in an immature state. The immature state refers to the state of pods that can be used as food raw materials defined in the food raw material DB provided by the Ministry of Food and Drug Safety, and refers to pods in a young (soft, before swelling) state.

Next, the soybeans from which moisture has been removed are cut into 0.3 to 5 cm, and dried at 50 to 120 ° C. for 1 to 96 hours.

In the present invention, by drying the pods containing water, the shell of the pods expands to promote the movement of water during the drying process and to enable long-term preservation. If the drying process is not performed, long-term storage is not possible, and the quality may deteriorate due to contamination of microorganisms during storage.

If the temperature and time during drying are less than the lower limit, the risk of microbial contamination may increase due to insufficient drying, and if the temperature and time exceed the upper limit, the soybean pods may be carbonized to generate benzopyrene, and quality such as flavor may deteriorate.

Next, an extraction process is performed after adding a solvent to the dried bean pod powder.

The solvent is added in an amount of 500 to 5,000 parts by weight, preferably 1,000 parts by weight, based on 100 parts by weight of the dried soybean pods. If the amount of the solvent added is less than the lower limit, the extraction yield may be lowered, and if the amount is greater than the upper limit, if the production efficiency is lowered, more time may be required in the subsequent unit process.

If the extraction time is less than the lower limit, the content may not be sufficiently extracted and the yield may be lowered. If the extraction time exceeds the upper limit, the bean pods are softened and excessive starch is eluted, and subsequent unit processes may take a lot of time.

The conditions of the centrifugation process are 1,000 to 10,000 g, preferably 8000 g, and the pore size of the filtration membrane is 0.01 to 10 μm, preferably 0.45 μm.

If the rotation g and pore size during centrifugation and filtration are less than the lower limit, starch may not be sufficiently removed, and if it exceeds the upper limit, process efficiency may decrease and production cost may increase.

The Dodu pod extract may be processed into an extract powder by spray drying, freeze drying, vacuum drying, or hot air drying. The extract powder is prepared in the form of powder or granules having a water content of 10% or less so as to have excellent solubility when rehydrated in water and excellent storage stability.

In addition, according to another aspect of the present invention, the present invention provides a Dodu pod extract with improved rutin content and functionality, prepared according to the above-described method.

In addition, according to another aspect of the present invention, the present invention provides a health functional food or food additive containing the Dodu pod extract as an active ingredient.

Dodu pod extract with increased rutin content and functionality, prepared by the manufacturing method according to the present invention, has enhanced antioxidant and anti-inflammatory activity, which is the unique functionality of Dodu pod, so it can be used as a food additive or health functional food material for this purpose. possible.

When the Dodu pod extract according to the present invention is used as an active ingredient additive for health functional food or general food, the Dodu pod extract according to the present invention can be added as it is or used together with other foods or food ingredients, and can be used according to a conventional method. can be used appropriately. The mixing amount of the active ingredient can be appropriately determined according to each purpose of use, such as prevention, health, or treatment.

In general, when preparing food or beverage, the Dodu pod extract according to the present invention may be added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be less than the above range, and since the present invention uses an extract from a natural product, there is no problem in terms of safety, so the above range The above amount can also be used.

The type of food is not particularly limited, and examples of food to which the substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, and dairy products including ice cream. , various soups, beverages, tea, drinks, alcoholic beverages and vitamin complexes, etc., and may include all foods in a conventional sense.

Among the health functional foods according to the present invention, beverages may contain various flavoring agents or natural carbohydrates as additional ingredients, like conventional beverages. The aforementioned natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used. The ratio of the natural carbohydrate may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the functional food according to the present invention.

In addition to the above, the health functional food containing the bean pod extract according to the present invention as an active ingredient contains various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH It may contain regulators, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, and these components may be used independently or in combination. The proportion of these additives is not limited, but is generally selected in the range of 0.01-0.1 parts by weight based on 100 parts by weight of the total composition of the present invention.

That is, the Dodu pod extract or extract powder of the present invention can be used in health foods or sauces.

The health functional food is a food prepared by adding soybean pod extract to food materials such as beverages, teas, spices, gum, confectionery, etc., or by encapsulation, powdering, suspension, etc., and when ingested, it has a specific effect on health It means coming, but unlike general drugs, it has the advantage of not having side effects that can occur when taking drugs for a long time by using food as a raw material. The health functional food of the present invention obtained in this way is very useful because it can be consumed on a daily basis. The amount of soy bean pod extract added to such health functional foods cannot be uniformly determined depending on the type of target health functional food, but it can be added within a range that does not impair the original taste of the food. It ranges from 0.01 to 50% by weight, preferably from 0.1 to 20% by weight. In addition, in the case of health functional foods in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of pills, tablets, capsules or beverages.

In addition, the sauces may include soybean sauce, dressing sauce, soybean paste sauce, and the like.

In addition, according to another aspect of the present invention, the present invention provides a cosmetic composition for anti-oxidation or anti-inflammatory, comprising the extract of Dodu pod with enhanced rutin content and functionality as an active ingredient.

The Dodu pod extract prepared by the manufacturing method according to the present invention can be used as a cosmetic material for this purpose because the unique function is preserved.

The extract in the present composition may be contained in an amount of 1 to 30% by weight, and may be contained in an amount of 5 to 20% by weight in the total weight ratio.

In addition, the cosmetic composition containing the extract of Dodu pods of the present invention may include conventional adjuvants and carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and flavors in addition to the extract, and may be formulated with a carrier. there is. The cosmetic composition of the present invention can be prepared in any formulation conventionally prepared in the art, for example, a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing , It may be formulated as an oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be prepared in formulations of flexible lotion, nutrient lotion, nutrient cream, massage cream, essence, eye cream, hair tonic, shampoo, rinse, cleansing cream, cleansing foam, cleansing water, pack, spray, and powder. .

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. can

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane / May contain a propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butyl glycol, glycerol aliphatic esters, polyethylene glycol or fatty acid esters of sorbitan.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Star cellulose, aluminum metahydroxide, bentonite, agar or tracanth and the like may be used.

When the formulation of the present invention is surfactant-containing cleansing, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters may be used.

Since the cosmetic composition of the present invention includes the above-described Dodu pod extract with enhanced rutin content and functionality, duplicate descriptions are omitted to avoid excessive complexity in the present specification.

In addition, according to another aspect of the present invention, the present invention provides a pharmaceutical composition for antioxidant and anti-inflammatory containing the above-described Dodu pod extract with enhanced rutin content and functionality as an active ingredient.

In a specific embodiment of the present invention, the present inventors analyzed the viability of macrophages in order to determine whether the Dodu pod extract with increased rutin content and functionality was cytotoxic, and as a result, it was confirmed that there was no cytotoxicity. In addition, the present inventors analyzed ABTS elimination, DPPH elimination, and total polyphenol content in order to confirm the antioxidant effect of the Dodu pod extract with enhanced rutin content and functionality, and as a result, it was confirmed that it exhibited increased antioxidant activity. In addition, the present inventors analyzed the amount of nitric oxide (NO) produced in order to confirm the anti-inflammatory effect of the rutin content and functionality of the Dodu pod extract, and as a result, it was confirmed that it exhibited a high nitric oxide production inhibitory effect.

Therefore, the extract of the Dodu pod with enhanced rutin content and functionality of the present invention can be usefully used as a pharmaceutical composition for preventing or treating inflammatory diseases as a pharmaceutical composition for antioxidant and anti-inflammatory.

The inflammatory disease may be selected from the group consisting of inflammatory bowel disease, glomerulonephritis, inflammatory skin disease, retinitis, gastritis, hepatitis, enteritis, arthritis, tonsillitis, sore throat, bronchitis, pneumonia, pancreatitis and nephritis.

The pharmaceutical composition according to the present invention may contain the extract of the present invention alone or may further contain one or more pharmaceutically acceptable carriers, excipients or diluents. In the above, "pharmaceutically acceptable" is a non-toxic composition that is physiologically acceptable and does not inhibit the action of the active ingredient when administered to humans and does not usually cause allergic reactions such as gastrointestinal disorders and dizziness or similar reactions. says

A pharmaceutically acceptable carrier may further include, for example, a carrier for oral administration or a carrier for parenteral administration.

Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like.

In addition, the carrier for parenteral administration may include water, suitable oil, saline, aqueous glucose and glycol, and the like, and may further include a stabilizer and a preservative. Suitable stabilizers include antioxidants such as sodium bisulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifier, a suspending agent, and the like in addition to the above components. Other pharmaceutically acceptable carriers and formulations may be referenced as described in the following literature (Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).

The composition of the present invention can be administered to mammals including humans by any method. For example, it can be administered orally or parenterally. Parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal administration can be

The pharmaceutical composition of the present invention may be formulated into a formulation for oral administration or parenteral administration according to the administration route as described above. In the case of preparations for oral administration, the composition of the present invention may be formulated into powders, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions, etc. using a method known in the art. can For example, preparations for oral use may be obtained by combining the active ingredient with a solid excipient, which is then milled and, after adding suitable auxiliaries, processed into a mixture of granules to obtain tablets or dragees. Examples of suitable excipients include sugars including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol, starches including corn starch, wheat starch, rice starch and potato starch, cellulose, Celluloses including methyl cellulose, sodium carboxymethylcellulose and hydroxypropylmethyl-cellulose, and the like, fillers such as gelatin, polyvinylpyrrolidone, and the like may be included. In addition, cross-linked polyvinylpyrrolidone, agar, alginic acid or sodium alginate may be added as a disintegrant, if desired. Furthermore, the pharmaceutical composition of the present invention may further include an anticoagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifier, and a preservative.

In the case of preparations for parenteral administration, they may be formulated in the form of injections, creams, lotions, external ointments, oils, moisturizers, gels, aerosols, and nasal inhalants by methods known in the art.

The total effective amount of the composition of the present invention can be administered to the patient in a single dose or by a fractionated treatment protocol in which multiple doses are administered over a long period of time. The pharmaceutical composition of the present invention may vary the content of the active ingredient according to the severity of the disease. Preferably, the preferred total dose of the pharmaceutical composition of the present invention may be about 0.01 μg to 1,000 mg, most preferably 0.1 μg to 500 mg per kg of patient body weight per day. However, the dose of the pharmaceutical composition is determined by considering various factors such as the formulation method, administration route, and number of treatments as well as the patient's age, weight, health condition, sex, severity of disease, diet, and excretion rate. Therefore, considering this point, those skilled in the art will be able to determine an appropriate effective dosage of the composition of the present invention. The pharmaceutical composition according to the present invention is not particularly limited in its formulation, administration route and administration method as long as it exhibits the effects of the present invention.

Since the pharmaceutical composition of the present invention includes the above-described Dodu pod extract with enhanced rutin content and functionality, duplicate descriptions are omitted to avoid excessive complexity in the present specification.

In addition, according to another aspect of the present invention, the present invention provides a feed additive for antioxidant and anti-inflammatory comprising the extract of the Dodu pod as an active ingredient.

The feed additive of the present invention contains the above-mentioned Dodu pod extract of the present invention as an active ingredient and is safe, so when used as a feed additive, it can contribute to improving the quality of the target animal.

For the feed additive of the present invention, the above-described Dodu pod extract may be used as it is or additionally known carriers such as grains and their by-products acceptable for the target animal, stabilizers, etc. may be added, and, if necessary, citric acid, fumalic acid, Organic acids such as adipic acid, lactic acid, and malic acid; phosphates such as sodium phosphate, potassium phosphate, acidic pyrophosphate, polyphosphate (polyphosphate), polyphenol, catechin, alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice Extracts, natural antioxidants such as chitosan, tannic acid, and phytic acid, antibiotics, antibacterial agents, and other additives may be added, and the form may be in a suitable state such as powder, granule, pellet, or suspension. In the case of use, the extract of the bean pod can be supplied alone or mixed with feed.

Since the feed additive of the present invention includes the above-described Dodu pod extract with enhanced rutin content and functionality, duplicate descriptions are omitted to avoid excessive complexity in the present specification.

The present invention relates to a method for preparing a Dodu pod extract with increased rutin content and functionality and an extract prepared thereby, wherein the Dodu pod extract with increased rutin content and functionality prepared according to the method of the present invention has excellent ABTS scavenging and DPPH scavenging And since it shows an effect of increasing the total polyphenol content and an excellent nitric oxide production inhibitory effect, it can be usefully used as an antioxidant and anti-inflammatory composition.

1 is a photograph showing the characteristics of soybean pods and soybean raw materials used in the present invention.
Figure 2a shows the HPLC-UV vis chromatogram HPLC at 280 nm, and Figure 2b shows the result of measuring the rutin content. Data are presented as mean ± SD (n = 3).
Figure 3a shows the effect of ISBP extract on ABTS radical scavenging activity, Figure 3b shows the effect of ISBP extract on DPPH radical scavenging activity, Figure 3c shows the effect of ISBP extract on total polyphenol content, , Figure 4d shows the effect of ISBP extract on total flavonoid content. VC, vitamin C; GA, gallic acid. Data are presented as mean ± SD (n = 5).
Figure 4a shows the effect of ISBP extract on cell viability in the LPS-induced RAW264.7 cell line, and Figure 4b shows the effect of ISBP extract on nitric oxide (NO) levels. Values are mean ± SD (n = 6). * p < 0.05 vs NC.

Hereinafter, the examples are only for explaining the present invention in more detail, and those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention It will be self-evident for

Experiment method

experimental material

The bean pods used in this example were grown and harvested in Hwasun-gun, Jeollanam-do in 2021, dried with hot air, and then pulverized and used.

reagent

DPPH, ABTS, L-ascorbic acid, Folin-Ciocalteu's phenol reagent, gallic acid, and Rutin used in the experiment were purchased from Sigma Chemical Co. (USA).

extract manufacturing

The soybean pods were dried with hot air at 55 ± 1 ° C for 2 days, ground, and stored at -20 ° C. 100 g of dried and pulverized samples were extracted with 30% and 50% fermented alcohol (I1-San Trading Co., Ltd., Haman, Korea) at 1000 mL, 100 ° C hot water and 1000 mL room temperature, respectively, for 24 hours. After extraction, centrifugation was performed at 8000 g for 20 minutes, and the supernatant was filtered through a 0.45 μm membrane filter to prepare an extract. The extract was prepared by concentrating with a vacuum concentrator.

Measurement of pH and soluble solids content

A pH meter (Orion Star A211 pH benchtop meter; Thermo Scientific?; Waltham, MA, USA) was used to measure pH, and a refractometer (Sccharometer, W.S.R.O-90, Atago, Co., Tokyo, Japan) was used to measure sugar content.

DPPH assay

The DPPH radical scavenging ability of hot water and alcohol extracts from Dodu pods was measured using the method of Blois MS (1958). 2 mL of 75 nM DPPH solution was mixed with 1 mL of each concentration sample, left at 37 ° C for 30 minutes, and then absorbance was measured at 517 nm using a UV / visible spectrophotometer. L-ascorbic acid was used as a control, and the DPPH radical scavenging activity was obtained by obtaining the standard curve of the concentration against the absorbance of the standard, and then obtaining the antioxidant capacity of the sample.

Antioxidant capacity (%) = 100-100*{(O.D-blank O.D of sample)/(O.D of O.D-blank of control}

ABTS assay

The evaluation of antioxidant activity through the ABTS radical scavenging ability of hot water and alcohol extracts from Dodu pods was measured using the method of Re R et al. (1999). 1.0 mM AAPH (2,2'-azobis-(2-amidinopropane)HCl) and 2.5 mM ABTS2-(2,2'-azino-bis (3-ethyl benzthiazoline-6-sulfonic acid) diammonium salt) in a water bath at 70 °C. After dissolving in 100ml of PBS, dilute at 734nm until the ABTS solution shows an absorbance of about 0.65. After filtering the solution, it is measured at 37℃. After measuring absorbance while reacting 20 mL of sample with 980 mL of ABTS solution for 10 minutes, antioxidant capacity is measured. After mixing 7 mM ABTS and 2.45 mM potassium persulfate at a ratio of 1:1 and leaving it in the dark at 30° C. for 12 hours, absorbance at 413 nm was diluted to 0.7 using 5 mM potassium phosphate buffer (pH 7.4). 40 μL of the sample prepared for each concentration was mixed with 4 mL of diluted ABTS reagent, reacted for 1 minute, and absorbance was measured at 413 nm. After obtaining the standard curve of the concentration against the absorbance of the standard, the antioxidant capacity of the sample is obtained.

Antioxidant capacity (%) = 100-100*{(O.D-blank O.D of sample)/(O.D of O.D-blank of control}

Total phenolic compound and flavonoids content

The total phenolics content was determined using the method of Folin and Denis [21] with minor modifications. 0.5 mL each of the diluted sample (50-fold dilution) extract and 25% Na2CO3 solution was dispensed into a test tube, reacted for 3 minutes, and then 0.25 mL of 0.2N Folin-Ciocalteu phenol reagent was added to develop color for 3 minutes. After centrifugation, the supernatant was measured at 720 nm, and the content was calculated by substituting it into a standard calibration curve prepared using gallic acid.

The total flavonoids content was determined by slightly modifying the method of Lee et al. [22]. 0.5 mL of the diluted sample (20-fold dilution) was dispensed into a test tube, mixed with 1 mL of diethylene glycol solution and 0.01 mL of 1 N NaOH, reacted in a constant temperature water bath (37 ° C) for 1 hour, and then measured at 420 nm using rutin. The content was calculated from the standard calibration curve prepared by

Rutin content measurement

The concentrated liquid sample was filtered through a 0.45 μm membrane filter and used as an analysis sample. The sample injection amount was 10 μL and the flow rate was 0.8 mL/min. SHISEIDO (4.6Х250 nm, 5 μm, SHISEIDO, Tokyo, Japan) was used as the HPLC column, and analysis was performed at a wavelength of 280 nm. As the mobile phase solvent, HPLC water (0.1% acetic acid) and HPLC acetonitrile were used as solvent A and solvent B, respectively, and the analysis conditions were 16 min-16%, 18 min-100%, 22 min-100%, 25 min-16%, 30 min-16% were analyzed.

Example 1. Selection of indicator substances in ISBP (Immature Sword Bean Pod)

In order to select the most important indicator substance in raw material standardization for the development of ISBP (Immature Sword Bean Pod) as a health functional food, the present inventors conducted a component investigation on 125 kinds of flavonoids and phenolic compounds, and Table 1 below sorted on.

[Table 1]

As a result, rutin, which was not analyzed in soybeans but was specifically detected only in pods, has the highest stability, has a high component content in soybean pods, and has the following formula 1, is selected as an indicator compound. did

[Formula 1]

Example 2. Confirmation of rutin content according to extraction conditions of soybean pods

In order to establish conditions capable of increasing the content of rutin, the indicator compound selected in Example 1, the present inventors extracted ISBP under various extraction conditions (hot water extraction, alcohol concentration of 30% and 50%) as shown in Table 2 below. After that, the rutin content was detected through HPLC UV-vis.

[Table 2]

As a result, as shown in FIGS. 2a and 2b, the content of rutin was significantly higher when 30% fermented alcohol (ethanol) was used as an extraction solvent for the bean pods, which was about 3 times more significant than that of the hot water extract. appeared to be high.

Example 3. Confirmation of antioxidant according to extraction conditions of soybean pods

In order to establish conditions capable of enhancing antioxidant as a functional effect, the present inventors extracted ISBP under various extraction conditions as shown in Table 2, and then confirmed the levels of ABTS elimination, DPPH elimination, total polyphenol content and total flavonoid content, respectively. .

As a result, as shown in Figures 3a to 3d, when extracting pods using 30% fermented alcohol (ethanol) as an extraction solvent, the antioxidant activity (ABTS, DPPH scavenging), polyphenol content, and flavonoid content were significantly the highest. appeared to be high.

Example 4. Confirmation of cytotoxicity and anti-inflammation according to extraction conditions of soybean pods

The present inventors, in order to establish conditions capable of enhancing cell viability as functional efficacy, extracted ISBP under various extraction conditions as shown in Table 2 above, and then confirmed the effects on cytotoxicity and anti-inflammatory in the RAW 264.7 cell line, respectively.

As a result, as shown in Figure 4a, none of the extracts showed cytotoxicity.

In addition, as a result, as shown in Figure 4b, when the nitric oxide (NO) level was confirmed after inducing inflammation with LPS (lipopolysaccharide) in the RAW 264.7 cell line, all ISBP extracts reduced NO production in a dose-dependent manner. Confirmed.

Overall, the extract method of the present invention not only significantly increased the rutin content as an indicator substance in the pods, but also showed excellent effects in enhancing functionalities such as antioxidant and anti-inflammatory properties.

Therefore, the present invention establishes an optimal extraction method for increasing the rutin content and functionality of soybean pods, a new raw material, and through this, a foundation that can be used as basic information for standardization of raw materials of soybean pods in the future is established. Through this, it can have high marketability in the food material industry and processing industry.

Examples of formulations of the extract of Dodu pod prepared according to the method of the present invention will be described below, but the present invention is not intended to limit them, but only to specifically describe them.

Formulation Example 1. Preparation of powder

Dodu pod extract powder 20 mg

Lactose 100 mg

Talc 10 mg

A powder is prepared by mixing the above ingredients and filling them in an airtight bag.

Formulation Example 2. Preparation of tablets

Dodu pod extract powder 10 mg

Corn Starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above ingredients, tablets are prepared by tableting according to a conventional tablet manufacturing method.

Formulation Example 3. Preparation of capsule formulation

Dodu pod extract powder 10 mg

3 mg of crystalline cellulose

Lactose 14.8 mg

Magnesium stearate 0.2 mg

Capsules are prepared by mixing the above ingredients and filling them into gelatin capsules according to a conventional capsule preparation method.

Formulation Example 4. Preparation of Injections

Dodu pod extract powder 10 mg

Mannitol 180 mg

Sterile Distilled Water for Injection 2974 mg

Na ₂ HPO ₄ , 12H ₂ O 26 mg

It is prepared with the above component content per 1 ampoule according to the conventional method for preparing injections.

Formulation Example 5. Preparation of liquid formulation

Dodu pod extract powder 20 mg

Isomerized sugar 10 g

5 g mannitol

Appropriate amount of purified water

According to the conventional manufacturing method of liquid formulations, each component is added to purified water to dissolve, lemon flavor is added in an appropriate amount, the above components are mixed, and purified water is added to adjust the total to 100, and then filled into a brown bottle for sterilization to prepare a liquid.

Formulation Example 6. Manufacture of health functional food

Dodu pod extract 1,000 mg

Appropriate amount of vitamin mixture

Vitamin A Acetate 70 μg

Vitamin E 1.0 mg

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

Vitamin B6 0.5 mg

Vitamin B12 0.2 μg

Vitamin C 10 mg

10 μg of biotin

Nicotinamide 1.7 mg

Folic acid 50 μg

Calcium Pantothenate 0.5 mg

Appropriate amount of mineral mixture

Ferrous sulfate 1.75 mg

Zinc Oxide 0.82 mg

Magnesium Carbonate 25.3 mg

Potassium Phosphate Monobasic 15 mg

Dibasic Calcium Phosphate 55 mg

Potassium citrate 90 mg

Calcium Carbonate 100 mg

Magnesium Chloride 24.8 mg

Although the composition ratio of the above vitamin and mineral mixture is a mixture of ingredients suitable for health functional food in a preferred embodiment, the mixing ratio may be arbitrarily modified, and the above ingredients are mixed according to the normal health functional food manufacturing method. Then, granules are prepared and can be used for preparing a health functional food composition according to a conventional method.

Formulation Example 7. Manufacturing of functional beverages

Dodu pod extract powder 1,000 mg

Citric Acid 1,000 mg

100 g of oligosaccharides

2 g plum concentrate

1 g of taurine

Add purified water to total 900 mL

After mixing the above ingredients according to the usual health drink manufacturing method, stirring and heating at 85 ° C. for about 1 hour, the resulting solution is filtered and collected in a sterilized 2 L container, sealed and sterilized, and then refrigerated. It is used for preparing the functional beverage composition of the present invention.

Although the composition ratio is a mixture of ingredients suitable for a relatively favorite beverage in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and ethnic preferences such as the class of demand, the country of demand, and the purpose of use.

Although the present invention has been described in terms of the preferred embodiments mentioned above, various modifications and variations are possible without departing from the spirit and scope of the invention. The appended claims also cover such modifications and variations as fall within the subject matter of this invention.

Claims (14)

A method for producing a Dodu pod extract with enhanced rutin content and functionality of Formula 1 below, comprising the step of extracting by adding a solvent to dried Dodu pod powder:
[Formula 1]

.
According to claim 1,
Characterized in that the functionalities are antioxidant activity and anti-inflammatory activity.
According to claim 1,
The method, characterized in that the solvent is at least one selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, and mixed solvents thereof.
According to claim 3,
The lower alcohol having 1 to 4 carbon atoms is at least one selected from the group consisting of methanol, ethanol, propanol, isopropanol, butanol and isobutanol.
According to claim 4,
The method, characterized in that the ethanol is 10% to 90% (v / v) fermented alcohol.
According to claim 1,
The method characterized in that the solvent is added in 500 to 5,000 parts by weight based on 100 parts by weight of the dry powder of soybean pods.
According to claim 1,
The extraction time is 1 to 100 hours, and the extraction temperature is 10 to 120 ° C.
According to claim 1,
Further comprising the step of filtering after centrifugation after the extraction step; The pore size of the filtration membrane in the filtration is characterized in that 0.01 to 10 ㎛, method.
According to claim 8,
Further comprising the step of drying and pulverizing the filtrate filtered in the filtration step; Characterized in that the powdering is carried out by a method selected from the group consisting of spray drying, freeze drying, vacuum drying and hot air drying methods.
An extract of Dodu pods with improved rutin content and functionality, prepared according to the method of any one of claims 1 to 9.
An antioxidant or anti-inflammatory health functional food comprising, as an active ingredient, a Dodu pod extract with enhanced rutin content and functionality, prepared according to the manufacturing method of any one of claims 1 to 9.
An antioxidant or anti-inflammatory cosmetic composition comprising, as an active ingredient, a Dodu pod extract with enhanced rutin content and functionality, prepared according to the manufacturing method of any one of claims 1 to 9.
An antioxidant and anti-inflammatory pharmaceutical composition comprising, as an active ingredient, a Dodu pod extract with enhanced rutin content and functionality, prepared according to the method of any one of claims 1 to 9.
An antioxidant and anti-inflammatory feed additive comprising, as an active ingredient, a Dodu pod extract with enhanced rutin content and functionality, prepared according to the manufacturing method of any one of claims 1 to 9.

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