WO2011043564A2 - Composition containing black ginseng extracts for preventing or treating liver cancer - Google Patents
Composition containing black ginseng extracts for preventing or treating liver cancer Download PDFInfo
- Publication number
- WO2011043564A2 WO2011043564A2 PCT/KR2010/006744 KR2010006744W WO2011043564A2 WO 2011043564 A2 WO2011043564 A2 WO 2011043564A2 KR 2010006744 W KR2010006744 W KR 2010006744W WO 2011043564 A2 WO2011043564 A2 WO 2011043564A2
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- WIPO (PCT)
- Prior art keywords
- ginseng
- black
- steaming
- panax
- black ginseng
- Prior art date
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61K36/258—Panax (ginseng)
-
- 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
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- 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
Definitions
- the present invention relates to a method for producing black ginseng by separating steamed ginseng, which is more than four years old, into the body and the rice, dried with white ginseng, and repeated several times.
- the present invention relates to a method for producing black ginseng having increased benzopyrene stability and Rg 3 component and having a prophylactic or therapeutic effect on liver cancer, and a composition thereof.
- Ginseng ( ⁇ , ginseng) is a perennial herb of the dicotyledon type Arboraceae, grows in deep mountain forests and grows as a medicinal plant.
- the root of ginseng is called ginseng because its shape is human. The shape of the roots varies with age, and harvest is done in 4-6 years. Ginseng as it is harvested is called ginseng, and dried ginseng is called white ginseng. Steaming red ginseng makes red ginseng, which is called red ginseng. When the red ginseng is steamed and dried several times, it becomes black ginseng.
- red ginseng and black ginseng is known to improve the active ingredients beneficial to health compared to fresh ginseng by changing the pharmacological components through steaming and drying process.
- the main bioactive components of red ginseng and black ginseng are known as Ginsenoside-Rb 1 , Rb 2 , Rc, Rd, Re, Rg 1 , Rg 2 , Rg 3 and Ro contained in saponin.
- Black ginseng is known to be more enhanced ginsenoside Rg 3 component than in particular red ginseng.
- Black ginseng is a newly developed product, and since it was recently developed, it is not listed as a regular food, but it contains ginsenoside Rg 3 and can be processed into pouches or concentrates and classified as red ginseng processed foods.
- Black ginseng has already been shown to be effective in boosting immunity, anticancer and sexual function. In addition, through the experience and research results of the present inventors, it was confirmed that it is effective in hepatitis, diabetes and cirrhosis.
- the present inventors have developed a method for increasing the ginsenoside component of black ginseng through Korean Patent No. 529475 and again invented a method for enhancing Rg 3 in the ginsenoside component.
- the present invention utilizes a new method that can further enhance the Rg 3 component as compared to Korean Patent Publication No. 840003, which is known to be able to enhance ginsenoside Rg 3 .
- the method of the present invention proposes a method for producing black ginseng under conditions that can remove benzopyrene, which is known to be produced in the heating process during the manufacture of black ginseng, to below a safety standard.
- Benzopyrene is a yellow crystalline solid belonging to the polycyclic aromatic hydrocarbons (PAHs) group, and it is known that carbohydrates, proteins and lipids are produced as incomplete combustion when decomposed at temperatures between 300 and 600 °C in foods. Recently, it is also reported to be produced in processed food below 100 °C.
- the International Agency for Research on Cancer (IARC) underwent a primary toxicity assessment in 1989 and re-evaluated in August 2006 to classify benzopyrene as the first group of human carcinogens.
- the present inventors confirmed that the black ginseng extract of the present invention has a selectively excellent effect on liver cancer.
- black ginseng extract has an anticancer effect, it is known about the effects on cancer or oxidation-related diseases of colorectal cancer of Korean Patent Application Laid-Open No. 2006-83301 and 2008-106077 Korean Patent Publication No. 2006-83301. It is not about selective effects.
- Still another object of the present invention is to provide a method for producing black ginseng body and rice ginseng, which are effective in preventing or treating liver cancer.
- the present invention is a method for producing black ginseng by repeating steaming and drying the ginseng,
- Step 1 washing the ginseng and separating the body and ginseng;
- Step 2 drying the washed ginseng body and the ginseng so that the moisture content is less than 14%;
- Step 4 drying the steamed body and rice ginseng at a temperature of 25 ⁇ 50 °C for 3 to 5 hours;
- Step 6 drying the black ginseng body and black rice ginseng after repeated steaming and drying of step 5 at a temperature of 25 to 50 ° C. to finally have 10% or less of moisture;
- Ginseng of the first step may be 4-6 years old ginseng.
- the moisture of the ginseng dried in the second step is preferably about 5 ⁇ 14%.
- the steaming temperature of the 3 step is preferably 80 ⁇ 90 °C, if less than 80 °C ginsenoside Rg 3 component does not increase much and benzopyrene is not removed well, even if it exceeds 90 °C Rg 3 component also increases Benzopyrene is not removed by the safety regulations.
- the total number of steaming after the completion of the process 5 to 3 is preferably 6 to 13 times by adding 5 to 12 additional steams to the first steam. If the total number of steams is less than six, the ginsenoside Rg 3 component is not increased much and benzopyrene is not removed as well as the safety regulations. As the total number of steams increases, Rg 3 increases but other ginsenosides fall off or disappear, so the number of steams and temperature must be well controlled.
- the black ginseng body of the present invention and the pulverized extract or solvent (purified water or ethanol) extract of the ginseng can be used when producing a health food or anti-cancer functional medicines, including tablets, capsules, pills, solutions, etc. .
- the extract of the black ginseng body and the rice ginseng may be prepared as a powder through lyophilization, hot air drying or spray drying, and may be added to pharmaceutical preparations or foods.
- Ginseng as defined in the present invention is a perennial plant belonging to the genus Panax, Panax ginseng, Panax quinquefolia, Panax Notoginseng, Panax vietnamensis, Panax vietnamensis, Panax elegathior (Panax elegatior), Panax wangianus or Panax bipinratifidus, preferably Panax ginseng or Panax quinquefolia.
- the pharmaceutical composition comprising the black ginseng body and the ginseng according to the present invention that can be used as an agent for treating cancer is freeze-dried, hot air-dried or spray-dried the extracts of the black ginseng body and the ginseng powder, granules, Oral formulations such as tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions.
- Carriers, excipients, and diluents that may be included in compositions comprising black ginseng body and rice ginseng include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium Phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
- Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin in the extract. The mixture is prepared.
- lubricants such as magnesium stearate and talc are also used.
- Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin.
- Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories.
- the non-aqueous solvent and suspending agent propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used.
- As the base of the suppository witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
- the dosage of the active ingredient will vary depending on the age, sex and weight of the subject to be treated, the particular disease or pathology to be treated, the severity of the disease or pathology, the route of administration and the judgment of the prescriber. Dosage determination based on these factors is within the level of skill in the art and generally dosages range from 0.001 mg / kg / day to approximately 2000 mg / kg / day. More preferred dosage is 1 mg / kg / day to 100 mg / kg / day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.
- the pharmaceutical composition for preventing and treating liver cancer-related diseases of the present invention comprises 0.1 to 50% by weight of the black ginseng extract based on the total weight of the composition.
- the extract of the present invention can be administered to mammals such as mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injections. Since the extract of the present invention has little toxicity and side effects, it is a drug that can be used safely even when taken for long periods of time.
- the present invention provides a health functional food for cancer treatment improvement comprising the black ginseng and black rice ginseng according to the present invention is a food supplement acceptable food supplement.
- the health functional food of the present invention includes the form of tablets, capsules, pills or liquids, and the food to which the compound of the present invention can be added, for example, various foods, beverages, gums, teas, vitamin complexes, etc. And functional foods.
- ginsenoside Rg 3 is significantly increased in the black ginseng prepared through the present invention than the black ginseng prepared in the prior art.
- the amount of ginsenoside Rg 3 increases sequentially as the number of steaming increases, and the formation of the amount is sensitive to temperature and drying conditions.
- Benzopyrene also known as a carcinogen, has been removed to below safety regulations by reducing its amount through repeated steaming at the temperature and steaming conditions of the present invention.
- benzopyrene is detected to be 1.0 ppb or more, which is higher than the prescribed value, but it can be confirmed that the amount of benzopyrene decreases below the prescribed value through the steaming process.
- benzopyrene is produced from pesticides remaining in ginseng as a raw material, and the benzopyrene thus produced is removed through the repeated steaming and drying conditions of the present invention.
- benzopyrene was also detected in the ginseng before steaming, benzopyrene was detected only when black ginseng was prepared using the ginseng, and no benzopyrene was detected in ginseng from the beginning.
- the black ginseng with the increased ginsenoside Rg 3 has an excellent anticancer effect against liver cancer. This has established a method for producing black ginseng that is effective in liver cancer-related diseases and safe from carcinogens.
- FIG. 1 is a flowchart illustrating a process of preparing black ginseng according to the method of the first preferred embodiment.
- Figure 2 is a graph showing the results of the MTT assay for the cell survival rate of hepatic cancer cell HepG2 of Experimental Example 2-1.
- Example 1 Preparation of black ginseng main body and rice ginseng and extracts using the same: steaming temperature 85 °C, drying temperature 50 °C, the total number of steaming 13 times>
- the dried white ginseng was changed to red ginseng and black ginseng to black ginseng, and the black ginseng body and rice were prepared, and the last drying process was completed until the water became less than 10%. Thereafter, the black ginseng body and the ginseng were ground to prepare an extract of the black ginseng body and the rice ginseng.
- the pulverized black ginseng body and rice ginseng were put into 100 l of purified water having a temperature of 80 ° C., respectively, and stirred for 8 hours at a rotational speed of 60 revolutions per minute, and then precipitated for 3 hours.
- the precipitated extract was filtered to remove solids, and the filtrate was lyophilized to finally prepare a black ginseng body and a ginseng extract as powder.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the steaming temperature was 80 ° C.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the steaming temperature was 90 ° C.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed only six times.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed only seven times.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed only eight times.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed only 9 times.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed only 10 times.
- the black ginseng body and the extract were extracted under the same conditions as in Example 1, but the total number of steaming was performed only 11 times.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed 12 times.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the drying temperature was 25 ° C.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the steaming temperature was 70 ° C.
- the black ginseng body and the rice extract were prepared under the same conditions as in Example 1, but the steaming temperature was 75 ° C.
- the black ginseng body and the rice extract were prepared under the same conditions as in Example 1, but the steaming temperature was 95 ° C.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed only once.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed twice.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but only three times of total steaming was performed.
- the black ginseng body and the extract were extracted under the same conditions as in Example 1, but the total number of steaming was performed only four times.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but only five times of total steaming was performed.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed 14 times.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but drying was performed at 20 ° C.
- the black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but drying was performed at 55 ° C.
- red ginseng root red ginseng body
- red ginseng extract were prepared using purified water at 80 ° C. in the same manner as the extraction method of Example 1.
- the ginsenoside components of the black ginseng extract and the red ginseng extract of Comparative Example 4 prepared according to the methods of Examples 1 to 4 and Comparative Examples 1 to 3 were analyzed. Since ginsenoside components are known to be more in the root portion, black and red ginseng were used in this experiment, respectively.
- ginsenosides The analysis of ginsenosides was carried out in accordance with Food Code [Chapter 4. Food Standards and Standards ⁇ 16.Ginseng Products ⁇ 16-10.Concentrated Red Ginseng (1) Red Ginseng Identification Test High-Speed Liquid Chromatography]. The senoside test method was used, and Table 1 is a table showing the analysis of ginsenoside content of black ginseng and red ginseng.
- the results of Table 1 confirm that the ginsenoside Rg 3 of black ginseng was significantly increased as the number of steaming was increased, and in particular, a large amount of ginsenoside Rg 3 was increased in at least 8 times and in ginsenosides at 9 or more times. It was confirmed that the Rg 3 content was maintained about the same.
- the steaming temperature of ginsenosides Rg 3 and Rh 1 was most increased between 80 and 90 ° C around 85 ° C. I could confirm it.
- the ginsenosides Rg 3 and Rh 1 contents of red ginseng showed significantly lower values.
- the total amount of irradiated black ginseng of Example 1 was analyzed to be 508.9 mg / g, and the irradiated amount of irradiated with the same amount of irradiated with red ginseng of Comparative Example 4 (72.1 mg / g) was increased during initial steaming. It was confirmed that the increase (not shown in the diagram).
- the cell survival rate of hepatic cancer cell HepG2 was measured using the black ginseng extract powder of Example 1 and the red ginseng extract powder of Comparative Example 4.
- Black ginseng was used as a representative of the extract of Example 1 having a high Rg 3 content and black and red ginseng as in Experimental Example 1.
- the black ginseng extract of Example 1 of the present invention can confirm the 50% inhibitory activity against liver cancer cell proliferation at the concentration of 8 ⁇ g / ml or less, the red ginseng extract of Comparative Example 3 to the liver cancer cell proliferation It can be seen that 50% inhibition against the high concentration of 1000 ⁇ g / ml or more. As a result, the black ginseng prepared by the production method of the present invention can be confirmed to have a selective and excellent anticancer effect on liver cancer cells.
- HepG2 cells to be transplanted were placed in cell culture flasks using DMEM (Gibco BLR) cell culture medium containing 100 U / ml penicillin, 100 g / ml streptomycin, 10% bovine serum and cultured in a 95% CO 2 37 ° C. cell culture medium. .
- DMEM Gibco BLR
- the cells were washed twice with HBSS (Hanks balanced salt solution, Gibco BRL) and collected with 0.2% trypsin.
- Experimental animals used 9-week old BALB / c nude mice (SLC inc., Shizuoka, Japan), each of which was 9 animals. Each extract was orally administered daily at 300 mg / kg and only purified water was administered to the control group.
- these nude mice were injected with 1 ⁇ 10 7 HepG2 cells to obtain tumor lines, which were then passaged to the passage group.
- the tumor mass was recovered in three passages, and then the tumor mass was transplanted to eight passages.
- Tumor sections were made by cutting the outer areas where rapid division occurred to a certain size (3 ⁇ 3 ⁇ 3 mm) at the expense of animals containing tumors growing rapidly with sufficient blood supply before tumor necrosis occurred.
- the tumor piece was placed at the end of the trocar and the end was reached to the trunk body side of the rear cell of the animal.
- the trocar was lightly and quickly rotated out of 360 ° to allow the tumor piece to settle in the desired location and the dissection site was disinfected.
- the experiment was terminated and the autopsy was performed to determine the weight of solid cancers extracted from each subject, and the actual volume was measured using digital plethysmomether.
- the black ginseng extract of the present invention was confirmed that the ability to inhibit the weight and tumor volume of the tumor in liver cancer animal model experiments than the red ginseng extract.
- black ginseng and red ginseng extracts were used in the same manner as in Experiment 1 and Experiment 2.
- the benzopyrene analysis method is the black ginseng body prepared in Examples 1 to 4 and Comparative Examples 1 to 3 according to the benzopyrene test method of the health functional food (so-called black ginseng) of December 31, 2007 4 g of the extract was extracted and concentrated with dichloromethane, dissolved in hexane, passed through a prolysyl cartridge, dried and dissolved in acetonitrile, and analyzed by high performance liquid chromatography (High Chromatograph). 3-methylcollanthene was used as an internal standard, and a fluorescence detector was used as a detector.
- the column was used in the reverse phase distribution type (C18 Supelco LC-PAH 4.6mm ⁇ 250mm I.D.) and the analysis conditions are shown in Table 4 below and are shown in Table 5 by repeating five times.
- Benzopyrene is less than 1.0 ppb below safety regulations.
- the amount of benzopyrene was also detected in ginseng before steaming, but as expected, the amount of benzopyrene was detected as 1.02 ppb. Because of this, clean ginseng that is not detected benzopyrene was steamed separately before steaming, and black ginseng steamed using ginseng that is not detected by benzopyrene was confirmed that benzopyrene was not detected unlike before (as shown in the diagram). Not).
- the black ginseng produced using the present invention may be added to medicines or foods and used as a cancer prevention agent, a cancer treatment agent, or the like.
- the present invention provides a high anticancer drug and a method for preparing a functional food containing a black ginseng extract using a black ginseng body and rice.
- the above ingredients are mixed and filled in an airtight cloth to prepare a powder.
- tablets are prepared by tableting according to a conventional method for preparing tablets.
- the above ingredients are mixed and filled into gelatin capsules to prepare capsules.
- composition ratio of the above-mentioned vitamin and mineral mixtures is composed of relatively suitable ingredients suitable for health foods in a preferred formulation example, but the compounding ratios may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method.
- the granules may be prepared and used for preparing a health food composition according to a conventional method.
- composition ratio is a composition suitable for a preferred beverage in a preferred formulation example
- the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and use purpose.
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Abstract
The present invention relates to a method for specifically increasing Rg3 ingredients from among the ginsenoside ingredients of black ginseng. The black ginseng extracts prepared by the method exhibit a high anticancer activity against human liver cancer cells. Further, it has been verified that the method of the present invention has the effect of decreasing the content of benzopyrene which is known to be mainly generated during heating.
Description
본 발명은 4년근 이상 된 수삼을 몸체와 미삼으로 분리하여 백삼으로 건조한 후 증숙과 건조 과정을 수차례 반복하여 흑삼을 제조하는 방법에 관한 것이다.The present invention relates to a method for producing black ginseng by separating steamed ginseng, which is more than four years old, into the body and the rice, dried with white ginseng, and repeated several times.
더욱 상세하게는 벤조피렌 안정성 및 Rg3 성분이 증가되고 간암 예방 또는 치료 효과가 있는 흑삼을 제조하는 방법 및 이의 조성물에 관한 것이다. More particularly, the present invention relates to a method for producing black ginseng having increased benzopyrene stability and Rg 3 component and having a prophylactic or therapeutic effect on liver cancer, and a composition thereof.
인삼(人蔘, ginseng)은 쌍떡잎식물 산형화목 두릅나무과의 여러해살이풀로 깊은 산의 숲속에서 자라며 약용식물로 재배한다. 인삼의 뿌리는 그 형태가 사람 형상이므로 인삼이라 한다. 뿌리의 형태는 나이에 따라 차이가 있고 수확은 4∼6년근 때에 한다. 수확된 상태 그대로의 인삼을 수삼이라고 하며 수삼을 건조한 것을 백삼이라고 한다. 수삼을 증숙하여 붉은색을 띄는 것을 홍삼(red ginseng)이라 하며 상기 홍삼을 수회에 걸쳐 증숙 및 건조하게 되면 검은색을 띄는 흑삼(black ginseng)이 된다. 이러한 홍삼 및 흑삼은 증숙 및 건조 과정을 통해 약리 성분이 변화되어 수삼과 비교하여 건강에 유익한 활성성분이 증진되는 것으로 알려져 있다. 홍삼과 흑삼의 주요 생리활성 성분은 사포닌 속에 함유되어 있는 진세노사이드(Ginsenoside)-Rb1, Rb2, Rc, Rd, Re, Rg1, Rg2, Rg3, Ro 등으로 알려져 있다. Ginseng (人蔘, ginseng) is a perennial herb of the dicotyledon type Arboraceae, grows in deep mountain forests and grows as a medicinal plant. The root of ginseng is called ginseng because its shape is human. The shape of the roots varies with age, and harvest is done in 4-6 years. Ginseng as it is harvested is called ginseng, and dried ginseng is called white ginseng. Steaming red ginseng makes red ginseng, which is called red ginseng. When the red ginseng is steamed and dried several times, it becomes black ginseng. These red ginseng and black ginseng is known to improve the active ingredients beneficial to health compared to fresh ginseng by changing the pharmacological components through steaming and drying process. The main bioactive components of red ginseng and black ginseng are known as Ginsenoside-Rb 1 , Rb 2 , Rc, Rd, Re, Rg 1 , Rg 2 , Rg 3 and Ro contained in saponin.
흑삼은 특히 홍삼과 비교하여 진세노사이드 Rg3 성분이 더욱 강화된 것으로 알려져 있다. 흑삼은 신개발 제품으로서 최근에 개발되었기 때문에 정식 식품으로 등재되어 있지는 않으나, 진세노사이드 Rg3가 함유되어 있어 파우치나 농축액으로 가공되어 홍삼가공식품류로 분류될 수 있다. 흑삼이 면역력 증강, 항암 및 성기능 개선 등에서 효과가 있음은 이미 소비자들의 경험을 통해 확인되고 있다. 또한 이 외에도 본 발명자의 경험 및 연구결과를 통해 간염, 당뇨 및 간경화 등에도 효과가 있음을 확인하였다.Black ginseng is known to be more enhanced ginsenoside Rg 3 component than in particular red ginseng. Black ginseng is a newly developed product, and since it was recently developed, it is not listed as a regular food, but it contains ginsenoside Rg 3 and can be processed into pouches or concentrates and classified as red ginseng processed foods. Black ginseng has already been shown to be effective in boosting immunity, anticancer and sexual function. In addition, through the experience and research results of the present inventors, it was confirmed that it is effective in hepatitis, diabetes and cirrhosis.
본 발명자는 한국등록특허 제529475호를 통하여 흑삼의 진세노사이드 성분을 증가시킬 수 있는 방법을 개발하였으며 이에 대해 다시 상기 진세노사이드 성분 중 특히 Rg3를 증강시킬 수 있는 방법을 발명하였다. 본 발명은 진세노사이드 Rg3를 증강시킬 수 있다고 이미 공지된 한국공개특허 제840003호와 비교해 Rg3 성분을 더 강화할 수 있는 새로운 방법을 이용한다. The present inventors have developed a method for increasing the ginsenoside component of black ginseng through Korean Patent No. 529475 and again invented a method for enhancing Rg 3 in the ginsenoside component. The present invention utilizes a new method that can further enhance the Rg 3 component as compared to Korean Patent Publication No. 840003, which is known to be able to enhance ginsenoside Rg 3 .
또한 본 발명의 방법은 흑삼을 제조하는 도중 가열 공정에서 생성된다고 알려져 있는 벤조피렌을 안전규정 수치 미만으로 제거할 수 있는 조건의 흑삼 제조 방법을 제안한다. 벤조피렌은 다환방향족탄화수소(PAHs, polycyclic aromatic hydrocarbons) 그룹에 속하는 황색의 결정성 고체로서, 식품에 있어 탄수화물, 단백질, 지질 등이 300∼600℃ 사이 온도에서 분해할 때 불완전연소로 생성되는 것으로 알려져 있는데 근래에는 100℃ 이하의 가공식품에서도 생성되는 것으로도 보고되었다. 국제암연구소(IARC, International Agency for Research on Cancer)의 경우 1989년에 1차 독성평가를 진행하였고, 2006년 8월 재평가를 하여 벤조피렌은 제 1 그룹 인체 발암 물질로 분류하고 있다. 흑삼 제조와 벤조피렌 저감에 대해서는 한국공개특허 제2009-93310호가 공지되어 있는데 상기 방법은 건조 온도를 조절하여 벤조피렌이 증가되지 않도록 하는 방법에 대한 기술이지만 본 발명의 방법은 증숙 과정을 증가할수록 벤조피렌이 제거되는 방법으로 상기 한국공개특허 제2009-93310호와는 다른 기술이다.In addition, the method of the present invention proposes a method for producing black ginseng under conditions that can remove benzopyrene, which is known to be produced in the heating process during the manufacture of black ginseng, to below a safety standard. Benzopyrene is a yellow crystalline solid belonging to the polycyclic aromatic hydrocarbons (PAHs) group, and it is known that carbohydrates, proteins and lipids are produced as incomplete combustion when decomposed at temperatures between 300 and 600 ℃ in foods. Recently, it is also reported to be produced in processed food below 100 ℃. The International Agency for Research on Cancer (IARC) underwent a primary toxicity assessment in 1989 and re-evaluated in August 2006 to classify benzopyrene as the first group of human carcinogens. It is known that black ginseng production and benzopyrene reduction is disclosed in Korea Patent Publication No. 2009-93310. The method is a technique for controlling the drying temperature so that the benzopyrene is not increased, but the method of the present invention increases the steaming process to remove benzopyrene. The technique is different from the Korean Patent Publication No. 2009-93310.
또한 본 발명자는 본 발명의 흑삼 추출물이 간암에 선택적으로 우수한 효과가 있는 것을 확인하였다. 현재 흑삼 추출물이 항암효과가 있다는 것에 대해서는 한국공개특허 제2006-83301호의 대장암과 한국공개특허 제2008-106077호의 암질환 또는 산화 관련 질환에 대한 효과에 대해서 공지되어 있지만 본 발명과 같이 간암에 대해 선택적 효과에 대한 것은 아니다.In addition, the present inventors confirmed that the black ginseng extract of the present invention has a selectively excellent effect on liver cancer. Although black ginseng extract has an anticancer effect, it is known about the effects on cancer or oxidation-related diseases of colorectal cancer of Korean Patent Application Laid-Open No. 2006-83301 and 2008-106077 Korean Patent Publication No. 2006-83301. It is not about selective effects.
본 발명의 목적은 4년근 이상의 수삼을 이용하여 증숙 온도 및 증숙 횟수를 조절하여 진세노사이드 Rg3가 증강되고 벤조피렌이 저감된 흑삼 몸체 및 미삼을 제조하는 방법을 제공하는 것이다.It is an object of the present invention to provide a method for producing black ginseng body and rice ginseng, in which ginsenoside Rg 3 is enhanced and benzopyrene is reduced by controlling steaming temperature and number of steaming using 4 g of ginseng or more.
본 발명의 또 다른 목적은 간암을 예방 또는 치료할 수 있는 효과가 있는 흑삼 몸체 및 미삼을 제조하는 방법을 제공하는 것이다.Still another object of the present invention is to provide a method for producing black ginseng body and rice ginseng, which are effective in preventing or treating liver cancer.
본 발명은 수삼을 증숙과 건조를 반복하여 흑삼을 제조하는 방법으로, The present invention is a method for producing black ginseng by repeating steaming and drying the ginseng,
(1공정) 인삼을 세척하고 몸체와 미삼으로 분리하는 단계; (Step 1) washing the ginseng and separating the body and ginseng;
(2공정) 세척된 인삼의 몸체와 미삼의 수분 함량이 14% 이하가 되도록 건조하는 단계; (Step 2) drying the washed ginseng body and the ginseng so that the moisture content is less than 14%;
(3공정) 건조된 인삼의 몸체와 미삼을 각각 80~90℃의 온도에서 8~10시간 동안 증숙하는 단계; (3 step) steaming the body and dried ginseng of dried ginseng at a temperature of 80 ~ 90 ℃ for 8 to 10 hours;
(4공정) 증숙된 인삼의 몸체와 미삼을 25~50℃의 온도로 3~5시간 동안 건조하는 단계; (Step 4) drying the steamed body and rice ginseng at a temperature of 25 ~ 50 ℃ for 3 to 5 hours;
(5공정) 상기 3공정의 증숙 단계과 4공정의 25~50℃ 건조 단계를 연속으로 5~12회 반복하는 단계; 및 (5 steps) repeating 5 to 12 times the steaming step of the three steps and the 25 ~ 50 ℃ drying step of the four steps in a row; And
(6공정) 5공정의 반복 증숙 및 건조가 완료된 흑삼 몸체와 흑미삼을 최종적으로 수분이 10% 이하가 되게 25~50℃의 온도에서 건조하는 단계; (Step 6) drying the black ginseng body and black rice ginseng after repeated steaming and drying of step 5 at a temperature of 25 to 50 ° C. to finally have 10% or less of moisture;
를 포함하여 흑삼을 제조할 수 있다. It can be prepared including black ginseng.
상기 1공정의 인삼은 4~6년근 인삼일 수 있다.Ginseng of the first step may be 4-6 years old ginseng.
상기 1공정에서 인삼의 몸체와 미삼을 분리할 때는 표피를 벗기지 않아서 인삼의 몸체와 표피 사이에 형성되어 있는 사포닌의 손실을 막아야 한다. When separating the body of the ginseng and the ginseng in the first step does not peel the epidermis to prevent the loss of saponin formed between the body and the epidermis of the ginseng.
상기 2공정에서 건조된 인삼의 수분은 적절하게는 5~14% 전후가 되는 것이 좋다.The moisture of the ginseng dried in the second step is preferably about 5 ~ 14%.
상기 3공정의 증숙 온도는 80~90℃가 바람직한데, 80℃ 미만이 되면 진세노사이드 Rg3 성분이 많이 증가되지 않고 벤조피렌도 잘 제거되지 않는데, 90℃를 초과하게 되어도 Rg3 성분이 역시 증가되지 않으면서 벤조피렌도 안전규정 수치만큼 제거되지 않는다. The steaming temperature of the 3 step is preferably 80 ~ 90 ℃, if less than 80 ℃ ginsenoside Rg 3 component does not increase much and benzopyrene is not removed well, even if it exceeds 90 ℃ Rg 3 component also increases Benzopyrene is not removed by the safety regulations.
상기 3공정에서 5공정을 완료한 뒤의 총 증숙 횟수는 최초 증숙 1회에 추가 증숙 5~12회를 더하여 6~13회가 바람직하다. 총 증숙 횟수가 6회 미만이 되면 진세노사이드 Rg3 성분이 많이 증가되지 않으며 벤조피렌이 안전규정 수치만큼 잘 제거되지 않는다. 총 증숙 횟수가 증가할수록 Rg3는 증가하나 다른 진세노사이드 성분이 떨어지거나 소멸되기 때문에 증숙 횟수와 온도를 잘 조절해야 한다.The total number of steaming after the completion of the process 5 to 3 is preferably 6 to 13 times by adding 5 to 12 additional steams to the first steam. If the total number of steams is less than six, the ginsenoside Rg 3 component is not increased much and benzopyrene is not removed as well as the safety regulations. As the total number of steams increases, Rg 3 increases but other ginsenosides fall off or disappear, so the number of steams and temperature must be well controlled.
또한, 본 발명의 흑삼 몸체와 미삼의 분쇄물 또는 용매(정제수 또는 에탄올) 추출물은 항암기능이 있는 건강 식품이나 정제, 캡슐제, 환제, 액제 등을 포함하는 기능성 의약품 등을 제조할 때 사용될 수 있다. In addition, the black ginseng body of the present invention and the pulverized extract or solvent (purified water or ethanol) extract of the ginseng can be used when producing a health food or anti-cancer functional medicines, including tablets, capsules, pills, solutions, etc. .
상기 흑삼 몸체와 미삼 추출물을 제조하기 위해서는, In order to manufacture the black ginseng body and the extract of misam,
건조된 흑삼 몸체 또는 미삼을 분쇄하는 단계; Grinding the dried black ginseng body or rice ginseng;
상기 분쇄된 흑삼 몸체 또는 미삼 1kg 당 70~90℃의 온도를 가지는 정제수 5~120ℓ를 상기 분쇄된 흑삼 몸체 또는 미삼과 혼합하고 1분 당 50~70 회전의 회전 속도로 5~10시간 동안 교반하는 단계; 5 to 120 L of purified water having a temperature of 70 to 90 ° C. per 1 kg of the ground black ginseng body or rice ginseng is mixed with the ground black ginseng body or rice ginseng and stirred for 5 to 10 hours at a rotational speed of 50 to 70 rotations per minute. step;
상기 교반된 추출물을 2~14시간 동안 침전하여 액상과 고형체로 분리하는 단계; 및 Precipitating the stirred extract for 2 to 14 hours to separate the liquid and solid body; And
상기 추출물을 여과하여 고형체를 제거한 여액을 수득하는 단계;Filtering the extract to obtain a filtrate from which solids are removed;
를 포함할 수 있다. It may include.
상기 흑삼 몸체와 미삼의 추출물은 동결건조, 열풍건조 또는 분무건조 등을 통해 분말로 제조되어 약학제제 또는 식품에 첨가될 수 있다. The extract of the black ginseng body and the rice ginseng may be prepared as a powder through lyophilization, hot air drying or spray drying, and may be added to pharmaceutical preparations or foods.
본 발명에서 정의되는 인삼은 파낙스(Panax)속에 속하는 다년생 식물로, 고려인삼(Panax ginseng), 화기삼(Panax quinquefolia), 전칠삼(삼칠, Panax notoginseng), 죽절삼(Panax vietnamensis), 파낙스 엘레가티오르(Panax elegatior), 파낙스 완지아누스(Panax wangianus) 또는 파낙스 피핀라티푸스(Panax bipinratifidus), 바람직하게는 고려인삼(Panax ginseng) 또는 화기삼(Panax quinquefolia)을 포함한다.Ginseng as defined in the present invention is a perennial plant belonging to the genus Panax, Panax ginseng, Panax quinquefolia, Panax Notoginseng, Panax vietnamensis, Panax vietnamensis, Panax elegathior (Panax elegatior), Panax wangianus or Panax bipinratifidus, preferably Panax ginseng or Panax quinquefolia.
암 치료용 제제로 사용될 수 있는 본 발명에 따른 흑삼 몸체와 미삼을 포함하는 약학 조성물은, 상기 흑삼 몸체와 미삼의 추출물을 동결건조, 열풍건조 또는 분무건조한 것을 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 흑삼 몸체와 미삼을 포함하는 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토오즈, 덱스트로오즈, 수크로오즈, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 제제화 할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 추출물에 적어도 하나 이상의 부형제 예를 들면, 전분, 탄산칼슘, 수크로오즈 또는 락토오즈, 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다. The pharmaceutical composition comprising the black ginseng body and the ginseng according to the present invention that can be used as an agent for treating cancer is freeze-dried, hot air-dried or spray-dried the extracts of the black ginseng body and the ginseng powder, granules, Oral formulations such as tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injectable solutions. Carriers, excipients, and diluents that may be included in compositions comprising black ginseng body and rice ginseng include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium Phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, gelatin in the extract. The mixture is prepared. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
상기 활성성분의 투여량은 치료받을 대상의 연령, 성별, 체중과, 치료할 특정 질환 또는 병리 상태, 질환 또는 병리 상태의 심각도, 투여경로 및 처방자의 판단에 따라 달라질 것이다. 이러한 인자에 기초한 투여량 결정은 당업자의 수준 내에 있으며, 일반적으로 투여량은 0.001㎎/㎏/일 내지 대략 2000㎎/㎏/일의 범위이다. 더 바람직한 투여량은 1㎎/㎏/일 내지 100㎎/㎏/일이다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다. The dosage of the active ingredient will vary depending on the age, sex and weight of the subject to be treated, the particular disease or pathology to be treated, the severity of the disease or pathology, the route of administration and the judgment of the prescriber. Dosage determination based on these factors is within the level of skill in the art and generally dosages range from 0.001 mg / kg / day to approximately 2000 mg / kg / day. More preferred dosage is 1 mg / kg / day to 100 mg / kg / day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.
본 발명의 간암 관련 질환의 예방 및 치료용 약학조성물은, 조성물 총 중량에 대하여 상기 흑삼 추출물을 0.1 내지 50 중량%로 포함한다.The pharmaceutical composition for preventing and treating liver cancer-related diseases of the present invention comprises 0.1 to 50% by weight of the black ginseng extract based on the total weight of the composition.
본 발명의 추출물은 쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관내 주사에 의해 투여될 수 있다. 본 발명의 추출물은 독성 및 부작용은 거의 없으므로 예방 목적으로 장기간 복용시에도 안심하고 사용할 수 있는 약제이다. The extract of the present invention can be administered to mammals such as mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injections. Since the extract of the present invention has little toxicity and side effects, it is a drug that can be used safely even when taken for long periods of time.
또한, 본 발명은 상기 본 발명에 따른 흑삼과 흑미삼은 식품학적으로 허용 가능한 식품보조 첨가제를 포함하는 암 치료 개선을 위한 건강기능식품을 제공한다. 본 발명의 건강기능식품은 정제, 캡슐제, 환제 또는 액제 등의 형태를 포함하며, 본 발명의 화합물을 첨가할 수 있는 식품으로는, 예를 들어, 각종 식품류, 음료, 껌, 차, 비타민 복합제, 건강기능성 식품류 등이 있다.In addition, the present invention provides a health functional food for cancer treatment improvement comprising the black ginseng and black rice ginseng according to the present invention is a food supplement acceptable food supplement. The health functional food of the present invention includes the form of tablets, capsules, pills or liquids, and the food to which the compound of the present invention can be added, for example, various foods, beverages, gums, teas, vitamin complexes, etc. And functional foods.
본 발명을 통하여 제조된 흑삼에서 종래기술로 제조된 흑삼보다 진세노사이드 Rg3가 뛰어나게 증가되는 것을 확인하였다. 또한 진세노사이드 Rg3의 양은 증숙 횟수가 더해갈수록 순차적으로 증가되는 것을 확인할 수 있으며 온도 및 건조 조건에 따라 그 양의 생성이 민감한 것으로 나타났다. It was confirmed that ginsenoside Rg 3 is significantly increased in the black ginseng prepared through the present invention than the black ginseng prepared in the prior art. In addition, it can be seen that the amount of ginsenoside Rg 3 increases sequentially as the number of steaming increases, and the formation of the amount is sensitive to temperature and drying conditions.
발암물질로 알려진 벤조피렌도 본 발명의 온도 및 증숙 조건에서 반복되는 증숙 과정을 거치면서 그 양이 줄어들어 안전규정 수치 이하로 제거되었다. 본 발명의 증숙 초기에 벤조피렌은 규정치 이상인 1.0ppb 이상으로 검출되지만 증숙 과정을 통해 벤조피렌의 양이 규정치 이하로 줄어드는 것을 확인할 수 있다. 이는 벤조피렌이 원재료인 인삼에 남아있는 농약에서 생성되는 것으로 추측되며 이렇게 생성된 벤조피렌은 본 발명의 반복되는 증숙 및 건조 조건을 통해 제거된다. 실제로 증숙 전의 인삼에서도 벤조피렌이 검출되었으며 상기 인삼을 이용하여 흑삼을 제조하였을 때만 벤조피렌이 검출되었고 처음부터 인삼에서 벤조피렌이 검출되지 않는 것은 흑삼의 제조 과정에서는 검출되지 않았다. Benzopyrene, also known as a carcinogen, has been removed to below safety regulations by reducing its amount through repeated steaming at the temperature and steaming conditions of the present invention. In the early stage of steaming of the present invention, benzopyrene is detected to be 1.0 ppb or more, which is higher than the prescribed value, but it can be confirmed that the amount of benzopyrene decreases below the prescribed value through the steaming process. It is assumed that benzopyrene is produced from pesticides remaining in ginseng as a raw material, and the benzopyrene thus produced is removed through the repeated steaming and drying conditions of the present invention. In fact, benzopyrene was also detected in the ginseng before steaming, benzopyrene was detected only when black ginseng was prepared using the ginseng, and no benzopyrene was detected in ginseng from the beginning.
또한 상기 진세노사이드 Rg3가 증가된 흑삼이 홍삼에 비해 간암에 대해 탁월한 항암 효과가 있는 것을 확인할 수 있었다. 이로써 간암 관련 질환에 효과가 있으면서도 발암물질에서 안전한 흑삼을 제조할 수 있는 방법을 확립하였다. In addition, it was confirmed that the black ginseng with the increased ginsenoside Rg 3 has an excellent anticancer effect against liver cancer. This has established a method for producing black ginseng that is effective in liver cancer-related diseases and safe from carcinogens.
도 1은 바람직한 실시예 1의 방법대로 흑삼을 제조하는 과정을 나타내는 순서도이다.1 is a flowchart illustrating a process of preparing black ginseng according to the method of the first preferred embodiment.
도 2는 실험예 2-1의 간암세포 HepG2의 세포생존율에 대한 MTT 어세이의 결과를 나타내는 그래프이다.Figure 2 is a graph showing the results of the MTT assay for the cell survival rate of hepatic cancer cell HepG2 of Experimental Example 2-1.
이하, 본 발명의 바람직한 실시예를 상세히 설명하기로 한다. 그러나, 본 발명은 여기서 설명되는 실시예에 한정되지 않고 다른 형태로 구체화될 수도 있다. 오히려, 여기서 소개되는 실시예는 개시된 내용이 철저하고 완전해질 수 있도록 그리고 당업자에게 본 발명의 사상이 충분히 전달될 수 있도록 하기 위해 제공되는 것이다. Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. Rather, the embodiments introduced herein are provided so that the disclosure may be made thorough and complete, and to fully convey the spirit of the invention to those skilled in the art.
<실시예 1 : 흑삼 본체와 미삼의 제조 및 이를 이용한 추출물 제조 : 증숙온도 85℃, 건조온도 50℃, 총 증숙 횟수 13회><Example 1: Preparation of black ginseng main body and rice ginseng and extracts using the same: steaming temperature 85 ℃, drying temperature 50 ℃, the total number of steaming 13 times>
6년근 고려인삼을 채취하여 흙 또는 이물질을 제거한 뒤 정제수로 세척한 뒤 수분을 제거하고 상기 수삼을 몸체와 미삼(뿌리)으로 분리하였다. 분리된 인삼의 몸체와 미삼 10kg씩을 각각 수분함량이 14% 이하가 되도록 자연 건조하여 인삼으로 제조하였다. 건조가 완료된 인삼은 몸체와 미삼을 따로 구분하여 85℃의 온도에서 10시간 동안 증숙한 다음 건조기에서 50℃의 온도로 4시간 동안 건조하였고 동일하게 증숙과 건조를 12번 반복하였다(총 13회 증숙). 증숙과 건조 과정을 반복하면서 건조된 백삼이 홍삼으로 홍삼이 흑삼으로 검게 변하면서 흑삼 몸체와 미삼이 제조되었고 마지막 건조과정은 수분이 10% 이하가 될 때까지 지속한 후 완료하였다. 이후 흑삼 몸체와 미삼의 추출물을 제조하기 위해 상기 흑삼 몸체와 미삼을 각각 분쇄하였다. 상기 분쇄된 흑삼 몸체와 미삼을 각각 80℃의 온도를 가지는 정제수 100ℓ에 넣어 1분당 60 회전의 회전 속도로 8시간 동안 교반한 뒤 3시간 동안 침전시켰다. 침전된 추출물을 여과하여 고형분을 제거하고 여과 여액은 동결건조를 하여 흑삼 몸체와 미삼 추출물을 최종적으로 분말로 제조하였다. Six years old Korean ginseng was collected and the soil or foreign matters were removed, washed with purified water, water was removed, and the fresh ginseng was separated into a body and rice ginseng (root). The body of separated ginseng and 10kg each of ginseng were naturally dried to have a water content of 14% or less, and then manufactured by ginseng. The dried ginseng was steamed for 10 hours at a temperature of 85 ℃, and then dried for 4 hours at a temperature of 50 ℃ in a dryer, and the same steaming and drying were repeated 12 times (total 13 steaming). ). Repeating the steaming and drying process, the dried white ginseng was changed to red ginseng and black ginseng to black ginseng, and the black ginseng body and rice were prepared, and the last drying process was completed until the water became less than 10%. Thereafter, the black ginseng body and the ginseng were ground to prepare an extract of the black ginseng body and the rice ginseng. The pulverized black ginseng body and rice ginseng were put into 100 l of purified water having a temperature of 80 ° C., respectively, and stirred for 8 hours at a rotational speed of 60 revolutions per minute, and then precipitated for 3 hours. The precipitated extract was filtered to remove solids, and the filtrate was lyophilized to finally prepare a black ginseng body and a ginseng extract as powder.
<실시예 2. 증숙온도>Example 2 Steaming Temperature
2-1. 증숙온도 80℃2-1. Steam temperature 80 ℃
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 증숙 온도를 80℃로 하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the steaming temperature was 80 ° C.
2-2. 증숙온도 90℃2-2. Steam temperature 90 ℃
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 증숙 온도를 90℃로 하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the steaming temperature was 90 ° C.
<실시예 3. 증숙 횟수>Example 3. Number of Steaming
3-1. 6회 증숙(반복 증숙 및 건조 5회)3-1. 6 steaming (5 times steaming and drying)
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 총 증숙 횟수를 6회만 수행하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed only six times.
3-2. 7회 증숙(반복 증숙 및 건조 6회)3-2. 7 steaming (6 repeated steaming and drying)
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 총 증숙 횟수를 7회만 수행하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed only seven times.
3-3. 8회 증숙(반복 증숙 및 건조 7회)3-3. 8 steaming (7 times steaming and drying)
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 총 증숙 횟수를 8회만 수행하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed only eight times.
3-4. 9회 증숙(반복 증숙 및 건조 8회)3-4. 9 steaming (8 repeated steaming and drying)
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 총 증숙 횟수를 9회만 수행하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed only 9 times.
3-5. 10회 증숙(반복 증숙 및 건조 9회)3-5. 10 steaming (9 repeated steaming and drying)
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 총 증숙 횟수를 10회만 수행하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed only 10 times.
3-6. 11회 증숙(반복 증숙 및 건조 10회)3-6. 11 steaming cycles (10 times steaming and drying)
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 총 증숙 횟수를 11회만 수행하였다. The black ginseng body and the extract were extracted under the same conditions as in Example 1, but the total number of steaming was performed only 11 times.
3-7. 12회 증숙(반복 증숙 및 건조 11회)3-7. 12 steaming (11 repeated steaming and drying)
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 총 증숙 횟수를 12회 수행하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed 12 times.
<실시예 4. 건조온도 25℃><Example 4. Drying temperature 25 ℃>
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 건조온도를 25℃로 하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the drying temperature was 25 ° C.
<비교예 1 : 증숙 온도> Comparative Example 1: Steaming Temperature
1-1. 증숙 온도 70℃1-1. Steam temperature 70 ℃
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 증숙 온도를 70℃로 하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the steaming temperature was 70 ° C.
1-2. 증숙 온도 75℃1-2. Steam temperature 75 ℃
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 증숙 온도를 75℃로 하였다. The black ginseng body and the rice extract were prepared under the same conditions as in Example 1, but the steaming temperature was 75 ° C.
1-3. 증숙 온도 95℃1-3. Steam temperature 95 ℃
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 증숙 온도를 95℃로 하였다. The black ginseng body and the rice extract were prepared under the same conditions as in Example 1, but the steaming temperature was 95 ° C.
<비교예 2 : 총 증숙 횟수>Comparative Example 2: Total number of steaming
2-1. 1회 증숙(반복 증숙 및 건조 없음)2-1. 1 steaming (no repeated steaming or drying)
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 총 증숙 횟수를 1회만 수행하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed only once.
2-2. 2회 증숙(반복 증숙 및 건조 1회)2-2. 2 steaming (1 time steaming and drying)
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 총 증숙 횟수를 2회만 수행하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed twice.
2-3. 3회 증숙(반복 증숙 및 건조 2회)2-3. 3 steaming (2 times steaming and drying)
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 총 증숙 횟수를 3회만 수행하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but only three times of total steaming was performed.
2-4. 4회 증숙(반복 증숙 및 건조 3회)2-4. 4 steaming (3 times steaming and drying)
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 총 증숙 횟수를 4회만 수행하였다. The black ginseng body and the extract were extracted under the same conditions as in Example 1, but the total number of steaming was performed only four times.
2-5. 5회 증숙(반복 증숙 및 건조 4회)2-5. 5 steaming (4 times steaming and drying)
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 총 증숙 횟수를 5회만 수행하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but only five times of total steaming was performed.
2-6. 14회 증숙(반복 증숙 및 건조 13회)2-6. 14 steaming (13 repeated steaming and drying)
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 총 증숙 횟수를 14회 수행하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but the total number of steaming was performed 14 times.
<비교예 3 : 건조온도>Comparative Example 3: Drying Temperature
3-1. 20℃3-1. 20 ℃
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 건조를 20℃에서 수행하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but drying was performed at 20 ° C.
3-2. 55℃3-2. 55 ℃
상기 실시예 1과 동일한 조건으로 흑삼 몸체와 미삼 추출물을 제조하되 건조를 55℃에서 수행하였다. The black ginseng body and the extract of the ginseng were prepared under the same conditions as in Example 1, but drying was performed at 55 ° C.
<비교예 4 : 홍삼과 홍미삼 추출물 제조>Comparative Example 4: Preparation of Red and Red Ginseng Extracts
시중에 유통되는 6년근 홍삼근(홍삼 몸체)과 홍미삼을 구입하여 실시예 1의 추출방법과 동일하게 80℃에서 정제수를 이용하여 홍삼근과 홍미삼 추출물을 제조하였다.Six years old red ginseng root (red ginseng body) and red ginseng were distributed in the market, and red ginseng root and red ginseng extract were prepared using purified water at 80 ° C. in the same manner as the extraction method of Example 1.
<실험예 1 : 흑삼의 성분 분석>Experimental Example 1: Component Analysis of Black Ginseng
상기 실시예 1 내지 실시예 4와 비교예 1 내지 비교예 3의 방법대로 제조한 흑삼 추출물 및 비교예 4의 홍삼 추출물의 진세노사이드 성분을 분석하였다. 진세노사이드 성분은 잔뿌리 부분에 더 많은 것으로 알려져 있기에 본 실험에서는 각각 흑미삼과 홍미삼을 사용하였다.The ginsenoside components of the black ginseng extract and the red ginseng extract of Comparative Example 4 prepared according to the methods of Examples 1 to 4 and Comparative Examples 1 to 3 were analyzed. Since ginsenoside components are known to be more in the root portion, black and red ginseng were used in this experiment, respectively.
진세노사이드의 분석은 식품공전 [제 4.식품별 기준 및 규격→16.인삼제품류→16-10.농축홍삼류의 시험방법 중 (1)홍삼성분 확인시험 고속액체크로마토그래피법]에 준하여 진세노사이드 검사방법을 사용한 것이며 표 1은 흑삼 및 홍삼의 진세노사이드 함량을 분석한 수치를 나타내는 표이다. The analysis of ginsenosides was carried out in accordance with Food Code [Chapter 4. Food Standards and Standards → 16.Ginseng Products → 16-10.Concentrated Red Ginseng (1) Red Ginseng Identification Test High-Speed Liquid Chromatography]. The senoside test method was used, and Table 1 is a table showing the analysis of ginsenoside content of black ginseng and red ginseng.
표 1의 결과를 확인하면 흑삼의 진세노사이드 Rg3는 증숙 횟수가 증가될수록 유의적으로 증가되었으며 특히 8회 이상에서 다량의 진세노사이드 Rg3가 증가된 것을 확인하였고 9회 이상에서는 진세노사이드 Rg3 함량이 거의 비슷하게 유지되는 것을 확인할 수 있었다. 또한 증숙 온도는 85℃를 전후로 80~90℃ 사이에서 진세노사이드 Rg3와 Rh1 함량이 가장 많이 증가하였으며 너무 낮은 온도(20℃)에서 건조하게 되면 전반적으로 품질이 좋지 않는 흑삼이 제조되는 것을 확인할 수 있었다. 이에 대해 홍삼의 진세노사이드 Rg3와 Rh1 함량은 현저하게 낮은 수치를 보여주었다.The results of Table 1 confirm that the ginsenoside Rg 3 of black ginseng was significantly increased as the number of steaming was increased, and in particular, a large amount of ginsenoside Rg 3 was increased in at least 8 times and in ginsenosides at 9 or more times. It was confirmed that the Rg 3 content was maintained about the same. In addition, the steaming temperature of ginsenosides Rg 3 and Rh 1 was most increased between 80 and 90 ° C around 85 ° C. I could confirm it. In contrast, the ginsenosides Rg 3 and Rh 1 contents of red ginseng showed significantly lower values.
참고로 실시예 1의 흑삼의 총 조사포닌 양은 508.9mg/g으로 분석되었으며 상기 조사포닌은 초기 증숙에는 비교예 4의 홍삼(72.1mg/g)과 비슷한 조사포닌 양을 가지다가 증숙회수가 증가하면서 증가하는 것을 확인할 수 있었다(도표에는 나타내지 않음). For reference, the total amount of irradiated black ginseng of Example 1 was analyzed to be 508.9 mg / g, and the irradiated amount of irradiated with the same amount of irradiated with red ginseng of Comparative Example 4 (72.1 mg / g) was increased during initial steaming. It was confirmed that the increase (not shown in the diagram).
<실험예 2 : 항암효과 분석>Experimental Example 2 Analysis of Anticancer Effect
2-1. in-vitro 항암효과 실험(세포 실험)2-1. in-vitro anticancer effect experiment (cell experiment)
흑삼의 항암 효과를 확인하기 위해 상기 실시예 1의 흑삼 추출물 분말, 비교예 4의 홍삼 추출물 분말을 이용하여 간암세포 HepG2에서 세포 생존율을 측정하였다. 흑삼은 Rg3 함량이 대표적으로 높은 실시예 1의 추출물을 대표로 사용하였고 실험예 1과 마찬가지로 흑미삼과 홍미삼을 사용하였다. In order to confirm the anticancer effect of black ginseng, the cell survival rate of hepatic cancer cell HepG2 was measured using the black ginseng extract powder of Example 1 and the red ginseng extract powder of Comparative Example 4. Black ginseng was used as a representative of the extract of Example 1 having a high Rg 3 content and black and red ginseng as in Experimental Example 1.
먼저 96웰-플레이트에 1×105개/㎖개의 세포를 100㎕씩의 세포배양 배지(100U/㎖ 페니실린, 100g/㎖ 스트렙토마이신, 10% 우혈청이 들어있는 DMEM)를 이용하여 37℃, 5% CO2 인큐베이터에서 24시간 동안 배양하였다. 시험물질인 흑삼 추출물과 홍삼 추출물을 최고 농도 1000μg/㎖에서 2μg/㎖까지 처리하고 24시간 동안 배양하였다. 각 웰당 새로운 배지를 200㎕씩 넣어준 후 MTT(thiazolyl blue, SIGMA co.)를 2μg/㎖의 농도로 준비하여 50㎕씩 첨가하여 3시간에서 4시간 동안 반응을 시켰다. 각 웰당 시험물질 220㎕을 제거하고 보라색 물질 30㎕만 남긴 후 DMSO(dimethyl sulfoxide)를 150㎕ 첨가하여 10분간 잘 혼합하여 침전물을 용해시킨 후 ELISA 플레이트 리더에서 540nm 흡광도로 OD(optical density)값을 측정하였다. 모든 시험결과는 세포를 배양하지 않은 웰에서 측정된 흡광도에 대해 보정한 값을 산출하였다. 표 2는 흑삼 추출물을 간암 세포에 처리하여 나타나는 세포 생존율에 대한 결과를 나타내는 표이며 도 2는 이를 나타낸 그래프다.First, 1 × 10 5 cells / ml in 96 well-plate at 37 ° C. using 100 µl cell culture medium (100 U / ml penicillin, 100 g / ml streptomycin, DMEM containing 10% bovine serum) Incubated for 24 hours in a 5% CO 2 incubator. Test substance black ginseng extract and red ginseng extract were treated from the highest concentration of 1000 μg / ㎖ to 2 μg / ㎖ and incubated for 24 hours. After 200 μl of fresh medium was added to each well, MTT (thiazolyl blue, SIGMA co.) Was prepared at a concentration of 2 μg / ml, and 50 μl was added to react for 3 to 4 hours. After removing 220 μl of test substance for each well, only 30 μl of purple substance was added, and 150 μl of DMSO (dimethyl sulfoxide) was added and mixed well for 10 minutes to dissolve the precipitate. Measured. All test results yielded corrected values for absorbance measured in wells in which the cells were not cultured. Table 2 is a table showing the results for the cell survival rate treated by liver cancer cells treated with black ginseng, Figure 2 is a graph showing this.
표 2 및 도 2의 결과를 확인하면 본 발명의 실시예 1의 흑삼 추출물은 8μg/㎖ 이하 농도에서 간암세포 증식에 대한 50% 억제활성을 확인할 수 있는데 비교예 3의 홍삼 추출물은 간암세포 증식에 대한 50% 억제는 1000μg/㎖ 이상의 높은 농도임을 확인할 수 있다. 이로써 본 발명의 제조방벙으로 제조된 흑삼은 간암세포에 대해 선택적이고 우수한 항암효과가 있는 것을 확인할 수 있다.Checking the results of Table 2 and Figure 2 the black ginseng extract of Example 1 of the present invention can confirm the 50% inhibitory activity against liver cancer cell proliferation at the concentration of 8μg / ㎖ or less, the red ginseng extract of Comparative Example 3 to the liver cancer cell proliferation It can be seen that 50% inhibition against the high concentration of 1000 μg / ㎖ or more. As a result, the black ginseng prepared by the production method of the present invention can be confirmed to have a selective and excellent anticancer effect on liver cancer cells.
2-2. in vivo 항암효과 실험(동물실험)2-2. in vivo anticancer effect experiment (animal experiment)
이식할 HepG2 세포를 100U/㎖ 페니실린, 100g/㎖ 스트렙토마이신, 10% 우혈청이 들어있는 세포배양 배지 DMEM(Gibco BLR)를 이용하여 세포배양 플라스크에 담아 95% CO2 37℃ 세포배양기에서 배양하였다. 세포가 유착되면 HBSS(Hanks balanced salt solution, Gibco BRL)로 2번 씻어내고 0.2% 트립신으로 세포를 수집하였다. 실험동물은 각 실험군마다 9마리씩 6주령된 BALB/c계 누드 마우스(SLC inc., Shizuoka, Japan)를 사용하였다. 각 추출물은 300mg/kg 단위로 매일 경구투여하였으며 대조군에는 정제수만을 투여하였다. 우선적으로 이들 누드 마우스에 1×107 개의 HepG2 세포를 주입하여 종양주를 획득한 후 계대군에 계대하였다. 3회에 걸친 반복계대로 원래의 고형암의 성질을 회복한 종양괴를 얻은 후 이 종양괴를 8마리의 계대군에 이식하였다. 종양의 중심괴사가 일어나기 전에 충분한 혈액공급으로 급속히 자라는 단계의 종양을 함유한 동물을 희생시켜 주로 급속한 분열이 일어나는 외곽부위를 일정한 크기(3×3×3㎜)로 잘라 종양절편을 만들었다. 투관침의 끝에 종양편을 올리고 동물의 좌측전지의 후방의 체간 측면부에 끝이 이르도록 하였다. 투관침을 가볍고 빠르게 360°를 회전시켜며 빼주어 종양편이 목표한 위치에 자리잡도록 해주고 절제부위는 소독하였다. 최초 투여후 24일이 경과하면 실험을 종료하고 부검하여 개체별로 고형암을 적출하여 중량을 측정하였고 digital plethysmomether를 이용하여 실부피를 측정하여 표 3에 나타내었다. HepG2 cells to be transplanted were placed in cell culture flasks using DMEM (Gibco BLR) cell culture medium containing 100 U / ml penicillin, 100 g / ml streptomycin, 10% bovine serum and cultured in a 95% CO 2 37 ° C. cell culture medium. . When the cells adhered, the cells were washed twice with HBSS (Hanks balanced salt solution, Gibco BRL) and collected with 0.2% trypsin. Experimental animals used 9-week old BALB / c nude mice (SLC inc., Shizuoka, Japan), each of which was 9 animals. Each extract was orally administered daily at 300 mg / kg and only purified water was administered to the control group. First, these nude mice were injected with 1 × 10 7 HepG2 cells to obtain tumor lines, which were then passaged to the passage group. The tumor mass was recovered in three passages, and then the tumor mass was transplanted to eight passages. Tumor sections were made by cutting the outer areas where rapid division occurred to a certain size (3 × 3 × 3 mm) at the expense of animals containing tumors growing rapidly with sufficient blood supply before tumor necrosis occurred. The tumor piece was placed at the end of the trocar and the end was reached to the trunk body side of the rear cell of the animal. The trocar was lightly and quickly rotated out of 360 ° to allow the tumor piece to settle in the desired location and the dissection site was disinfected. After 24 days after the initial administration, the experiment was terminated and the autopsy was performed to determine the weight of solid cancers extracted from each subject, and the actual volume was measured using digital plethysmomether.
표 3의 결과를 통해 본 발명의 흑삼 추출물이 홍삼 추출물보다 간암 동물모델 실험에서도 종양의 무게와 종양 부피 억제 하는 능력이 뛰어나다는 것을 확인할 수 있었다.Through the results of Table 3, the black ginseng extract of the present invention was confirmed that the ability to inhibit the weight and tumor volume of the tumor in liver cancer animal model experiments than the red ginseng extract.
<실험예 3 : 벤조피렌 검출 분석>Experimental Example 3: Benzopyrene Detection Analysis
본 실험에서도 상기 실험예 1 및 실험예 2와 동일하게 흑삼과 홍삼의 미삼 추출물을 이용하였다. 벤조피렌 분석방법은 2007년 12월 31일자 식품의약품안전청 '건강기능식품(소위 흑삼)중 벤조피렌 시험법 지침'에 따라 실시예 1 내지 실시예 4 및 비교예 1 내지 비교예 3에서 제조된 흑삼 몸체(추출물 아님) 4g을 디크로로메탄으로 추출 및 농축한 후, 헥산에 녹여 프로리실카트리지를 통과·건조시켜 아세토나이트릴에 녹여 액체크로마토그래프(High Performance Liquid Chromatograph)로 분석하였다. 내부 표준물질로는 3-메틸콜란트젠을 사용하였으며, 검출기는 형광검출기(Fluorescence detector)을 사용하였다. 또한 칼럼은 역상분배형(C18 Supelco LC-PAH 4.6mm × 250mm I.D.)를 사용하였고 분석조건은 하기의 표 4과 같으며 5회 반복하여 표 5에 나타내었다. 벤조피렌은 1.0 ppb 이하를 안전규정 미만으로 하고 있다. In this experiment, black ginseng and red ginseng extracts were used in the same manner as in Experiment 1 and Experiment 2. The benzopyrene analysis method is the black ginseng body prepared in Examples 1 to 4 and Comparative Examples 1 to 3 according to the benzopyrene test method of the health functional food (so-called black ginseng) of December 31, 2007 4 g of the extract was extracted and concentrated with dichloromethane, dissolved in hexane, passed through a prolysyl cartridge, dried and dissolved in acetonitrile, and analyzed by high performance liquid chromatography (High Chromatograph). 3-methylcollanthene was used as an internal standard, and a fluorescence detector was used as a detector. In addition, the column was used in the reverse phase distribution type (C18 Supelco LC-PAH 4.6mm × 250mm I.D.) and the analysis conditions are shown in Table 4 below and are shown in Table 5 by repeating five times. Benzopyrene is less than 1.0 ppb below safety regulations.
표 5의 결과로부터 본 발명의 흑삼 제조에 있어서 초기 증숙 조건에서는 벤조피렌의 양이 오히려 많지만 증숙 횟수가 증가하면서 벤조피렌 검출량이 점점 줄어들고 있는 것을 확인할 수 있었고 상기에 언급된 대로 벤조피렌의 양이 1.0ppb 이하일 때 안전한 것으로 규정되기 때문에 상기 실시예 1 내지 실시예 4에서 제조된 흑삼은 안전한 제품임을 확인할 수 있었다. 또한 흑삼을 건조하는 온도도 본 발명의 조건보다 높게 되면 벤조피렌이 1.0ppb 이하로 완전히 저감되지는 않았다. 이로써 본 발명의 방법은 흑삼 제조 공정에서 발생할 수 있는 벤조피렌을 오히려 제거할 수 있는 방법임을 확인할 수 있다. From the results of Table 5, in the production of black ginseng of the present invention, the amount of benzopyrene was increased in the initial steaming condition, but the amount of benzopyrene was gradually decreased as the number of steaming was increased. Since it is defined as safe, the black ginseng prepared in Examples 1 to 4 was confirmed to be a safe product. In addition, when the temperature of drying black ginseng is higher than the conditions of the present invention, benzopyrene is not completely reduced to 1.0 ppb or less. As a result, it can be seen that the method of the present invention can remove benzopyrene, which may occur in the black ginseng manufacturing process.
참고로 증숙 전의 인삼에서도 벤조피렌 양을 검출하였는데 예상했던 바와 달리 벤조피렌의 양이 1.02ppb로 검출되었다. 이 때문에 증숙 전에 벤조피렌이 검출되지 않는 깨끗한 인삼을 별도로 구분하여 증숙을 하게 되었는데 상기 벤조피렌이 검출되지 않는 인삼을 이용하여 증숙한 흑삼은 이전과 달리 벤조피렌이 검출되지 않는 것을 확인할 수 있었다(도표로 나타내지는 않음). For reference, the amount of benzopyrene was also detected in ginseng before steaming, but as expected, the amount of benzopyrene was detected as 1.02 ppb. Because of this, clean ginseng that is not detected benzopyrene was steamed separately before steaming, and black ginseng steamed using ginseng that is not detected by benzopyrene was confirmed that benzopyrene was not detected unlike before (as shown in the diagram). Not).
본 발명을 이용하여 생성된 흑삼은 의약품이나 식품에 첨가되어 암 예방제, 암 치료제 등으로 제조되어 사용될 수 있다. 구체적인 예로서 본 발명은 흑삼 몸체와 미삼을 이용한 흑삼 추출물이 포함된 항암성이 높은 약제와 기능성 식품의 제조방법을 제공한다. The black ginseng produced using the present invention may be added to medicines or foods and used as a cancer prevention agent, a cancer treatment agent, or the like. As a specific example, the present invention provides a high anticancer drug and a method for preparing a functional food containing a black ginseng extract using a black ginseng body and rice.
<사용예 1: 약제의 제조예> <Use Example 1: Preparation of Pharmaceuticals>
1-1. 산제의 제조1-1. Manufacture of powder
실시예 1의 흑미삼 추출물 분말.................. 300 mgBlack ginseng extract powder of Example 1 ... 300 mg
유당 ......................................... 100 mgLactose ......................................... 100 mg
탈크 ......................................... 10 mgTalc ......................................... 10 mg
상기의 성분들을 혼합하고 기밀포에 충진하여 산제를 제조한다.The above ingredients are mixed and filled in an airtight cloth to prepare a powder.
1-2. 정제의 제조1-2. Manufacture of tablets
실시예 1의 흑미삼 추출물 분말.................. 300 mgBlack ginseng extract powder of Example 1 ... 300 mg
옥수수전분 .................................... 100 mgCorn starch ..................... 100 mg
유당 ......................................... 100 mgLactose ......................................... 100 mg
스테아린산 마그네슘 .......................... 2 mgMagnesium Stearate ......................................... 2 mg
상기의 성분들을 혼합한 후 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조한다.After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.
1-3. 캅셀제의 제조1-3. Manufacture of capsule
실시예 1의 흑미삼 추출물 분말.................. 300 mgBlack ginseng extract powder of Example 1 ... 300 mg
옥수수전분 ................................... 100 mgCorn starch ..................... 100 mg
유당 ......................................... 100 mgLactose ......................................... 100 mg
스테아린산마그네슘 ........................... 2 mgMagnesium Stearate ........................... 2 mg
통상의 캡슐제 제조방법에 따라 상기의 성분을 혼합하고 젤라틴 캡슐에 충전하여 캡슐제를 제조한다.According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare capsules.
<사용예 2: 건강식품의 제조예> <Use Example 2: Production Example of Health Food>
2-1. 건강식품의 제조2-1. Manufacture of health food
실시예 1의 흑미삼 추출물 분말.................. 200 ㎎Black rice ginseng extract powder of Example 1 .. 200 mg
비타민 혼합물.................................. 적량Vitamin Blend ...........
비타민 A 아세테이트 .......................... 70 ㎍Vitamin A Acetate ............... 70 μg
비타민 E ..................................... 1.0 ㎎Vitamin E ..................................... 1.0 mg
비타민 B1 .................................... 0.13 ㎎Vitamin B1 ......................................... 0.13 mg
비타민 C ..................................... 10 ㎎Vitamin C ......................................... 10 mg
비오틴 ........................................ 10 ㎍Biotin ......................................... 10 μg
니코틴산아미드 ............................... 1.7㎎Nicotinamide ......................................... 1.7mg
엽산........................................... 50 ㎍Folic acid ........................... 50 ㎍
판토텐산 칼슘 ................................. 0.5 ㎎Calcium Pantothenate ..................... 0.5 mg
황산제1철...................................... 1.75 ㎎Ferrous Sulfate ............... 1.75 mg
산화아연....................................... 0.82 ㎎Zinc Oxide ............... 0.82 mg
탄산마그네슘 .................................. 25.3 ㎎Magnesium Carbonate ......................................... 25.3 mg
제1인산칼륨.................................... 15 ㎎Potassium monophosphate ............... 15 mg
제2인산칼슘.................................... 55 ㎎Dicalcium Phosphate Dibasic ......................................... 55 mg
구연산칼륨..................................... 90 ㎎Potassium Citrate ............... 90 mg
탄산칼슘 ...................................... 100 ㎎Calcium Carbonate ......................... 100 mg
염화마그네슘 .................................. 24.8 ㎎Magnesium Chloride ......................................... 24.8 mg
상기의 비타민 및 미네랄 혼합물의 조성비는 비교적 건강식품에 적합한 성분을 바람직한 제제예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 건강식품 제조방법에 따라 상기의 성분을 혼합한 다음, 과립을 제조하고, 통상의 방법에 따라 건강식품 조성물 제조에 사용할 수 있다.The composition ratio of the above-mentioned vitamin and mineral mixtures is composed of relatively suitable ingredients suitable for health foods in a preferred formulation example, but the compounding ratios may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. The granules may be prepared and used for preparing a health food composition according to a conventional method.
2-2. 건강 음료의 제조2-2. Manufacture of healthy drinks
실시예 1의 흑미삼 추출물 분말................... 500 ㎎Black rice ginseng extract powder of Example 1 ... 500 mg
구연산 ......................................... 100 ㎎Citric Acid ......................................... 100 mg
올리고당 ....................................... 100 gOligosaccharide ......................................... 100 g
매실농축액 ..................................... 2 gPlum concentrate ..................................... 2 g
타우린 ......................................... 1 gTaurine ......................................... 1 g
정제수를 가하여 전체 .......................... 900 ㎖Purified water is added to the whole ........... 900 ㎖
통상의 건강 음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1시간 동안 85℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2ℓ용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 건강음료 조성물 제조에 사용한다. After mixing the above components in accordance with a conventional method for preparing a healthy beverage, and then stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilized and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.
상기 조성비는 비교적 기호음료에 적합한 성분을 바람직한 제제예로 혼합 조성하였지만 수요계층이나, 수요국가, 사용 용도 등 지역적, 민족적 기호도에 따라서 그 배합비를 임의로 변형 실시하여도 무방하다.Although the composition ratio is a composition suitable for a preferred beverage in a preferred formulation example, the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and use purpose.
Claims (5)
- (1공정) 인삼을 세척하고 몸체와 미삼으로 분리하는 단계; (Step 1) washing the ginseng and separating the body and ginseng;(2공정) 세척된 인삼의 몸체와 미삼의 수분 함량이 14% 이하가 되도록 건조하는 단계; (Step 2) drying the washed ginseng body and the ginseng so that the moisture content is less than 14%;(3공정) 건조된 인삼의 몸체와 미삼을 각각 80~90℃의 온도에서 8~10시간 동안 증숙하는 단계; (3 step) steaming the body and dried ginseng of dried ginseng at a temperature of 80 ~ 90 ℃ for 8 to 10 hours;(4공정) 3공정의 증숙된 인삼의 몸체와 미삼을 25~50℃에서 3~5시간 동안 건조하는 단계; (Step 4) drying the body and steamed ginseng of three steps of steamed ginseng at 25 to 50 ° C. for 3 to 5 hours;(5공정) 3공정의 증숙 단계와 4공정의 건조 단계를 연속으로 5~12회 반복하는 단계; (5 steps) repeating the steaming step of the three steps and the drying step of the fourth step 5 to 12 times in succession;(6공정) 5공정의 반복 증숙 및 건조가 완료된 흑삼 몸체와 미삼을 최종적으로 25~50℃에서 수분이 10% 이하가 되게 건조하는 단계; 및(Step 6) drying the black ginseng body and the rice ginseng, which has been repeatedly steamed and dried in step 5, to finally have water of 10% or less at 25 to 50 ° C .; And(7공정) 6공정의 흑삼을 용매 추출하여 얻은다음 용매를 제거 건조하여 흑삼 추출물을 제조하는 단계;(7 step) to obtain a black ginseng extract obtained by solvent extraction of the black ginseng of step 6 and then dried to remove the black ginseng;를 특징으로 하는 것으로 Rg3 함량이 130mg/g 이상이고 벤조피렌 검출량이 1.0ppb 이하를 함유하는 흑삼 추출물의 제조방법. A method of producing black ginseng extract, characterized in that the Rg 3 content is 130mg / g or more and the benzopyrene detection amount is 1.0ppb or less.
- 제 1 항에 있어서,The method of claim 1,상기 1공정의 인삼은 4~6년근 인삼인 것을 특징으로 하는 흑삼의 제조방법.The ginseng of the first step is a method for producing black ginseng, characterized in that 4 to 6 years old ginseng.
- 제 2 항에 있어서,The method of claim 2,상기 인삼은 파낙스(Panax)속에 속하는 다년생 식물로, 고려인삼(Panax ginseng), 화기삼(Panax quinquefolia), 전칠삼(삼칠, Panax notoginseng), 죽절삼(Panax vietnamensis), 파낙스 엘레가티오르(Panax elegatior), 파낙스 완지아누스(Panax wangianus) 또는 파낙스 피핀라티푸스(Panax bipinratifidus)을 포함하는 것을 특징으로 하는 흑삼의 제조방법.The ginseng is a perennial plant belonging to the genus Panax, Panax ginseng, Panax quinquefolia, Panax ginseng, Panax notoginseng, Panax vietnamensis, Panax elegatior. , Panax wanianus (Panax wangianus) or Panax bipinratifidus (Panax bipinratifidus) characterized in that it comprises a method of producing black ginseng.
- 제 1 항의 흑삼 추출물을 유효성분으로 포함하는 간암 예방 및 개선용 건강기능식품.Health functional food for liver cancer prevention and improvement comprising the black ginseng extract of claim 1 as an active ingredient.
- 제 1 항의 흑삼 추출물을 유효성분으로 포함하는 간암 예방 또는 치료용 약학 조성물.A pharmaceutical composition for preventing or treating liver cancer comprising the black ginseng extract of claim 1 as an active ingredient.
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