KR20120096764A - Functional food composition for the oral purpose with anti-inflammatory activity and it's method of manufacturing - Google Patents

Abstract

PURPOSE: An oral purpose functional food composition having anti-inflammatory function, and a producing method thereof are provided to offer prevention and treating effects on inflammation diseases and joint inflammation. CONSTITUTION: A producing method of an oral purpose functional food composition comprises the following steps: crushing morus alba, and extracting the morus alba using 0-99% ethanol water solution at 30-80 deg C for 2-8 hours for 1-4 times, and cooling and filtering the obtained morus alba extract; decompress-concentrating the morus alba extract; freeze-drying the concentrate to obtain morus alba extract powder; washing glycine max with purified water, extracting the glycine max using 0-80% of alcohol water solution at 30-100 deg C for 2-8 hours for 1-4 times, and cooling the filtering the obtained glycine max extract; decompress-concentrating the glycine max extract, and freeze-drying the concentrate to obtain glycine max extract powder; washing onions without peeling with the purified water, and extracting the onion using 10-100% alcohol water solution at 60-100 deg C for 2-8 hours for 1-4 times, and cooling the filtering the obtained onion extract; decompress-concentrating the onion extract, and freeze-drying the concentrate to obtain onion extract powder; and mixing the morus alba extract powder, the glycine max extract powder, and the onion extract powder in a weight ratio of 1:0.1-5;0.1-50.

Description

Functional food composition for anti-inflammatory oral administration and method for producing the functional food composition {Functional food composition for the oral purpose with anti-inflammatory activity and it's method of manufacturing}

The present invention relates to a functional food composition for oral administration and to a method for preparing the functional food composition for oral administration, in particular, a method for extracting and purifying active ingredients from onions, lettuce and frost, and an anti-inflammatory oral containing the extracts. A functional food composition for administration.

The present invention is a mixture of onion extract, lettuce extract, and frost extract in an appropriate ratio analgesic effect, inhibitory effect on the production of inflammation-causing factors (nitrogen monoxide, prostaglandin), inflammation-induced cytokines (TNF-α, IL-1β, IL-6) Inhibits production, inhibits the activity of inflammation-related enzymes (COX2, iNOS, Lipoxygenase, Type I Collagenase), inhibits the expression of inflammation-related enzymes (iNOS, COX-2), antioxidants (DPPH scavenging, H2O2 scavenging, superoxide radical scavenging) The present invention relates to a functional food composition for anti-inflammatory oral administration for anti-inflammatory analgesics, arthritis prevention and symptom ameliorating agents.

In general, onion (Allium cepa L.) is also called yanggun and can be eaten as a root vegetable in the classification of raw materials of food industry. Compared to other root vegetables, it contains more carbohydrates, proteins, vitamins and minerals (potassium, iron, phosphorus, sodium, etc.), galactan, xylan, methyl pentose and hemicellulose ) Spicy ingredients include allyl n-propyl disulfide, allicin, and dihydrogen sulfide, which are vaporized or decomposed when heated to give 50 times sweeter sugar. To form a mercaptan (propylmercaptan). Quercetin (C15H10O7) is found in the skin of onions (including red onions or black onions), and is known to prevent fat loss and prevent high blood pressure. The leaves contain vitamin I 5,000 IU, vitamin C 45 mg, calcium 80 mg, magnesium 24 mg and potassium 220 mg in 100 g. In general, dihydrogen sulfide, mercaptan, alcohol, alcohol, disulfide, allyldisulfide, trisulfide, aldehyde, etc. Known as the main fragrance ingredient in. [References: 1. Primary Color Korea Plant Book, Kyohaksa Co., pp67 (1996) 2. The Natural Plant Book, Hyangmunsa, pp92 (1979) 3. The Primary Color Korea Plant Book, Academy Book, pp122 (1996)]

The mulberry is also the fruit of the mulberry tree (Morus alba L.), also called the lettuce, the mulberry, the nausea, and the black heart. It is an animal and plant that can be used as the main ingredient (fruit, leaf, sprig) in food festival. Are classified. Small fruits are gathered and consist of one lump, long round, 1 ~ 2㎝ long, 0.5 ~ 0.8㎝ in diameter. Yellow, magenta or dark purple with short stems.

In addition, according to the classical literature, such as herbal wood, small diameter, jinjanggi, main breakfast, etc., the nature is cold, the taste is sweet, there is no poison, quenches thirst, relaxes the five intestines, benefits diabetes and joints, If you eat long, you can forget hunger. The components of the loser include Albafuran A, Albafuran B, Albafuran C, Albanol A, Albanol B, Alboctalol, alpha-Pinene (+,-), Artecanin, Astragalin, Bergapten, beta-Amyrin, beta-Amyrin acetate, 3-beta- Hydroxystigmast-5-en-7-one, beta-Resorcylaldehyde, beta-Sitosterol, Betulinic acid Mulberrofuran T, omega-Hydroxymoracin N, Oxydihydroresveratrol, Oxyresveratrol (E-form), Quercetin, Rutin, Resorcinol, Resveratrol, Sanggen Con Sang Don Con , Sanggenon E, Sanggenon P, Scopoletin, Stigmasterol, Umbelliferone and Zeatin. [Reference: 1. Medicinal Plants of Korea, Kyohaksa Co., Ltd., pp73, (2000) 2. Ingredients and Utilization of Herbs, Ilwol Seokgak, pp180 (1976) 3. Collection of Herbal Medicines (Medicinal Herbs) Standards, Korea Medical Index, pp202 (1998) 4. Food Ingredients, Korea Food and Drug Administration, Ingredients-Mulberry Search Results]

On the other hand, Glycine max (L.) Merr. Is called black bean or black beans and is classified as bean in the food ingredient classification. Nutritional value per 100 g: energy 378 kcal, water 11.7 g, protein 34.3 g, lipid 18.1 g, sugar 26.5 g, fiber 4.0 g, ash 5.4 g, calcium 224 mg, phosphorus 629 mg, iron 7.8 mg, sodium 5 mg, potassium 1,539 mg, vitamin B1 0.34mg, vitamin B2 0.22mg, niacin 1.9mg, rich in unsaturated fatty acids (linoleic acid), vitamin E, potassium, etc., and bioactive substances include Amino acid (aspartic acid, glutamic acid), Carotene, Vitamin (B1, B2, B12), Isoflavone, Daidzin, Genistin, Choline, Organic acid (folic acid, pantothenic acid), Lecithin, Saponins (aglycon of soyasapogenol A, B, C, D, E).

The epidermis also contains delphinidin-3-o-β-D-glucoside, cyanidin-3-o-β-D-glucoside, and petuinidin-3-o-β-D-gluc-oside. According to the classical literature, Suritae has been used as a cough potion since ancient times because it has an effect of strengthening the bronchus and strengthening the mucous membrane of the intestine. Eating frosted sweet sugar with black sugar or honey is effective for bronchial or asthma as well as coughing. Boiled frosted water is good for each disease, stomach ulcer and kidney disease. Seoritae has a detoxifying effect, so in Oriental medicine, it is roasted and dried to make poisonous powder. [References: 1. Growth Characteristics and Composition of General Components of Soybean Sprouts and Soybean Varieties, Journal of the Soybean Research Association, 18th Annual Meeting, pp4 ~ 7 (1992) 2. Regional Variation of Seed Contents of Soybean High Protein System : pp157 ~ 162 (1993) 3. A Study on the Protein and Fat Content of Korean Soybeans, Han Yuji, 7: pp40 ~ 44 (1975) 4. Food Ingredients, Korea Food and Drug Administration, Raw Materials Name-Suritae Search Results]

Pain is an unpleasant sensation that the mechanism of stimulating the cerebral cortex and the systemic region through the afferent neuropathy from pain receptors to nerve fibers. Depending on the duration of pain, there are acute pain, subacute pain, recurrent pain, persistent cancer pain, chronic pain and chronic pain syndrome. Pain-related substances include prostaglandin, leukotriene, thromboxane and serotonin, in which arachidonic acid is produced by the cyclooxygenese and lipooxigenase enzymes. ), Catecholamines, protons, histamine, potassium ions, and the like.

In addition, pain therapy includes physical therapy (cold therapy, heat therapy, percutaneous electrical neurostimulation therapy, exercise therapy), drug therapy (painkillers, non-narcotic analgesics, nonsteroidal anti-inflammatory drugs, antidepressants), injection therapy (mainly local to the muscle pain area). Injections, but each treatment usually has side effects, making it difficult to expect adequate and complete treatment.

To date, the mechanism of inflammation is the release of histamine and kinin due to cell damage, resulting in vasodilation, increased capillary permeability, and aggregation of macrophages into inflammatory sites, resulting in increased blood flow, swelling, immune cells and antibodies at the site of infection. Movement, pain, and fever occur.

In addition to the peptide-inducing factor, inflammation-causing factors include prostaglandins, leukotrienes, lipid factors such as platelet activators, inflammatory synthase, free radicals such as NO (nitric oxide), cell adhesion molecules, immune system, and coagulation. Factors are involved. Currently, synthetic drugs such as ibuprofen, antihistamines, steroids, cortisones, immunosuppressants, and immunosuppressive drugs are used, but there are many side effects such as temporary or simple symptom relief, hypersensitivity reactions, and immune system deterioration. Fundamental treatment is difficult.

Degenerative arthritis, also called osteoarthritis (degenerative joint disease), is sometimes caused by trauma, disease, or malformations, but the majority of patients cause inflammation and pain mainly due to degenerative damage to cartilage that protects the joints. to be.

In addition, although aging generally increases the incidence of degenerative arthritis, sex, genetic factors, obesity, and lifestyle are also associated. In severe cases, degenerative arthritis is treated by arthroscopic removal of vitreous and synovial membranes, synovial resection, curettage, multiple perforation, arthroplasty and pharmacotherapy.

The exact cause of rheumatoid arthritis is not known yet, but autoimmunity is known as a major mechanism. Inflammation of the synoviium begins to spread to surrounding cartilage and bones, leading to deformation of the joints (primarily the fingers, wrists, toes, and joints of the elbows, shoulders, ankles, and knees as the disease progresses). In addition, complications such as subcutaneous nodules, vasculitis, pulmonary fibrosis, anemia and skin ulcers occur. Various drugs are used to treat the disease, but no drug has been developed to cure rheumatoid arthritis.

Among the drugs used for arthritis, synthetic drugs include anti-inflammatory analgesics such as aspirin and acetaminophen, non-steroidal anti-inflammatory drugs (NSAIDs) such as cerebrex and viox, and tumor necrosis factor blockers-rheumatoid arthritis. ), But it is impossible to cure the disease, and serious side effects of the digestive system and blood coagulation have been reported.

In addition, products such as joins tablets, adroda capsules, imoton tablets, seimi tablets, atrene tablets, glucosamine, and chondroitin sulfate are currently used in natural medicines, but the products are insignificant or have undergone rigorous clinical research to prove their effectiveness. rare. Indeed, taking glucosamine for the treatment of arthritis in Korea has become a social epidemic, but a study conducted by the US FDA to investigate the therapeutic effect of a large number of patients showed no effect at all.

In view of the above-mentioned problems, the present invention invents an inflammatory disease, painful disease and arthritis symptom improving agent using natural plant extracts, so that there is no concern of side effects compared to other synthetic drugs, arthritis more than simple symptom relief. It is an object of the present invention to provide a functional food composition for anti-inflammatory oral administration and a method for preparing the functional food composition for anti-inflammatory oral administration having an excellent effect on the improvement and prevention of fundamental symptoms.

In addition, if most of the anti-inflammatory and analgesics developed so far focused on inhibiting the activity of proinflammatory enzymes such as COX-2, Lipoxygenase, iNOS, the present invention not only inhibits the activity of these enzymes, but also in vivo It is an object of the present invention to provide a functional food composition for anti-inflammatory oral administration which aims at improving the root symptom of inflammation and pain by inventing a natural extract that also inhibits biosynthesis (expression).

In addition, the present invention by developing a substance that inhibits the activity of Type I collagenase and antioxidant activity to prevent the destruction of articular chondrocytes before and after the disease, degenerative of elderly people in the high-risk group of disease treatment and disease development Another object is to provide a functional food composition for anti-inflammatory oral administration that prevents the development of arthritis.

Functional food composition for anti-inflammatory oral administration of the present invention for achieving the above object contains a mixed plant composition of onion extract, lettuce extract and frost extract as a main component, prevention of inflammatory diseases and pain diseases and arthritis It is characterized by improving the symptoms.

The mixed plant composition is preferably to be mixed in the weight ratio of 1: 1 (0.1 ~ 5): (0.1 ~ 50) of the lettuce extract, frost extract and onion extract.

Functional food composition for anti-inflammatory oral administration of the present invention for achieving the above object, containing the plant composition mixed with the extract of the loser and frost extract as a main component, to prevent and symptomatic symptoms of inflammatory diseases and pain diseases and arthritis It is characterized by improving.

The mixed plant composition is preferably to be mixed in the weight ratio of 1: 1 (0.1 ~ 5) of the lettuce extract and frost extract.

Functional food composition for oral administration of the present invention for achieving the above object, by containing the plant composition mixed with the lettuce extract and onion extract as a main component, it is characterized in that the prevention and symptoms of inflammation, pain, arthritis .

The mixed plant composition is preferably to be mixed in the weight ratio of the lettuce extract and onion extract 1: 1: (0.1 ~ 50).

In addition, the functional food composition for anti-inflammatory oral administration of the present invention for achieving the above object contains a plant composition mixed with frost extract and onion extract as a main component, to prevent inflammation, pain, arthritis and improve symptoms It is characterized by.

The mixed plant composition is preferably such that the frost extract and onion extract is mixed in a weight ratio of 1: (0.1 ~ 50).

Functional food for oral administration to which the anti-inflammatory functional food composition is administered is preferably one of capsules, pills, granules, powders, candy, gum, tablets, beverages, and syrups.

The preparation method for preparing a lettuce extract among the extracts contained in the anti-inflammatory functional food composition of the present invention is 2 to 8 at 30 to 80 ° C. with an aqueous solution of 0 to 99% ethanol in a weight ratio of 4 to 10 times after grinding the mulberry. A first step of extraction, cooling and filtration for 1 to 4 times; A second step of concentrating the filtrate obtained in the first step at 60 to 100 ° C. under reduced pressure; And a third step of lyophilizing the extract obtained in the second step and preparing a powder extract, wherein the index material extracted through the process is set to 0.1 to 20%.

The preparation method for obtaining the frost extract in the extract contained in the functional food composition for anti-inflammatory oral administration of the present invention, after washing twice with purified water 30 to 30% by 0 ~ 80% alcoholic aqueous solution of 4 to 10 times the weight ratio. A first step of extraction, cooling and filtration at 100 ° C. for 2 to 8 hours, 1 to 4 times; A second step of concentrating the filtrate obtained in the first step at 60 ° C. under reduced pressure; And a third step of lyophilizing the extract obtained in the second step and preparing a powder extract, wherein the total saponin, which is an indicator material extracted from the process, is 0.2 to 10%. .

Preparation method for obtaining onion extract of the extract contained in the functional food composition of the present invention, after washing the onion twice with purified water without separating the skin peel 4 to 10 times the weight ratio of 10 to 100% alcoholic aqueous solution 60 to 100 A first step of extracting 1 to 4 times at 2 ° C. for 2 to 8 hours, cooling and filtration; A second step of concentrating the filtrate obtained in the first step at 60 ° C. under reduced pressure; And a third step of lyophilizing the extract obtained in the second step and preparing a powder extract, wherein the index material extracted from the process is quercetin (0.1).

According to the functional food composition for anti-inflammatory oral administration of the present invention, by using a composition of natural plant extracts showing synergistic effects, antioxidant activity (DPPH radical scavenging activity, SOD-like activity, H 2 O 2 scavenging activity), inflammatory enzyme (COX-2) , Lipoxygenase, iNOS, Type I collagenase) inhibits the activity and inhibits the expression in vivo, in vivo production of inflammation-causing substances (nitrogen monoxide, prostaglandin, TNF-α, IL-1β, IL-6), inhibitory inflammatory edema By producing an excellent functional plant composition exhibits a useful effect as a prophylactic and symptomatic agent for inflammatory diseases, painful diseases and arthritis.

1 is a graph showing the SC50 value in ppm concentration of 50% scavenging DPPH radicals of the mixed extracts prepared by Reference Examples 1 to 3 and Example 1 of the present invention.
Figure 2 is a graph showing the SC50 value showing the SOD-like activity of the mixed extract prepared by Reference Examples 1 to 3 and Example 1 in a concentration of ppm.
Figure 3 is a graph showing the SC50 value in ppm concentration of the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention 50% H2O2.
Figure 4 is a graph showing that the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inhibits the production of nitrogen monoxide induced by LPS in mouse macrophages.
Figure 5 is a graph showing the IC50 value in ppm concentration of the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inhibits COX-1 enzyme by 50%.
Figure 6 is a graph showing the IC50 value in ppm concentration of the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inhibits COX-2 enzyme by 50%.
Figure 7 is a mixed extract prepared according to Reference Examples 1 to 3 and Example 1 of the present invention, the group treated with lipooxygenase enzyme activity in the blood of chronic animal model of chronic inflammation by MIA (mono sodium iodoacetate) A graph showing the rate of inhibition in comparison with.
8 is a graph showing the amount of production of TNF-α in the foot portion where the edema of the acute inflammatory animal model caused by carrageenan occurred in the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention.
9 is a graph showing the amount of production of TNF-α in the knee joint site of the mixed extract prepared according to Reference Examples 1 to 3 and Example 1 of the present invention chronic inflammation induced animal model by MIA.
10 is a graph showing the amount of production of IL-1β in the foot portion of the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention caused carrageenan swelling.
Figure 11 is a graph showing the amount of IL-1β production in the knee joint site where the mixed extract prepared according to Reference Examples 1 to 3 and Example 1 of the present invention inflamed chronic animal model by MIA.
12 is a graph showing the amount of IL-6 produced in the foot portion of the mixed extract prepared according to Reference Examples 1 to 3 and Example 1 of the present invention caused by carrageenan swelling animal model.
Figure 13 is a graph showing the amount of IL-6 produced in the knee joint site where the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inflamed chronic animal model by MIA.
14 is a graph showing that the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inhibits the production of prostaglandin induced by LPS in mouse macrophages.
15 is a graph showing that the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inhibits the production of prostaglandin in the knee joint site where inflammation of the chronic inflammation animal model by MIA occurred.
Figure 16 is a graph showing the IC50 value in ppm concentration of the mixed extract prepared by Reference Examples 1-3 and Example 1 of the present invention inhibits Type I collagenase by 50%.
17 is a mixture extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention to inhibit the expression of iNOS, COX-1, COX-2 enzymes in the foot area where edema of acute inflammatory animal model by carrageenan occurred (Ibu: Ibuprofen, Thread 1-100: Example 1 100 mg / weight Kg treatment, Thread 1-400: Example 1 400 mg / weight Kg, Joi: Joining tablet 400 mg / weight 1 Kg treatment)
18 is a mixture extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inhibits the expression of iNOS, COX-1, COX-2 enzymes induced expression in the inflammatory response of mouse macrophages by LPS (Ibu: Ibuprofen, Thread 1-200: Example 1 200ppm treatment, Thread 1-1K: Example 1 1000ppm treatment, Joi: Joining tablet 1000ppm treatment)
19 is a view showing the effect of inhibiting the paw edema of the animal model of acute inflammation caused by carrageenan of the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention.

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

The present invention is mixed with onion extract, lettuce extract and frost extract in an appropriate ratio, by optimizing in a predetermined amount of analgesic effect than each extract, Type I collagenase, lipoxygenase, COX-1 and COX- 2 inhibitory effect, inflammatory edema alleviation effect, inhibitory effect of production of inflammatory factors nitrogen monoxide (NO) and prostaglandins (inhibition effect of in vivo expression of inflammatory enzymes iNOS, COX-1 and COX-2, inflammation Inflammatory disease that is more prominent by inhibiting the production of the induced cytokines TNF-α, IL-1β and IL-6, as a potential inducer of arthritis, and having a radical free radical scavenging effect and an antioxidant effect that are considered as a serious factor of the disease The present invention relates to a functional food composition for anti-inflammatory oral administration useful for the prevention and improvement of symptoms of painful diseases and arthritis.

Such a composite food composition was used to prepare the composition by extracting onion (including red onions or black onions), lettuce and frosted with water or alcoholic aqueous solution, respectively.

1) First, the method of preparing the composition by extracting the onion is as follows.

After washing the onion twice with purified water without separating the skin, grind it for 10 to 20 minutes at 500 to 700 rpm using a food grinder, and then use it for 4 to 10 times at 10 to 100% ethanol aqueous solution at 60 to 100 ° C for 2 to 8 hours. Extraction, cooling 1 to 4 times, and a first step of filtration with a stainless steel strainer of 0.125mm (80mesh) specifications. The second step of concentrating the filtrate obtained in the first step to 60 brix at 60 ° C. under reduced pressure. The extract obtained in the second step is lyophilized and a third step of preparing a powder extract is performed.

In particular, the onion powder extract obtained by the above method contains quercetin in an amount of usually 0.1 to 40% by weight.

2) Next, the method of preparing a composition by extracting the loser is as follows.

After grinding the heartache at 500 ~ 1000rpm using a food grinder, extract 4 ~ 10 times with 0 ~ 99% ethanol aqueous solution at 30 ~ 80 ℃ for 2 ~ 8 hours, 1 ~ 4 times, and then cool it. A first step of filtration is carried out using a stainless steel strainer of mesh size.

The second step of concentrating the filtrate obtained in the first step under reduced pressure to 20brix at 20 ~ 100 ℃. The extract obtained in the second step is lyophilized and a third step of preparing a powder extract is performed.

The core powder extract obtained by the above method contains, in particular, rutin in an amount of 0.01 to 20% by weight.

3) Next, the method of preparing the composition by extracting the frost is as follows.

After washing twice with purified water, extracted 2-4 times at 30-100 ℃ for 2 ~ 4 hours with 4 ~ 10 times of water or 5 ~ 80% aqueous ethanol solution, and cooled it to 0.074mm (200 mesh) standard. Perform the first step of filtration with a stainless steel strainer.

The second step of concentrating the filtrate obtained in the first step under reduced pressure up to 15brix at 60 ° C. The extract obtained in the second step is lyophilized and a third step of preparing a powder extract is performed.

The frosted powder extract obtained by the above method contains, in particular, 0.2 to 10% by weight of saponin.

In the above 1) to 3) process of the onion extract, lettuce extract and frost extract is mixed in a weight ratio of 1: 0.2 ~ 1 is contained in the rutin 0.01 to 10% by weight.

For reference, the origin of the onion used in the present invention is Changnyeong, Gyeongsangbuk-do, and the native species was used, and the origin of the heartache was harvested in early summer, Gochang, Jeollabuk-do. Harvest was used. However, the scope of the present invention is not limited because of the origin or variety of each plant.

The plant extract may be used as such, but food-acceptable moisture absorbents, excipients, forming agents, diluents, carriers, and the like, for preparing powders, tablets, granules, or gels may be used. Can be mixed together.

In addition, the present invention is an inflammatory disease comprising as a main ingredient a combination selected from the group consisting of onions, lettuce and frost extract, obtained from the above method, and a combination selected from the group consisting of a vitamin group and quercetin, etc. Provided are foods containing other food ingredients in addition to functional compositions for painful diseases and arthritis.

The food includes beverages, special nutritional supplements, health supplements, functional foods and other foods.

Meanwhile, the form of the food includes powder, granule, tablet, capsule, liquid or beverage form.

In addition, the present invention, the food additives in the food, such as fungicides, spices, seasonings, various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, such as colorants and neutralizing agents (cheese, chocolate, etc.) ), Pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, or the like as essential raw materials for food materials It provides a method of using a food additive characterized in that. At this time, the food additive may be added to the food by dipping, spraying or mixing the food, the ratio of these additives is not so important but selected from the range of 0 to about 20% by weight per 100% by weight of the functional composition of the present invention Is common.

It may also be added to foods or beverages for the purpose of preventing and improving symptoms of inflammatory diseases, painful diseases and arthritis. In this case, the amount of the composition in the food or beverage is generally 10 to 80% by weight, preferably 20 to 70% by weight of the total food weight in the case of the health functional food composition of the present invention, 0.1? Based on 100 ml 30 g, preferably 1? It can be added at a rate of 30 g.

Hereinafter, the present invention will be described in detail with reference to the following examples, but the scope of the present invention is not limited by the following examples.

Reference Example 1 Preparation of Onion Extract

Grind the onion for 10-20 minutes at 500 ~ 700rpm using a food grinder, extract and cool 2 to 8 hours, 1 to 4 times at 60 to 100 ℃ in 10 to 100% aqueous ethanol solution of 4 to 10 times, 0.125 After filtering with a stainless steel strainer of mm (80mesh) standard, the filtrate obtained here is concentrated under reduced pressure to 20brix at 60 ~ 100 ℃. The extract obtained in the concentration step is lyophilized to prepare a powder extract.

REFERENCE EXAMPLE 2 Preparation of a Lettuce Extract

The fresh heart washed with purified water was crushed for 20 to 60 minutes at 500 to 1000 rpm using a food grinder, and then extracted 4 to 10 times at 0 to 35% aqueous ethanol solution for 2 to 8 hours and 1 to 4 times at 50 to 60 ° C. After cooling and filtering with a stainless steel strainer of 0.125mm (80mesh) standard, the filtrate obtained here is concentrated under reduced pressure to 20brix at 60 ~ 100 ℃. The extract obtained in the concentration step is lyophilized to prepare a powder extract.

Reference Example 3: Preparation of Seoritae Extract

The frosted washed with purified water was extracted and cooled 2-4 times at 60-100 ℃ for 2 ~ 4 hours with 4 ~ 10 times of water or 20 ~ 50% ethanol aqueous solution, and filtered by 0.074mm (200mesh) stainless steel strainer. And the filtrate obtained here is concentrated under reduced pressure at 60 ℃ to 15brix. The extract obtained in the concentration step is lyophilized to prepare a powder extract.

Example 1: Preparation of Mixed Extract

The onion extract obtained from Reference Example 1, Reference Example 2, and Reference Example 3, and the extract from the lettuce and the frost extract were mixed using a powder mixer in a 5: 5: 1.2 weight ratio.

Experimental Example 1 DPPH radical scavenging activity

DPPH (2,2-Diphenyl-1-picrylhydrazyl, Sigma, MO, USA), a very stable free radical, was used to determine the electron donating ability of each sample by using a hydrogen proton-radical scavenger that discolors purple to yellow diphenylpicrylhydrazine. It was. The results are as shown in FIG. 1 and Table 1.

1 is a graph showing the SC50 value in ppm concentration of 50% scavenging DPPH radicals of the mixed extracts prepared according to Reference Examples 1 to 3 and Example 1 of the present invention.

Experimental Method

After dissolving each sample by concentration, 75 ml of each was mixed with 750 ml of DPPH (0.2 mM in ethanol) and 675 ml of tertiary purified water. After reacting for 30 minutes at room temperature, 200 ml of the reaction solution was transferred to a 96 well plate, and the absorbance was measured at a wavelength of 520 nm with an ELISA reader device (Molecular Devices, CA, USA). The DPPH radical scavenging activity (%) was calculated using the formula ((absorbance of 1-sample added / absorbed sample-free)) X100. The SC50 value represents the minimum concentration, in ppm, required to eliminate 50% of the generated radicals.

TABLE 1

As shown in Table 1 and Figure 1, it can be seen that the scavenging ability of the DPPH radical for Example 1 prepared by the present invention is superior to the case of using each extract or the joining tablet of SK Pharmaceuticals. . Vitamin C was used as a positive control.

Experimental Example 2: SOD-like activity experiment

The scavenging activity of oxygen radicals liberated from xanthine by xanthine oxidase was compared to determine the scavenging activity of the superoxide radical of each plant extract composition. As shown in FIG. 2 and <Table 2> which were attached.

Figure 2 is a graph showing the SC50 value in ppm concentrations showing the SOD-like activity of the mixed extract prepared according to Reference Examples 1 to 3 and Example 1 of the present invention.

Experimental Method

A coloring reagent consisting of xanthine solution (0.4mM) and NBT solution (0.24mM, nitro blue tetrazolium) solution in xanthine oxidase solution (0.045U / ml) dissolved in buffer solution (0.1M phosphate buffer, pH 8.0) at 37 ℃ After adding the sample solution dissolved in each concentration, the reaction was stopped for 20 minutes, and the reaction was stopped by the addition of 1 ml of SDS (70 mM) solution, and then the degree of discoloration due to oxidation of cytochrome-c was measured at 560 nm.

<Table 2>

As shown in Table 2 and Figure 2, it can be seen that the SOD-like activity for Example 1 prepared by the present invention is superior to the case of using the respective extracts or using the joining tablet of SK Pharmaceutical. Vitamin C was used as a positive control.

Experimental Example 3 H2O2 Scavenging Capacity Experiment

In general, the Fenton reaction is used to measure the antioxidant activity of converting hydrogen peroxide, a product generated after the decomposition of superoxide dismutase (SOD) into safer water and oxygen. The hydroxyl radical was measured by the degree of absorption of MDA (malondialdehyde) generated by oxidizing 2-deoxyribose at 520nm and the results are shown in Figure 3 and Table 3.

Figure 3 is a graph showing the SC50 value in ppm concentration of the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention to remove H2O2 by 50%.

Experimental Method

To 10 ml of FeSO4-7H2O (50mM) -EDTA (50mM) solution and 10ml of 2-deoxyribose (50mM) solution, add 20 ml of each sample dissolved in each concentration, and then add 0.2 ml sodium phosphate buffer (pH 7.0). ) 30 ml, 10 ml of H2O2 (50 mM) and 20 ml of distilled water were added and reacted at 37 ° C. for 4 hours. Then, 100 ml of 2.8% trichloroacetic acid and 100 ml of 1% 2-thiobarbituric acid dissolved in NaOH (50 mM) solution were added. After heating and rapid cooling, the absorbance was measured at 530 nm using a spectrophotometer. Hydroxy radical scavenging activity (%) was calculated by the formula (1-absorbance of sample added / absorbance of sample free) X 100.

<Table 3>

As shown in Table 3 and Figure 3, it can be seen that the H 2 O 2 scavenging ability for Example 1 prepared by the present invention is superior to the case of using each extract or the joining tablet of SK Pharmaceutical. Vitamin C was used as a positive control.

Experimental Example 4: LPS-induced NO production inhibition experiment

The effect of inhibiting the production of NO (nitric oxide), which is an inflammation-causing substance, was measured by Griess reaction of nitric oxide concentration in culture medium produced by treating LPS (lipopolysaccharide) on mouse macrophages. The results are shown in FIG. 4 and <Table 4>.

4 is a graph showing that the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inhibits the production of nitric oxide induced in LPS in mouse macrophages.

Experimental Method

NO concentration was measured by Griess reaction of nitric oxide concentration in the culture. First, dispensing RAW 264.7 cells derived from 1 × 10 4 cells / well of mouse macrophages into a 96 well culture plate using DMEM medium containing FBS and antibiotics, and then 24 hours at 37 ° C. and a 5% CO 2 incubator. Incubated for After 24 hours, the medium used for the previous culture was removed and fresh DMEM medium containing no FBS and antibiotics was dispensed and the plant composition prepared in Example 1 was treated by concentration (mg / mL). After 1 hour, 1 mg / mL LPS was treated and incubated for 24 hours. NO produced during the cultivation was measured by the total concentration of NO2- existing in the cell culture medium using the Griess reagent, which was reacted for 10 minutes in a 96 well plate by mixing 50 μL of the cell culture supernatant and 50 μL of the Griess reagent. After absorbance was measured at 540nm using an ELISA reader. Nitric oxide scavenging ability (%) was expressed using the formula ((absorbance of the reaction group without the 1-sample / absorbance of the control group without the sample) × 100.

TABLE 4

As shown in Table 4 and Figure 4, the use of the plant extract composition prepared according to the present invention was found to significantly inhibit the production of NO, one of the LPS-induced inflammatory substances in mouse macrophages. In particular, when Example 1 was treated, NO production was found to be excellent.

Experimental Example 5: Inhibition of activity of COX-1 and COX-2

To determine the efficacy of each sample to inhibit the activity of the pro-inflammatory enzymes COX-1 (cyclooxygenase-1) or COX-2 (cyclooxygenase-2), the substrates of these enzymes, using arachidonic acid, The phenomenon of discoloration by using oxidation was used, and the results are shown in FIGS. 5 to 6 and Table 5.

Figure 5 is a graph showing the IC50 value in ppm concentration of the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inhibits COX-1 enzyme by 50%,

Figure 6 is a graph showing the IC50 value in ppm concentration of the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inhibits COX-2 enzyme by 50%.

Experimental Method

COX-1 or COX-2 inhibitory effect was measured in a 100-well Tris-HCl buffer (40 μl) with 60 units / ml of COX-1 or 30 units / ml of COX-2 per well in a 96 well plate. pH 8.0) 90μl, 30μM EDTA 20μL, 150μM hematin 20μl and sample 20μl were mixed and reacted at 25 ° C for 5 minutes, 5mM TMPD 5μl and 20mM arachidonic acid were added for 5 minutes at 25 ° C. Absorbance was measured at 590 nm. The percentage inhibition of COX-1 or COX-2 was expressed as a 50% inhibitory concentration (IC50) using the equation COX inhibition rate (%) = [(absorbance of the control group-absorbance of the experimental group) / absorbance of the control group] × 100

<Table 5>

As shown in Table 5 and Figures 5 to 6, the use of the plant extract composition prepared according to the present invention was found to significantly inhibit the activity of inflammation-related enzymes COX-1 and COX-2, particularly In case of treatment of Example 1, the inhibition of COX-1 and COX-2 activity was very good.

Experimental Example 6 Manufacture of MIA (Mono sodium iodoacetate) induced degenerative arthritis animal model

Intra-articular injection of MIA into SD rats inhibits metabolism of chondrocytes, causing cartilage, ligament, and tendon damage to osteoarthritis Was used to generate the principle. The results are as shown in Table 6.

Experimental Method

Inject 50 ml of MIA (40mg / ml) using a 1ml syringe with a 27 gauge needle into the joint cavity of SD rats weighing more than 300g. Each sample, 10 saline, or Ibuprofen The dose was administered orally at daily doses for 14 days.

<Table 6>

Experimental Example 7: Lipooxygenase (LOs, Lipoxygenases) inhibitory activity

Lipogenasease using Lipoxygenase Inhibitor Screening Assay Kit (Cayman Chemical Com, MI, USA) to measure the efficacy of each sample to inhibit lipooxygenase, an inflammation and pain-causing enzyme, in a degenerative arthritis animal model using MIA. The principle of measuring the amount of hydrogen peroxide as a result of the lipoxygenation reaction by using was used, the results are shown in Figure 7 and Table 7.

Figure 7 shows the ratio of the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inhibits Lipoxygenase enzyme activity in the blood of chronic inflammatory animal model by MIA compared to the untreated group of samples. It is a graph figure.

Experimental Method

After 10 μl of sample, 90 μl of 5-LO (220 units / ml) and 10 μl of 1 mM linoleic acid were reacted at room temperature for 5 minutes, and then 100 μl of chromogen was added for 5 minutes at room temperature, followed by 490 nm wavelength using an ELISA reader. Absorbance was measured at. The results were expressed as 50% inhibitory concentration (IC50) and the 5-LO inhibition rate (%) was expressed as IC50 using [(absorbance of control-absorbance of control group) / absorbance of control group] × 100.

<Table 7>

As shown in Table 7 and Figure 7, when using the plant extract composition prepared according to the present invention was found that the activity of the inflammation-related enzyme lipooxygenase is inhibited, especially in the case of treating Example 1 Showed a very good inhibition of the activity of lipooxygenase. In addition, Ibuprofen, which was used as a positive control, did not appear to affect the inhibition of lipooxygenase activity.

Experimental Example 8: TNF-α production inhibition experiment

To evaluate the efficacy of each sample in reducing the expression of TNF-α, an inflammation-inducing factor, the TNF-α sandwich ELISA kit (eBiosience, Vienna, Austria) was studied in carrageenan-induced edema and MIA-induced degenerative arthritis ), And the results are shown in FIGS. 8-9 and <Table 8-9>.

FIG. 8 is a graph showing the amount of TNF-α production in the foot region in which the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention caused swelling of an acute inflammatory animal model caused by carrageenan.

9 is a graph showing the amount of production of TNF-α in the knee joint where the mixed extract prepared according to Reference Examples 1 to 3 and Example 1 of the present invention caused edema of a chronic inflammation animal model by MIA.

Experimental Method

Each sample was sacrificed by carrageenan-induced edema animal model and MIA-induced degenerative arthritis animal model, which were orally administered once daily at the period and dose shown in Tables 8 to 9, and blood and plasma were separated. The concentration of -α was measured by the method suggested by the manufacturer of the ELISA kit. First, TNF-α in plasma was bound to anti-rat TNF-α antibody coated on a 96 well plate, and each well was washed five times, followed by biotin-conjugated anti-rat TNF-α. After the reaction of the antibody, each well was washed five times, treated with streptavidin-HRP, and finally colored with tetramethyl-benzidine, and the absorbance was measured at 450 nm using an ELISA reader. The amount of TNF-α was calculated arithmetic proportionally using the response standard curve of the standard recombinant TNF-α included in the ELISA kit.

<Table 8>

<Table 9>

As shown in Tables 8 to 9 and 8 to 9, TNF is an inflammation-related factor when the plant extract composition prepared by the present invention is used in an acute inflammatory animal model by carrageenan and a chronic inflammatory animal model by MIA. In vivo production of -α was suppressed. In particular, when Example 1 was treated, it was found that the inhibition of in vivo production of TNF-α was excellent. In addition, Ibuprofen used as a positive control was found to have only a slight effect on the in vivo production of TNF-α.

Experimental Example 9 IL-1β Production Inhibition Experiment

To evaluate the efficacy of each sample to reduce the in vivo expression of IL-1β, an inflammation-inducing factor, the IL-1β sandwich ELISA kit (eBiosience, Vienna, Austria) was studied in carrageenan-induced edema and MIA-induced degenerative arthritis. ), And the results are shown in FIGS. 10 to 11 and <Table 10 to 11>.

FIG. 10 is a graph showing the amount of IL-1β production in the foot region in which the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention caused swelling of an acute inflammatory animal model caused by carrageenan,

11 is a graph showing the amount of IL-1β production in the knee joint where the mixed extract prepared according to Reference Examples 1 to 3 and Example 1 of the present invention caused edema of a chronic inflammation animal model by MIA.

 Experimental Method

IL-1β present in plasma after sacrificing blood collection and plasma separation after sacrificing carrageenan-induced edema animal model and MIA-induced degenerative arthritis animal model orally administered once daily with the periods and doses shown in Tables 10-11. The concentration of was measured by the method suggested by the manufacturer of the ELISA kit. First, IL-1β in plasma is bound to anti-rat IL-1β antibody coated on a 96 well plate, and each well is washed five times, followed by biotin-conjugated anti-rat IL-1β. After the reaction of the antibody, each well was washed five times, treated with streptavidin-HRP, and finally colored with tetramethyl-benzidine, and the absorbance was measured at 450 nm using an ELISA reader. The amount of IL-1β was calculated arithmetic proportionally using the standard reaction curve of the standard recombinant IL-1β included in the ELISA kit.

<Table 10>

<Table 11>

As shown in Tables 10 to 11 and FIGS. 10 to 11, IL-related inflammation factors when the plant extract composition prepared according to the present invention is used in an acute inflammatory animal model by carrageenan and a chronic inflammatory animal model by MIA It was found that the in vivo production of -1β was suppressed. In particular, when Example 1 was treated, the inhibition of in vivo production of IL-1β was very excellent. In addition, Ibuprofen, used as a positive control, was found to have only a minor effect on the in vivo production of IL-1β.

Experimental Example 10 IL-6 Production Inhibition Experiment

IL-6 sandwich ELISA kit (eBiosience, Vienna, Austria) for carrageenan-induced edema and MIA-induced degenerative arthritis animal models ), And the results are shown in FIGS. 12 to 13 and <Table 12 to 13>.

12 is a graph showing the amount of IL-6 production in the foot region where edema of acute inflammatory animal model caused by carrageenan occurred in the mixed extract prepared according to Reference Examples 1 to 3 and Example 1 of the present invention.

FIG. 13 is a graph showing the amount of IL-6 production in the knee joint where the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention caused edema of a chronic inflammation animal model by MIA.

Experimental Method

IL was present in plasma after sacrificing blood collection and plasma separation at the expense of carrageenan-induced edema animal model and MIA-induced degenerative arthritis animal model, which were orally administered once daily at the periods and doses shown in Tables 12-13. The concentration of -6 was measured by the method suggested by the manufacturer of the ELISA kit.

First, IL-6 in plasma was bound to anti-rat IL-6 antibody coated on a 96 well plate, and each well was washed five times, followed by biotin-conjugated anti-rat IL-6. After reacting the antibody, streptavidin-HRP was treated, and finally, tetramethyl-benzidine was developed and the absorbance was measured at 450 nm using an ELISA reader. The amount of IL-6 was calculated arithmetic proportionally using the standard reaction curve of the standard recombinant IL-6 included in the ELISA kit.

<Table 12>

TABLE 13

As shown in Tables 12 to 13 and Figures 12 to 13, IL is an inflammation-related factor when a plant extract composition prepared by the present invention is used in an acute inflammatory animal model by carrageenan and a chronic inflammatory animal model by MIA. In vivo production of -6 was found to be suppressed. In particular, when Example 1 was treated, it was found that the inhibition of in vivo production of IL-6 was very excellent. In addition, Ibuprofen, used as a positive control, was found to have only a minor effect on the in vivo production of IL-6.

Experimental Example 11: Prostaglandin (PGs, Prostaglandins) production inhibition experiment

To evaluate the efficacy of each sample to inhibit the production of prostaglandins, inflammation and pain-causing factors, in inflammatory cell models using LPS and animal models of degenerative arthritis using MIA, Ptostaglandin Screening EIa Kit (Cayman Chemical Co., MI, USA) using the principle of prostaglandin and tracer (prostaglandin artificially coupled to acetylcholinesterase) competitively binds to the anti-PGs-antibody, the results are shown in Figures 14-15 and Tables 14-15 As shown.

14 is a graph showing that the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inhibits the production of prostaglandin induced by LPS in mouse macrophages,

15 is a graph showing that the mixed extract prepared according to Reference Examples 1 to 3 and Example 1 of the present invention inhibits the production of prostaglandin in the knee joint site where edema of a chronic inflammation animal model by MIA occurred.

Experimental Method

RAW 264.7 cells derived from mouse macrophages were cultured in a 96 well culture plate using DMEM medium containing FBS and antibiotics for 1 hour after dispensing 1 × 10 4 cells / well in 37 ° C., 5% CO 2 incubator for 24 hours. It was. After 24 hours, the medium used for the previous culture was removed and fresh DMEM medium containing no FBS and antibiotics was dispensed and each sample was treated by concentration. After 1 hour, 1 mg / mL LPS was treated and incubated for 24 hours. Prostaglandin AChE Tracer and antiserum were added to a 96-well plate coated with anti-PGs antibody, and 50 μl of standard prostaglandin or each sample-treated medium was added and reacted at room temperature for 18 hours. The wells are washed five times, 200 ml of Ellman's Reagent is added and developed for 70 minutes, and the absorbance is measured at a wavelength of 420 nm using an ELISA reader. The amount of prostaglandin was arithmetic proportionally calculated using the standard curve included in the ELISA kit.

TABLE 14

TABLE 15

As shown in Tables 14 to 15, when the plant extract composition prepared according to the present invention is used, the production of prostaglandin, which is one of LPS-induced inflammatory substances in mouse macrophages, is inhibited. In Example 1, the inhibition of prostaglandin production was found to be excellent.

Experimental Example 12: Type I Collagenase Inhibition Experiment

PZ-peptide (4-phenylazobenzyloxycarbonyl-Pro-Lue-Gly-), a substrate of recombinant Collagenase enzyme (Sigma, MO, USA), was used to examine the effect of each sample on the activity of Type I Collagenase, which has a major adverse effect on articular cartilage health. Pro-D-Arg, Fluka Chemie GmbH, Buchs, Switzerland) was measured using the discoloration phenomenon when decomposing the results are shown in Figure 16 and Table 16.

Figure 16 is a graph showing the IC50 value of the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inhibiting Type I collagenase 50% by the concentration of ppm.

Experimental Method

After dissolving collagenase enzyme at a concentration of 0.2mg / ml and diluting PZ-peptide in buffer at a concentration of 0.3mg / ml, vortex after mixing 100ml of sample by concentration, 150ml of collagenase and 250ml of PZ-peptide, and vortex After reacting for 30 minutes, 0.5 ml of citric acid (25 mM) is added, ethyl acetate 1.5 ml is added, and then reacted at room temperature for 10 minutes. Absorbance is measured at 320 nm using a quartz cuvette.

TABLE 16

As shown in Table 16 and Figure 16, it can be seen that Type I Collagenase inhibitory ability for Example 1 prepared by the present invention is superior to the case of using each extract or the joining tablet of SK Pharmaceuticals. .

Experimental Example 13: In vivo expression inhibition experiment of iNOS, COX-1, COX-2

Western blot was used to determine the efficacy of each sample to inhibit the in vivo expression of pro-inflammatory enzymes iNOS, COX-1, and COX-2 in inflammatory cell models using LPS and acute inflammatory animal models using carrageenan. The results are as shown in Figs. 17-18.

17 is a mixture extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention to inhibit the expression of iNOS, COX-1, COX-2 enzymes in the foot area where edema of acute inflammatory animal model by carrageenan occurred (Ibu: Ibuprofen, Thread 1-100: Example 1 100 mg / weight Kg treatment, Thread 1-400: Example 1 400 mg / weight Kg, Joi: Joining tablet 400 mg / weight Kg treatment),

18 is a mixture extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention inhibits the expression of iNOS, COX-1, COX-2 enzymes induced expression in the inflammatory response of mouse macrophages by LPS (Ibu: Ibuprofen, Thread 1-200: Example 1 200ppm treatment, Thread 1-1K: Example 1 1000ppm treatment, Joi: Joining tablet 1000ppm treatment)

Experimental Method

As shown in <Table 17 ~ 18>, the protein expression level caused by the inflammatory response was prepared by carrageenan-induced acute inflammation animal model and LPS-induced inflammatory cell model. Quantify the total protein collected, electrophoresis (SDS-PAGE) of 20 mg total protein per well, transfer to nitro cellulose membrane, and then iNOS (Santa Cruz Biotechnology, CA, USA), COX- 1 (Cell Signaling Technology, MA, USA), COX-2 (Cell Signaling Technology, MA, USA) primary and secondary antibodies (Santa Cruz Biotechnology, CA, USA) and enhanced chemiluminescence (ECL, Pierce, IL, USA) was expressed on the X-ray film and the identity of the total protein was verified by the amount of β-actin (Santa Cruz Biotechnology, CA, USA).

TABLE 17

TABLE 18

As shown in Tables 17 to 18 and 17 to 18, it can be seen that the expression of iNOS, COX-2, which is an inflammation-related enzyme for Example 1 prepared by the present invention, was inhibited. Ibuprofen also inhibited the expression of iNOS but did not affect the expression of COX-1 and COX-2.

Experimental Example 14 edema inhibition experiment for carrageenan-induced acute inflammation

In the carrageenan-induced acute inflammatory response by the plant composition prepared in Example 1, the degree of inhibition of foot edema was observed by time period compared to the control group, and the results are shown in Table 17 and FIG. 19.

19 is a view showing the effect of inhibiting the paw edema of the animal model of acute inflammation caused by carrageenan of the mixed extract prepared by Reference Examples 1 to 3 and Example 1 of the present invention.

Experimental Method

Spraque-Dqwley (SD) rats were inoculated with oral administration at the concentration of 10 in the following <Table 19>, and 1% carrageenan-dissolved saline solution was injected into the left plantar portion in an amount of 100 ml to induce foot edema. Then, the degree of foot edema was measured at each hour for 6 hours using a Digimatic micrometer (Mitutoyo Corporation, Japan).

TABLE 19

Preparation Example: Although the preparation example of the food or health functional food containing the extract of the present invention will be described, the present invention is not intended to limit this, but is intended to explain in detail.

Preparation Example 1 Preparation of Powder

300 mg of functional composition of Example 1, 100 mg of lactose, 10 mg of talc

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Preparation Example 2 Preparation of Tablet

50 mg of functional composition of Example 1, 100 mg of corn starch, 100 mg of lactose, 2 mg of magnesium stearate

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Preparation Example 3 Preparation of Capsule

50 mg of functional composition of Example 1, 100 mg of corn starch, 100 mg of lactose, 2 mg of magnesium stearate

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.

Preparation Example 4 Preparation of Liquid

100 mg of functional composition of Example 1, 10 g of isomerized sugar, 5 g of mannitol, appropriate amount of purified water

According to the conventional method of preparing a liquid solution, each component is added and dissolved in purified water, lemon flavor is added to the appropriate amount, the above components are mixed, purified water is added, and the whole is adjusted to 100 ml by adding purified water to a brown bottle. Fill and sterilize to prepare a liquid.

Preparation Example 5 Preparation of Health Functional Food

1000 mg of the functional composition of Example 1, vitamin mixture proper amount, 70 μg of vitamin A acetate, 1.0 mg of vitamin E, vitamin B10 13 mg, vitamin B20 15 mg, vitamin B6 0.5 mg, vitamin B12 0.2 μg, vitamin C 10 mg, biotin 10 μg, nicotinic acid amide 1.7 mg, folic acid 50 μg, calcium pantothenate 0.5 mg, mineral mixture appropriate amount, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, primary potassium phosphate 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-mentioned vitamin and mineral mixture is comparatively mixed with a composition suitable for health food as a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional method for producing healthy foods , Granules can be prepared and used in the manufacture of health food compositions according to conventional methods.

Preparation Example 6 Preparation of Healthy Drink

1000 mg of the functional composition of Example 1, 1000 mg of citric acid, 100 g of oligosaccharide, 2 g of plum concentrate, 1 g of taurine, and purified water were added to adjust the total amount to 900 ml.

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was heated at 85 DEG C for about 1 hour with stirring, and the solution thus prepared was filtered to obtain a sterilized 2-liter container, which was sealed and sterilized, &Lt; / RTI &gt;

Although the compositional ratio is relatively mixed with a component suitable for a favorite drink, it is also possible to arbitrarily modify the compounding ratio according to the regional or national preference such as the demand class, the demanding country, and the use purpose.

Claims (12)

A functional food composition for anti-inflammatory oral administration, comprising a plant composition, which is a mixture of onion (Allium cepa L.) extract, Morus alba L. Mulberry extract, and Glycine max (L.) Merr extract. The method according to claim 1,
The mixed plant composition is a functional food composition for oral administration of anti-inflammatory oral, characterized in that the mixture of the loser extract, frost extract and onion extract in a weight ratio of 1: (0.1 to 5): (0.1 to 50), respectively. A functional food composition for anti-inflammatory oral administration, comprising a plant composition in which a lettuce extract and a frost extract are mixed. The method according to claim 3,
The mixed plant composition is functional food composition for oral administration of anti-inflammatory oral, characterized in that the mixture of the loser extract and frost extract 1: 1 (0.1 to 5) by weight ratio. A functional food composition for anti-inflammatory oral administration, which comprises a plant composition in which a lettuce extract and an onion extract are mixed. The method according to claim 5,
The mixed plant composition is functional food composition for oral administration of anti-inflammatory oral, characterized in that the mixture of the lettuce and onion extract in a weight ratio of 1: (0.1 to 50). A functional food composition for anti-inflammatory oral administration, comprising a plant composition in which frost extract and onion extract are mixed. The method of claim 7,
The mixed plant composition is a functional food composition for anti-inflammatory oral administration, characterized in that the frost extract and onion extract is mixed in a weight ratio of 1: (0.1 to 50). The method according to any one of claims 1 to 8,
Functional food composition for oral administration to which the anti-inflammatory functional food composition is administered is any one of capsules, pills, granules, powders, candy, gum, tablets, beverages, syrups. . In the method for producing a functional food composition for anti-inflammatory oral administration to prepare the extract of the loser according to any one of claims 1, 3, 5,
A first step of extracting 2 to 8 hours, 1 to 4 times at 30 to 80 ° C. with an aqueous solution of 0 to 99% ethanol at a weight ratio of 4 to 10 times after grinding the cores, and cooling and filtering;
A second step of concentrating the filtrate obtained in the first step at 60 to 100 ° C. under reduced pressure; And
And a third step of lyophilizing the extract obtained in the second step and preparing a powder extract.
Method of producing a functional food composition for oral administration of anti-inflammatory, characterized in that the rutin (rutin) which is the index material extracted through the process is 0.1 to 20%. In the method for producing an anti-inflammatory oral food composition for preparing the frost extract according to any one of claims 1, 3, 7,
Washing the frosted with purified water twice and then extracting 1 to 4 times at 30 to 100 ° C. for 2 to 8 hours with 4 to 10 times the weight ratio of 0 to 80% alcoholic aqueous solution, followed by cooling and filtration;
A second step of concentrating the filtrate obtained in the first step at 60 ° C. under reduced pressure; And
Lyophilizing the extract obtained in the second step, and preparing a powder extract;
Method for producing a functional food composition for anti-inflammatory oral administration, characterized in that the total saponin (saponin) which is the indicator material extracted from the process is 0.2 to 10%. In the method for producing a functional food composition for anti-inflammatory oral administration for producing the onion extract according to any one of claims 1, 5, 7,
After washing the onion twice with purified water without separating the skin peeled and extracted 2 to 8 hours, 1 to 4 times at 60 to 100 ℃ with 10 to 100% alcoholic aqueous solution of 4 to 10 times the weight ratio, the first to cool and filter Steps;
A second step of concentrating the filtrate obtained in the first step at 60 ° C. under reduced pressure; And
And a third step of lyophilizing the extract obtained in the second step and preparing a powder extract.
Quercetin (quercetin), which is an indicator material extracted from the process, is 0.1 to 40%, characterized in that the method for producing a functional food composition for oral administration of anti-inflammatory.

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