KR100893779B1 - The extracting method of Angelica gigas Nakai that has effect of scavenging action on free radical - Google Patents

Abstract

The present invention relates to a method for extracting the Angelica gigas extract having a free radical scavenging effect.

Charcoal Angelica extract with toxic free radical scavenging effect is a liquid concentrate composed of more than 90% of the major components of the decancin and decosinol angelate, the present invention is that the Angelica Angelica extract is in the form of water that can be administered to the human body Extracted only with and ethanol, and also using a purification method using the difference in temperature and solubility for high purity.

 Angelica gigas Nakai, Decursin, Decursinol angelate, antioxidant, free radical

Description

The extracting method of Angelica gigas Nakai that has effect of scavenging action on free radical}

1 is an extract-purified concentrated purified Angelica gigas of 96% purity

Figure 2 Angelica Angelica HPLC Analysis and Structure of Decosinol, Decosin and Decosinol Angelate Extracted and Purified from gigas Nakai)

Champagne Angelica has been used as a Korean medicine as a blood donor as a country of origin, and recently, it has been reported that Decacin, a Korean indicator, and Decacinol, a major ingredient, have blood flow improvement and anti-Helicobacter action, and the inventors have revealed pure water. It has been shown in animal experiments that purified decosin is effective in inhibiting kidney toxicity, preventing renal failure due to diabetic complications, and diabetic hypertension.

( PCT / KR99 / 00632 ).

     A study on the antioxidant effect of Angelica gigas extract disclosed by "Kang" was published. In this case, the antioxidant effect of the extract using hot water extract and three times using 70% methanol were extracted and then concentrated. . (Korean Journal of Food and Nutrition Science, Vol. 33, No.7, page 1112 ~ 1118, 2004, 2004 Poster presented at the Korean Society of Community Nutrition Conference Spring Conference)

     The research results of “Kang” and others were tested for the antioxidant effect by simply concentrating hot water extract or 70% methanol extract. The results showed that 1mg / ml of Cangdong Angelica hot water extract had a free radical scavenging effect of 66.3%, and 70% methanol extract 1mg. / Ml scavenging effect was reported as 81.5%, but the analysis results for the active ingredient was not shown. In other words, decursin and decursinol angelate were not highly purified and tested.

      In addition, the inventors of the present invention prepared and secured a concentration method that is simpler than the existing method and secured a large amount of extraction, and it was confirmed that the solution was easily absorbed in the intestine when administered orally to the experimental animals.

      Decusin and decosinol angelate have not been well known as structural isomers until now, but the present inventors have studied the action and have also established and analyzed the analysis method (Korean Patent Registration No .: 0509843).

      Decasin may be present as a solid at room temperature, but decosinol angelate is present as a liquid even at minus 70 degrees, and in the Angelica gigas, the ratio of decosinol to decansinol angelate is about 3: 2. After purifying through this study, the exact content and ratio through mass analysis showed that Decursin 50 ~ 54.5%, Decursinol angelate 40 ~ 42%, Decursinol 0.1 ~ 0.3%, Decosinol derivatives about 2-3 Purified material consisting of% and other 1-2% was secured.

      These results indicated that the ratio of Deckersin and Dekersinol Angelate was 57:43, which was slightly different from that of 60 to 40, that is, 3: 2. In addition, the content of deckerin was above 90%, which resulted in more than expected results. The existing alcohol extraction method or 70% alcohol extraction method has been known that the content of decusin present in Angelica gigas is about 35% of the extract concentrate and the amount is not more than 50%, but as a result of this study, more than 99% alcohol alcohol And when the pharmacopeia ethanol was used, the concentrated content was high, resulting in containing up to 75% or more and 88%. (Korean patent registration number: 0509843)

      This study aims to develop a method to improve the purity by using the difference of temperature and solubility by improving the existing method.

The present invention provides purity 90 using practical ethanol, which can be used directly in medicine or foods, as the active extract of Angelica gigas extract, which is an active ingredient of decusin and decosinol angelate, which has an excellent effect on eliminating harmful free radicals. It is an object of the present invention to provide a method for producing a Angelica gigas extract concentrate containing decansin and decosinol angelate as high purity of more than%.

In order to achieve the above object, the present invention improves the purification method already developed, and this time the purification method developed by using ethanol and water and temperature, solubility so that the main component content can be more than 90% The present invention is characterized by producing water, and the present invention is characterized in that the extract extracted from Angelica gigas extract has excellent efficacy in eliminating harmful free radicals and is used as an excellent anti oxidant in terms of stability. It is done.

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      Conventional purification methods require the use of organic solvents for the purification of more than 90%. Organic solvents have residual, toxic, and adverse odors, but the developed method is a simple and economical method that does not cause this problem. Can be used directly for medicine or medicine. It is considered that the free radicals can be effectively removed by the combined use of decansin and decosinol angelate, which is a concentrated extract mixture of Angelica gigas.

       It is claimed in the present invention to prepare and use a complex containing deckercin and decosinol angelate as a high purity product having a purity of 90% or more using practical ethanol which can be used directly in medicine or food. In addition, by using the Angelica Angelica extract as the active ingredient of decusin and decosinol angelate, the antioxidant effect is excellent and safety is intended to be used as an antioxidant (anti oxidant).

Common Korean Angelica Angelica Much research has been conducted on the effects of decursin, decursinol angelate, decursinol, and / or two or more of the main components of gigas Nakai. , Angelica ( Angelica Extracts from gigas Nakai have traditionally been used to treat anemia, sedatives, painkillers, etc. (DJ Jung, A. Porzel, S. Huneck, Gigasol and other coumarins from Angelica gigas.Phytochemistry, 30, 710-712, 1991 )

Despite the fact that various biological activities of the decursin and decosinol angelate derivatives have been reported, the lack of information on their physiological dynamics has not been achieved through the technology of purely separating and purifying them. This is because it was difficult.

      Hazardous free radicals generated in cells change the biomolecules such as lipids, proteins, and nucleic acids, leading to aging. These harmful free radicals are produced in small amounts in normal metabolic processes, and can be produced excessively during smoking, bacterial infections, pollution, disease or excessive stress, and UV exposure. The excess free radicals produced in the body break down the antioxidant defense mechanisms of the living body, resulting in lipid peroxidation, DNA modification, and pathological physiological abnormalities. Therefore, solidifying these antioxidant mechanisms can be said to maintain health and delay aging.

The free radical scavenging effect of the extract of Angelica gigas extract was carried out. As a result, the Angelica gigas extract concentrate had a sample concentration (SC ₅₀ ) of 0.047 weight that was required to reduce 50% of the ethanol concentration of 1,1-diphenyl-2-picrylhydrazyl. It was about%, which means that SC ₅₀ value of S. mushroom extract (Patent No. 10-0206538) is known to be excellent in antioxidant effect and 0.23% by weight of SC ₅₀ and SC of chondroitin (Purity 90%) (Patent No. 10-0365242) _It was found to be very good compared to the ₅₀ value of 0.56% by weight and the SC ₅₀ value of the soybean saponin (Patent Publication 10-0566857) about 0.34% by weight. Therefore, as a result of the present invention, the antioxidant effect of the extract of Angelica gigas extract was superior to any other substances.

( Example )

Hereinafter, the actual experimental example of the present invention will be described in detail.

The following experimental example relates to an extraction method containing 90% or more, more preferably 95% or more, of the main components of decancin and decosinol angelate from Angelica gigas and illustrates the results of the antioxidant activity using the extract. will be.

[ Example  One] Decker  Solubility output

Solubility was calculated using purely synthetic purified decusin. 100 mg of decusin was added to 10 ml of 60% ethanol, mixed well, and the undissolved material was precipitated by centrifugation at 10,000 rpm for 1 minute, the supernatant was removed, and the amount of precipitated material was calculated. The solubility as a result of subtracting the amount precipitated from 100 mg was found to be 6.6 mg / mL (60% ethanol). In addition, the solubility in 100% ethanol calculated by the same method was about 4,870 mg / ml. It is said to be internal and external. This property was used for recrystallization to increase the purity.

[ Example  2] high content Concentrate  Manufacturing method

As a method of separating and concentrating Decusin and Decusinol Angelate from Angelica gigas, it was carried out using Korean Angelica gigas, Korean Angelica gigas. Originating in China, originating in Japan, and originating in Vietnam are of low content and are not economically valuable.

For the food and medicinal purposes, the extraction solvent was ethanol and alcohol alcohol, and purified water was used when using water. Grind the raw Angelica Angelica to less than 40 mesh (40 mesh), dry it to 5% or less of moisture, and add 2 to 4 times the amount of Angelica Angelica pulverized with medicinal alcohol or alcohol (hereinafter referred to as 'ethanol'). After 12 hours, the extract was sufficiently extracted and filtered. In this case, the content of decusin and decosinol angelate in the concentrate was 33%.

This concentrate was evaporated to dryness to obtain a highly viscous material. 1 liter of ethanol per kilogram of this substance was dissolved to dissolve the main component, and the resulting precipitate was left at 20 ° C for 10 hours to remove the precipitate, which was then centrifuged to obtain a supernatant. The supernatant was evaporated to dryness to give a concentrate.

50 liters of 60% ethanol (diluted ethanol) was added to this concentrate and eluted. In this case, the concentrations of Decusin and Decusinol Angelate contained in the concentrate were 330 grams and the amount of the main component that could be dissolved in 50 liters of 60% ethanol was also used.

After elution, centrifugation gave only a 60% ethanol layer. This was evaporated to dryness to give a concentrate. This was dissolved in 99% alcohol and the supernatant was taken to give the final concentrate. (See Fig. 1)

Representative results of the liquid chromatography (HPLC) are as follows, the content of decosin 54.52%, decosinol angelate 41.26%, decosinol 0.299%, the total content was 96% and the decosin derivative 2.5 Showed%. Confirmation test was carried out using a mass spectrophotometer as the molecular weight. Each molecular weight was MW 328.35, MW 328.35, MW 246.25. (See Figure 2)

Example 3 Hazardous Free Radical Scavenging Effect

The chaff extract extracted from Example 2 was used. In other words, the extract of Angelica gigas was used as a main component of the decansin, decosinol angelate, decosinol having a purity of 96%. Toxic free radical scavenging effect was observed.

To 0.5 ml of 0.2 mM 1,1-diphenyl-2-picrylhydrazyl ethanol solution, 1 ml of various concentrations prepared by using ethanol as a solvent, Absorbance was measured at 517 nm after standing for a minute. As a result, the concentration (SC ₅₀ ) that reduces the harmful free radicals and the free radicals by 50% was calculated as follows. (Free radical scavenging ability of the sample can be expressed as scavenging activity (SC ₅₀ ), where SC ₅₀ is the concentration of DPPH (1,1-diphenyl-2-picrylhydrazyl) Results in a concentration of sample required to decrease by 50%)

In this Example, SC ₅₀ was found to be 0.047 wt% (Table 1) . The value of SC ₅₀ means the point where the value of DPPH Inhibition ratio (%) shows 50%.

This is the SC ₅₀ the value of the antioxidant effect is excellent and known linteus extract SC ₅₀ value is 0.23% by weight as chondroitin (Purity 90%) (Patent Registration Publication No. 10-0365242 of the Patent Registration No. 10-0206538 Bulletin) It was found to be very good compared to 0.56% by weight and the SC ₅₀ value of the weak bean saponin (Patent Publication 10-0566857) was about 0.34% by weight. Therefore, as a result of the present invention, the antioxidant effect of the extract of Angelica gigas extract was superior to any other substances.

TABLE 1

DPPH (1,1-diphenyl-2-picrylhydrazyl) Inhibition ratio (%)

           = [1- (E -B) / C] x 100

      . B (Blank)-0.5 ml of EtOH + 1 ml of test sample

      . C (Control)-0.5 ml of 0.2 mM DPPH + 1 ml of solvent

      . E (Experimental)-0.5 ml of 0.2 mM DPPH + 1 ml of test sample

As can be clearly seen through the above embodiment, Angelica Angelica provided by the present invention ( Angelica The concentrate purified from gigas Nakai) is a high-purity refinement using only ethanol and water, which is very useful when developing into pharmaceuticals or foods, and contains high purity decansin and decusinol angelate as main ingredients. High purity material was obtained using the difference in solubility and the difference in solubility according to the mixing ratio of ethanol and water. Angelica Angelica Concentrates purified from gigas Nakai (deckerin, decosinol angelate, decosinol) have become useful inventions that can be used industrially as anti-oxidants through toxic free radical scavenging. will be.

Claims (7)

delete delete delete delete delete delete Grind the raw Angelica Angelica to less than 40 mesh and dry it to less than 5% water content, and extract it under shaking by adding 2 to 4 times more medicinal alcohol or alcohol to the ground Angelica Angelica. And concentrating to obtain a highly viscous material; Adding 1 liter of medicinal alcohol or spirit alcohol per kilogram of this substance to dissolve the main ingredient, and leave the precipitate at 10 ° C. for 10 hours to remove the resulting precipitate by centrifugation to obtain a supernatant and to evaporate and dry the supernatant to obtain a concentrate. Solubilizing the concentrate with 60% ethanol using the solubility of the main component in the concentrate; Centrifuging and removing impurities insoluble in 60% ethanol; Take 60% ethanol layer and evaporate to dryness: Take the supernatant by dissolving evaporated material in 99% ethanol: The supernatant is re-evaporated to dryness, and the method of producing a extract of Angelica gigas extract, characterized in that it contains at least 90% by weight of decancin, decansinol, and decansinol angelate.

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