KR101852032B1 - Beverages for preventing hyperlipidemia containing Platycodon grandiflorum and its preparing method - Google Patents

Abstract

The present invention relates to a beverage containing bellflower having a hyperlipidemia preventing or improving activity and a process for producing the same. According to the present invention, the sweet potato-containing beverage according to the present invention can be effectively used as a food for preventing or improving hyperlipidemia, because it reduces blood cholesterol concentration and is effective for hyperlipidemia and reduces fat accumulation to prevent anti-obesity. In addition, there is no cytotoxicity, toxicity to the drug, and side effects are avoided, so that they can be used safely even when taken for a long period of time and have a stable effect in the body.

Description

Technical Field [0001] The present invention relates to a beverage containing a bellflower having an effect of preventing hyperlipidemia containing bellflower as an active ingredient and a method for producing the same,

The present invention relates to a bellflower-containing beverage having the activity of preventing or ameliorating hyperlipidemia, which comprises bellflower as an active ingredient, and a process for producing the same.

Hyperlipidemia is an important risk factor for ischemic heart disease or cerebrovascular disorder, which is an arteriosclerotic disease, because fat metabolism such as triglyceride and blood cholesterol is not properly performed and the amount of fat in the blood is increased. The risk of hyperlipidemia due to changes in dietary habits and lifestyle habits is very high, so adults and adolescents are increasing the problem of hyperlipemia, and hyperlipemia causes complications such as diabetes and hypertension. Complex drugs are emerging to increase the therapeutic effect of existing cardiovascular treatment drugs such as the statin family. Some hyperlipidemia medicines have side effects that cause side effects such as hyperlipidemia medicines are urgently needed. As with hypertension medicines, it is time to develop medicines and natural products that have few side effects as lifelong medicines. Examples of therapeutic agents for hyperlipidemia include cholesterol biosynthesis inhibitors and cholesterol absorption inhibitors. Statins, fibrates, and ACAT inhibitors are used as therapeutic agents. Among them, statin drugs are widely used. This drug acts as an HMG-CoA reductase inhibitor to inhibit the synthesis of cholesterol. It decreases the concentration of LDL-cholesterol in the blood, I will. It also increases HDL-cholesterol, but only slightly increases the effect, because it increases only about 5-10% of the original value. Since myopathy may occur rarely, it is necessary to measure blood creatine kinase (CK) levels if myalgia is present at the time of drug administration.

Obesity is one of the most prevalent diseases in which the pattern of disease is rapidly changing to a developed country type as the hygiene environment is improved due to the improvement of the living standard and the average life span is extended due to the westernization of the diet. Thus, adult diseases have emerged as the biggest medical problem today, and the obesity, which is a major cause of these adult diseases, is increasing rapidly. Obesity refers to the phenomenon that extra calories are accumulated in the body in the form of fat by consuming excessive calories compared to the calories consumed. Obesity is thought to be caused by various causes such as genetic influences, environmental influences due to westernized diet, and psychological effects due to stress. However, the precise cause and mechanism of obesity has not been established yet. However, since obesity can act not only as a problem of obesity itself, but also as a cause of diseases such as cardiovascular disease or diabetes (Manson et al., New England J. Med., 333, pp677-685, 1995; JAMA, 282, pp1523-1529, 1999). There is a growing interest in the treatment of obesity worldwide.

In addition, studies on the regulation of the human immune system against the natural extract have been actively conducted because the immune system abnormality breaks health and causes various diseases. Therefore, the development of immunomodulators for safe natural products that can minimize side effects can be a very important material for the development of food, cosmetics and pharmaceuticals. For example, mulberry bark and shiitake mushrooms are used as immunostimulants, starfish, longevity, and mushroom as anti-cancer adjuvants. Licorice, licorice glycyrrhizin, sea cucumber, Research is underway on cell differentiation. The development of immunomodulatory agents using these natural materials has attracted many people's interest and interest because they can safely treat cancer and chronic inflammatory diseases without side effects.

Accordingly, the present applicant has developed a drink containing a platycodon which has the activity of preventing and improving hyperlipidemia from natural extracts and a method for producing the same.

Korean Patent No. 10-1162710 Korean Patent No. 10-0495315

Accordingly, the inventors of the present invention have completed the present invention by producing a beverage having a bronchial disease and hyperlipidemia-inhibiting effect, wherein the bellflower-containing beverage has a preventive effect on hyperlipemia.

Accordingly, an object of the present invention is to provide a method for manufacturing a frozen edible product, Mixing the freeze-dried bellflower and the sub ingredient and obtaining an extract using purified water; And adding an enzyme-treated stevia to the extract, wherein the method comprises the step of:

Another object of the present invention is to provide a beverage containing bellflower having the activity of preventing or improving hyperlipidemia, which is produced by the above-mentioned method.

It is another object of the present invention to provide a method for preparing a microorganism which comprises lyophilizing in-vitro cultured diploid or tetraploid platelets; Extracting freeze-dried bellflower with purified water, filtering and concentrating to obtain an extractive filtrate; Lyophilizing the extracted filtrate to obtain a solid fraction; And a step of mixing the solid component and the distilled water to obtain an extract. The present invention also provides a method of manufacturing a drink containing the balloon, which has the activity of preventing or improving hyperlipidemia.

Another object of the present invention is to provide a drink containing the bellflower having the activity of preventing or improving hyperlipidemia, which is produced by the above-mentioned method.

In order to accomplish the above object, the present invention provides a method for manufacturing a freeze- Mixing the freeze-dried bellflower and the sub ingredient and obtaining an extract using purified water; And adding an enzyme-treated stevia to the extracted liquid. The present invention also provides a method of manufacturing a beverage containing the bellflower having the activity of preventing or improving hyperlipidemia.

In an embodiment of the present invention, the bellflower and the supplementary material may be used in an amount of 50 to 70 parts by weight, more preferably 6 to 10 parts by weight, mulberry Chinese cabbage, 6 to 10 parts by weight, 6 to 10 parts by weight, To 10 parts by weight and a dry weight of 6 to 10 parts by weight.

The present invention provides a drink containing bellflower having the activity of preventing or improving hyperlipidemia, which is produced by the above method.

The present invention relates to a method for preparing a microorganism, comprising the steps of: lyophilizing in-vitro cultured diploid or tetraploid bellflower; Extracting freeze-dried bellflower with purified water, filtering and concentrating to obtain an extractive filtrate; Lyophilizing the extracted filtrate to obtain a solid fraction; And a step of mixing the solid content and the distilled water to obtain an extract. The present invention also provides a method of manufacturing a drink containing a flower crucible having the activity of preventing or improving hyperlipidemia.

In one embodiment of the present invention, there is provided a beverage containing bellflower having the activity of preventing or improving hyperlipidemia, which is produced by the above-described method.

The present invention relates to beverages containing a bellflower having a preventive effect on bronchial diseases and hyperlipemia and a process for producing the same. According to the present invention, the sweet potato-containing beverage according to the present invention is effective for reducing hypercholesterolemia by decreasing the blood cholesterol concentration, and also for reducing the fat accumulation and preventing the anti-obesity, thereby being useful as a food for preventing or improving hyperlipemia and bronchial diseases. In addition, there is no cytotoxicity, toxicity to the drug, and side effects are avoided, so that they can be used safely even when taken for a long period of time and have a stable effect in the body.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a schematic diagram showing a sample derivatization and HR CG / TOF analysis process for comparatively analyzing useful components of diploid and tetraploid Dorrissa. FIG.
FIG. 2 is a graph showing the results of chromatographic analysis of diploid and tetraploid moraines. FIG.
FIG. 3 is a schematic diagram showing the component differences of diploid and tetraploid moratoria using the orthogonal partial least squares discriminant analysis (OPLS-DA) method.
FIG. 4 is a schematic diagram showing an HR CG / TOF analysis process for detecting an efficacy substance according to an extraction solvent of diploid and tetraploid Dorago.
FIG. 5 is a graph showing the results of chromatographic analysis for detecting an efficacy substance according to an extraction solvent of diploid and tetraploid.
FIG. 6 is a schematic view showing a process for producing the present invention of the present invention and a spout drink using the same.
Fig. 7 is a schematic view showing a process for producing a tea-borne tea drink according to the present invention.
Fig. 8 shows the effect of the corynebacterium diphtheriae (Left picture) and bellflower beverage (right picture) according to the contents of the bellflower.
A: beverage containing 50% bellflower, B: beverage containing 55% bellflower, C: beverage containing 60% bellflower, D: beverage containing 65% bellflower, E: beverage containing 70%
2: bellflower tea beverage, 3: bellflower mixed drink, 4: extract beverage
Figure 9 is a graph showing the effect of the bronchial-induced bacteria Streptococcus pyogenes (Left picture) and bellflower beverage (right picture) according to the contents of bellflower.
A: beverage containing 50% bellflower, B: beverage containing 55% bellflower, C: beverage containing 60% bellflower, D: beverage containing 65% bellflower, E: beverage containing 70%
2: bellflower tea beverage, 3: bellflower mixed drink, 4: extract beverage
Fig. 10 is a graph showing the effect of Klebsiella pneumoniae subsp . pneumoniae (Left picture) and bellflower beverage (right picture) according to the contents of bellflower.
A: beverage containing 50% bellflower, B: beverage containing 55% bellflower, C: beverage containing 60% bellflower, D: beverage containing 65% bellflower, E: beverage containing 70%
2: bellflower tea beverage, 3: bellflower mixed drink, 4: extract beverage
Fig. 11 shows the results of the activity of inhibiting the alpha -amylase activity of the drug-containing beverage.
A: beverage containing 50% bellflower, B: beverage containing 55% bellflower, C: beverage containing 60% bellflower, D: beverage containing 65% bellflower, E: beverage containing 70%
2: bellflower tea beverage, 3: bellflower mixed drink, 4: extract beverage
12 is a graph showing the results of inhibiting activity of? -Glucosidase in the platycodon containing beverage.
A: beverage containing 50% bellflower, B: beverage containing 55% bellflower, C: beverage containing 60% bellflower, D: beverage containing 65% bellflower, E: beverage containing 70%
2: bellflower tea beverage, 3: bellflower mixed drink, 4: extract beverage
Fig. 13 shows the result of inhibiting the procine pancreatic lipase activity of the bellflower-containing beverage.
A: beverage containing 50% bellflower, B: beverage containing 55% bellflower, C: beverage containing 60% bellflower, D: beverage containing 65% bellflower, E: beverage containing 70%
2: bellflower tea beverage, 3: bellflower mixed drink, 4: extract beverage
Fig. 14 shows the results of measurement of SOD enzyme activity of the edible beverage.
A: beverage containing 50% bellflower, B: beverage containing 55% bellflower, C: beverage containing 60% bellflower, D: beverage containing 65% bellflower, E: beverage containing 70%
2: bellflower tea beverage, 3: bellflower mixed drink, 4: extract beverage
Fig. 15 shows the result of measuring the CAT activity of the bellflower-containing beverage.
B: 55% large-containing beverage, C: 60% bellflower-containing beverage, D: 65% bellflower-containing beverage, E: 70%
2: bellflower tea beverage, 3: bellflower mixed drink, 4: extract beverage
Fig. 16 shows the result of measuring the APX activity of the bellflower-containing beverage.
B: 55% large-containing beverage, C: 60% bellflower-containing beverage, D: 65% bellflower-containing beverage, E: 70%
2: bellflower tea beverage, 3: bellflower mixed drink, 4: extract beverage
Fig. 17 shows the result of measuring the POD activity of the edible beverage.
B: 55% large-containing beverage, C: 60% bellflower-containing beverage, D: 65% bellflower-containing beverage, E: 70%
2: bellflower tea beverage, 3: bellflower mixed drink, 4: extract beverage

The present invention is characterized by providing a method of manufacturing a drink containing a platycodon which has an activity of preventing or improving hyperlipidemia.

As the material of the beverage having the activity of preventing or improving hyperlipidemia of the present invention, the 'bellflower' is also called Gakgyeong, Dorat, Gilbyeol, Chewyeop, Jeonyeong, Roots are thick, stems grow straight, white juice comes out. The height is 40 ~ 100cm. Leaves are alternate phyllotaxis, long ovate or broad basal, serrate on edge, no petiole. The tip of the leaf is sharp and the base is wide. The front side of the leaf is green, the back side is grayish blue, no hairs, 4 ~ 7cm long, 1.5 ~ 4cm wide. Flower is white or purple in July ~ August, bell-shaped bloom with 4 ~ 5cm in diameter and 5 pieces in tip. The calyx is also divided into five, and the branch is bulky. There are 5 stamens, 1 pistil, 5 ovaries and 5 pistils. The fruit is ovate with ovate and ripened with calyx. Breeding is good with seeds. In spring and autumn, roots are picked and eaten raw or eaten as herbs. The main component of the bellflower is saponin. Kikyoku of herbal medicine is peeled off or dried as it is, and it is used for heat treatment, waste heat, tonsillitis, diarrhea in the oriental medicine. It is called for. Albiflorum for the white flower, double flower for the double flower, and double flower for the white flower. Flower language is eternal love. It is distributed in Korea, Japan, and China.

The inventors of the present invention have confirmed through experiments that the above-mentioned bellflower-containing beverage has the activity of preventing or improving hyperlipemia, and in particular, the bellflower can be easily ingested by being made into beverage.

More specifically, according to one embodiment of the present invention, the ingredients of the bellflower were analyzed to prepare the bellflower-containing beverage, and the useful components of the diploid and tetraploid bellows were analyzed by applying HR GC / TOF (See FIG. 1). As a result of analysis, it was confirmed that the dihydrate and tetraploid bellflower differ in their usefulness depending on their purity. (See FIGS.

In order to detect the efficacious substances according to the extraction solvent, the detection efficiencies of the components of the five kinds of extraction solvents were compared with those of methanol, ethanol (70:30 v: v) mixed solvent, methanol, ethanol, methylene chloride, hexane (70:30 v: v) mixed solvent. (Refer to Figs. 4 and 5).

In order to prepare the beverage of the present invention, the bellflower and the raw material were washed to remove soil and foreign matter, and each material was cut into an appropriate size and lyophilized. The mixture was mixed with the ratios shown in Table 6 and extracted with purified water. Treated Stevia extract was added thereto to prepare a blend-containing mixed drink (see Fig. 6).

The 'bellflower' for preparing the mixed beverage is not limited to the bellflower, but may be selected from the group consisting of a bellflower, an in-vitro cultured bellflower or an in-vitro cultured bellflower.

In order to prepare beverages using the diploid or tetraploid drosophila roots of the present invention, bellflowers were first selected and lyophilized and then pulverized into powder form. Then, the dried powder was extracted, filtered and concentrated to obtain a solid fraction . The obtained solid component is mixed with distilled water to prepare an extract, wherein the solid content is 2% and the distilled water is 98%. A small amount of juice was added to the extract to prepare a tea beverage-containing tea beverage composition, which was sterilized and packed in a PET bottle (see FIG. 7).

The inventors of the present invention conducted an experiment to verify the antimicrobial activity against the bronchial disease bacteria of the platycodon containing beverage according to another embodiment of the present invention. The platycodon beverage according to the content of the platycodon, Respectively. Corynebacterium , a bronchial disease-causing bacterium diphtheriae , Streptococcus pyogenes , Klebsiella pneumoniae subsp . pneumoniae (Fig. 8 to 10). As shown in Fig. 8 to 10, the antimicrobial activity of the bellflower was significantly inhibited regardless of the blending ratio of the bellflower.

Further, the present inventors tried to verify the anti-hyperlipidemic effect of the drug-containing beverage through another embodiment. As a result of analyzing the activity of HMG-CoA reductase, which is a human immune cell, (See Table 8). ≪ tb > < TABLE >

Furthermore, the inventors of the present invention evaluated the α-amylase inhibitory activity, α-glucose inhibitory activity and pancreatic lipase inhibitory activity of the bellflower-containing beverage. In the present invention, the beverage containing the bellflower of the present invention inhibited α-amylase, α-glucose and pancreatic lipase (See Figs. 11 to 13).

Meanwhile, the inventors of the present invention measured the antioxidative activity of the bellflower-containing beverage by using SOD (Superoxide Dismutase), CAT (Catalase), APX (Ascorbate Peroxidase) and POD (Peroxidase) 17).

Accordingly, the present invention is characterized by providing a method for preparing a food containing a balloon, which has an activity of preventing or improving hyperlipidemia.

In the present invention, 'hyperlipidemia' includes, but is not limited to, hypercholesterolemia, hypertriglyceridemia, or low density lipoprotein cholesterolemia.

In the present invention, 'anti-obesity' refers to the ability of cells to inhibit α-amylase, α-glucose, pancreatic lipase inhibition and adipocyte differentiation, and the concentration of the composition for anti-obesity may be 10 to 1000 μg / ml .

In the present invention, 'obesity' means that the amount of fat in the body is excessively increased. Obesity is caused by an imbalance in the synthesis and degradation of fat in adipocytes. Obesity, which is caused by excess fat accumulation, increases the risk of developing chronic degenerative diseases and increases the risk of coronary artery disease, hypertension, stroke, hyperlipemia and diabetes .

In the present invention, " prevention " means all actions that inhibit obesity or delay progression by administration of the composition of the present invention.

&Quot; Improvement " in the present invention means all the actions of improving or ameliorating symptoms of obesity by administration of the composition of the present invention.

In the present invention, " treatment " means, unless otherwise indicated, reversing, alleviating, inhibiting or preventing the disease or disorder to which the term applies, or one or more symptoms of the disease or disorder And the term " treatment " as used herein refers to an act of treating when " treating " is defined as described above.

The term " administration " as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

In addition, the present invention can prevent the diseases related to the inhibition of adipocyte differentiation and the carbohydrate inhibitory activity.

Therefore, it can be usefully used as a beverage containing bellflower as an active ingredient for the prevention of obesity and hyperlipemia.

The composition of the present invention may be a food composition. The food composition may contain various flavors or natural carbohydrates such as an ordinary food composition, as well as an effective ingredient, such as bellflower, as an additional ingredient.

Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And polysaccharides, for example, conventional sugars such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. The above-described flavors can be advantageously used as natural flavorings (tau martin), stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.).

The food composition of the present invention can be formulated in the same manner as the above pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolates, foods, confectionery, pizza, ram noodles, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes, .

In addition, the food composition may contain flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring agents and thickening agents (cheese, chocolate etc.), pectic acid and its salts, Alginic acid and its salts, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks and the like. In addition, the food composition of the present invention may contain natural fruit juice and pulp for the production of fruit juice drinks and vegetable drinks.

As an effective ingredient of the present invention, edible balloon is a natural substance with little toxicity and side effects, and therefore can be safely used for long-term use for the purpose of preventing hyperlipemia and anti-obesity.

The health functional food of the present invention can be manufactured and processed in the form of tablet, capsule, powder, granule, liquid, ring and the like for the purpose of preventing hyperlipemia and anti-obesity.

In the present invention, the term "health functional food" refers to a food prepared and processed by using raw materials or ingredients having useful functions in accordance with Law No. 6727 on Health Functional Foods, and the nutritional control on the structure and function of the human body Or for the purpose of obtaining a beneficial effect for health use such as physiological action.

The health functional foods of the present invention may contain conventional food additives and, unless otherwise specified, whether or not they are suitable as food additives are classified according to the General Rules for Food Additives approved by the Food and Drug Administration, Standards and standards.

Examples of the food items included in the above food additives include chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as persimmon extract, licorice extract, crystalline cellulose, high color pigment and guar gum; L-glutamic acid sodium preparations, noodle-added alkalis, preservative preparations, tar coloring preparations and the like.

For example, the health functional food in the form of tablets may be prepared by granulating a mixture of the active ingredient of the present invention with an excipient, a binder, a disintegrant, and other additives by a conventional method, , The mixture can be directly compression molded. In addition, the health functional food of the tablet form may contain a mating agent or the like if necessary.

The hard capsule of the capsule-type health functional food can be prepared by filling a hard capsule, which is an effective ingredient of the present invention, with a mixture obtained by mixing the drug with an additive such as an excipient. The soft capsule is prepared by mixing the dresser with an excipient such as an excipient Can be prepared by filling a mixture into a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a coloring agent, a preservative and the like, if necessary.

The ring-shaped health functional food can be prepared by molding the mixture of the active ingredient of the present invention, the edible part of the present invention, excipient, binder, disintegrant and the like by a conventionally known method, and if necessary, Or the surface may be coated with a material such as starch, talc.

The health functional food of the granular form can be prepared by granulating the mixture of the active ingredient of the present invention and the excipient, the binder, the disintegrant and the like in a granular form by a conventionally known method, and if necessary, a flavoring agent, ≪ / RTI >

The health functional food may be a beverage, a meat, a chocolate, a food, a confectionery, a pizza, a ramen, a noodle, a gum, a candy, an ice cream, an alcoholic beverage, a vitamin complex and a health supplement food.

Hereinafter, the present invention will be further described by way of examples. These examples are for further illustrating the present invention, and the scope of the present invention is not limited to these examples.

< Example  1> Analysis of unknown substance in bellflower

To analyze unknown substances other than the known major components for the diploid and tetraploid material, we used high resolution gas chromatography / ion flight mass spectrometer (HR GC / TOF) to analyze unknown substances Respectively.

1-1. Diploid , Tetraploid  Comparison analysis of useful components of bellflower

(GC GC / TOF) was used to analyze unknown substances for the analysis of unrecognized components of dodecahedra and quadrate platycodon. The lyophilized samples were extracted with a solvent of ethanol: methanol = 7: 3, and each extract was derivatized with pyridine + methoxyamine hydrochloride solution for 1 hour 30 minutes and then with MSTFA for 30 minutes. HR GC / TOF-MS (See Fig. 1).

As shown in Table 1, a large amount of organic acids and saccharides were detected in diploid bellflower, and thus the compositional characteristics were largely different according to the drainage property (See Table 2). After confirming this fact, diploid bellflower and tetraploid bellflower were used in the experiment of the present invention.

Comparative analysis of useful components of dibasic bellflower Amino acids One I-Alanine 9 Phenylalanine 2 L-Valine 10 L-Asparagine 3 Serine 11 Xylitol 4 L-threonine 12 N-methylgluconamide 5 3-Aminopiperidin-2-one 13 L-Tyrosine 6 L-Aspartic acid 14 5-allyl-1-methoxy-2,3-dihydroxybenzene 7 5-oxo-L-proline 15 Inositol 8 L-glutamic acid 16 9,12-Octadecadienoic acid

Comparative analysis of useful components of 4-platycodon Organic acids, Sugars One lactic acid (2-hydroxypropanoic acid) 2 Butanedioic acid 3 malic acid 4 Oleic acid 5 10'-Apo-.beta.-carotenoic acid, 5,6-dihydro-5,6-dihydroxy-, methyl ester 6 a-D-glucopyranosyl-D-Glucose

1-2. In extraction solvent  Comparison of efficacy detection

(HR GC / TOF) was used to analyze unknown substances for the analysis of useful components for the tetraploid, tetraploid, and tetraploid material. The lyophilized samples were extracted with a solvent of hexane, methylene chloride, ethanol, methanol, ethanol: methanol = 7: 3, and then the extracts were extracted with pyridine + methoxyamine hydrochloride solution for 1 hour and 30 minutes, And confirmed by HR GC / TOF-MS (see FIG. 4).

As a result of the comparison of useful components of diploid and tetraploid bellflower using five kinds of extraction solvents, the extraction efficiency was high in the order of methanol and ethanol (70:30 v: v) mixed solvent, methanol, ethanol, methylene chloride and hexane In particular, it was confirmed that the component detection efficiency in methanol and ethanol mixed solvent was the most excellent (see FIG. 5).

<Example 2> Improvement of bronchial disease and antihyperlipidemia Derived optimum dosage of effective beverage

In order to carry out experiments related to antibacterial and antihyperlipidemic efficacy against bronchial disease bacteria, the blending ratio of beverage containing bellflower was designed.

2-1. Bellflower mixed drink

The blended beverage containing bellflower was prepared by blending blended bean japonica (cabbage chaji), omija, gugija, ginko, ginko, and dried bean as optimum functional ingredients, The optimum blending ratios of the compositions were designed in five blending ratios as described in Table 6. &lt; tb &gt; &lt; TABLE &gt;

The extract beverage is a concentrated ampule-form beverage which is prepared so that it can be mixed with the above-mentioned bellflower-containing mixed beverage at the same compounding ratio and manufacturing process, and diluted for drinking. That is, the beverage of the ampoule formulation prepared by concentrating the blender-containing mixed drink (containing 50% of the bellflower) is referred to as an extract drink in the present invention.

Mixed beverage composition material Formulation ratio (%) A B C D E Bellflower 50 55 60 65 70 Chinese cabbage 10 9 8 7 6 Schisandra 10 9 8 7 6 Ginkgo 10 9 8 7 6 Gugija 10 9 8 7 6 Dry mu 10 9 8 7 6 system 100.0 100.0 100.0 100.0 100.0

Five kinds of specimens were prepared by freeze-drying, and then mixed with each other at the mixing ratios shown in Table 3. For the preparation of the mixed drink with bellflower, the five kinds of specimens such as pine nut, cabbage, omija, ginkgo, % Was added to the extract to prepare a mixed drink and packaged in a spout pouch of capacity (see FIG. 6).

2-2. Bellflower tea drink

Dragee - containing tea beverages were prepared by lyophilization of Dipodachlorophylla or In - cabbage cultivated Diptera Rhizoma root extract powder by lyophilization, followed by extraction with purified water and filtration to produce an extract. The extract was concentrated and lyophilized to prepare a solid. The solid was used in an amount of 2 parts by weight based on 100 parts by weight (see FIG. 7).

&Lt; Experimental Example 1 > Antibacterial effect of beverage containing bellflower on bronchial disease bacteria

(Meat extract 1%, tryptone 1%, NaCl 0.5%, glucose 0.2%, and the like) in the beverage prepared in Example 2 to test the efficacy of the bellflower- yeast extract 0.3%, pepton 0.1%, pH 7.3). Bacterial culture medium was modified with blood agar base (Difco 0045), and antimicrobial activity assay was measured by disk agar plate diffusion method. That is, the soft agar cultured at a concentration of 10 ⁷ CFU / mL in a medium suitable for each test microorganism was dispensed into a petri dish and placed on a paper disc. 40 μL of the sample extract was added to each well, (See Table 7). When an inhibitory loop was generated within 24 hours, the antimicrobial activity was determined to be positive, and the diameter of the inhibitory loop was measured and compared (see FIG. 8).

The strains used for the experiment to inhibit bronchial disease and the list used for the culture conditions Strains Cultivarion conditions Corynebacterium diphtheriae
(KCCM 40413)
Klebsiella pneumoniae subsp.
pneumoniae (KCCM 41285)
Streptococcus pyogenes
(KCCM 11817) 37 ° C, Trypticase Soy Agar (BBL211043) with 5% defibrinated Sheep blood
37 ° C, Nutrient Agar

37 ° C, Trypticase Soy Agar (BBL211043) with 5% defibrinated sheep blood

Corynebacterium , a bronchial disease-causing bacterium diphtheriae , Streptococcus pyogenes, Klebsiella pneumoniae subsp . As a result of the antibacterial activity test of paddy - rice mixed drink against pneumoniae , all strains showed a significant effect regardless of the blend ratio.

As a result of the analysis of the formulation of bellflower-containing beverages, the extracts showed a rather high activity in extract beverages, and no significant effect was observed in tea beverages and mixed beverages. do.

&Lt; Experimental Example 2 >

The efficacy of the prototype for anti-hyperlipidemic was evaluated by measuring the activity of HMG-CoA reductase, which specifically inhibits the synthesis of cholesterol. The HMG-CoA Reductase Assay Kit (Sigma) was used for the analysis. HMG-CoA reductase is a rate-limiting enzyme for the synthesis of cholesterol. It has been reported that lowering the activity of HMG-CoA reductase in the liver increases the activity of hepatic LDL-receptor and thereby decreases the blood cholesterol concentration.

 Evaluation of HMG-CoA reductase inhibitory activity of the present invention of the present invention Samples Specific activity (Unit / mg protein / min) Inhibition rate (%) Pravastatin (con) 38.13 89.42 ± 3.21 A 65.72 58.03 + - 4.52 B 64.91 50.92 ± 3.65 C 71.40 46.01 + - 5.33 D 63.29 52.15 + - 2.86 E 62.47 52.76 ± 3.78 2 76.00 34.97 ± 3.15 3 71.14 23.93 ± 1.93 4 75.46 42.94 + 5.06 * Mixed beverage containing bellflower (A: 50%, B: 55%, C: 60%, D: 65%, E: 70%).
2: Drinking tea drink with bellflower 3: Mixed drink with bellflower 4: Drinking drink

The inhibitory activity of HMG-CoA reductase was found to be 58.03% in the mixture of 50% blend (A) and the highest inhibitory activity. In addition, inhibitory activity of each product formulation showed a relatively high inhibitory activity of 42.94% in extract beverage (see Table 8).

&Lt; Experimental Example 3 > Analysis of fat and carbohydrate inhibitory activity of the bellflower-containing beverage

3-1. α-Amylase inhibitory activity

Α-Amylase inhibitory activity was evaluated in order to investigate the blood glucose lowering effect of bellflower-containing beverages. The α-Amylase inhibitory activity was modified by the method of Lim et al. The samples were diluted by concentration, and 25 μL of α-amylase (0.25 U / mL) diluted with 2.5 μL of sample and 50 mM phosphate buffer (pH 7.0) was preincubated at 37 ° C. for 10 minutes. 25 μL was added and reacted at 37 ° C for 10 minutes. Then, the reaction was stopped by heating at 100 ° C for 5 minutes. To the reaction solution was added 150 μL of a solution of 3, 5-dinitrosalicylic acid (DNS) and heated at 100 ° C for 5 minutes to develop color. The absorbance was measured at 540 nm using a 96-well microplate reader. The inhibition rate of α-amylase was calculated by the following equation.

Inhibition rate (%) = [1- (sample enzyme activity / control enzyme activity)] x 100

As a result of assaying the α-amylase inhibitory activity of the bellflower-containing beverage, a relatively high inhibitory activity was exhibited in a 55% mixed drink composition with a blend ratio of 60%, and the product formulation exhibited a high inhibitory activity in extract beverage (see FIG. 11) .

3-2.A-Glucosidase inhibitory activity

The inhibitory activity of α-glucosidase was evaluated in order to examine the blood glucose lowering effect of the bellflower-containing beverage. The α-glucosidase inhibitory activity was determined by measuring the absorbance at 405 nm by mixing a sample of 10 mg / mL concentration, 50 μL of 0.15 U / mL α-glucosidase enzyme and 360 μL of 0.2 M potassium phosphate buffer (pH 6.8) And 50 μL of 5 mM 4-nitrophenyl-α-D-glucopyranoside (pNPG) was added and reacted at room temperature for 10 minutes. The absorbance was measured at the same wavelength and enzyme inhibition activity was calculated from the change in absorbance.

As a result of assaying the? -Glucosidase inhibitory activity of the beverage composition according to the mixing ratio of the bellflower and the beverage containing the bellflower according to the formulation, it was confirmed that the 65% mixed drink composition and the extract have a relatively high inhibitory activity.

3-3.Pancreatic lipase inhibitory activity

In order to investigate the anti - obesity effect of bellflower - containing beverages, lipase inhibitory activity, which plays an important role in fat absorption, was evaluated. Porcine pancreatic lipase (PPL, type II) activity was measured using p-nitrophenyl butyrate (p-NPB) as a substrate. First, the PPL stock solution was prepared at a concentration of 1 mg / mL in 0.1 mM potassium phosphate (pH 6.0), and stored at -20 ° C. To measure lipase inhibitory activity, 25 μL of extract (20 mg / mL) and 25 μL of PPL solution were added to 950 μL of 0.1 mM potassium phosphate (pH 7.2, 0.1% Tween 80) and incubated at 30 ° C. for 1 hour. NPB was added and reacted at 30 ° C for 5 minutes, and the absorbance was measured at 405 nm.

     Inhibition activity (%) = [1- (B-C) / A] 100)

     A: Absorbance without added sample

     B: Absorbance with addition of sample

     C: Absorbance without addition of enzyme

As a result of the porcine pancreatic lipase inhibitory activity test on the beverage composition according to the blend ratio of the blender and the blender-containing beverage according to the formulation, relatively high inhibitory activity was observed in the mixed blend composition of 50% blend ratio and the product formulation showed a high inhibitory activity (See Fig. 13).

&Lt; Experimental Example 4 > Antioxidant enzyme activity of bellflower-containing beverage

4-1. SOD (Super Oxide Dismutase)

SOD enzyme activity assays were performed using an assay kit (Sigma, 19160). That is, the photochemical NBT method was used to test the ability of SOD enzyme to inhibit the reduction of NBT (Nitroblue tetrazolium). The reaction solution was adjusted to 50 mM carbonic buffer (pH 10.2), 0.1 mM EDTA, 0.1 mM Xanthine and 0.025 mM NBT, and the inhibition of NBT reduction was measured at 450 nm. Each sample was repeated 3 times, and SOD enzyme activity was calculated by the following equation.

SOD activity (NBT reduction inhibition rate,%) =

[(Ablank1-Ablank3) - (Asample-Ablank2)] / (Ablank1-Ablank3) x100

The SOD activity of the beverage composition and the drug-containing beverage according to the blend ratio of the blend was found to be 70% or higher without any significant difference between the blend ratios. (See Fig. 14).

4-2. CAT (Catalase)

CAT activity was measured by the method of Aebi et al. (1984). The reaction solution was 50 mM potassium phosphate buffer (pH 7.0) and 10 mM H 2 O 2. CAT activity was measured by adding the extract to the reaction solution and then changing the absorbance at 240 mM for 2 minutes.

CAT activity of the beverage composition and the beverage containing the bellflower according to the mixing ratio of the bellflower showed the highest activity in the blend composition of 50% of the bellflower, and the product showed relatively high activity in the beverage of the extract (see FIG. 15).

4-3. APX (Ascorbate Peroxidase)

The activity of APX was measured by the Nacano and Asada (1981) method and the degree of oxidation of ascorbate was measured at 290 nm for 2 minutes. The reaction solution of APX was 100 mM potassium phosphate buffer (pH 7.5) supplemented with 0.5 mM ascorbate and 0.2 mM H 2 O 2.

The APX activity of the bellflower-containing beverage showed the highest activity in the 50% blend composition, and in the blend-containing beverage according to the formulation, the mixed beverage and the extract had relatively high activity (see FIG. 16).

4-4. POD (Peroxidase)

POD activity was measured by the method of Egley et al. (1983). The reaction mixture was adjusted to a final concentration of 40 mM K-PO4 buffer (pH 6.9), 1.5 mM guaiacol, and 6.5 mM H2O2. The POD activity was measured by measuring the absorbance at 470 nm for 2 minutes using a spectrophotometer.

The POD activity of the beverage composition and the beverage containing starch according to the blend ratio was found to be relatively high in the blend composition of 55% and 70%, respectively, See FIG. 17).

The present invention has been described with reference to the preferred embodiments. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is defined by the appended claims rather than by the foregoing description, and all differences within the scope of equivalents thereof should be construed as being included in the present invention.

Claims (2)

Lyophilizing the bellflower and the sub ingredient;
Mixing the freeze-dried bellflower and the sub ingredient and obtaining an extract using purified water; And
And adding enzyme-treated stevia to the extract,
Wherein the bellflower contains 50 to 65 parts by weight based on 100 parts by weight,
Characterized in that said ingredient is a mixture of 6 to 10 parts by weight of pale blue cabbage, 6 to 10 parts by weight of omija, 6 to 10 parts by weight of ginkgo fruit, and 6 to 10 parts by weight of dry matter. A method of manufacturing a beverage. A beverage containing a bellflower having the activity of preventing or improving hyperlipidemia, which is produced by the method of claim 1.

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