KR101740136B1 - Natural liposome comprising red ginseng for improving blood circulation, process for the preparation thereof and food or pharmaceutical composition comprising the same - Google Patents
Natural liposome comprising red ginseng for improving blood circulation, process for the preparation thereof and food or pharmaceutical composition comprising the same Download PDFInfo
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- KR101740136B1 KR101740136B1 KR1020150122071A KR20150122071A KR101740136B1 KR 101740136 B1 KR101740136 B1 KR 101740136B1 KR 1020150122071 A KR1020150122071 A KR 1020150122071A KR 20150122071 A KR20150122071 A KR 20150122071A KR 101740136 B1 KR101740136 B1 KR 101740136B1
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- KR
- South Korea
- Prior art keywords
- natural
- red ginseng
- extract
- liposome
- blood circulation
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/10—Foods or foodstuffs containing additives; Preparation or treatment thereof containing emulsifiers
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
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- A23L3/3472—Compounds of undetermined constitution obtained from animals or plants
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
- A23L5/30—Physical treatment, e.g. electrical or magnetic means, wave energy or irradiation
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/25—Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K9/127—Liposomes
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/326—Foods, ingredients or supplements having a functional effect on health having effect on cardiovascular health
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
- A23V2250/21—Plant extracts
- A23V2250/2124—Ginseng
Abstract
The present invention relates to a liposome comprising a natural emulsifier, a solvent, a natural coating agent and a natural preservative; And a red ginseng extract as a capturing substance, a method for producing the same, and a food or pharmaceutical composition containing the same.
Description
The present invention relates to a natural liposome containing red ginseng for improving blood circulation, a method for producing the same, and a food or pharmaceutical composition containing the same.
Ginseng is a perennial herbaceous perennial belonging to the genus Oga and Ginseng, and it is known that saponin contained in ginseng has a beneficial effect. Particularly, saponin contained in red ginseng processed ginseng is very mild, has no toxicity due to excessive administration, has a variety of effects such as improving blood circulation and controlling immune function.
Saponin contained in ginseng is also called ginsenoside because of its chemical structure and its different effect from saponin found in common plants. These ginseng saponins are known to be decomposed by intestinal microorganisms and then absorbed by the biosynthetic enzymes even after ingestion because of their high molecular weight. However, when ginseng or ginseng saponin is ingested, there is a difference in kind, concentration, and activity of intestinal microorganisms for each person ingested, so that the drug efficacy by saponin can not be uniformly obtained.
In recent years, in order to increase the absorption of ginseng saponin into the body, studies using a drug carrier such as liposome are actively under way.
Liposome is an artificially produced vesicle composed of a lipid bilayer, and has a structure similar to that of a vital cell membrane. The liposomes are biocompatible and composed of a phospholipid, which is a major component of a vital cell membrane, and exist in the form of capsules in which a hydrophilic and hydrophobic substance in an aqueous solution is surrounded by a lipid bilayer of liposome. Such liposomes are capable of storing desired useful substances (drugs, nutrients, etc.) in the bilayer structure, and are capable of transporting unstable and difficult-to-handle substances into cells. However, in spite of these advantages, the liposome has a problem that not only the formulation is unstable, the collection efficiency is very low, but also the solvent and preservative used for liposome production cause skin irritation. In addition, the cholesterol used as a raw material of the liposome has a problem that it forms a liposome lipid membrane firmly and hinders the release of the trapping material and lowers the cell membrane permeability of the trapping material.
To solve these problems, many studies have been conducted on liposomes containing red ginseng extract (saponin). For example, Korean Unexamined Patent Publication No. 2002-0023876 discloses a cosmetic composition containing ginsenoside Re and Rh2 complex derived from red ginseng or ginseng, and a method for producing the same. Korean Patent Laid-Open Publication No. 2003-0060017 discloses microemulsified particles containing ginsenoside F1 by nanoemulsion technology and a composition for external application for skin using the same. However, the aforementioned prior art documents describe a cosmetic composition comprising a synthetic liposome, and no liposome made of a natural substance is disclosed, and no food or pharmaceutical composition containing the liposome is disclosed at all.
In order to solve the above-mentioned problems, it is an object of the present invention to provide a natural liposome containing red ginseng for improving blood circulation that can enhance the cell uptake of a red ginseng extract while being harmless to human body, a method for producing the same, and a food or pharmaceutical composition containing the same. .
The present invention relates to a liposome comprising a natural emulsifier, a solvent, a natural coating agent and a natural preservative; And a red ginseng extract as a capturing material.
The present invention also relates to a method for producing an aqueous emulsion, comprising: (a) a first step of mixing a natural emulsifier with a solvent, followed by ultrasonic treatment at 35 to 80 ° C; (b) a second step of adding the red ginseng extract to the mixture of the first step followed by ultrasonic treatment at 35 to 80 캜; (c) a third step of adding a natural coating agent to the mixture of the second step and ultrasonication at 35 to 80 ° C; And (d) adding to the mixture of the third step a solvent warmed at 35 to 80 캜 and a natural preservative, and subjecting the mixture to ultrasonic treatment. The present invention also provides a method for producing the natural liposome containing red ginseng for blood circulation improvement.
Further, the present invention provides a food or pharmaceutical composition comprising the natural liposome containing red ginseng for blood circulation improvement.
The natural liposome containing red ginseng for blood circulation improvement according to the present invention can absorb the red ginseng extract without any individual difference in the intestines, and thus the efficacy can be uniformly displayed. At this time, the food or pharmaceutical composition containing the red ginseng-containing natural liposome may absorb the active ingredient of red ginseng into the body when ingested, thereby exhibiting an excellent blood circulation improving effect.
1 is a schematic view of a method for producing a natural liposome containing red ginseng for improving blood circulation according to an embodiment of the present invention.
FIG. 2A is a graph showing the particle size according to Experimental Example 1 of the present invention, and FIG. 2B is an image showing the particle size and morphology according to Experimental Example 1 of the present invention.
3 is a graph showing cell permeability according to Experimental Example 2 of the present invention.
4 is an image showing blood ginsenoside according to Experimental Example 3 of the present invention.
5A to 5E are graphs showing blood circulation improving functionalities according to Experimental Example 4 of the present invention.
Hereinafter, a natural liposome containing red ginseng for improving blood circulation according to the present invention, a method for producing the same, and a food or pharmaceutical composition containing the same will be described in detail with reference to the accompanying drawings. However, these descriptions are provided only to illustrate the present invention, and the scope of the present invention is not limited by these exemplary explanations.
<Natural liposome containing red ginseng for blood circulation improvement>
The present invention relates to a liposome comprising a natural emulsifier, a solvent, a natural coating agent and a natural preservative; And a red ginseng extract as a capturing material.
1. Liposome constituent material
Liposomes according to one embodiment of the present invention include natural emulsifiers, solvents, natural coatings and natural preservatives. Liposomes made of such natural substances are not toxic to the human body and can be ingested. In addition, the liposome can form a stable phospholipid bilayer, and can transfer the capture material stably to the intestines and into the intestinal cells.
(1) Natural emulsifiers
Natural emulsifiers contain phospholipids and fatty acids derived from natural materials and constitute a lipid bilayer.
Examples of the phospholipid include phosphatidylcholine, lysophosphatidylcholine, phosphatidylethanolamine, hydrogenated phophatidylcholine (derived from soybean), phosphatidylserine, phosphatidylglycerol, and phosphatidylinositol ) May be used. Hydrogenerated phosphatidylcholine, which is preferably excellent in oxidation stability, may be used.
The fatty acid may be at least one selected from the group consisting of palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid (derived from soybean) .
The natural emulsifier may be 70 to 90% by weight of the phospholipid and 1 to 5% by weight of the fatty acid based on the weight of the whole natural emulsifier.
These natural emulsifiers may be used in combination of two or more species. For example, two kinds of natural emulsifiers or three kinds of natural emulsifiers can be mixed. The first natural emulsifier may be selected from the group consisting of phosphatidylcholine, lysophosphatidylcholine, phosphatidylethanolamine, palmitic acid, stearic acid, oleic acid, linoleic acid, acid, and linolenic acid (derived from soybean). As the second natural emulsifier, hydrogenated phosphatidylcholine (derived from soybean) may be used. As the third natural emulsifier, cetearyl olivate cetearylolivate and sorbitanolivate (from olive) can be used.
In addition, the natural emulsifier may further include a natural fatty acid. As the natural fatty acid, evening primrose oil may be used. The natural fatty acid may be 0.1 to 5 wt% based on the weight of the total natural emulsifier.
The content of the natural emulsifier is preferably 0.1 to 10% by weight based on the weight of the whole red ginseng-containing natural liposome .
(2) Solvent
The solvent causes the lipid bilayer of the natural emulsifier to be in the form of a capsule surrounding the capture material.
As the solvent, at least one selected from the group consisting of distilled water, natural butylene glycol, propanediol, glycerin and fermented alcohol can be used. Preferably, distilled water, glycerin can be used, and more preferably distilled water can be used.
It is preferable that the content of the solvent is a residual amount excluding the natural emulsifier, the natural coating agent, the natural preservative and the collecting material based on the weight of the whole red ginseng-containing natural liposome.
(3) Natural coatings
Natural coatings are materials that coat the surface of the liposome to maintain the capsule form of the lipid bilayer surrounding the capture material. When a natural liposome containing red ginseng coated with a natural coating agent is ingested, the natural coating agent is decomposed by the intestinal enzyme, and the lipid bilayer binds with the cell membrane, and the substance is absorbed by the cell.
As the natural coating agent, at least one natural coating agent selected from the group consisting of maltodextrin, cellulose and pectin may be used as the polysaccharide decomposed by intestinal enzymes. Preferably maltodextrin can be used.
The content of the natural coating agent is preferably 0.01 to 5% by weight based on the weight of the whole red ginseng-containing natural liposome.
(4) natural preservatives
Natural preservatives prevent the decay of liposome constituents (natural emulsifiers, solvents, natural coatings) and trapping substances present in the liposomes, thus maintaining the stability of the final red ginseng-containing natural liposome.
The natural preservative may be at least one natural extract selected from the group consisting of citrus extract, citron extract, eucalyptus extract, ginseng extract, clove extract and grapefruit extract as natural preservatives that can be added to foods. The natural preservative is generally distinguished from chemical preservatives such as dehydroacetic acid, sorbic acid, benzoic acid, p-hydroxybenzoic acid esters, propionic acid and salts thereof, and is harmless to the human body because of its toxicity.
The content of the natural preservative is preferably 0.001 to 3% by weight based on the weight of the whole red ginseng-containing natural liposome.
2. Collecting material
The capturing material according to the present invention is a substance trapped in the lipid bilayer of the liposome containing the natural emulsifier, the solvent, the natural coating agent and the natural preservative. As the capturing material, there can be used a ternary extract derived from ginseng, red ginseng, white ginseng and ginseng.
In addition to anticancer and antiallergic, the triple extract is known to have excellent effects such as improvement of blood circulation, central inhibition and mental stability, analgesia, improvement of memory, inhibition of hypertrophy of protein and kidney glomeruli, antidiabetic, inhibition of platelet aggregation and chronic renal failure. Especially, red ginseng processed with ginseng has various kinds of saponin, which is more effective than general ginseng, and contains a high amount of saponin. Therefore, the capturing material is preferably red ginseng extract. At this time, the red ginseng extract can be reduced in molecular size by ultrasonic treatment to increase the absorption rate in the body while preventing the drug substance from being destroyed.
The content of the red ginseng extract is preferably 0.1 to 30% by weight based on the weight of the whole red ginseng-containing natural liposome.
As described above, the natural liposome containing red ginseng for blood circulation improvement of the present invention is in the form of capsules in which a low molecular weight red ginseng extract having a drug effect is enclosed by a lipid bilayer, and can increase the cell uptake of red ginseng extract. When the natural liposome containing red ginseng for blood circulation improvement is ingested, the cell membrane and the liposome bind to each other in the intestine and the cell absorption rate of the red ginseng extract is increased. Therefore, the effect of improving red blood ginseng in the body can be expected.
<Production method of natural liposome containing red ginseng>
The present invention provides a method for preparing a natural liposome containing red ginseng for blood circulation by using an ultrasonic wave crusher, an emulsifying device, and the like. However, the present invention is not limited to such a production method, and the steps of each process may be modified or optionally mixed as necessary.
As shown in FIG. 1, a method for preparing a natural liposome containing red ginseng for blood circulation improvement according to an embodiment of the present invention comprises: (a) a first step of ultrasonication at 35 to 80 after mixing a natural emulsifier with a solvent; (b) a second step in which the red ginseng extract is added to the mixture of the first step and then ultrasonicated at 35 to 80; (c) a third step of adding a natural coating agent to the mixture of the second step and then ultrasonifying the mixture at 35 to 80; And (d) adding to the mixture of the third step a solvent heated to 35-80 and a natural preservative and ultrasonically treating the mixture. After the fourth step, it may further include a fifth step of repeatedly performing ultrasonic processing while gradually lowering the temperature.
Hereinafter, the manufacturing method will be described separately for each process step as follows.
Step 1: After mixing at least one natural emulsifier with a solvent, At 80 ° C Ultrasonic treatment.
As the natural emulsifier, at least one natural emulsifier containing a phospholipid and a fatty acid derived from a natural substance may be used.
Examples of the solvent include distilled water, natural butylene glycol, propanediol, glycerin, fermented alcohol, and the like.
The ultrasonic treatment is a work for homogenizing a mixture of a natural emulsifier and a solvent, and an ultrasonic liquid processor may be used.
The natural emulsifier and the distilled water are mixed and then heated to 35 to 80 ° C, preferably 40 to 70 ° C, and then subjected to ultrasonic treatment to homogenize the phospholipid and the fatty acid of the natural emulsifier in the distilled water.
Step 2: After adding the red ginseng extract to the mixture of the first step, At 80 ° C Ultrasonic treatment.
The red ginseng extract can be extracted at room temperature or by heating under the condition that the active ingredient is not destroyed. At this time, a method for extracting red ginseng extract may be any conventional extraction method known in the art. For example, extracts in liquid or powder form can be obtained using extraction methods such as cold extraction, ultrasonic extraction, reflux cooling extraction, and hot water extraction.
The red ginseng extract may be mixed with an extraction solvent. When distilled water is used as the extraction solvent, the mixing ratio of the red ginseng extract to the distilled water is in the range of 0.1 to 5: 1, preferably 1: 1, based on the weight ratio.
After mixing the red ginseng extract with the mixture of the first step, the mixture is heated to 35 to 80 ° C, preferably 40 to 60 ° C, and then subjected to ultrasonic treatment to reduce the size of the red ginseng extract and form a liposome containing the same do.
Step 3: After adding the natural coating agent to the mixture of the second step, At 80 ° C Ultrasonic treatment.
The natural coating agent is coated so that the shape of the lipid bilayer containing the red ginseng extract formed in the second step is stabilized so that the red ginseng extract of the final red ginseng-containing natural liposome is absorbed only in intestinal cells.
The surface of the lipid bilayer containing the red ginseng extract is coated by mixing the natural coating agent with the mixture of the second step and then heating the mixture to 35 to 80 ° C, preferably 40 to 60 ° C, and then performing ultrasonic treatment.
Step 4: adding to the mixture of the third step 35 - At 80 ° C Warmed Solvent and natural preservative are added and sonicated.
As the solvent, the same solvent as that used in the first step may be used.
The natural preservative prevents the decay of the liposome constituent material (natural emulsifier, solvent, natural coating agent) and the trapping substance present in the liposome, thereby maintaining the stability of the natural liposome containing red ginseng. In addition, the natural preservative is derived from a natural substance, has no toxicity, is harmless to the human body, and does not damage the natural liposome containing red ginseng.
The mixture of the natural preservative and the third step is mixed with a solvent heated to 35 to 80 ° C, preferably 40 to 60 ° C, and then subjected to ultrasonic treatment to obtain a natural liposome of a lipid bilayer containing a red ginseng extract To form particles.
Step 5: After the fourth step, the temperature is gradually lowered and repeatedly subjected to ultrasonic treatment.
The natural liposome containing red ginseng prepared by the above-described method is subjected to ultrasonic treatment at a temperature of 35 to 70 DEG C for a final time of 1 to 10 minutes, preferably 3 to 8 minutes while gradually lowering the temperature, Thereby producing a smaller and uniform final red ginseng-containing natural liposome.
The ultrasonic treatment may be repeatedly carried out to produce a natural liposome containing red ginseng having a desired particle size, preferably 1 to 3 times.
As described above, the process for preparing a natural liposome containing red ginseng for blood circulation according to the present invention can form a natural liposome containing a low-molecular red ginseng extract to increase the absorption rate of red ginseng extract in the intestines.
<Natural red ginseng Liposomes Containing food or pharmaceutical composition &
The natural liposome containing red ginseng for improving blood circulation according to the present invention binds with the cell membrane to increase the cell uptake rate of the red ginseng extract so that the active ingredient can act effectively in the human body.
These red ginseng extracts and natural liposomes containing the same are safe for human body, and when the natural liposome containing red ginseng for blood circulation improvement is ingested, the active ingredient can be exhibited by the red ginseng extract absorbed in the intestines, .
Specifically, the composition containing the natural liposome containing red ginseng can be used as a food such as various foods, beverages, gums, tea, vitamin complex, functional foods, and the like.
In addition, the composition containing the red ginseng-containing natural liposome has been found to be excellent in blood circulation improvement through cell experiments, and can be used as a pharmaceutical composition.
The food or pharmaceutical composition containing such a natural liposome containing red ginseng can improve arteriosclerosis by alleviating the arteriosclerosis or alleviate symptoms of ongoing arteriosclerosis.
Hereinafter, the present invention will be described concretely with reference to Examples. However, the following Examples are intended to illustrate one embodiment of the present invention, but the scope of the present invention is not limited by the following Examples.
[ Example 1] Red ginseng containing natural Liposome
The natural liposome containing red ginseng was prepared according to the composition shown in Table 1 below.
First, a natural emulsifier (Lipoid s75, Lipoid GMBH) was mixed with distilled water, heated to 65, and disrupted using an ultrasonic liquid processor. The red ginseng extract and the distilled water were mixed at a weight ratio of 1: 1 to prepare a capturing material. When the natural emulsifying agent was homogenized, the prepared capturing material was added, followed by ultrasonic disruption and mixing. When the natural emulsifier and the capturing material were completely homogenized, maltodextrin was added and the homogenization was performed by ultrasonic disintegration. Then, the remaining distilled water heated to 50 < 0 > C and grapefruit extract were added and ultrasonically disrupted to form liposome type particles. The temperature was gradually lowered, and finally ultrasonic disruption was repeated three times for 2 minutes and 30 seconds to homogenize, thereby preparing a natural liposome containing red ginseng.
[ Comparative Example 1] Red ginseng extract
The red ginseng extract used in Example 1 was used.
[ Comparative Example 2] Red ginseng content synthesis Liposome
The synthetic liposome containing red ginseng was prepared according to the composition shown in Table 2 below.
First, lecithin, cholesterol, ethanol and propylene glycol as synthetic emulsifiers were mixed with a part of distilled water. The synthetic emulsifier was mixed with stirring using a homomixer at 1300 rpm at 50 DEG C for 10 minutes. Then, when the synthetic emulsifier was homogenized, red ginseng extract was added and mixed again with emulsification equipment. When the synthetic emulsifier and red ginseng extract were mixed, the remaining distilled water, maltodextrin, and potassium sorbate were added to prepare a synthetic liposome. The resulting synthetic liposome was homogenized by ultrasonication to prepare a synthetic liposome containing red ginseng.
[ Comparative Example 3] Natural Liposome
Natural liposomes were prepared in the same manner as in Example 1, except that distilled water was used instead of red ginseng extract.
[ Experimental Example 1] Characteristic analysis
(1) Particle size analysis using laser particle size analyzer
The particle size of the natural liposome containing red ginseng of Example 1, the red ginseng extract of Comparative Example 1 and the red ginseng containing red ginseng of Comparative Example 2 was measured using a laser particle size analyzer. The results are shown in FIG.
As shown in FIG. 2A, the particle size of the natural liposome containing red ginseng (Example 1) was smaller than that of the red ginseng extract (Comparative Example 1) and the synthetic liposome containing red ginseng (Comparative Example 2).
(2) Electronic scanning microscope Particle size and morphology analysis
The particle size and shape of the natural liposome containing red ginseng of Example 1, the red ginseng extract of Comparative Example 1 and the red ginseng containing red ginseng of Comparative Example 2 were analyzed using a scanning electron microscope. The samples prepared in Example 1 and Comparative Examples 1 and 2 were frozen at -70 ° C for 2 hours in a freezer and then lyophilized using a freeze dryer. Then, the coating was spread out thinly and coated once with a PdPt catalyst, and then its surface and cross section were observed with a scanning electron microscope. The results are shown in FIG. 2B.
As shown in FIG. 2B, the natural liposome containing red ginseng (Example 1) was smaller in particle size and uniform in shape than the red ginseng extract (Comparative Example 1) and the synthetic liposome containing red ginseng (Comparative Example 2).
(3) Capture Measure efficiency
2 ml of the natural liposome containing red ginseng of Example 1 and the red ginseng containing red ginseng of Comparative Example 2 was filtered through a filter (0.45 um, 25 mm syringe filter) to remove the non-captured substances in the liposome. After removing the non-captured substances, the natural liposomes or synthetic liposomes were packed in the distilled water in the amount of the removed liposomes or synthetic liposomes and centrifuged at 5000RCF for 40 minutes in a centrifuge filter. The separated supernatant was filled with distilled water and recovered. The absorbance was measured at 280 nm, the maximum absorption wavelength of the red ginseng extract. The initial concentration of all the substances and the measured values of the concentrations of the collected substances were checked to confirm the collection efficiency. The results are shown in Table 3 below.
(Absorption efficiency (%) = absorbance of liposome containing red ginseng through filter / absorbance of liposome containing first red ginseng × 100)
As shown in Table 3, the natural liposome containing red ginseng (Example 1) showed higher collection efficiency than the synthetic liposome containing red ginseng (Comparative Example 2).
[ Experimental Example 2] Confirm cell permeability
A permeability test was carried out to predict the intestinal absorption rate of the human body.
When the permeable membrane of Transwell (12 well) was coated with collagen and dried, Caco-2 cells, which are human colon epithelial cells, were dispensed at 2 × 10 5 cells / well. Cells were cultured for 14 days and when the monolayer was formed, the medium was removed and replaced with a transport medium. After the transportation medium was maintained for 30 minutes, the apical chamber of each well was treated with distilled water, the natural liposome containing red ginseng of Example 1, and the red ginseng extract of Comparative Example 1, respectively. After 2 hours, the medium was recovered from the basolateral chamber and the absorbance was measured at the maximum absorption wavelength (280 nm) of red ginseng. The results are shown in FIG.
As shown in FIG. 3, the treatment of the natural liposome containing red ginseng (Example 1) showed significantly higher cell permeability than the treatment of red ginseng extract (Comparative Example 1).
[ Experimental Example 3] Confirming blood ginsenoside
Six - week - old SD rats were used to stop the diets except water supply for 24 hours before the experiment. Each of the liposomes containing the red ginseng of Example 1, the red ginseng extract of Comparative Example 1 and the red ginseng containing the red ginseng of Comparative Example 2 was orally administered to the oral cavity in an amount of 1% relative to body weight and was collected 3 hours later. As a standard sample, ginsenosides Rb1, Rb2, Rg3, and Compound K, purchased from Embo Laboratories, were used and a total of 10 ul was dispensed. As the developing solvent, chloroform: methanol: purified water was mixed at a ratio of 65:35:10 and only the down phase was separated. 10% sulfuric acid solution was sprayed and heated at 110 DEG C for 10 minutes to develop color. The results are shown in FIG.
As shown in Fig. 4, when the natural liposome containing red ginseng (Example 1) was fed, ginsenoside Rb2 was detected in the blood.
[ Experimental Example 4] Confirming functional improvement of blood circulation
(1) Human Aortic smooth muscle cells ( HASMC ) Toxicity experiment
Containing ginseng of after dispensing the
As shown in FIG. 5A, when the natural liposome containing red ginseng (Example 1) was treated, the cell survival rate showed almost no change.
(2) Human Aortic smooth muscle cells (HASMC) MMP -2 activity inhibitory effect
An appropriate number of HASMCs were cultured in a culture dish together with the medium. When the cells were filled in the culture dish, the culture medium was changed to a medium containing distilled water, the natural liposome containing red ginseng in Example 1, the red ginseng extract in Comparative Example 1 and the red ginseng containing red ginseng in Comparative Example 2, followed by culturing for 24 hours. The medium was loaded onto 10% SDS-polyacrylamide gel containing 0.05% gelatin, and the gel was stained. The results are shown in FIG. 5B.
As shown in FIG. 5B, when treated with the natural liposome containing red ginseng (Example 1), when the red ginseng extract (Comparative Example 1) was fed, compared to the treated red ginseng synthetic liposome (Comparative Example 2) -2 activity was inhibited.
(3) atherogenic agent MMP -1 Protein Expression Inhibitory Effect
The appropriate number of HASMC cells were cultured with the medium in a culture dish. When the cells were filled in the culture dish, the culture medium was changed to a medium containing the samples to treat distilled water, the natural liposome containing red ginseng of Example 1, the red ginseng extract of Comparative Example 1, and the synthetic liposome containing red ginseng of Comparative Example 2, Induced factor (PDGF-BB). After culturing for 24 hours, the medium was collected and concentrated 4 times by vacuum. The amount of protein in the concentrated medium was quantified and mixed in SDS loading buffer. It was boiled for 5 minutes and then cooled on ice for 15 minutes. Loaded onto 10% SDS-polyacrylamide gel and transferred to a PVDF membrane. The membrane was reacted for 2 hours in a 5% skim milk-TBST solution and then reacted overnight at 4 ° C in a MMP-1 antibody diluted in a 5% skim milk-TBST solution. The solution was shaken for 30 minutes in TBST solution, and then attached with secondary antigen. MMP-1 expression was confirmed using a Western blot film by reacting secondary antigens with an ECL solution in a dark room, and the results are shown in FIG. 5c.
As shown in FIG. 5C, when treated with the red ginseng extract (Comparative Example 1), the treatment of the natural liposome containing the red ginseng (Example 1) was more effective than the treatment of the synthetic liposome containing red ginseng (Comparative Example 2) -1 expression was suppressed.
(4) Human Aortic smooth muscle cells (HASMC) Confirm movement inhibition effect
HASMCs were cultured in 6 wells of Smooth muscle cell medium (SMCM) (Lonza, USA) or normal MCDB 131 medium. When the cells were tightly packed, the cells were exchanged with serum-free media, cultured for 24 hours, and treated with 25 ug / ml of mitomycin C for 30 minutes. After the medium was removed, the HASMC cells were scratched with a tip. After washing once with PBS, distilled water, the natural liposome containing red ginseng of Example 1, the red ginseng extract of Comparative Example 1 and the red ginseng containing red ginseng of Comparative Example 2 were each pretreated for 1 hour. Thereafter, the cells were cultured in an incubator for 24 hours after treatment with 10 ng / ml of atherogenic factor (PDGF-BB), and the cells were confirmed with a microscope. The results are shown in FIG. 5d.
As shown in FIG. 5D, in the case of treating the natural liposome containing red ginseng (Example 1), when treated with the red ginseng extract (Comparative Example 1), compared with the case of treating the synthetic liposome containing red ginseng (Comparative Example 2) Of the patients.
(5) Human Aortic smooth muscle cells (HASMC) Confirmation of inhibition of vascular infiltration
The transwells were coated with collagen and matrigel and dried. (24 wells) were each prepared by adding distilled water, a natural liposome containing red ginseng of Example 1, a red ginseng extract of Comparative Example 1, and a synthetic liposome containing red ginseng of Comparative Example 2 and a medium (BSA + serum-free medium) And the cells were cultured in an incubator for 12 hours. The transwells were then immersed in methanol for 1 minute, in hematoxylin for 10 minutes, and then washed with distilled water to remove water. The transwells were again stained with eosin for 4 minutes, washed with distilled water, and then all cells inside the transwells were wiped off and dried for 1 hour or more. The collagen layer of Transwell was cut with a razor blade, washed with xylene, placed on a slide glass, and then observed with a microscope. The result is shown in Fig. 5E.
As shown in FIG. 5E, when the natural liposome containing red ginseng (Example 1) was treated and the red ginseng extract (Comparative Example 1) was treated, compared to the case where the red lip gum containing the red ginseng (Comparative Example 2) Of the patients.
Claims (11)
Liposome constituents comprising natural emulsifiers, solvents, natural coatings and natural preservatives; And
Red ginseng extract
Containing red ginseng for improving blood circulation.
Wherein the natural emulsifier comprises at least one phospholipid selected from the group consisting of phosphatidylcholine, lysophosphatidylcholine, phosphatidylethanolamine, hydratidine phosphatidylcholine, phosphatidylic acid, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, Natural red liposome containing red ginseng.
Wherein the natural emulsifier comprises at least one fatty acid selected from the group consisting of palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid.
Wherein the solvent comprises at least one solvent selected from the group consisting of distilled water, natural butylene glycol, propanediol, glycerin and fermented alcoholic beverages.
Wherein the natural coating agent comprises at least one natural coating agent selected from the group consisting of maltodextrin, cellulose and pectin.
Wherein the natural preservative comprises at least one natural extract selected from the group consisting of citrus extract, citron extract, eucalyptus extract, ginseng extract, clove extract and grapefruit extract.
Wherein the natural liposome containing red ginseng contains 0.1 to 10% by weight of a natural emulsifier, 0.01 to 5% by weight of a natural coating agent, 0.001 to 3% by weight of a natural preservative, 0.1 to 30% by weight of a red ginseng extract, Natural red liposome containing red ginseng.
(b) a second step of adding the red ginseng extract to the mixture of the first step followed by ultrasonic treatment at 35 to 80 캜;
(c) a third step of adding a natural coating agent to the mixture of the second step and ultrasonication at 35 to 80 ° C; And
(d) adding to the mixture of the third step a solvent warmed at 35 to 80 DEG C and a natural preservative, and ultrasonifying
7. The method for producing a natural liposome containing red ginseng for blood circulation according to any one of claims 1 to 7.
And a fifth step of repeating ultrasonic treatment while gradually lowering the temperature after the fourth step.
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